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and  Laboratory  Methods.  1349 

bacillus  which  the  authors  regard  as  the  true  coli  bacillus.  Finding  this  bacillus 
in  such  large  numbers,  the  authors  were  led  to  experiment  with  it,  and  suc- 
ceeded in  demonstrating  that  cultures  of  the  bacillus  were  pathogenic  for  the 
horse.  By  the  use  of  these  cultures,  partly  with  food  and  partly  by  intravenous 
inoculation,  they  succeeded  in  reproducing  the  disease  in  experimental  animals. 
They  are  of  the  conclusion,  therefore,  that  the  widespread  coli  bacillus  is 
occasionally  the  cause  of  serious  and  fatal  epidemics  among  horses. 

H.  w.  c. 

Bienstock.    Du  role  des  Bacteries  de  I'intestin.      The    author    gives    a    very   suggestive 
Ann.  de  I'lnst.  Past.  XIV,  p.  71:0,  iqoo.  .1        r  •  r    ^1 

paper  upon  the  functions  of  the  or- 
dinary intestinal  bacteria.  He  has  previously  shown  in  the  intestine  of  animals 
the  presence  of  a  Bacillus  putrificus,  which  produces  a  putrefying  action  on 
proteids.  He  now  finds  that,  under  normal  conditions,  such  putrefaction  of  the 
contents  of  the  intestine  does  not  occur.  This  fact  seems  surprising,  inasmuch 
as  B.  putrificus  is  constantly  present  in  the  intestine,  and  the  conditions  are 
apparently  proper  for  its  growth.  Bienstock  is  of  the  opinion  that  putrefactive 
action  is  checked  by  the  presence,  in  the  intestine,  of  certain  aerobic  bacteria, 
such  as  lactic  and  the  coli  bacilli.  Experiments  show  that  the  putrefaction 
produced  by  B.  putrificus  does  not  take  place  when  a  quantity  of  these  aerobic 
bacteria  are  present.  The  author  concludes,  therefore,  that  these  aerobic  bac- 
teria, which  are  uniformly  found  in  the  normal  intestine,  are  of  direct  value  to 
the  human  body  in  preventing  the  putrefaction  of  the  intestinal  contents.  He 
points  out  the  fact  that  sterilized,  and  even  pasteurized,  milk  is  not  so  readily 
digested  as  raw  milk,  especially  by  persons  with  intestinal  disturbances,  and 
this  he  attributes  to  the  fact  that  since  the  heat  has  destroyed  the  lactic  organ- 
isms, these  organisms  are  not  present  in  the  intestine  to  prevent  the  putrificus 
from  producing  putrefaction.  In  short,  the  author  concludes  that  the  reason 
why  ordinary  micro-organisms  are  needed  in  the  intestine  is  to  prevent  the 
putrefaction  which  would  otherwise  occur  in  the  intestinal  contents,  owing  to  the 
presence  of  certain  putrefying  micro-organisms  which  are  always  found. 

H,  w.  c. 

Reed,  Carroll  and  Agramonte.    The  Etiology      These  authors  have  presented  a  further 
of  Yellow  Fever.     Med.  Rec,  Feb.  16, 190 1.      ^gp^j.^  ^p^^  their  conclusions  in  regard 

to  the  relation  of  yellow  fever  to  mosquitoes.  The  results  reached  are  of 
immense  importance  and  are  too  numerous  to  be  summarized.  The  most 
important  are,  that  the  disease  is  transmitted  from  yellow  fever  patients  to 
healthy  persons  by  the  bites  of  mosquitoes,  there  being  a  period  of  incubation 
from  41  hours  to  5  days.  They  have  repeatedly  succeeded  in  reproducing 
the  disease  by  allowing  mosquitoes  (culcx  fasciatus)  to  bite  patients  and,  subse- 
quently, healthy  individuals.  They  find  that  an  attack  of  yellow  fever  conveyed 
by  a  mosquito  bite  confers  immunity  against  disease.  A  house  is  only  infested 
with  yellow  fever  when  containing  no  mosquitoes.  Yellow  fever  is  not  dis- 
tributed by  soiled  articles  of  clothing  or  bedding,  as  has  been  supposed.  The 
spread  of  yellow  fever  may  be  most  effectually  prevented  by  protecting  the 
patient  from  mosquito  bites.     These  conclusions,  which  represent  only  a  few  of 


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Journal  of 


Applied  Microscopy 


and 


Laboratory  Methods 


Volume  IV 


V 


B  O  T  A  N  I  C   *.  L    .^' 


JANUARY  TO  DECEMBER 

1901 


ROCHESTER,  N.  Y. 
Issued  by  the  Publication  Department  of  the  Bausch  &  Lomb  Optical  Co» 


,  c^3^3^ 


\J 


■^ 


INDEX. 


Absorbent  cotton  in    the   study   of    Infusoria, 

1580. 
Absorption  of  iron  through  the  intestine,  1537. 
Absorption  of  proteids,  1426. 
Acanthocephali,  collection  and  preservation  of, 

1582. 
Accelerating  effect  of  heat  upon  growth,  cause 

of,  1 22 1. 
Accessory  adrenal  body  in  the  broad  ligament, 

1342. 
Achromatic  spindle  in  the  spore  mother  cells 

of  Osmunda  regalis,  1255. 
Acid  resisting  bacteria  in  lower  animals,   1391. 
Actinomycosis  in  man,  142S. 
Action  of  anilin   stains  on   animal   and  plant 

cells,  1334. 
Acute  and  chronic  laryngitis,   1544. 
Acute  interstitial  nephritis,  character  of  exuda- 
tion in,  1422. 
Adaptation  of  bacteria  to  alexines  of  the  blood, 

II43- 
Agglutination  of  the  tubercle  bacillus  by  the 
various  exudations  of    tuberculous  animals, 
1142. 
Alchemilla  arvensis,  fertilization  in,   1381. 
Alexines  in  the  blood,  1429. 
Alumosihcates,  1589. 

Aluminum,  micro-chemical  analysis  of,  1 530. 
Amitotic  division,  1336. 

Amitotic  division  in  the  cells  of  Sertoli,   1303. 
Ammonium,  micro-chemical  analysis  of,  11 89. 
Amphipoda,  phototaxis  in,  1547. 
Amphithce  longimana,  habits  and  natural   his- 
tory of,  1 2 19. 
Anatomy  of  the  Cat,  1420. 
Animal  life  in  the  deep  sea,  1541. 
Anthrax  in  food,  virulence  of,   1551. 
Anurida    maritima,    development    of    mouth 

parts  in,   12 10. 
Apochromat  ( 5  mm.)  after  Prof.  C.  S.  Hastings 

in  the  photography  of  diatoms,  1442. 
Apparatus : 

An  improvised  microtome,  1162. 
Combined    condenser   and   polarizer   for 

petrographical  microscopes,   11 55. 
Combined  ureometer  and    saccharometer, 

1286. 
Cone  net,  Birge,  1405. 
Cone  net,  Birge,  modification  of,  1407. 
Damp  chamber  for  use  on  the  klinostat, 

1499- 
Device  for  paraffin  embedding,  1595. 
Device  for  supporting  Pasteur  flasks,  1 1 57. 
Fermentation    tubes    for     bacteriological 

investigations  of  fermentation,  1286. 
For  photographing  diatoms,  1439. 
For  phototaxis  work,   1 184. 
Hand  "crab"  used  in  dredging,  1232. 


Improved    automatic  microtomes,   131 7. 

Improved  photo-microgTaphic  apparatus, 
1366. 

Laboratory  camera  stand,  1202. 

New  freezing  microtome  for  use  with  car- 
bon dioxide  tanks,  1320. 

Normal  electrode  for  physiological  work, 
1471. 

Sealing  stone  jar  for  zoological  laborato- 
ries, 1 261. 

Simple  washing  device,  1297. 

Staining  large  numbers  of  sputum  speci- 
mens, 1594. 

Stereo-photo-micrographic  camera,  11 13. 

U-cell  for  filtration  in  study  of  Infusoria, 

1579- 
Ureometer,  1 156. 
Ventilated  dish  for  bacteria  cultures,  1197. 

Arrangement  of  cilia  on  Paramecium,  1566. 

Arthropod  vision,  1540. 

Arthopoda,  depigmenting  the  eyes  of,  1580. 

Aspergillus  oryxse,  1536. 

Atlas  of  medulla  and  mid-brain,  1543. 

Bacillus  coli  as  the  cause  of  intestinal  troubles 
in  horses  in  west  Prussia,  1348. 

Bacillus  coli  in  water,  1552. 

Bacteria,  relation  of,  to  other  fungi,   1391. 

Bacteria,  role  of,  in  the  intestine,  1349. 

Bacteria  concerned  in  urea  fermentation,  1347. 

Bacteria  in  a  swimming  bath,  1350. 

Bacteria  in  milk  of  cows  suffering  from  mas- 
titis, 1 187. 

Bacterial  self-purification  of  streams,  1266. 

Bacterial  treatment  of  London  sewage,   1430. 

Bacteriology  of  healthy  organs,  1266. 

Barium,  micro-chemical  analysis  of,  1293,1323. 

Beale's    carmine   for   staining   isolated    nerve 
cells,  1564. 

Biological  Laboratories  of  Ripon  College,  1 149. 

Biology  wall  charts,  1172. 

Bird  Life :  a  guide  to  the  study  of  our  com- 
mon birds,  1418. 

Birge  collecting  net,  1405. 

Birge  collecting  net,  modification  of,  1407. 

Blood  examination,  1514. 

Blood  forming  organs,  investigation  of,  1462. 

Blood  vessels  in  Annelids,  structure  of,  1213. 

Botanical  Laboratory  and  Botanical  Garden  of 
the  Tokyo  Imperial  University,  Japan,  1477. 

Botany  at  the  Biological  Laboratory  at  Wood's 
HoU,  i486. 

Brain  of  the  turtle,  function  of,  1546. 

Bronchitis,  bacteriology  of,  114 1. 

Cadmium,    micro-chemical    analysis    of,   1456, 
1499. 

Calcite,  crystallization  of,  from    copper   mines 
of  Lake  Superior,  131 3. 

Calcites  from  the  Bad  Lands,  1 144. 


INDEX. 


Calcium,  micro-chemical  analysis  of,  1242. 
Calliphora     erythrocephala,    development    of, 

1578. 
Camera,  stereo-photo-micrographic,  11 13. 
Cancer,  etiology  of,  11 36. 
Cancer,  miscellaneous  studies  in,  1428. 
Casts   found  in  urine,  how  to  preserve  as    per- 
manent specimens,  1476. 
Cell  division  in  Protozoa,  13S2. 
Cell  plate  in  higher   plants,    development    of 

function  of,   1 132. 
Centrosome,   the    kinetic   center   of   the   cell, 

1302. 
Centrosome  and  sphere  in  Crepidula,  1505. 
Centrosomes  in  the  higher  plants,  1461. 
Ceramothamnion  codii,  a  new  rhodophyceous 

alga,  14 1 2. 
Cesium  and  rubidium,  1125. 
Cestodes,    collection     and     preservation    of, 

1581. 
Cestodes,  development  of,  1340. 
Chalcopyrite,  experiments  on,  1394. 
Champignons,  sexual  reproduction  in,   1535. 
Changes  that  take  place  in  the    bacterial    con- 
tents of  water  during  transportation,  1553. 
Charcot-Leyden  crystals  associated  with    eosi- 
nophilic cells,  1387. 
Chemical   analysis   of   the  sexual  products  of 

sea  urchin,  1 1S5. 
Chemical  quantitative  analysis   of   proteids  in 

stomach  contents,  12 16. 
Chemical  relationship  of   colloid,  mucoid,  and 

amyloid  substances,  1426. 
Cholera  bacillus,  spores  of,  1392. 
Chrome-silver  impregnation  for   nerve   tissue, 

1560. 
Chromosomes  of  the  germ  cells  of  Metazoa, 

1578. 
Cirrhosis  of  the  Liver,  nature  and  distribution 

of.  new  tissue  in,  1 263. 
Cleistogamous  flowers,  1536. 
Columbia  University,  growth  in  ten  years,  1567. 
Combined  condenser  and  polarizer  for   petro- 

graphical  microscopes,  11 55. 
Combined  slide  and  cover-glass  forceps,    1436. 
Combined  ureometer  and  saccharometer,  1286. 
Cone  net,  Birge,  1405. 
Connecting    threads  in  the    tissues    of    Pinus 

sylvestris,  1502. 
Constant  current  on  organisms,  effect  of,  1140. 
Contact  irritability  produced  by  salt  solutions, 

1584. 
Contributions  to  our    knowledge   of   color   in 

photo-micrography,  1 489. 
Corundum,  formation  of,  1144. 
Course  in  biology  in  the    Horace  Mann  High 

School,  1249. 
Course  of  study  in  invertebrate  zoology  in  the 

Marine    Biological    Laboratory   at    Wood's 

HoU,   1 48 1. 
Crocodilians,  Lizards,  and    Snakes    of    North 

America,  1340. 
Crystallography  of  cadmium  and  zinc,  1224. 
Current    Bacteriological    Literature,    reviews, 
1141,1186,   1222,    1266,    1310,    1347,    1391, 
1427,    1472,    154S,   1587. 
Current    Botanical    Literature,  reviews,    1131, 
1 174,    1206,   1255,   1299,   1332,    1381,    141 1, 
1460,  1502,  1535,   1573- 


Current  Zoological  Literature,  reviews,  1212, 
1260,  1339,  1385,  1418,  1465,  1506,  1541.  1579. 

Cycas  revoluta,  tuber-like  rootlets  of,  1334. 

Cytology,  Embryology  and  Microscopical 
Methods,  reviews,  1133,  1176,  1208,  1257, 
1301,  1334,  13S2,  1414,  1462,  1504,  1537, 
1576. 

Damp  chamber  for  use  with  klinostat,  1499. 

Daphnia  magna,  reactions  of,  to  certain  changes 
in  its  environment,   11 40. 

Demarcation  current  of  a  resting  nucleus,  1425. 

Demonstration  of  reactions  of  unicellular  or- 
ganisms, 1222. 

Demonstration  of  reticulate  vessels,   1356. 

Description  of  the  new  wing  of  the  laboratory 
of  hygiene  at  the  University  of  Pennsylvania, 

1234- 

Development  of  embryo  of  mouse  up  to  time 
it  is  encapsuled  in  the  decidua  reflexa,  1386. 

Development  of  pollen  grain  of  Carex,  117^. 

Device  for  leveling  the  microscope,  1458. 

Device  for  rearing  dragon  flies,  1277. 

Device  for  supporting  Pasteur  flasks,  11 57. 

Diatoms,  modern  conception  of  the  structure 
and  classification  of,   14 13. 

Difflugia,  experiments  on,   1464. 

Digestive  processes  and  functions  of  the  an- 
terior part  of  the  alimentary  tract  in  selach- 
ians, 1470. 

Diphtheria  bacillus  in  the  mouths  of  healthy 
individuals,  1 145. 

Directing  marks  in  microscopical  sections  for 
the  purpose  of  reconstruction  by  wax  plate 
modeling,  1537. 

Directions  for  Beginners  in  Bacteriology,  13 10. 

Double  fertilization  in  maize,  141 1. 

Drones  from  the  unfertilized  eggs  of  bees,  1577. 

Easy  method  of  mounting  and  preserving 
mosquitoes,  11 29. 

Echinococcus  alveolaris  and  multilocularis, 
1544. 

Ehrlich's    triacid    mixture  for    staining  blood, 

I3'4- 
Ehrlich-Weigert  anilin    methyl-violet    method 

for  staining  tubercle  bacilli,  1395. 
Elastic  fibers  of  the  skin,  method  of    staining, 

1423- 

Elastic  tissue  in  the  lung,  methods  of  demon- 
strating,  1338. 

Electrical  conductivity  of  the  blood,   1423. 

Embryochemical  studies,  1426. 

Embryo  sac  of  Peperomia,   1299. 

Endurance  of  pest  bacilli  in  pure  cultures,  1472. 

Enzymes  in  cheese,  11 86. 

Eosinophilic  cells,  origin  of,  1422. 

Eosinophilic    cells,   staining,    1417. 

Eosinophilic  leucocytes  in  tumors,  1 508. 

Epilepsy  parasite,  1188. 

Epithelial  cells  in  mouth  and  pharynx  of  sal- 
amander larvoe,  1383. 

Erigenia  bulbosa,  morphological  and  anatom- 
ical study  of,  1175. 

Essentials  of  Practical  Bacteriology,  13 10. 

Examination  of  blood,  1593. 

Exclusion  and  elimination  of  pathogenic  bac- 
teria from  sewage,  1551. 

External  osmotic  pressure  upon  green  algs, 
effect  of,  1 574. 

Eye  spot  of  Euglena  viridis,  1 133. 


INDEX. 


Eyes  of  hydromedusae,  structure  of,   12 13. 
Eyes  of  lower  vertebrates,  histology  and  physi- 
ology of,  1586. 
Fermentation  of  bread,  abnormal,  1222. 
Fermentation  tube  adapted  to  rapid  handling 

in  routine  work,  1404. 
Fermentation  tubes  for  bacteriologic  investiga- 
tions of  fermentation,  1286. 
Flagellata  in  Engler  &  Prantl's  "Die  Naturlichen 

Pfianzenfamilien,"  1261. 
Flattening  and  fixing  paraffin  sections  on  slide, 

1196. 
Formalin  as  a  preservative  of    zoological    ma- 
terial, 1339. 
Formalin  as  a  reagent  in  blood  studies,  121 1. 
Formalin  solutions,  table  of,  1435. 
Formol  as  a  fixing  fluid,   1257. 
Formulae : 

Culture  medium  for  germination  of  spores, 

1 148. 
Decolorizing  solution  for  staining   in  toto 

with  Delafield's  hasmatoxylin,  1172. 
Ehrlich's  tracid  mixture  for  staining  blood, 

1314- 
Fluid  for  pure  cultures  of  Infusoria,  1221. 
Fluids  for  fixing  and  preserving  zoological 
specimens     and    anatomical    material, 
15S2. 
Hydrochloric  acid  alcohol,   11 46. 
Kresylechtviolett,  1492. 
Kultschitzki's  hematoxylin,  11 79. 
Mallory's  stain  for  tumors,  1137. 
Nutrient  gelatin,  1165. 
Nutrient  salt  solution,  114S. 
Phloxin-methylen  blue  for  blood,  1305. 
Preservative  for  Crustacea,  1341. 
Saturated  solutions  of  anilin  stains,  1397. 
Unna-Tanzer's  orcein  solution,  1146. 
Fresh  water  micro-fauna  and  flora,  treatise  on, 

1212. 
Gabbett's  method  for  staining  tubercle  bacilli, 

1395- 

General  methods  for  the  study  of  the  nervous 
system,  1557. 

General  Physiology,  reviews,  1138,  1182,  12 18, 
1264,  1307,  1343,  1389,  1423,  1470,  1510, 
1546. 

General  Physiology  for  high  schools,  based 
upon  the  nervous  system,  1346. 

Gill  clefts  in  the  lizard,  origin,  growth,  disap- 
pearance and  derivations  of,  1419. 

Ginkgo  biloba,  fertilization  in,  1332. 

Gland-like  carcinoma  of  epidermic  origin,  1 180. 

Glands  of  stomach  in  different  stages  of  activ- 
ity, 1504. 

Glucinum,  micro-chemical  analysis  of,  1373, 
1497- 

Glycogen  in  parasitic  worms,  1346. 

Golden  calcite,  1313. 

Golgi's  rapid  method  for  chrome-silver  impreg- 
nation, 1561. 

Gram's  method  for  staining  diphtheria  bacilli, 
1476. 

Gregarines,  cellular  changes  caused  by,   1506. 

Growing  tip  of  Allium,  1396. 

Gymnosperms,  morphology  and  development 
of,  1503,  1573. 

Haemolymph  glands,  normal  histology  and 
-  pathology  of,  1387. 


Haine's  list  for  sugar  in  urine.  1353. 

Hair-cells  and  ciliated-cells,  1133. 

Handbook  of  systematic  botany,  1460. 

Harlequin  fly,  structure  and  life  history  of, 
1506. 

Harriman  Alaska  expedition,  1385. 

Helosis  guayanensis,  embryo  sac  of,  1461. 

High  power  photo-micrography,  1 1 58. 

Hippolyte  varians,  color  changes  in,  1182. 

Home  life  of  wild  birds,  1419. 

Home  made  wall  charts,  1195. 

Hydroids  of  the  Wood's  Holl  region,  1541. 

Illinois  Gulch  meteorite,  1352. 

Immersion  oil  in  collapsible  tubes,  1567. 

Immunity,  Buchner  on,  1427. 

Improved  automatic  microtomes,   1317. 

Improvised  microtome,  1162. 

Indium  in  Tungsten  minerals,  1394. 

Individuality  of  the  germ  nuclei  during  the 
cleavage  of  the  egg  of  Crepidula,  1577. 

Inflammation,  histology  of,  1421. 

Infusoria,  acclimatisation  of,  to  chemical  media, 
1220. 

Infusoria,  ciliate,  galvanotaxis  and  chemotaxis 
of,  1 5 10. 

Inhibition  by  artificial  section  of  the  fission 
plane  in  Stenostoma,  130S. 

Inoculation  against  rinderpest,  11 43. 

Intravascular  growth  of  certain  endothelio- 
mata,  1469. 

Introduction  to  Practical  Bacteriology  for  Stu- 
dents and  Practitioners  of  Comparative  and 
Human  Medicine,  1142. 

Introduction  to  Study  of  Zoology  for  use  of 
High  Schools  and  Academies,  1578. 

Invertebrates  of  North  America,  Synopses  of, 
1420. 

Iodine  compounds  in  the  tissues  after  the  ad- 
ministration of  potassium  iodide,  1426. 

Iron  haematoxylin  and  orange-G,  staining 
nerve  tissue  with,  1557. 

Irritable  phenomena  in  living  organisms,  1425. 

Isopods  of  Atlantic  coast  of  North  America, 
1420. 

Kaiserling  method  for  the  preservation  of 
pathological  specimens,  1592. 

Karyokinetic  spindle  in  pollen-mother-cells  of 
Lavatera,  development  of,  1131. 

Koch's  new  views  concerning  bovine  and 
human  tuberculosis,  1548. 

Kresofuchsin,  a  new  stain,  1414. 

Kresylichtviolett,  a  new  general  stain,  1492. 

Labeling  mounted  specimens,  1566. 

Laboratory  camera  stand,  1202. 

Laboratory  courses   by  correspondence,  1437. 

Laboratory  equipment  of  the  "  Bahama  Expe- 
dition "  from  the  University  of  Iowa,   1229. 

Laboratory  Guide  in  Elementary  Bacteriology, 
1310. 

Laboratory  photography : 

An  improved  photo-micrographic  appara- 
tus, 1366. 
Contributions  to  our  knowledge  of  color  in 

photo  micrography,  1489. 
High  power  photo-micrography,  1158. 
Laboratory  camera  stand,  1202. 
Photographing  diatoms,  1439. 
Photo-micrography :  I,  Introductory,  1399 ; 
II,  Apparatus,  1525. 


VI 


INDEX. 


Processes  of  photomicrography,  1199. 

Stereo-photo-micrography,  11 13. 

The  ortol  developer,  1487. 

The   photo-micrography   of    tissues    with 

simple  apparatus,  1283. 
The  value   of   the   telephoto   lens,    1241, 
1568. 
Lepra  bacilli  in  nasal  cavities  of   lepers,  1391. 
Leucocytes,  methods  for  fixation  and  differen- 
tiation of,  1 177. 
Leucocytes  in  the  thymus  in  bony  fishes,  origin 

of,  1 30 1. 
Lichens,  association   of  alga  and  fungus  in, 

1208. 
Lime  cysts  on  the  surface  of  the  kidney,  1306. 
Lithium,  micro-chemical  analysis  of,  1191. 
Lymph  cells  in  the  marrow  of  the  bones,  1258. 
Magnesium,  micro-chemical  analysis  of,  1376, 

1495,  1497- 
Magnifiers,  1448. 
Malarial  parasitology,  1304. 
Marine  Biological  Laboratory  at  Cold  Spring 

Harbor,  L.  I.,  1279. 
Melonite,  1226,  135 1. 
Mercury  in  urine,  estimation  of,  11 46. 
Mesogamy  in  Cucurbita,  1413. 
Metallic  sodium  in  blowpipe  analysis,  1393. 
Method  for  collection  and  preservation  of  par- 
asitic worms,    1580. 
Method  for  rearing  Amoeba,  1566. 
Method  of  determining  the  comparative  grav- 
ity of  alcohol  when  dehydrating  by  osmosis, 
1409. 
Method  of  injecting  small  vessels,  1282. 
Methods  for  use  in  the  study  of  Infusoria,  1 579. 
Methods  : 

Beale's   carmme   for   isolated  nerve  cells, 

1564. 
Chenzinski's  method    for  staining  gono- 

coccus,  1515. 
Chrome-silver     impregnation    for     nerve 

tissue,  1560. 
Clinical  quantitative  analysis  of  proteids 

in  stomach  contents,  12 16. 
Contrast    staining    of    blood    corpuscles, 

1335- 

Cultivating  and  staining  the  diphtheria 
bacillus,  1 3 14. 

Demonstrating  arrangement  of  cilia  on 
Paramecium,  1566. 

Detection  and  determination  of  sugar  in 
urine,  1353. 

Determining  the  comparative  gravity  of 
alcohol  when  dehydrating  by  osmosis, 
1409. 

Differentiation  of  blood  of  various  ani- 
mals, 1268. 

Double  staining  of  cartilage,  1337. 

Embedding  in  celloidin,  1 539. 

Estimation  of  mercury  in  urine,  1146. 

Examination  of  blood,  1 145. 

Experiments  upon  the  formation  of  en- 
zymes by  bacteria,   1587. 

Finishing  laboratory  desk  tops,  1240. 

Flattening  and  fixing  paraffin  sections  on 
slides,  1 196. 

Germination  of  spores,  1147. 

Golgi's  rapid  method  for  chrome-silver  im- 
pregnation, 1 561. 


Gram's    method    for    staining  diphtheria 

bacilli,  1476. 
Hardening  nerve  tissue  with  formaldehyde, 

1562. 
Injecting  small  vessels,  1282. 
Kresylichtviolett,  staining  with,  1492. 
Labeling  mounted  specimens,  1566. 
Making    preparations     of   bone   marrow, 

1259. 
Making  slides  of  Amoeba,  1450. 
Malarial  parasitology,  1304. 
Measuring  crystals,  1225. 
Methylen  blue  and  erythrosin  method    for 

nerve  cells,  1560. 
Mountingand  preserving  mosquitoes,  1 129. 
Mounting  mouth  parts  of  cray  fish,  1435. 
Neisser's  method  of  staining  gonococcus, 

1515. 
Nissl's  method   for   staining   nerve   cells, 

1558. 
Orienting  and  cutting  small  opaque  ob- 
jects, 1505. 
Osmic  acid  for  staining   peripheral   nerve 

fibers,  1564. 
Phenyl-hydrazin  test  for  sugar,  1268. 
Preparing  permanent  specimens  to  dem- 
onstrate structure  of  earthworms,  1409. 
Preservation    of    pathological  specimens, 

1592. 
Preservation  of  sputum  for  microscopical 

examination,   1268. 
Preserving   bacteria   for    museum    speci- 
mens, 1268. 
Preserving  Crustacea,  1341. 
Quantitative  test  for  bacteria  in  milk  with 

low  magnifying  power,  1596. 
Rearing  Amoeba,  1566. 
Schiitz  method   of  staining   gonococcus, 

1515. 
Simultaneous   staining    of   blood  smears 

with  eosin  and  methylen  blue,  1227. 
Staining  bacteria  in  root  tubercles  of  legu 

minous  plants,  1528. 
Staining    connective    and    elastic    tissue, 

1262. 
Staining  echinococcus,  1545. 
Staining  elastic  fibers  of  the  skin,  1423. 
Staining  elastic  fibers  in  sputum,  1146. 
Staining  eosinophile  cells,  141 7. 
Staining  fat,  11 78,  1227,  1508. 
Staining  flagella,  1472. 
Staining  gonococcus,  1515. 
Staining  in  toto  with   Delafield's  haenia- 

toxylin,  1 172. 
Staining  Infusoria,  1435. 
Staining  large  numbers  of    sputum    speci- 
mens, 1594. 
Staining  malaria  parasite,  1435. 
Staining  nerve  cells,  1 180. 
Staining  nerve  tissue,  1557. 
Staining  sections  for  class  work,  1194. 
Staining  sections  of  tumor,  1137. 
Staining  sections  while  embedded  in  paraf- 
fin, 121 2. 
Staining  tubercle  bacilli,  1395,  1472. 
Studying  Infusoria,   1579. 
Tallqvist's  method  of  haemoglobin  estima- 
tion, 1596. 
Testing  for  B.  coli  in  water,  1403. 


INDEX. 


Weigert's    myelin    stain  for  nerve  fibers, 

1563- 
Widal  test  in  diagnosis  of  typhoid  fever, 

1434- 
Methods  in  Plant  Histology,  1412. 
Micro-chemical  analysis : 

X.     Potassium,  1121. 
XI.     Ammonium,  1189. 
XII.     The  analytical  reactions  of  group, 
II;  calcium,   1242. 

XIII.  Strontium,  barium,  1289. 

XIV.  Barium,  continued,  1323. 

XV.     Magnesium      group ;     glucinum, 

XVI.     Zinc,  1 45 1. 
XVII.     Magnesium    group    separations ; 
glucinum,      magnesium,      zinc, 
cadmium,  1495. 
XVIII.     Aluminum,  1529. 
Microtome,  an  improvised,  1162. 
Microtome,  automatic  wheel,  131 7. 
Microtome,  new  freezing,  for  use  with  carbon 

dioxide  tanks,  1320. 
Microtome,  precision,   13 18. 
Mitosis  in  the  spore-mother-cells  of  Lilium,i  175 

1206. 
Mitotic  division  in  unfertilized  Arbacia  eggs, 

1139- 
Modern  Photography  in  Theory  and   Practice, 

1204. 
Mollusks,   monograph    on   the  Tectibranches, 

1507- 

Mosquitoes,  1465. 

Mosses  with  a  Hand  Lens,  1299. 

Mucous  plug  in  surface  epithelium  of  stomach, 
origin  of,  1 134. 

Multiple  primary  malignant  tumors,    1583. 

MultipUcation  of  nuclei  in  striated  muscle  of 
vertebrates,  1336. 

Muscle  fibers,  methods  in  study  of,  121 1. 

Muscles  in  frogs  of  both  sexes  at  different  sea- 
sons of  the  year,  1309. 

Mycetozoa,  preliminary  study  of,  in 9. 

Myeloma,  case  of  multiple,  1 181. 

Myocarditis,  relati6n  of,  to  sclerosis  of  coro- 
nary arteries,  1341. 

Naias  major,  fertilization  in,  1575. 

Natrolite,  1144. 

Nature  and  distribution  of  the  new  tissue  in 
cirrhosis  of  the  liver,  1263. 

Nelumbo,  embryogeny  of,  1575. 

Nematodes,  collection  and  preservation  of, 
1581. 

New  biological  laboratories  of  Ripon  Col- 
lege, 1 1 49. 

New  blood  stain,  1305. 

New  freezing  microtome  for  use  with  carbon 
dioxide  tanks,  1320. 

New  medical  laboratories  of  the  University 
of  Pennsylvania,  1445. 

New  meteoric  iron,  131 2. 

New  stain  for  fibrin,  1227. 

New  test  for  chlorine  for  use  with  blow  pipe, 

1352- 
New  thermo-regulator,  1449. 
New  York  botanical  garden,  1 1 15. 
Nissl's  method  for  staining  nerve  tissue,  1558. 
Nodule  organisms  in  leguminous  plants,  1551. 
Non-nucleated  organisms,  cytology  of,  1207. 


Normal  and  Pathological  Histology,  reviews, 
1 136,  1 180,  1214,  1262,  1304,  1341,  1387, 
1421,  1468,  1508,  1544. 

Normal  electrode  for  physiological  work,  1471. 

Nostoc  commune,  heterocysts  in,  1381. 

Notes  on  examination  of  blood,  1145. 

Notes  on  Recent  Mineralogical  Literature,  re- 
views, 1 144,  1224,  1312,  1350,  1393,  1431, 
1474,  1513,  1553. 

Notes  on  testing  for  B.  coli  in  water,  1403. 

Nuclear  division  in  Protozoa,  1464. 

Nucleoproteids  of  the  brain,  1426. 

Nucleus,  function  of,  1135. 

Oedogoniacea;,  morphology,  and  taxonomy  of, 

1535- 
Oligocheta,  anatomy  and  histology  of  nervous 

system  of,  1542. 

Organography  of  plants,  1131. 

Orcein  in  the  demonstration  of  elastic  fibers  in 
sputum,  1 146. 

Organisms  of  nitrification,  1347. 

Orientation  of  organisms,  1425. 

Origin  of  the  cones  of  the  multipolar  spindle 
in  Gladiolus,  1413. 

Ortol  developer,  1487. 

Osmic  acid  for  staining  peripheral  nerve  fibers, 
1564. 

Osmotic  pressure  as  a  factor  in  biological  phe- 
nomena, 1345. 

Otter  seine  for  the  exploration  of  the  deeper 
seas,  1542. 

Pachymeningitis  hsemorrhagica  interna.    12 14. 

Parthenogenesis  in  Antennaria  alpina,  1255. 

Pellia,  nuclear  studies  on,  1333. 

Pelomyxa,  notes  on  rearing,  1260. 

Pelomyxa,  some  results  of  feeding,  1260. 

Permeability  of  red  blood  corpuscles  for  differ- 
ent substances,  1424. 

Pheln's  method  of  staining  the  malaria  para- 
site, 1435. 

Phenyl-hydrazen  test  for  sugar  in   urine,  1353. 

Photo-micrography,  1399,  1525. 

Photo-micrography  of  tissues  with  simple  ap- 
paratus, 1283. 

Phototropism  in  some  arthropods,  1390. 

Physiological  characteristics  of  the  cell,  1307. 

Physiological  studies  on  the  blood  of  animals 
deprived  of  the  adrenals,  1586. 

Physiology  in  the  Detroit  High  School,  1585. 

Physiology  of  growth,  1343- 

Pigment  of  brown  induration  of  the  lung, 
1584. 

Pigmentation  cirrhosis  of  the  liver  in  hasmo- 
chromatosis,  1342. 

Plan  for  a  ureometer,  1 156. 

Planaria  maculata,  physiology  of,  with  special 
reference  to  regeneration,  1265. 

Planarians,  with  and  without  eyes,  reactions  of, 
1 138. 

Plimmer's  bodies  in  carcinoma,  1138. 

Pneumonia  fibrosa  chronica,  1262. 

Point  in  the  technique  of  blood  counting,  1226. 

Poisonous  effects  of  pure  NaCl  solution,  1221. 

Polarity  among  marine  Algje,  1333. 

Polarity  of  the  ovocyte,  egg  and  larva  of 
Strongylocentrotus  lividus,  1576. 

Potassium,  micro-chemical  analysis  of,  1121. 

Preliminary  study  of   Mycetozoa,  in 9. 

Pre-Medical  course  at  Purdue  University,  1591. 


INDEX. 


Preservation  of  sputum  for  microscopical  exam- 
ination, 1268. 

Primary  endotheliomata  of  the  left  superior 
pulmonary  vein,  1468. 

Protozoa,  morphology,  taxonomy  and  ecology 
of,  1 4 18. 

Pseudo  diphtheria  bacilli,  varieties  of,  1430. 

Pseudomonas  hyacinthi  (Wakker),  hyacinth 
germ,  1267. 

Pseudo  tetanus  bacillus,  1473. 

Pulmonary  gangrene,  bacilli  of,  1223. 

Pupation  of  larvae  of  insects  under  limited 
amount  of  air,  1 546. 

Purdy's  method  for  quantitative  determination 
of  sugar  in  urine,  1353. 

Pus,  causes  for  formation  of,   1588. 

Pycnogonids,  habits  of,  1308. 

Pycnopodia  helianthoides,  multiplication  of  the 
rays  and  bilaterial  symmetry  in,  1214. 

Quantitative  test  for  bacteria  with  low  magni- 
fying power,  1 596. 

Quartz,  1145. 

Queen  torch,  1228. 

Quick  and  simple  method  of  fixing  the  blood 
corpuscles  for  differential  staining,   1306. 

Rapid  method  of  making  slides  of  Amoeba, 
1450. 

Reaction  of  Eritomostraca  to  stimulation  by 
light,  1 1 83. 

Regeneration  in  Cystopteris,  1300. 

Relative  susceptibility  of  domestic  animals  to 
the  contagion  of  human  and  bovine  tuber- 
culosis, 1310. 

Respiration  of  Desmognathus,  1135. 

Rheumatic  fever,  etiology  of,  1473. 

Rhythmic  activity  of  the  oesophagus,  influence 
of  various  media  on,  1585. 

Ringing  slides,  1595. 

Root  tubercles  in  leguminous  plants,  1348. 

Rubidium  and  cesium,  1 125. 

Scarlet  fever,  etiology  of,  1142. 

Schizea  pusilla,  life  history  of,  1206. 

Scrofulous  glands,  study  of,  1428. 

Sealing  stone  jar  for  zoological  laboratories, 
1261. 

Secretion  and  absorption,  morphological 
changes  in  the  intestinal  epithelium  during, 
1415. 

Separation  of  alumina  from  molten  magmas, 
1 144. 

Septic  vibrio  and  symptomatic  anthrax,  compar- 
ative study  of,  1429. 

Short  method  for  the  Widal  test,  1565. 

Silage,  causes  operative  in  formation  of,  1186. 

Silicate,  theory  of,  1553. 

Silicates,    action  of   ammonium    chloride  on, 

1393- 

Simple  washing  device,  1297. 

Smegma  bacillus,  1223 

Soft  rot  on  carrot  and  other  vegetables,   1267. 

Source  of  leucocytes  and  the  true  function  of 

the  thymus,  1208. 
Species  hepaticarum,  1535. 
Spermatogenesis  in  Batrachoseps,  1 540. 
Spermotogenesis  in  Staphylinus,  1577. 
Spermatozoa  of  ferns,  physiology  of,  1 536. 
Spematozoa   in    man,    domestic   animals,  and 

rodents,   1360. 
Sphalerite  crystals  from  Galena,  Kansas,  1352. 


Spiral  swimming  of  organisms,  151 2. 

Sporodinia,  nuclei  of  the  zygospores  in,   1332. 

Stain  for  simultaneous  staining  of  blood  smears 
with  eosin  and  methylen  blue,  1227. 

Staining  bacteria  in  the  root-tubercles  of  legu- 
minous plants,  1528. 

Staining  in  toto  with  Delafield's  haematoxylin, 
1172. 

Staining  the  diphtheria  bacillus,  1476. 

Staining  sections  for  class  work,  1194. 

Staining  nerve  tissue,  1557. 

Staphylococci,  production  of  toxines  by,    1589. 

Staphylococcus  albus,  morphology  of,    11 43. 

Static  diffusion  of  gases  and  liquids  in  relation 
to  the  assimilation  of  carbon  and  transloca- 
tion in  plants,  1256. 

Stereo-photo-micrography,  11 13. 

Sterilized  air,  physiological  effects  of,  on  ani- 
mals, 1346. 

Stokesite,  1 145. 

Striated  muscles,cancers  and  sarcomas  in,  12 15. 

Striated  muscles,  normal  and  pathological  his- 
tology of,  12 1 5. 

Strontium,  micro-chemical  analysis  of,  1289. 

Study  of  bacteria  in  the  public  schools,  1164. 

Studying  and  photographing  the    wild    bird, 

1517- 

Substitutes  for  hydrochloric  acid  in  testing  car- 
bonates, 1352. 

Sulvanite,  a  new  mineral,  1352. 

Swarm  spore  formation  in  Hydrodictyon  utricu- 
latum,  1300. 

System  der  Bacterien,  Migula's,  1347. 

Table  of  specific  gravities  of  saturated  solu- 
tions and  solubilities  of  anilin  stains,  1397. 

Tallqvist's  method  of  haemoglobin  estimation, 
1596. 

Technic  for  malaria  blood,  1382. 

Technic  of  blood  preparations,  1524. 

Technic  of  the  Widal  test,  1434. 

Telephoto  lens  in  photography,  value  of,  1241, 
1568. 

Tetrad  division  in  a  hybrid  plant,  11 74. 

Tetrad  formation  in  the  ovule  of  Larix,  11 74. 

Theory  of  phototactic  response,  1264. 

Thigmotaxis  in  Protozoa,    1184. 

Thyroid  gland,  experiments  on,  1583. 

Thyroid  gland,  pathology  and  diseases  of,  1 509. 

Tobacco,  fermentation  of,  1186. 

Tourmaline,  constitution  of,  1312. 

Tracheal  cells  of  the  Oestridse,  1176. 

Transmission  of  tuberculosis  by  postage 
stamps,  1429. 

Trematodes,  collection  and  preservation  of, 
1580. 

Tropical  dysentery,  etiology  of,  1141. 

Tube  filter  for  concentrating  Infusoria  in 
small  amount  of  liquid,  1579- 

Tubercle  bacilli  in  dairy  products,  131 1. 

Tubercle  bacillus,  life  duration  of,  in  cheese, 
1 187. 

Tubercle  bacillus,  thermal  death  point  of,  1223. 

Tuberculosis  of  the  heart,  1215. 

Tubularia  mesembryanthemum,  regulatory  pro- 
cesses of,  1 218. 

Tulipa  gesneriana,  life  history  of,    1381. 

Tunicate,  alternation  in  direction  of  heart  beat 
in,  1387. 

U-cell,  for  filtration  in  study  of  Infusoria,  1579. 


INDEX. 


University  of  Montana  Biological  Laboratory, 

1269,  1354, 
Ureometer,  plan  for,  11 56. 
Vaccinia  and  variola,  etiological  agent  in,  1429. 
Value  of  methylen  blue  as  an  intravitam  stain 

in  the  Tunicata,  1357. 
Value  of  the  telephoto  lens,  1241. 
Ventilated  dish  for  bacteria  cultures,  1197. 
Vertebrates,  development  of,  1212. 
Veterinary  College  at  Cornell,  fire  in,  iiii. 
Walnut  bacteriosis,  1587. 
Weigert's  Myelin  stain  for  nerve  fibres,  1563. 
Weigert's  neuroglia  method  in  demonstration 

of  fat  necrosis,  1416. 


Welsbach  light  in  high  power  microscopy, 
1356. 

White,  Moses  C,  life  of,  1109. 

Widal  test,  short  method  for,  1565. 

Widal  test,  technic  of,  1434. 

XylariacesE,  Minnesota,  1535. 

Yarn  siphon,  for  separating  Infusoria  from  cul- 
ture water  and  debris,  1579. 

Yellow  fever,  etiology  of,  1349. 

Ziehl-Neelson  method  for  staining  tubercle 
bacilli,  1395. 

Zinc,  micro-chemical  analysis  of,  1451,  1498. 

Zoologisches  Addressbuch,  14 19. 


INDEX. 


INDEX  OF  AUTHORS. 


Bardeen,  Charles  R. 

New  Freezing  Microtome  for  use  with  Car- 
bon-dioxide Tanks,  1320. 
Beal,  W.  J. 

Demonstration     of     Reticulate     Vessels, 
1356- 
Bessey,  Charles  E. 

Home  Made  Wall  Charts,   1195. 
Birge,  E.  a. 

The  Cone  Net,  1405. 
Boston,  L.  Napoleon. 

Spermatozoa  of  Man,  Domestic    Animals 
and  Rodents,  1360. 
Buxton,  B.  H. 

An  Improved  Photo-micrographic  Appara- 
tus,  1366. 
Chamberlain,  Charles  J. 

Current     Botanical    Literature,     reviews, 
1 131,  1 174,  1206,  1255,  1299,  1332,  1381, 
141 1,  1460,  1502,  1535,  1573. 
Laboratory  Courses    by    Correspondence, 
1437- 
Chamot,  E.  M. 

Micro-Chemical  Analysis : 
X.  Potassium,  1 121. 
XL  Ammonium,  1189. 
XI L  Analytical    Reactions   of     Group 
II,  1242. 

XIII.  Strontium,  Barium,  1289. 

XIV.  Barium  (continued),  1323. 
XV^.   Magnesium  Group,  1373. 

XVI.  Zinc,  Cadmium,   1451. 
XVII.  Magnesium    Group    Separations, 

1405. 
XVIII.  Aluminum,  1529. 
Claypole,  Agnes  M. 

Cytology,  Embryology  and    Microscopical 
Methods,    reviews,     1133,     1176,    1208, 
1257,  1301,  1334,  1382,  1414,  1462,  1504, 
1537,  I5^>7. 
Coi.E,  Leon  J. 

A  Method  for    Injecting    Small    Vessels, 
1282. 
CoXN,  H.  W. 

Current  Bacteriological  Literature.reviews, 
1 141,  1 186,  1222,  1266,  1310, 1347,  1391, 
1427,  1472,  1548,  1587. 
CouK,  Mel  T. 

Method  for  Rearing  Amoeba,  1^66. 

Davis,  B.  M. 

Flattening  and    Fi.xing    Paraffin    Sections 
on  .Slide,  1 196. 

De.\rness,  J. 

Magnifiers,  1448. 


Dennis,  D.  W. 

Photo- Micrography :   Introduction,    1399; 
An  Apparatus  adapted  to  All  Kinds  of 
Work,  1525. 
Dodge,  Charles  Wright. 

A  Short  Method  for  the  Widal  Test,  1565. 
Immersion  Oil  in  Collapsible  Tubes,  1567. 
The  Arrangement  of  Cilia  on  Paramecium, 
1566. 
Elrod,  Morton  J. 

The    University    of     Montana    Biological 

Station,  1269. 
The  Value  of  the  Telephoto    Lens,    1241. 
Further  Note  on    the    Use  of  the   Tele- 
photo  Lens,  1568. 
Evans,  Newton. 

Staining  in  Toto  with    Delafield's  Hnema- 

toxylin,  1172. 
Staining  Sections  for  Class  Work,   1194. 
Flexner,  Simon. 

The    New    Medical    Laboratories    of   the 
University  of  Pennsylvania,   1445. 
French,  G.  H. 

The  Epilepsy  Parasite,  1188. 

Fulton,  W.  A. 

Note  on  Examination  of  Blood,  1145. 
Gage,  S.  H. 

Fire  in  the  Veterinary  College  at  Cornell, 

mi. 
Moses  C.  White,  11 09. 
Gage,  Stephen  DeM. 

Notes  on  Testing  for  B.    Coli    in    Water, 
1403. 
Golden,  Katherine  E. 

A  Device  for  Supporting  Pasteur  Flasks, 
1157. 
Grave,  Caswell. 

The  Course  of  Study  in  Invertebrate  Zool- 
ogy in  the  Marine  Biological  Laboratory 
at  Wood's  Holl,   14S1. 
IIerrick,  Francis  H. 

Studying   and     Photographing    the    Wild 
Bird,  1517. 
Higgins,  F.  W. 

A  Point  in  the  Technic  of    Blood    Count- 
ing,  1226. 
IIousER,  Gilbert  L. 

General  Methods    for   the    Study    of   the 
Nervous  System,   1557. 
Howard,  Orson. 

Biology  Wall  Charts,  1172. 
Hunter,  George  W.,  Jr. 

The  Value  of  Methylen  Blue  as  an  Intra- 
vitam  Stain  in  the  Tunicata,  1357. 


INDEX. 


KoFoiD,  Charles  A. 

Current    Zoological    Literature,    reviews, 
121 2,  1260,  1339,  13S5,  1418,  1465,  1506, 
1541,  1578. 
Langenbeck,  Clara. 

Preliminary  Study  of  Mycetozoa,  1 1 19. 
Latham,  V.  A. 

Easy  Method    of  Mounting  and    Preserv- 
ing Mosquitoes,   1129. 
Leavitt,  Robert  G. 

Simple  Washing  Device,  1297. 
Leroy,  Louls. 

Table  of  Specific  Gravities    of    Saturated 
Solutions    and     Solubilities    of    Anilin 
Stains,  1397. 
MacDougal,  D.  T. 

The  New  York  Botanical  Garden,   11 15. 
Marsh,  C.  Dwight. 

The  New  Biological  Laboratories  of  Ripon 
College,   1 1 49. 
McClung,  C.  E. 

High  Power  Photo-Micrography,  11 58. 
The  Processes  of  Photo-Micrography,  1 1 99. 
Minot,  Charles  S. 

Improved  Automatic  Microtomes,  13 17. 
Miyake,  Kiichi. 

The  Botanical  Laboratory  and  the  Botan- 
ical   Garden    of   the   Tokyo    Imperial 
University,  Japan,  1477. 
Morse,  Ralph  L. 

Kresylechtviolett,  1492. 
Moses,  Alfred  J.  and  Luquer,  Lea  McI. 
Notes  on  Recent  Mineralogical  Literature, 
reviews,  1144,  1224,    1312,    1350,    1393, 
1431.  1474,  1513.  1553.  1589- 
Myers,  P.  C. 

Photographing  Diatoms,   1439. 
Nichols,  J.  B. 

A  Plan  for  a  Ureometer,  11 56. 
Nutting,  C.  C. 

The  Laboratory  Equipment  of  the  "  Ba- 
hama Expedition  "  from  the  University 
of  Iowa,  1229. 

Palmer,  Thos. 

A  New  Thermo-Regulator,  1449. 

Patterson,  W.  L. 

A  Combined  Condenser  and  Polarizer  for 
Petrographical  Microscopes,  1155. 

Peabody,  James  E. 

The  Study  of  Bacteria  in  the  Public 
Schools,   1 164. 


Pearce,  Richard  M. 

Normal  and    Pathological    Histology,    re- 
views,  1 136. 
Pearl,  Raymond. 

General  Physiology,  reviews,    1138,    1182, 
1218,  1264,  1307,  1343,  1398,  1423,  1470, 
1 5  ID,  1546,  1584. 
Peirce,  George  J. 

Staining  Bacteria  in  the  Root-tubercles  of 
Leguminous  Plants,  1528. 
Powers,  Irwin  LaV. 

An  Improvised  Microtome,  1162. 
Pratt,  H.  S. 

The    Marine     Biological     Laboratory    at 
Cold  Spring  Harbor,  L.  I.,  1279. 
Pratt,  Joseph  H. 

Normal  and    Pathological    Histology,    re- 
views,   1 1 80,    1 214,    1262,    1304,    1 34 1, 
1387,  1421,  1468,  1508,   1544,  1583. 
Rawlins,  B.  L. 

A  Few  Remarks  on  the  Technic  of  Blood 
Preparations,  1524. 
Reed,  Howard  S. 

A  Damp  Chamber  for  Use  on    the  Klino- 
stat,  1499. 
Reynolds,  T.  O. 

Device  for  Leveling  the  Microscope,  1458. 
Robin,  A. 

Combined  Ureometer  and  Saccharometer, 
1286. 
Rolfs,  P.  H. 

A  Laboratory  Camera  Stand,  1202. 
Shaw,  C.  H. 

Botany  at  the    Biological    Laboratory   at 
Wood's  Holl,  i486. 
Walmslp:y,  W.  H. 

The  Photo-Micrography  of   Tissues    with 
Simple  Apparatus,  1283. 
Whipple,  George  C. 

A  Ventilated  Dish  for    Bacteria  Cultures, 
1197. 
WiLLCOX,  M.  A. 

A  Rapid  Method  of  Making  Slides  of 
Amoeba,  1450. 

WOLCOTT,  ROBT.    H. 

A  Modification    of    the    Birge    Collecting 
Net,  1407. 
Woodford,  R.  P. 

A  Method  of  Determining  the  Compara- 
tive Gravity  of  Alcohol  when  Dehydrat- 
ing by  Osmosis,  1409. 

The  Ortol  Developer,  1487. 


INDEX. 


INDEX  OF  AUTHORS  REVIEWED. 


AliUUTT,   II.   S. 

Modern  Photography  in  Theory  and  Prac- 
tice, 1204. 

AiutoTT,  M.  E. 

Pigmentation  Cirrhosis  of  the  Liver  in  a 
case  of  IIa;mochromatosis,  lij42. 

Aknoldi,  W. 

Heilrrige  zur  Morphologic  einiger  Clym- 
nospermen,  150;],  Ihl'^. 

Arrico,  G. 

Ueber  die  Gegenvvart  und  iiber  die    Pha- 
sen    des    Kochschen    Bacillus    in    den 
sogenannten  skrophulosen  Lymphdrus- 
scn,  1428. 
Atkinson,  EMZAiiKTii  A. 

Indium  in  Tungsten  Minerals,  1394. 
Babcock  and  Russel. 

Causes    Operative   in    the    Formation    of 

Silage,  118G. 
Relation  of  the    Enzymes    of    Rennet    to 
Ripening  of  Cheddar  Cheese,   1180. 

Bakdkkn,  C.  R. 

On  the  Physiology  of  the  Planaria   macu- 
lata  with  especial  reference  to    the  Phe- 
nomena of  Regeneration,   12(i5. 
Barrows,  A.  S. 

Respiration  of  Desmognathus,  118."). 

Ba'I'aii.i.on,  E. 

La  pression  osmotique  et  les  grands  prob- 
Icmes  de  la  Biologie,  i:!45. 

Baum,  E. 

Ueber  die  punktformigen  Kalkkorperchen 
(sogen-verkalkte  Glomerule)  der  Nier- 
enrinde,  180(). 

Baum,  J. 

Beitrjige  zur  Kenntniss    der    Muskel-spin- 
deln,  1211. 
Beard,  J. 

The  Source  of  Leucocytes  and    the    True 
Function  of  the  Thymus,    1208. 
Bker,  Til. 

Ueber  primitive  Sehorgane,  loSO. 
Behrens. 

Ueber    die     oxydierenden    Bestandtheile 
und  die    Fermentation    des    Ueutschen 
Tabaks,  1180. 
Beijerinck. 

Anhiiufungsversuche  mit  Ureumbakterien, 
i;U7. 

Benda,  C. 

Eine  Makro-  und  Mikrochcmische  Reac- 
tion der  Fettegewebsnecrose,  1410. 


Benedici,  a.  L. 

Clinical  Quantitative  Analysis  of  Proteids 
in  Stomach  Contents,  121(5. 
Berg  II,  R.  S. 

Bietriige  zur  Vergleichende  Histologic  II. 
Ueber  den    Bau    der   Gefasse   bei    den 
Anneliden,  12l;J. 
Kleinere      histologische      Mittheilungen, 
i:]85. 

Bernard,  Cu. 

Recherches  sur  les  spheres  attractives 
chez  Lilium  candidum,  Helosis  guaya- 
nensis,  etc.,  1401. 

Bessey,  Charles  E. 

The  Modern  Conception  of  the  Structure 
and  Classification  of  Diatom.s,  with  a 
Revision  of  the  Tribes  and  a  Rearrange- 
ment of  the  North  American  Genera, 
14i;-5. 

BlCKKL,  A. 

Beitrage  zur  Gehirnphysiologie  der  Schild- 
krote,   ir)4(). 

BlEI.SCIIOWSKY  AND   PlIEN. 

Zur  Technik  der  Nervenzellenftirbung, 
1180. 

B  IE N stock. 

Du  role  des  Bacteries  de  I'intestin,  KMit. 
Bliesener. 

Bietrag  zur  I^ehre  von  Sporenbildung,  1392. 

Bock,  M.  de. 

Observations  Anatomiques  et  Ilistologi- 
ques  sur  les  Oligochetes  specialement 
sur  leur  Systcme  Musculaire,  1542. 

Bonnier,  P. 

L'Orientation,  1425. 

BosioN,  L.  Napoleon. 

How  to  Preserve  as  Permanent  Specimens 
Casts  found  in  Urine,    147(). 

BOVKRI,    Th. 

Die  Polaritiit  von  Ovocyte,  Ei  und  Larve 
des  Strongylocentrotus  lividus,  1570. 

Brand,  F. 

Bemerkungen  iiber  Grenzzellen  und  id^er 
spontanrothe  Inhaltskorper  der  Cyano- 
phycea:,  13SI. 

Britton,  Elizabeth  G.,  and  Taylor,  Alex- 
andria. 

Life  History  of  Schizea  pusilla,  1200. 

Brown,  II.  T.,  and  Escomhe,  F. 

Static  Diffusion  of  Gases  and  Liquids 
in  Relation  to  the  Assimilation  of  Car- 
bon ami  Translocation  in   Plants,  1256. 


INDEX. 


BUCHNER. 

Immunitat,  1427. 
BULLER,  A.  H.  R. 

Contribution   to    our    Knowledge   of   the 

Physiology     of     the     Spermatozoa    of 

Ferns,  1536. 

BURCKHARD,    G. 

Die  Implantation  des  Ei  der  Mans  in  die 
Uterusschleimhaut  und  die  Umbildung 
derselben  zur  Decidua,   1386. 

BUSQUET. 

Transmission  de    la   tuberculose    par    les 
timbrespost,    1429. 
Butters,  F.  K. 

A  Preliminary  List    of    Minnesota    Xylar- 
iaceae,  1535. 
Byxbee,  Edith  S. 

The    Development    of    the    Karyokinetic 
Spindle   in    the     Pollen-mothercells  of 
Lavatera,  1131. 
Cade,  A. 

Les  Elements  Secreteurs  des  glandes  gas- 
triques    du   fond  chez  les  mammiferes, 
1504. 
Campbell,  D.  H. 

The  Embryo-sac  of  Peperomia,  1299. 
Carlgren,  O. 

Ueber  die  Einwirkung  des  constanten  gal- 
vanischen  Stromes  auf   niedere    Organ- 
ismen,  1140. 
Chamberlain,  Charles  J. 

Methods  in  Plant  Histology,  1412. 
Chapman,  F.  M. 

Bird  Life :  A  Guide  to  the  Study  of    Our 
Common  Birds,  1418. 
Chodat,  R.,  and  Bernard,  C. 

Sur   le     sac    embryonnaire    de    I'Helosis 
guayanensis,  1461. 
Clarke,  F.  W. 

The  Constitution  of  Tourmaline,  1312. 
Clarke,  F.  W.,  and  Steiger,  G. 

Experiments  Relative  to  the  Constitution 
of    Pectolite,    Pyrophyllite,     Calamine, 
and  Analcite,  1513. 
The  Action  of  Ammonium  Chloride  upon 

Analcite  and  Leucite,  1393. 
The  Action  of  Ammonium  Chloride  upon 
Natrolite,  Scolecite,  Prehnite,  and    Pec- 
tolite, 1393. 
Cloetta,  M. 

Kann  das  Medicamentose    Eisen    nur   im 
Duodenum  resorbirt  werden  ?  1537. 
Clowes  and  Houston. 

The  Bacterial  Treatment  of  London  Sew- 
age, 1430.    ^ 
CoE,  W.  R. 

Papers  from  the  Harriman  Alaska    Expe- 
dition, XX,  The  Nemerteans,  1385. 
Cole,  L.  J. 

Notes  on  the  Habits  of  Pycnogonids,  1308. 

CONKLIN,  E.  G. 

Centrosome  and  Sphere  in  the  Matura- 
tion, Fertilization,  and  Cleavage  of 
Crepidula,  1505. 


Individuality  of  the  Germ  Nuclei  during 
the  Cleavage  of  the  Egg  of   Crepidula, 
1577. 
Conn,  H.  W. 

How  can   Bacteria  be  Satisfactorily  Pre- 
served for  Museum  Specimens  ?    1268. 
Cope,  E.  D. 

The  Crocodilians,  Lizards,  and  Snakes  of 
North  America,  1340. 
Coulter,  Stanley. 

Pre-medical  Course  at  Purdue  University, 
1.591. 
Courmant. 

L'agglutination  der  bacille  de   Koch  des 
epauchements  tuberculeux,  1142. 
COURTADE,   D. 

LTrritabilite  dans  la  Serie  Animale,  1425. 
Curtis,  H.  J. 

Essentials  of  Bacteriology,  1310. 
Dains. 

A  Pseudo  Tetanus  Bacillus,  1473. 

Dale,  H.  H. 

Galvanotaxis  and   Chemotaxis  of  CiHate 
Infusoria,  1510. 
Dangeard,  p.  a. 

La   reproduction  sexuelle  des    Champig- 
nons,   1535. 
Nuclear  division  in  Protozoa,  1464. 
Davis,  Bradley  M. 

Nuclear  Studies  on  Pellia,  1333. 
Delage,  Y.,  and  Delage,  M. 

Sur  les  relations  entre  le  constitution 
chimique  des  produits  sexuels  et  celle 
des  solutions  capables  de  determiner 
la  parthenogenese,  1185. 

Dewitz,  J. 

Verhinderung  der  Verpuppung  bei  Insek- 
tenlarven,  1546. 

Dinwiddie. 

The  Relative  Susceptibility  of  Domestic 
Animals  to  the  Contagion  of  Human 
and  Bovine  Tuberculosis,  1310. 

Dixon,  H.  H. 

On  the  first  mitosis  of  the  spore-mother- 
cells  of  Lilium,  1206. 

DOFLEIN,    F. 

Cell  division  in  Protozoa,  1382. 

Driesch,  H. 

Studien  liber  das  Regulationsvermogen 
der  Organismen,  1218. 

Du  Sablon,  Leclerc. 

Recherches  sur  les  fleurs  cleistogames, 
1536. 

Eastes,  G.  L. 

Note  on  the  Phenyl-Hydrazin  Test  for 
Sugar,  1268. 

ECKLES. 

An  Abnormal  Fermentation  of  Bread, 
1222. 

Edmunds,  N. 

The  Pathology  and  Diseases  of  the  Thy- 
roid Gland,  1509. 


INDEX. 


ElJKMANN. 

Ueber  Enzyme  der  Bakterien  und  Schim- 
melpilzen,  1587. 
ElSEN,  G. 

The    Spermatogenesis   of    Batrachoseps, 
1540. 

ElSMOND,  J. 

Ueber  die  Natur  der  Sogenannten  Kinet- 
ischen  Centren  der  Zellen,  1302. 
Erben,  F.,  and  Ceipek,  L. 

Analyse  der  Albits  von  Amelia,  1514. 
Ernst,  A. 

Beitrage  zur  Kenntniss  der  Entwickelung 
des    Embryo-sackes   und   des    Embryo 
(Polyembryonie)     von    Tulipa    Gesner- 
iana,  1381. 
EwiNG,  J. 

Malaria]  Parasitology,  1304. 
Farrington,  O.  C. 

Publications   Field    Columbian    Museum, 
1313. 
Feldbausch,  F. 

Ueber  das  Vorkommen  von  eosinophilen 
Leucocyten  in  Tumoren,  1508. 
Fisher,  Alfred. 

Die    Empfindlichkeit    der    Bakterienzelle 
und  das  baktericide  Serum,  1429. 
Fletcher,  L. 

On   a   Mass  of    Meteoric   Iron  from   the 
neighborhood  of  Caperr,  Rio  Senguerr, 
Patagonia,  1226. 
Flexner. 

The  Etiology  of  Tropical  Dysentery,  1141. 
Flexner,  S. 

Nature  and  Distribution  of  the  New  Tis- 
sue in  Cirrhosis  of  the  Liver,  1263. 
FOLSOM,  J.  W. 

The  Development   of  the  Mouth  Parts  of 
Anurida  maritima,  1210. 
Foote,  K.,  and  Strobell,  E.  C. 

Egg  of  Allolobophora  foetida,  1338. 
Foote,  W.  M. 

Note  on  New  Meteoric  Iron,  1312. 
Ford. 

The  Bacteriology  of  Healthy  Organs,1266. 
FOUQUE,   F. 

Contribution   a  I'etude  des  mineraux  de 
group  de  la  meiilite,  1351. 
Fraenkel. 

Zur  Kenntniss  der  Smegma  bacillus,  1223. 
Friedel,  G. 

Nouveaux  essais  sur  les  Zeolites,  1144. 
Frost,  W.  D. 

A  Laboratory  Guide  in   Elementary  Bac- 
teriology, 1310. 
FucHS,  E. 

Beitrage  zur   Kenntniss    der    Entstehung 
des   Vorkommens  und    der  bedeutung 
"  eosinophiler  "  Zellen,  mit  besonderer 
Beriicksichtigung  des  Sputums,  1422. 
Fujinami. 

Ueber  das  Histologische  Verhalten  des 
Quergestreiften  Muskels  an  der  Grenze 
bbsartiger  Geschwiilste,  1215. 


Ueber  die  Beziehungen  der  Myocarditis 
zu  den  Erkrankungen  der  Arterienwan- 
dungen,  1341. 
Fulleborn. 

Ueber  Formalingonservierung,  1339. 

FUNCK. 

A    Preliminary  Note    on    the    Etiological 
Agent  in  Vaccinia  and  Variola,  1429. 
FURST,   C.   M. 

Haarzellen  und  Flimmerzellen,  1133. 
Galloway,  T.  W. 

Studies  on  the  Cause  of  the  Accelerating 
Effect  of  Heat  upon  Growth,  1221. 
Gamble,  F.  W.,  and  Keeble,  F.  W. 

Hippolyte    varians ;     a    Study   in    Color 
Changes,  1182. 
Gareiss,  a. 

Ueber   Pseudomorphosen  nach   Cordierit 
&  Tschermak's,  1475. 
Gaule,  J. 

Ueber  den  Einfluss  der  Jahreszeit  auf  das 
Gewicht    der    Muskehi    bei    Froschen, 
1309. 
Glinski,  L.  K. 

Zur    Kenntniss  des   Nebenpankreas   und 
verwandter  Zustande,  1387. 
Godlewski,  E. 

O,  rozmnazanin  jader  w  niesniach  prazko- 

wanych  zwierzat  kregowych,  1336. 
O  wpywie   tlenu  na  rozwoj    organismow    i 
o  wymianie   gazow  w  pierwszych    stad- 
yach  rozwoju  zarodka  u  Rana  tempor- 
aria,  1586. 
GOEBEL,    K. 

Organographie  der  Pftanzen  insbesondere 
der  Archegoniaten  und  Samenpflanzen, 
1131. 
Golden,  Katherine  E. 

Aspergillus  oryzse,  1536. 

Goldhorn  L.  B. 

A  Rapid  Method  of  staining  the  Chroma- 
tin of  the  Malaria  Parasite,  1306. 

Goldschmidt,  V. 

Chrysoberyllzwilling  von  Ceylon,  1433. 
Ueber  Erkennung  eines  Zwillings,  1225. 
Ueber  Trogerit  und   kiinstlechen  Uranoc- 

pinit,  1394. 
Zur  Theorie  der  Zwillings-  und   Viellings- 

bildungen,  1433. 

Gonnard,  F. 

Etude  cristallographique    du    quartz   des 

geodes    des    marnes    oxfordiennes    de 

Meylan,  1145. 
Sur  un  groupe  de  cristaux  de  quartz  de 

Striegan,  1145. 

GoGDER,  G.   A. 

Sulvanite,  a  new  mineral,  1352. 

Greenough,  R.  B. 

On  the  Presence  of  the   So-called  "  Plim- 
mer  Bodies"  in  Carcinoma,  1138. 

Grimsley,  G.  p.,  AND  Bailey,  E.  H.  S. 

Report  on  Gypsum  and  Gypsum  Cement, 
1394. 


INDEX. 


Gromakowsky. 

Varieties    of    Pseudo    Diphtheria    Bacilli, 
1430. 
Grout,  A.  J. 

Mosses  with  a  Haiid-lens,  1299. 

Gruber,  Eduard. 

Ueber  das  Verhalten  der  Zellkerne  in  den 
Zygosporen  von  Sporodinia  grandis, 
Link.,  1332. 

Grunling,  Fr. 

Ueber  die  Mineralvorkommen  von  Cey- 
lon, 1432. 

Guiart,  J. 

Les  Mollusques  Tectibranches,  1507. 

GUIGNARD,  L. 

La    Double    Fertilization    dans    le    Mai's, 

1411. 
La   double    ficondation     dans    le    Naias 

Major,  1575. 

GURWITSCH,  A. 

Die  Vorstufen  der  Flimmerzellen  und  ihre 
Beziehungen  zu  Schleimzellen,  1383. 

Hall,  H.  O. 

The  Etiology  of  Scarlet  Fever,  1142. 

Harris,  H.  F. 

A  New  Method  of  Staining  Elastic  Tis- 
sue, 1422. 

Harrison. 

Die  Lebensdauer  des  Tuberkel  Bacillus 
im  Kase,  1187. 

Hartley,  E.  G.  G. 

Communications  from  the  Oxford  Miner- 
alogical  Laboratory  on  the  Constitution 
of  the  Natural  Arsenates  and  Phos- 
phates, 1513. 

Harvey,  N.  A. 

Introduction  to  the  Study  of  Zoology  for 
the  use  of  High  Schools  and  Acade- 
mies, 1578. 

Hauck,  L. 

Untersuchungen  zur  Normalen  und  Path- 
ologischen  Histologie  der  Quergestreif- 
ten  Musculatur,  1215. 

Heidenhain,  M. 

Ueber  die  erste  Entstehung  der  Schleimp- 
fropfe  beim  Oberflachenepithel  des 
Magens,  1134. 

Henninp,  C. 

Depigmenting  the  Eyes  of  Arthropoda, 
1578. 

Herrick,  F.  H. 

The  Home  Life  of  Wild  Birds;  a  New 
Method  of  the  Study  and  Photography 
of  Birds,  1419. 

Hill,  A.  W. 

The  Distribution  and  Character  of  Con- 
necting Threads  in  the  Tissues  of  Pinus 
sylvestris  and  other  allied  species,  1502. 

HiLLEBRANI),  M.  F. 

Mineralogical  Notes  :  Melonite,  Colorado- 
ite,  Petzite,  Hessite,  1226. 


HiLSUM. 

Bakteriologische    Untersuchung    e  i  n  e  s 
Schwimmbades  in  Bezugauf  Selbstrein- 
igung,  1350. 
Hiltner. 

Ueber  die  Ursachen,  welche  die  Grosse, 
Zahl,  Stellung  und  Wirkung  der  Wur- 
zelknollchen    der    Leguminosen    bedin- 
gen,  1.348. 
Hinterberger. 

Eine    Modifikation    des    Geisselfarbungs- 
verfahrens  nach  Ermengen,  1472. 
HiRN,  Karl  E. 

Monographie  und  Iconographie  derOedo- 
goniaceen,  1535. 

HiRSCHMANN,  A. 

Pathologisch -anatomische  Studien  iiber 
acute  u.  chronische  laryngitis  nicht- 
specifischen  Ursprungs  nebst  Bemer- 
kungen  iiber  Vorkommen  von  Plasma- 
und  Mastzellen,  1.544. 
His,  W. 

Ueber  Sogenannte  Amitosis,  1336. 
Hoffman. 

Die  Rolle  des  Eisen   bei  der  Blutbildung. 
Zugleich  ein  Beitrag  zur  Kentniss  des 
Wesens  der  Chlorose,  1462. 
Hoffman,  R.  W. 

Ueber  das  Orientiren  und  Schneiden  mik- 
roskopisch      kleiner,     undurchsichtiger 
und  dotterreicher  Objecte,  1505. 
Holm,  Theodore. 

Erigenia  bulbosa,  Nutt.,  1175. 
Holmes,  S.  J. 

Observations  on  the  Habits  and  Natural 
History  of  Amphithoe  longimana.  Smith, 
1219. 
Phototaxis  in  the  Amphipoda,  1547. 
Holmgren,  N. 

Ueber  den  Bau  der  Hoden  und  die  Sper- 
matogenese  von  Staphylinus,  1577. 
Holt,  E.  B.,  and  Lee,  F.  S. 

The     Theory   of    Phototactic    Response, 
1264. 
Howard,  L.  O. 

Mosquitoes,  1465. 
Howard,  W.  T. 

Observatiohs  on  the  Character  of  the 
Cells  in  the  Exudation  in  Acute  Inter- 
stitial Nephritis,  with  Special  Reference 
to  the  Presence  of  Cells  with  Eosino- 
philic Granulations,  1422. 
Hutchinson,  A. 
Stokesite,  1145. 

Ikeno,   S. 

Contribution  a  I'etude  de  la  fecondation 
chez  le  Ginkgo  biloba,  1332. 

Jennings,  H.  S. 

Demonstrations  of  the  Reactions  of 
Unicellular  Organisms,  1222. 

On  the  Significance  of  the  Spiral  Swim- 
ming of  Organisms,  1512. 

Jensen. 

Studien  iiber  die  Enzyme  im  Kase,  1186. 


INDEX. 


Johnston,  J.  B. 

A  Sealing  Stone  Jar  for  Zoological  Lab- 
oratories, 1261. 

Jolly,  M.  J. 

Rech arches  sur  la  division  indirecte  des 
cellules  lymphatique  granuleuses  de  la 
mole  des  os,  1258. 

Jones. 

Soft  Rot  on  Carrot  and  other  Vegetables, 
1267. 
Jordan. 

Some    Observations   upon    the    Bacterial 
Self-purification  of  Streams,  1266. 
JuDD,  Hidden,  and  Pratt. 

On  a  New  Mode  of  Occurrence  of  Ruby 
in  North  Carolina,  1514. 

JUCKENACK,  A. 

Beitrag  zur  Kenntniss  des  fadenziehenden 
Brotes,  1222. 

JUEL,  H.  O. 

Beitrage  zur  Kenntniss  der  Tetradenbild- 

ung,  1174. 
Vergleichende  Unterschungen  liber  typis- 
che  und  parthenogenetische  Fortpflan- 
zung  bei  der  Gattung  Antennaria,  1255. 

Karlinskl 

Zur  Kenntniss  der  saurefesten  Bakterien, 
1391. 
Keibel,  F.,  AND  Abraham,  K. 

Normentafel  zur  Entwicklungsgeschichte 
des  Huhnes,  1212. 
Kijanitzin,  J.  J. 

Weitere  Untersuchungen  iiber  den  Ein- 
fiuss  sterilisirter  Luft  auf  Thiere,  1346. 

KlZER,  E.  J. 

Formalin  as  a  Reagent  in  Blood  Studies, 
1211. 

KOBEF. 

The  Presence  of  Diphtheria  Bacilli  in  the 
Mouths  of  Healthy  Individuals,  1145. 

KOCKEL. 

New  Stain  for  Fibrin,  1227. 

KoRN,  Otto. 

Weitere  Beitrage  zur  Kenntniss  der  saure- 
festen Bakterien,  1223. 
Kreibich. 

Ueber  bakterien  frei  Eiterung  beim    Men- 
schen,  1588. 
Krompecher. 

Glandlike  Carcinoma  of  Epidermic  Origin, 
1180. 

Lameris  and  Harrevelt. 

Bakterienbefund  in  Kuhnmilch  nach  alge- 
heilter  Mastitis,  1187. 

Lawson,  a.  a. 

Origin  of  the  Cones  of  the  Multipolar 
Spindle  in  Gladiolus,  1413. 

Lechainche  and  Vallee. 

Etude  comparee  du  vibrion  septique  et  de 
la  bacteries  du  charbon  symptomatique, 
1429. 

Levene,  p.  a. 

Embryo-Chemical  Studies,  1426. 


Iodine  Compounds  in  the  Tissues   after 
the  Administration  of  Potassium  Iodide, 
1426. 
On  the  Absorption  of  Proteids,  1426. 
On  the  Nucleoproteids  of  the  Brain,  1426. 
The   Chemical    Relationship    of    Colloid, 
Mucoid,  and  Amyloid  Substances,  1426. 
Levin,  J. 

Physiological   Studies   on    the    Blood   of 
Animals  deprived  of  the  Adrenals,  1586. 
Lewinson,  J. 

Zur  Methode  der  Fettfarbung,  1178,  1227. 
Lewy. 

Die    Beziehungen   der   Charcot-Leydenn- 
ischen    Krystalle   zu    den    eosinophilen 
Zellen,  1387. 
Life,  A.  C. 

The  Tuber-like  Rootlets  of    Cycas  revo- 
luta,  1334. 
LiNKo,  Alex. 

Ueber  den  Bau  der  Augen  bei  den  Hydro- 
medusen,  1213. 
Linser,  P. 

Ueber  den  Bau  und  die  Entwicklung  des 
Elastischen  Gewebes  in  der  Lung,  1338. 
Livingston,  B.  E. 

On  the  Nature  of  the  Stimulus  which 
Causes  the  Change  of  Form  in  Poly- 
morphic Green  Algae,  1574. 

LOEB,  J. 

On  an  apparently  New  Form  of  Muscular 
Irritability  (  Contact  Irritability  ?  )  pro- 
duced by  Solutions  of  Salts  whose  An- 
ions are  Liable  to  form  Insoluble  Cal- 
cium Compounds,  1584. 

LONGO,    B. 

La  mesogamia  nella  commune  Zucca, 
1413. 

Looss,  A. 

Zur  Sammal-  und  Conservierungstechnik 
von  Helminthen,  1580. 

Lyon,  H.  L. 

Observations  on  the  Embryogeny  of  Ne- 
lumbo,  1.575. 

Macallum,  a.  B. 

On  the  Cytology  of  Non-nucleated  Organ- 
isms, 1207. 

MacCallum,  W.  G. 

On  the  Intravascular  Growth  of  Certain 
Endotheliomata,  1469. 

Macy,  M.  L.,  and  Norris,  H.  W. 

A  General  Physiology  for  High  Schools 
Based  upon  the  Nervous  System,  1346. 

Malkes,  J. 

Estimation  of  Mercury  in  Urine,  1146. 

Mallet,  F.  R. 

On  Langbeinite  from  the  Punjab  .Salt 
Range,  122(i. 

Marpmann,  G. 

Eine  neue  Vorschrift  zum  Konservirenen 
von  Zoologischen  und  Anatomischen 
Praparaten,  1582. 


INDEX. 


Martin,  Fr. 

Ueber  Scheinbar   spaltbaren    Quarz    von 
Karlsbad,  1514. 
Matthews,  A.  P. 

Some  Ways  of  Causing  Mitotic  Division 
in  Unfertilized  Arbacia  Eggs,  lloO. 

May,  Richard. 

The  Use  of  Orcein  in  the  Demonstration 
of  Elastic  Fibers  in  the  Sputum,  1140. 

Meek,  E.  R. 

Method  of  Staining  the  Elastic  Fibers  of 
the  Skin,  1423. 

Melczer,  G. 

Ueber  einige  Mineralien  vorwiegend    von 
Ceylon,  14o2. 
Melnikow-Rasweden  KOW. 

Pachymeningitis  Haemorrhagica    Interna, 

1214. 
Studien  iiber  den  Echinococcus  alveolaris 
sive  multilocularis,  1544. 

MiALL,  L.  C,  and  Hammund,  a.  R. 

Structure  and  Life-history  of  the  Harle- 
quin Fly,  1506. 

MlCH.^LIS,    L. 

Ueber  Fett-Farbstoffe,  1.508. 

MiGULA,  W. 

System  der  Bakterien,  lo47. 

Moll,  A. 

Zur  Histochemie  der  Korpels,  IM:]". 

Montgomery,  T.  H. 

A  Study  of  the  Chromosomes  of  the  Germ 
Cells  of  Metazoa,  157S. 

Moore,  A. 

Further  Evidence  of  the  Poisonous  Effects 
of  a  Pure  NaCl  Solution,   1221. 

Moore,  Veranus  A. 

An  Introduction  to  Practical  Bacteriology 
for  Students  and  Practitioners  of  Com- 
parative and  Human  Medicine,  1142. 
Directions  for  Beginners  in  Bacteriology, 
1310. 

Morgan,  Leonard  P.,  and  Smith,  E.  F. 
Experiments  on  Chalcopyrite,  1394. 

Moser,  a. 

Tuberculo.sis  of  the  Heart,  1215. 

Mi'lHLMANN,  M. 

Ueber  die  Ursache  des  Alters.  Grundziige 
der  Physiologie  des  Wachsthums  mit 
besonderer  Beriicksichtigung  des  Men- 
schen, 1343. 

Muller. 

Uber  Tuberkelbacillen  und  Sporenfar- 
bung  unter  Anwendung  von  Kalium  per- 
karbonat  und  Wasserstoffsuperoxyd, 
1472. 

MURBACH,  L. 

Physiology  in  the  High  School,  1585. 

MURBECK,  S. 

Ueber  das  Verhalten  des  PoUenschlauches 
bei  Alchemilla  arvensis  und  das  Wesen 
der  Chalazoganiie,  1381. 


Murray. 

A  Preliminary  Report    on     acid   resisting 
bacilli  with    special    reference    to    their 
occurrences  in  the  lower  animals,  1391. 
Neisser  and  Wechsberg. 

Ueber  das  Staphylotoxin,  1589. 
Neumann,  E. 

Das   Pigment  der  braunen  Lungenindura- 
tion,  1584. 
Neuwirth,  V. 

Titanit  von  der  Hiittellehne    bei    Werms- 
dorf  un  Mahren,  1514. 
Newkeld. 

Beitrag  zur  Kenntniss  der    Smegma  bacil- 
lus, 1223. 
Nichols,  E.  H. 

On  the  Etiology  of  Cancer,  1136. 
Nichols,  Henry  W. 

A  New  Test  for  Chlorine  for  Use  with  the 
Blow-pipe,  1352. 

NiKOLSKY. 

Charbon  chez  des  animaux  nourris    avec 
leur  ailments  habituel,  meles  de    spores 
charbonneuses,  1551. 
Noack,  W. 

Beitrage  zur  Entwicklungsgeschichte    der 
Musciden,  1-578. 
Nceoske,  H. 

Eosinophile    Zellen    und     Knochenmark, 
insbesondere    bei    chirurgischen    Infec- 
tionskrankheiten     und     Geschwiilsten, 
1417. 
Noll,  F. 

Ueber  die  Umkehrungsversuche  mit  Bryo- 
psis,    nebst    Bemerkungen    iiber   ihren 
zelligen  Aufbau  (Energiden),  1333. 
NussBAUM,  J.,  u.  Prymak,  T. 

Zur  Entwickelungsgeschichte  der  eympho- 
iden  Elemente  der  Thymus  bei  dem 
Knochenfischen,  1301. 

Nutting,  C.  C. 

The  Hydroids  of  the  Wood's  Holl  Re- 
gion, 1541. 

Obermuller. 

Ueber  neuere  Untersuchung  des  Vorkom- 
men  echter   Tuberkuloseereger    in   der 
Milch  und  dem  Molkereiprodukten  be- 
treffend,  1311. 
Oker-Bi.om,  M. 

Eine  Normal  Electrode  fiir  physiologische 

Zwecke,  1471. 
Thierische  Safte  und  Gewebe  in    physika- 
lisch-chemischer  Beziehung,  1423. 

Overton,  E. 

Studien  iiber  die  Aufnahme  der  Anilinfar- 
ben  durch  die  lebende  Zelle,    1334. 

Palache,  Charles. 

The  Crystallization  of  Calcite,  from  the 
Copper  Mines  of  Lake  Superior,  1313. 

Palisa,  J. 

Die  Entwickelungsgeschischte  der  Regen- 
erationsknospen,  welche  an  den  Grund- 
stiicken  isolirter  Wedel  von  Cystop- 
teris-Arten  entstehen,  1300. 


INDEX. 


Pappenheim,  a. 

Ueber  das  Vorkomnien  einkerniger  Zellen 
eingonorrhoischen  Urethralsecret,  1421. 
Parker,  G.  H.,  and  Burnett,  F.  L. 

The    Reactions   of    Planarians   with  and 
without  Eyes,  1138. 
Parsons,  Charles  L. 

The  Use  of  Metallic  Sodium  in  Blow-pipe 
Analysis,  1393. 

Peirce,  G.  J. 

The  Nature  of  the  Association  of  Alga 
and  Fungus  in  Lichens,  1208. 

Penard,  E. 

Experiments  on  Difflugia,  1464. 
Penfield,  S.  L.,  AND  Ford,  W.  E. 

Calcites  ( Siliceous)  from  the  Bad  Lands, 
1144. 

Peters,  A.  W. 

Some  Methods  for  Use  in  the  Study  of 
Infusoria,  1579. 

Peters,  K. 

Mittheilungen  zur  Entwicklungsgeschichte 
der  Eidechse,  1419. 

Petersen,  C.  G.  J. 

An  Otter-seine  for  the  Exploration  of  the 
Deeper  Seas,  1542. 

Petroff,  M. 

Neue  Farbungmethod  zur  rothe  Blutkor- 
perchen  in  Schnittpreparaten,  1335. 

Petrunkewitsch,  a. 

Die  Richtungskorper  und  ihr  Schicksal 
im  befruchteten  und  umbefruchteten 
Bienenei,  1577. 

Pierce,  Newton  B. 

Walnut  Bacteriosis,  1587. 

Piorhowska. 

The  Staining  of  Diphtheria  Organisms, 
1476. 

PiORKOwsKi  and  Jess. 

Bacterium  coli  als  Ursache  eines  seuchen- 
artigen  Pferdesterbens  in  Westpreussen, 
1348. 

Poynton  and  Paine. 

The  Etiology  of  Rheumatic  Fever,  1473. 

Pratt,  J.  H. 

On  the  Separation  of  Alumina  from    Mol- 
ten Magmas,  and  the  Formation  of  Cor- 
undum, 1144. 
On  the  Crystallography  of  the  Rubies  from 
Macon  Co.,  N.  C,  1514. 

Prenant,  A. 

Cellules  Tracheales  des  Oestres,  1176. 

Preston,  H.  L. 

Illinois  Gulch  Meteorite,  1352. 

Prior,  G.  T.,  and  Spencer,  L.  J. 

The  Identity  of  Binnite  with  Tennantite, 
and  the  Chemical  Composition  of  Fah- 
lerz,  1513. 

Putter,  A. 

Studien  iiber  Thigmotaxis  bei  Protisten, 
1184. 


Rabinowitsch,  L. 

Befund  von  saurefesten  Tuberkelbacillen 
ahrlichen  Bakterien  bei  Lungengangran, 
1223. 
Ueber  die  Gefahr  der  Uebertragung  der 
Tuberkulose  durch  Milch  und  Milch- 
produkte,  1311. 
Radl,  Em. 

Arthropod  Vision,  1540. 
Ueber  den  Phototropism    einiger   Arthro- 
poden,  1390. 
Reed,  Carroll,  and  Agramonte. 

The  Etiology  of  Yellow  Fever,  1349. 
Regaud,  Cl. 

Quelques  details  sur  la  division  amitotique 
des  Noyaux  de  Sertoli  chez  le  rat,  1303. 
Reichenbach. 

Ueber  Verzweigiing  bei  Spirillen,  1391. 
Reighard,  J.,  AND  Jennings,  H.  S. 
Anatomy  of  the  Cat,  1420. 

Retterer,  E. 

Transformation  de  la  cellule  cartilagineuse 
en  tissue  conjunctiv  reticule,  1335. 

Reuter,  K. 

Zur  Frage  der  Darmsresorption,  1415. 

Richards,  H.  M. 

Ceramothamnion  codii,  a  new  Rhodophy- 
ceous  alga,  1412. 
Richards,  Joseph  W.,   and    Powell,   Nor- 
man S. 

Substitutes  for  Hydrochloric  Acid  in  Test- 
ing Carbonates,  1352. 
Richardson,  Harriet. 

Key  to  the  Isopods  of  the  Atlantic  Coast 
of  North  America  with  descriptions    of 
new  and  little  known  species,  1420. 
Synopses   of     North    American    Inverte- 
brates, VIII,    The  Isopoda,  1420. 

Ritchie. 

The  Bacteriology  of  Bronchitis,  1141. 
Ritter,  W.  E.,  and  Congdon,  Edna  M. 

On  the  Inhibition  by  Artificial  Section  of 
the  Normal    Fission    Plane    of    Stenos- 
toma,  1308. 
Ritter,  W.  E.,  and  Crocker,  G.  R. 

Multiplication  of  the  Rays  and  Bilateral 
Symmetry  in  the  20-Rayed  Starfish, 
Pycnopodia  helianthoides,  1214. 

Robin,  A. 

A  Contribution  to  theTechnic  of  the   Wi- 

dal  Test,  1434. 
Preservation  of   Sputum  for    Microscopic 
Examination,  1268. 

Rogers. 

Schutzimpfung  gegen  Rinderpest,  1143. 

Rogers,  A.  F. 

Sphalerite  crystals  of  a  pecuhar  habit  and 
with  one  new  form  from  Galena,  1352. 

Rosenberger,  R.  C. 

A  New  Blood  Stain,  1305. 

ROTHIG,  p. 

Ueber  einen  neuen  Farbenstoff  Namens 
Kresofuchsin,  1414. 


INDEX. 


Russell  and  Hastings. 

The  Thermal    Death    Point   of   Tubercle 
BacilU,  1223. 
Sabin,  Florence  R. 

An  Atlas  of  the  Medulla   and    Mid-brain, 
1543. 
Sailer,  J. 

Primary  Endothelioma  of  the  Left  Super- 
ior Pulmonary  Vein,  1468. 
Saint-Remy,  G. 

Contribution  a  I'etude  du    developpement 
des  Cestodes,  1340. 
Sand,  Rene. 

fitude  Monographique  sur  le    Groupe  des 
Infusoires  tentaculiferes,  1418. 
Saitl. 

Beitrage  zur  Morphologie    des    Staphylo- 
coccus albus,  1143. 
Sayce,  O.  a. 

A  Method  of  Preserving  Crustacea,  1341. 
Schenck,  F. 

Physiologische    Characteristik    der   Zelle, 
1307. 
Schenck,  R. 

Ueber  die  Dynamik  der  Krystalle,  1432. 
Schonichen,  W.,  und  Kalberlah,  a. 

B.  Eyferth's  Einfachste  Lebensformen  des 
Tier-  und  Pflanzenreiches,  1212. 

SCHULTZ. 

Ueber  die  Lebensdauer  von  Bacillus  pes- 
tis  hominis  in  Reinkulturen,  1472. 
SCHULTZE,  L.   S. 

Untersuchungen  iiber  den  Herzschlag  der 
Salpen,  13S9. 
Seeliger,  O. 

Tierleben  der  Tiefsee,  1.541. 
Senn,  G. 

Flagellata   in    Engler   and     Prantl    "  Die 
Naturlichen  Pflanzenfamilien,"  1261. 
Siedlecki,  M. 

Contribution  a  I'etude  des  changemente 
cellulaires  provoques  par  les  Gregarines; 
1506. 

SjbBRING,  N. 

Ueber  das  Formol  als  Fixirungsfliissigkeit. 
Allgemeines  iiber  den  Bau  der  lebenden 
Zellen,  1257. 

Smith. 

The  Nodule  Organism  of  the  Leguminosae, 
1551. 

Smith,  G.  F.  H. 

A  Three-circle  Goniometer,  1433. 
Note  on  the  Identity  of  Paralaurionite  and 
Rafaelite,  1433. 

Smith,  R.  Wilson. 

The  Achromatic  Spindle  in  the  Spore- 
mother-cells  of  Osmunda  regalis,  1255. 

Smith,  S. 

Note  on  Staining  of  Sections  while  Im- 
bedded in  Paraffin,  1212. 

Stassano,  H. 

Function  of  the  Nucleus,  1135. 


Stepanow,  E.  M. 

Eine  neue  Einbettungsmethod  in  Celloidin 
1539. 
Stephani,  F. 

Species  Hepaticarum,  1535. 
Stephens,  J.  W.  W.,  and  Christopher,  R. 

S.  R. 

Technique  for  Malaria  Blood,  1382. 
Sternberg,  Carl. 

Zur  Kenntniss  des  Aktinomycespilzes, 
1428. 

Stiles,  P.  G. 

On  the  Rhythmic  Activity  of  the  Oesopha- 
gus and  the  Influence  upon  it  of  Vari- 
ous Media,  1585. 

Stober,  F. 

Sur  un  procede  pour  tailler  des  grains 
minerau.x  en  lames  minces,  1226. 

Stolc,  a. 

Beobachtungen  und  Versuche  iiber  die 
Verdauung  und  Bildung  der  Kohlenhy- 
drate  bei  einem  amobenartigen  Organ- 
ismus,  Pelomyxa  palustris  Greef,  1260. 

Stutzer. 

Die  Organismen  der  Nitrifikation,  1347. 

Sultan,  C. 

Beitrag  zur  Kenntniss  der  Schilddriisen 
Function,  1583. 

ten  Siethoff,  E.  G.  a. 

Eine  einfache  Construction  dersogenannte 
Interferenzkreuzes  der  zweiaxigen-Kry- 
stalle,  1431. 

Termier,  p. 

Nouvelle  contribution  a  I'etude  crystallo- 
graphique  du  cadmium  et  du  zinc  met- 
alliques,  1224. 

Sur  la  composition  chimique  et  les  prop- 
erties optique  de  la  leverrierite,  1312. 

Timberlake,  H.  G. 

Swarm  Spore  Formation  in  Hydrodictyon 

utriculatum  Roth.,  1300. 
The  Development  and    Function    of   the 
Cell  Plate  in  Higher  Plants,  1132. 

ToBLER,  Maria. 

Beitrag  zur  Frage  des  Vorkommens  von 
Tuberkel  bacillen  und  anderen  Saure- 
festen  Bacillen  in  der  Marktbutter,  1311. 

Trommsdorff. 

Ueber  Gewohnung  von  Bakterien  an  Alex- 
in, 1143. 

Uhlenhuth. 

Method  for  the  Differentiation  of  the 
Blood  of  various  Animals  with  especial 
Reference  to  the  Demonstration  of  Hu- 
man Blood,  1268. 

Vater,  Heinrich. 

Ueber  den  Einfluss  der  Losungenossen 
auf  die  Krystallization  des  Calcium- 
carbonates,  1394. 

Vernadsky,  W. 

Zur  Theorie  der  Silicate,  1553,  1589. 


INDEX. 


Viola,  C. 

Ueber   das  "  Glaukisiren  "    verschiedener 

Feldspathe,  1850. 
Ueber  optische  Erscheinung  am  Quaiz und 

am  Turmalin  von  Elba,  1474. 
Zur  Kenntniss  des  Anorthits   vom  Vesuv, 

1894. 

VOGEL,  K. 

Zur  Histologie  der  Pneumonia  fibrosa 
chronica,  1262. 

Wager,  H. 

The  eye  spot  of  Euglena  viridis,  1183. 

Wakker. 

Wakker's  Hyacinth  Germ,  Pseudomonas 
hyacinthi  (Wakker),  1267. 
Wallerant,  F. 

Perfectionnement  au    refractometre   pour 
les  cristau.x  microscopiques,  1226. 
Ward,  H.  L. 

New  Meterorite  from  Murphy,  1144. 
Notice  of  an  Aerolite  that  recently  fell  at 
Allegan,  Mich.,  1894. 
Warren,  E. 

On     the     Reaction    of    Daphina    magna 
(Straus)  to  certain    changes   in   its   en- 
vironment, 1140. 
Warthin. 

Accessory    Adrenal    Body   in    the    Broad 
Ligament,  1342. 
Warthin,  A.  S. 

A  Contribution  to  the  Normal    Histology 
and     Pathology     of    the    Haemolymph 
Glands,  1387. 
Wasmann,  E. 

Nervenphysiologie    und  Tierpsychologie, 
1587. 
Weinland,  E. 

Ueber  den  Glykogengehalt    einiger    para- 

sitischer  Wiirmer,  1346. 
Zur  Magenverdauung  der  Haifische,  1470. 

Wei.senfeld. 

Der  Befund  des  Bakterium  coli  in  Wasser 
und  das  Thierexperiment  sind  keine 
brauchbaren  Hilfsmittel  fur  die  hygien- 
ische  Beurteilung  des  Wassers,  1552. 


Wells,  H.  G. 

Multiple     Primary     Malignant    Tumors  ; 
Primary  Sarco-carcinoma  in  the  Thyroid 
of  a  Dog,  with  Mixed  Sarcomatous  and 
Carcinomatous  Metastasis,  1583. 
We'itstein,  R.  von. 

Handbuch  der    Systematischen    Botanik, 
1460. 
Whipple,  Geo.  C. 

Changes  that  Take  Place  in  the  Bacterial 
Contents  of  Waters  during  Transporta- 
tion, 1553. 
Whitney,  W.  F. 

A  Quick  and  Simple  "Method    for   Fixing 
the  Blood    Corpuscles   for   Differential 
Staining,  1806. 
Wili.ebrand,  E.  H. 

Stain  for  Simultaneous  Staining  of  Blood 
Smears  with  Eosin  and  Methylen  Blue, 
1227. 
Wilson,  H.  V. 

Notes  on  a  Species  of  Pelomyxa,  1260. 
Wilson,  J.  T. 

A  New  System  of  Obtaining  Directing 
Marks  in  Microscopical  Sections  for  the 
Purpose  of  Reconstruction  by  Wax-plate 
Modeling,  1537. 

WoROBIEFF,  V.  VON. 

Krystallographische  Studien  iiber  Turma- 
lin   von    Ceylon    und    einiger   anderen 
Vorkommen,  1433. 
Wright,  J.  H. 

A  Case  of  Multiple  Myeloma,  1181. 
WClfung,  E.  a. 

Ueber  die  Lichtbewegung    im    Turmalin, 
1474. 
Vasuda,  a. 

Studien  iiber  die  Anpassungs  fahigkeit  ein- 
iger Infusorien  an  concentrirte    Losun- 
gen,  1220. 
Yerkes,  R.  M. 

Reactions  of  Entomostracato  Stimulation 
by  Light,  1 183. 
Zollikofer,  R. 

Kammerfjirbung  der  Leucocyten,  1177. 


Journal  of 


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UEW   YORK 

O  T  A  rj  5  C  A  L 


Applied  Microscop 

and 

Laboratory  Methods. 


VOLUME   IV. 


JANUARY,   1901, 


Number  1 


MOSES  C.  WHITE. 


(1109) 


1110  Journal  of  Applied  Microscopy 


Moses  C.  White. 

Moses  Clark  White,  born  in  Paris,  Oneida  county,  N.  Y.,  July  24,  1819, 
died  in  New  Haven,  October  24,  1900.  It  will  be  seen  from  this  brief  sum- 
mary that  Dr.  White  was  in  his  eighty-second  year.  His  life  is  one  pleasant  to 
reflect  upon.  Like  the  career  of  so  many  Americans,  it  was  full,  and  showed 
the  vitality  of  this  new  world.  In  1840  he  went  to  the  Cazenovia  Seminary — 
the  seminary  which  has  given  a  start  to  so  many  noble  men  and  women  in  the 
central  part  of  New  York  State.  Here  he  prepared  for  college  and  entered 
Wesleyan,  graduating  with  the  class  of  1845.  For  the  next  two  years  he 
studied  medicine  and  theology  at  Yale,  and  in  1847  went  as  a  medical  mission- 
ary to  Foo  Chow,  China.  Here  he  took  charge  of  a  public  dispensary,  and 
gained  the  confidence  of  all  classes  of  the  people.  Owing  to  illness  in  his  fam- 
ily he  was  compelled  to  return  to  America  in  1853.  He  settled  as  a  physician 
in  New  Haven,  Ct.,  and  in  1857  became  a  teacher  in  the  Yale  Medical  School, 
and  at  the  time  of  his  death  still  held  an  honored  place  in  the  faculty.  His 
work  in  this  school,  dealing  with  the  microscopic  structure  and  pathology  of  the 
body,  naturally  made  him  one  of  the  ardent  advocates  of  the  microscope  in 
medicine,  and  his  work  outside  the  college  had  much  dependence  on  the  micro- 
scope as  the  instrument  of  research  or  demonstration.  Thus  from  18G9  to 
1875  we  find  him  giving  lectures  in  his  alma  mater,  Wesleyan,  on  the  micro- 
scopic structure  of  animals  and  plants.  He  was  naturally  led  to  consider 
various  medico-legal  questions  in  which  the  microscope  played  a  principal  role. 
When  the  great  Reference  Hand-Book  of  the  Medical  Sciences  appeared  some 
ten  years  ago,  one  of  its  most  accomplished  articles  was  the  one  on  "  Blood- 
Stains,"  by  Dr.  White.  Since  that  time  he  has  written  one  or  more  monographs 
on  blood  and  the  determination  of  the  corpuscles  of  different  animals.  He 
has  also  presented  papers  before  the  American  Microscopical  Society  on  various 
topics,  in  which  especially  difficult  phases  of  the  subject  were  handled  with  rare 
skill  and  success.  Even  at  the  last  meeting  of  the  society  in  New  York  he  pre- 
sented a  paper  which  gave  in  the  clearest  manner  the  difticulties  of  photograph- 
ing absorption  bands  in  certain  parts  of  the  spectrum.  His  exposition  was  an 
inspiration  to  the  younger  members,  for  it  showed  how  the  human  mind  could 
triumph  over  difiiculties  by  intelligent  persistence.  Not  only  has  he  presented 
admirable  papers  before  the  Microscopical  Society,  but  his  discussion  of  the 
papers  of  his  fellow  members  was  always  full  of  interest  and  sympathy,  and  it 
was  rare  that  he  did  not  add  some  exceedingly  good  suggestion  which  helped 
the  writer  of  the  paper  and  impressed  all  with  the  fertility  of  his  mind  and  its 
thorough  grounding  in  experience  as  well  as  in  fundamental  principles. 

The  men  who  built  the  foundations  of  American  science  are  fast  passing 
away.  Dr.  White  has  an  honorable  share  in  that  relating  to  microscopy.  "  He 
assisted  largely  in  the  preparation  and  publication  of  Silliman's  Physics,  and 
wrote  the  chapter  on  optics."  His  efforts  to  make  clear  to  classes  the  micro- 
scopic structure  of  organisms  led  him  naturally  to  try  to  so  improve  the  projec- 
tion niicroscope  that  all  could  see  at  once,  and  the  teacher  be  able  to  point  out 


and  Laboratory  Methods. 


1111 


exactly  what  feature  he  wished  to  be  observed.  For  this  he  conceived  of 
special  projection  lenses,  as  one  can  see  by  consulting  p.  194  of  the  first  volume 
of  this  Journal.  The  projection  microscope  will  ultimately  be  a  perfect  instru- 
ment by  the  loyal  and  intelligent  investigation  of  the  problem,  such  as  he  gave. 
It  would  be  cruel  to  begrudge  the  repose  which  a  full  and  noble  life  has 
earned ;  but  we  can  rightfully  hold  fast  to  the  inspiration  which  his  earnest, 
helpful  life  gives,  and  like  him  strive  to  advance  knowledge,  and  "  lend  a  hand." 
Cornell  University.  S.  H.  Gage. 


Fire  in  the  Veterinary  College  at  Cornell. 

November  loth,  in  the  early  morning,  the  New  York  State  Veterinary  Col- 
lege took  fire  and  the  Bacteriological  and  Histological  laboratories  situated  on 
the  third  floor  were  completely  destroyed.  Pictures  of  these  laboratories  were 
published  in  the  Journal  of  Applied  Microscopy,  Vol.  1,  p.  23. 


The  origin  of  the  fire  is  supposed  to  have  been  the  extinguishment  of  the 
gas  owing  to  low  gas  pressure  in  some  of  the  incubators.  Upon  an  increased 
pressure  the  room  was  filled  with  gas  and  ignited  by  the  flame  of  the  incubator, 
which  did  not  go  out.     This  is  simply  hypothesis,  however. 

The  two  pictures  show  very  well  the  conditions  existing  Tuesday  forenoon. 
In  the  laboratory,  the  twisted  girders  which  supported  the  roof,  and  numer- 
ous people  engaged  in  clearing  the  wreck  or  students  trying  to  discover  some  of 
their  lost  property.  The  other  picture  shows  the  east  side  of  the  building 
before  the  fire  was  extinguished. 

The  slow  burning  construction  enabled  the  fire  company  to  hold  the  flames 


1112 


Journal  of  Applied  Microscopy 


to  the  middle  part  of  the  third  floor,  the  two  ends  of  this  floor  being  injured 
only  by  smoke  and  water,  and  the  lower  floors  only  by  water.  Only  the  build- 
ing was  insured.  The  material,  microscopes  and  movable  furniture  were  not 
insured.  Over  forty  microscopes,  each  completely  equipped  with  two-thirds, 
one-eighth  dry  and  one-twelfth  oil  immersion  objectives,  triple  nose-piece,  Abbe 
condenser,  and  two  oculars,  were  completely  destroyed.  The  material  for  the 
courses  in  Pathology,  Bacteriology,  Histology  and  Embryology  were  all  burned, 
besides  much  valuable  material  for  research  which  had  been  collected  with 
much  care  and  no  little  expense  during  the  last  ten  years. 


While  much  was  lost,  much  more  was  saved.  Fortunately  the  most  valuable 
microscopes  and  apparatus  were  stored  in  the  wings  or  ends  of  the  main  build- 
ing, and  these  escaped  except  some  blackening  by  the  dense  smoke. 

By  Friday  evening  a  temporary  roof  had  been  put  over  the  burned  part,  and 
by  utilizing  the  museum  space  on  the  first  floor  for  a  laboratory,  the  work  was 
in  full  progress  the  next  Monday  morning.  The  professors  in  charge  wish  to 
express  their  grateful  appreciation  to  their  colleagues  all  over  the  country  for 
their  generous  offers  of  assistance  ;  they  are  also  grateful  to  the  manufacturers 
and  optical  companies  which  supplied  them  immediately  with  needed  apparatus, 
or  repaired  damaged  instruments.  S.  H.  Gage. 

Cornell  University. 


A  recent  writer  on  Fat-necrosis  finds  alcohol  with  celloidin  imbedding  prefe- 
rable to  formaldehyde  (4  per  cent,  solution),  Miiller's  fluid,  Flemming's  solution 
or  osmic  acid  as  a  fixative  for  necrotic  adispose  tissue  and  specimens  of  pan- 
creas.    Haematoxylin  and  eosin  were  found  the  most  satisfactory  for  staining. 


and  Laboratory  Methods. 


Ill:} 


LABORATORY  PHOTOGRAPHY. 

STEREO  PHOTO-MICROGRAPHY. 

Mr.  John  G.  Baker  has  sent  us  a  number  of  very  interesting  micro-stereographs 
of  insects  and  the  following  description  of  the  apparatus  and  methods  employed 
in  making  them.  The  work  was  first  publicly  exhibited  and  described  June  6th, 
1899,  before  the  Photographic  and  Microscopical  Branch,  Chemical  Section,  of 
the  Franklin  Institute. 


Figure    1. 

This  camera  was  constructed  for  the  purpose  of  making  stereoscopic  pictures 
of  small  objects. 

My  first  attempt  was  in  fitting  up  a  stereoscopic  camera  for  the  purpose,  but 
the  result  was  not  at  all  satisfactory,  although  it  made  some  very  fair  negatives. 
The  camera  proved  to  be  very  much  too  short,  and  the  lens  and  object  had  to 
be  changed  from  one  side  to  the  other,  all  of  which  made  it  very  inconvenient. 

The  next  attempt  is  embodied  in  the  instrument  shown  in  the  illustration.  It 
was  originally  a  lantern  slide  camera,  which  was  altered  to  what  you  now  see. 
The  shutter  used  is  a  4  x  5  "  Victor."  To  the  front  of  this  was  fitted  an  attach- 
ment to  carry  the  lens  and  also  to  hold  a  reflector  for  properly  illuminating  the 
object.  In  the  rear  of  the  shutter,  instead  of  a  lens,  a  ring  was  placed  to  cut 
off  any  reflected  light.  The  rear  end  of  the  camera  has  been  fitted  up  to  receive 
a  5  X  7  plate-holder,  but  in  such  a  way  that  it  may  be  used  in  two  positions,  so 
that  each  end  of  the  plate  may  be  exposed  independently  of  the  other.  The 
plate-holder  rests  against  a  partition  with  an  opening  in  it  of  a  size  just  sufficient 
to  cover  one-half  of  the  plate. 

The  lenses  for  very  small  objects  are  achromatic  objectives  that  are  used  in 
the  microscope,  but  for  this  work  are  changed  somewhat,  to  better  answer  the 
requirements. 

The  trouble  found  with  them  for  the  work  was  their  narrow  angle  of  view 
and  extremely  small  depth  of  focus,  and  each  of  these  faults  had  to  be  remedied 
before  it  was  possible  to  make  a  satisfactory  negative.  It  was  also  found  that 
the  rays  of  light,  in  passing  through  the  lens,  had  a  tendency  to  fog  the  plate  by 
coming  in  contact  with  fiat  surfaces,  even  when  these  were  blackened  with  the 


1114  Journal  of  Applied  Microscopy 

greatest  care.  This  trouble  was  overcome  satisfactorily  by  dispensing  with  the 
flat  surfaces ;  i.  e.,  by  making  them  on  a  bevel,  with  only  the  sharp  edge  to 
reflect  the  light. 

To  do  away  with  the  difliculty  arising  from  the  small  depth  of  focus,  the  only 
way  found  was  to  stop  down  the  lens. 

As  the  depth  of  some  objects  is  very  great  in  proportion  to  the  focal  length 
of  the  lens,  it  necessitates  the  use  of  a  very  small  stop.  The  smallest  stop  used 
by  me  for  this  work  has  a  diameter  of  y^f  „  of  an  inch,  and  the  edges  of  the 
opening  are  made  nearly  sharp  and  carefully  blackened.  The  rear  of  the  lens 
has  also  to  be  guarded  to  prevent  reflections  which  in  this  work  would  be  very 
serious.  Of  course,  the  time  of  exposure  requires  to  be  lengthened  in  proportion 
to  the  size  of  stop ;  many  times  the  exposure  has  taken  over  thirty  minutes, 
and  as  each  exposure  must  be  made  separately  on  the  plate,  the  time  will  be 
doubled. 


Figure  "2. 

Suppose  the  object  to  be  photographed  is  a  small  living  insect.  It  is  placed 
under  a  tumbler  which  has  a  small  hole  drilled  through  the  bottom.  Through 
this  opening  is  injected  a  small  quantity  of  ether.  This  soon  places  the  insect 
in  a  condition  to  be  handled.  We  then  set  it  up  on  its  feet  in  a  position  as  nearly 
life-like  as  possible  on  a  small  piece  of  opalin  glass,  and,  to  hold  it  in  position, 
each  of  its  feet  is  fastened  down  by  means  of  wax.  This  is  done  by  using  a  very 
small  tool,  heated  in  the  flame  of  a  spirit  lamp.  After  the  feet  are  fastened 
properly,  the  insect  is  placed  in  strong  fumes  of  cyanide  of  potassium  to  end  its 
life.  The  surplus  wax  is  now  carefully  removed  by  scraping  it  away  with  a  fine 
pointed  knife. 

The  object  is  now  ready  for  the  camera,  and  upon  the  pedestal  in  front  of  the 
lens  the  mounted  object  is  made  fast.  The  pedestal,  upon  which  the  mounted 
object  is  fastened,  has  a  rack  and  pinion  movement,  so  as  to  elevate  the  object 
to  the  required  height,  and  has  also  a  ball-and-socket  joint  on  the  top,  so  that 
the  object  can  be  placed  in  any  desired  position.     The  image  on  the  focusing 


and  Laboratory  Methods. 


1115 


screen  is  brought  in  position  horizontally  by  sliding  the  lens  and  board,  which 
can  be  done  by  turning  the  milled  head  on  the  top  of  the  camera. 

To  make  the  exposure,  place  the  object  in  its  best  position  and  focus 
as  sharply  as  possible.  With  the  image  in  the  proper  place  on  the  screen,  fasten 
front  and  rear  of  camera  by  means  of  the  clamp  screws  at  the  side,  run  in  the 
plate-holder  until  it  drops  into  the ^rsf  groove.  Set  the  camera  in  position,  with 
the  reflector  facing  a  northern  sky,  and  make  the  first  exposure. 


Figure  8. 

After  the  first  exposure  has  been  made  the  object  has  to  be  rotated.  Upon 
the  pedestal  will  be  found  a  graduated  circle,  divided  into  parts  of  five  degrees 
each,  and  also  a  pointer.  The  insect  has  now  to  be  rotated  one  of  the  gradu- 
ations (five  degrees),  from  left  to  right,  and  then  the  image  on  the  screen  is 
again  placed  in  position,  plate-holder  is  returned  and  run  back  as  far  as  possible 
until  it  drops  into  the  second  groove,  the  exposure  repeated  for  the  other  end  of 
the  plate.  By  revolving  the  object  in  the  direction  just  mentioned,  the  negative 
itself  is  made  stereoscopic,  and  can  be  placed  in  the  stereoscope  and  examined 
to  see  if  it  is  perfect. 


The  New  York  Botanical  Garden. 

"  The  advancement  of  botanical  science  and  knowledge  and  the  prosecution 
of  original  researches  therein  and  kindred  subjects  "  are  some  of  the  primary 
purposes  set  forth  in  the  charter  of  the  New  York  Botanical  Garden,  and  it  may 
be  of  interest  to  botanists  and  biologists  in  general  to  note  to  what  extent  and 
in  what  manner  research  work  may  be  prosecuted  in  this  institution. 

A  prerequisite  to  all  successful  botanical  work  consists  in  the  possession  of 
typical  specimens,  and  in  experimental  work  it  is  important  that  normal  con- 
ditions of  growth  should  be  available,  as  well  as  facilities  for  producing  and 
controlling  experimental  factors. 


11(1 


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and  Laboratory  Methods.  1117 

The  herbarium  contains  nearly  a  milUon  specimens,  and  is  composed  largely 
of  plants  which  have  been  subjected  to  critical  study,  and  are  consequently  well 
identified.  Some  very  notable  collections  are  embraced,  and  it  is  especially  rich 
in  American  forms,  and  in  ferns,  fungi,  and  mosses.  In  addition  to  being  the 
primary  means  of  research  bearing  upon  the  natural  affinities  of  plants,  it  is 
invaluable  as  a  reference  collection  in  morphological  studies.  The  herbarium 
is  increasing  at  the  rate  of  fifty  to  a  hundred  thousand  specimens  annually. 

The  living  plants  include  the  species  native  to  the  Garden  tract,  the  intro- 
duced forms  from  the  temperate  zone  in  the  herbaceous  grounds,  pinetum, 
fruticetum,  arboretum,  viticetura,  nurseries  and  boundary  plantations,  and  the 
tropical  and  desert  forms  in  the  horticultural  and  propagating  houses,  amounting 
to  about  six  thousand  species. 

Before  beginning  an  investigation  of  any  botanical  subject  it  is  of  the  greatest 
importance  that  the  worker  should  familiarize  himself  with  its  botanical  history 
to  learn  what  other  botanists  may  have  written  concerning  it.  To  this  end  he 
must  search  the  volumes  in  the  library.  Periodicals,  books,  pamphlets,  and 
manuscripts  must  be  examined,  and  the  extent  of  known  facts  gotten  well  in 
mind.  The  library  of  the  Garden  now  contains  nearly  nine  thousand  volumes, 
and  is  increasing  at  the  rate  of  over  fifteen  hundred  volumes  annually. 

The  facilities  of  the  Garden  are  open  only  to  students  who  have  demonstrated 
their  ability  to  carry  on  independent  research  work,  and  no  attention  is  given  to 
elementary  instruction.  The  intending  investigator,  having  complied  with  the 
regulations  of  the  institution  and  secured  a  table,  is  placed  in  consultation  with 
the  member  of  the  staff,  or  other  attending  botanists  most  familiar  with  the  sub- 
division of  the  subject  in  which  his  problem  lies,  from  whom  he  receives  only  so 
much  help  and  advice  as  may  be  necessary  to  enable  him  to  carry  his  work  to  a 
successful  end.  The  student  is  free  to  offer  the  results  of  his  work  in  the  form 
of  a  thesis  to  any  university  at  which  he  might  become  a  candidate  for  a  degree. 

The  actual  arrangement  and  extent  of  laboratory  space  and  organization  of 
the  equipment  of  the  laboratories  have  been  carefully  worked  out.  The  upper 
floor  of  the  museum  building,  with  an  area  of  nineteen  thousand  square  feet,  and 
some  special  rooms  in  the  basement,  are  devoted  to  research  work.  The  library 
is  housed  under  the  dome  and  in  a  stack  room  extension  to  the  rear.  The 
physiological  and  morphological  laboratories  occupy  the  western  end,  and  the 
taxonomic  laboratories  and  herbarium  the  eastern  end.  The  laboratories  include 
a  suite  of  fourteen  rooms,  giving  separate  facilities  for  work  in  the  main  divisions 
of  the  subject.  The  equipment  includes  a  supply  of  the  apparatus  necessary  for 
research.  Microscopes  of  the  most  approved  patterns  of  Bausch  &  Lomb,  Leitz, 
and  Zeiss,  with  batteries  of  objectives  of  a  wide  range,  are  found  to  meet  the 
needs  of  the  workers  who  have  used  the  laboratories  to  this  time.  The  photo- 
graphic room  contains  professional  stands  with  the  best  anastigmatic  lenses  of 
Zeiss,  Goerz,  and  Leitz,  projecting  and  photomicrographic  apparatus,  field 
cameras  and  accessories ;  space  is  also  afforded  for  the  precision  balances  of 
the  chemical  laboratories.  The  physiological  dark-room  is  constant  to  tempera- 
tures between  16°  and  21  °C.  The  chemical  laboratories  are  as  yet  only  supplied 
with  the  more  elemental  apparatus.    The  experimental  room  has  an  aquatic  tank, 


1118  Journal  of  Applied  Microscopy 

is  skylighted  and  has  a  cemented  floor ;  its  contiguity  to  the  other  laboratories 
is  a  great  advantage.  A  constant  temperature  room  in  the  basement  has  been 
found  to  furnish  a  satisfactory  thermographic  curve,  although  it  has  not  yet 
been  used  in  research  work. 

A  compartment  in  the  propagating  houses,  comprising  about  a  thousand  feet 
of  floor  space,  is  equipped  for  experimental  work  in  connection  with  the  labora- 
tories, and  ample  space  is  afforded  in  the  plantations  for  the  same  purpose. 

It  is  to  be  seen  that  the  Garden  affords  opportunity  for  research  in  all  of  the 
broader  questions  of  botany,  inclusive  of  climatological  influences,  acclimati- 
zation, history  of  species,  development  of  races  and  varieties,  hybridization  and 
horticultural  practice,  development,  general  morphology,  embryology,  physiology 
and  environmental  relationships  in  general,  and  natural  affinities  of  species  and 
groups. 

The  presence  of  a  number  of  investigators  in  different  phases  of  the  subject 
has  a  most  stimulating  effect  upon  the  individual  student,  and  the  mutual  inter- 
change of  views  does  much  to  counteract  the  tendency  to  over-specialization.  The 
number  of  registered  students  using  the  laboratories,  library,  or  herbarium  during 
the  past  year  was  twenty-eight,  and  most  of  them  were  graduates  of  colleges  and 
universities.  Botanists  from  other  institutions  using  the  facilities  of  the  insti- 
tution, for  periods  from  a  day  to  over  a  month,  numbered  more  than  a  score. 

An  especially  profitable  feature  exists  in  the  weekly  conventions,  at  which 
the  worker  gives  an  account  of  his  own  results,  a  review  of  some  recent  book 
or  article,  or  a  visiting  botanist  gives  an  address  upon  some  subject  of  general 
interest.     Subjects  have  been  recently  presented  as  follows  : 

"A  Summer's  Work  at  the  Royal  Herbarium  at  Kew,"  by  Professor  L.  M. 
Underwood. 

"  Life-history  and  Development  of  the  Gametophyte  of  Schizaea  pusilla,"  by 
Mrs.  Elizabeth  G.  Britton  and  Miss  A.  Taylor. 

"  The  Genus  Lycopodium,"  by  Professor  F.  C.  Lloyd. 

"  Confervae,"  by  Dr.  Tracy  Hazen. 

"  Marine  Flora  of  Bermuda,"  by  Dr.  M.  A.  Howe. 

"  Some  Features  of  the  Flora  of  the  Great  Plains,"  by  Professor  C.  E.  Bessey. 

"  Effect  of  Low  Temperature  upon  the  Growth  of  Sterigmatocystis  nigra," 
by  Miss  Ada  Watterson. 

"  Plants  and  Poisons,"  by  Dr.  R.  H.  True. 

"  Spore  Dissemination  in  the  Sordariaceae,"  by  Dr.  David  Griffiths. 

"  Flora  of  Montana  and  Yellowstone  National  Park,"  by  Dr.  P.  A.  Rydberg. 

"  Anatomy  of  the  Flowers  of  Certain  Grasses,"  by  Mr.  G.  V.  Nash. 

"  Mycorhizas  of  Monotropa,"  by  Dr.  D.  T.  MacDougal. 

"  Embryology  of  Viburnum,"  by  Miss  Nellie  Hewins. 

"Vegetative  Reproductions  of  the  Hepaticae,"  by  Dr.  M.  A.  Howe. 

"  Substances  Isolated  from  Cocoanuts,"  by  Mr.  J.  E.  Kirkwood. 

The  following  outline  shows  the  special  subjects  in  which  investigations  may 
be  carried  on,  together  with  the  name  of  the  person  under  whose  guidance  the 
work  may  be  done.  It  is  to  be  said,  however,  that  almost  any  problem  in  botany 
may  be  taken  up  by  trained  botanists  of  sufficient  experience  who  may  resort  to 


and  Laboratory  Methods.  1119 

the  laboratories  with  the  expectation  of  finding  the  material  facilities  for  their 
work.  The  laboratories  never  close,  and  the  worker  may  find  here  opportunity 
for  work  during  the  summer  vacation  season. 

The  guidance  of  research  work  is  distributed  as  follows : 

Physiology  of  the  Cell— Doctor  MacDougal. 

Ecology — -Professor  Lloyd. 

Morphology  of  Algae — Doctor  Howe,  Doctor  Richards. 

Morphology  of  Fungi — Professor  Underwood. 

Morphology  of  Bryophyta — Professor  Underwood,  Mrs.  Britton. 

Morphology  of  Pteridophyta — Professor  Underwood. 

Morphology  of  Spermatopyta — Doctor  Rydberg. 

Experimental  Morphology — Professor  Lloyd,  Doctor  MacDougal. 

Taxonomy  of  Algae — Doctor  Howe. 

Taxonomy  of  Fungi — -Professor  Underwood. 

Taxonomy  of  Bryophyta — Professor  Underwood,  Mrs.  Britton. 

Taxonomy  of  Pteridophyta — Professor  Underwood. 

Taxonomy  of  Spermatophyta — Doctor  Britton,  Doctor  Small,  Doctor  Rydberg. 

Taxonomy  of  Graminae — Mr.  Nash. 

Embryology  of  Spermatophyta — Professor  Lloyd. 

Special  Taxonomy  (critical  study  of  a  family  or  genus) — Professor  Under- 
wood, Doctor  Britton,  Doctor  Howe,  Doctor  Small,  Doctor  Rydberg,  Mr.  Nash, 
Mrs.  Britton,  Professor  Burgess. 

Regional  Botany — Professor  Underwood,  Doctor  Britton. 

Physiology  of  Nutrition — Doctor  Richards. 

Ecological  Physiology — Doctor  MacDougal,  Doctor  Curtis. 

Physiological  Anatomy — Doctor  Curtis. 

General  Physiology — Doctor  MacDougal,  Doctor  Curtis. 

D.  T.  MacDougal. 


Preliminary  Study  of  Mycetozoa. 

As  a  preliminary  to  the  study  of  slime  moulds,  as  suggested  in  the  article  of 
T.  H.  MacBride'^^  reviewed  in  the  September  number  of  this  Journal,  a  modi- 
fication of  the  method  given  by  Caspar  O.  Miller^ ^^  may  be  of  interest,  especially 
as  by  it  slime  moulds  in  all  phases  of  development  can  be  obtained  at  any  season 
desired,  and  in  such  a  form  as  to  be  suited  to  study  in  the  laboratory. 

Miller  discovered  that  plasmodia  were  developed  in  all  cultures  made  by 
filling  a  beaker  half  full  of  hay  and  covering  the  hay  with  ordinary  tap  water. 
Care  must  be  taken  to  allow  some  of  the  stalks  of  hay  to  project  above  the  water, 
to  serve  as  a  support  upon  which  the  plasmodia  may  climb.  The  beaker  was 
covered  with  a  cotton  plug  to  prevent  the  dust  in  the  air  from  entering.  The 
ordinary  moulds  which  appear  after  a  few  days  were  removed  with  sterilized 
forceps,  care  being  taken  to  loosen  the  under  layers  of  hay,  so  that  some  of  the 


(1)  Jour.  N.  Eng.  Bot.  Club,  2  :  1900. 

(2)  Quar.  Jour.  Min.  Science,  Vol.  41,  N.  S.,  p.  43. 


1120 


Journal  of  Applied  Microscopy 


Fig.  1. 


stalks  always  projected  above  the  water.  After  five  or  six  weeks,  plasmodia  from 
several  centimeters  to  several  inches  in  length  were  seen  to  spread  out  upon  the 
surface  of  the  beaker.  They  seem  to  prefer  the  smooth  surface  of  the  glass, 
perhaps  because  it  offers  the  only  large  surface  above  the  water  upon  which  the 
Plasmodia  can  spread.  From  two  to  twelve  days  after  their  appearance  on  the 
glass  above  the  water,  the  protoplasm  collects  at  one  or  a  number  of  the  points 
at  the  periphery  of  the  network  and  forms  sporangia,  leaving  behind  the 
so-called  hypothallus. 

Acting  upon  this  suggestion,  a  series  of  cultures  were  made  by  partly  filling 
beakers  with  hay,  then  slipping  glass  slides  between  the  hay  and  the  surface 
of  the  beaker,  and  adding  water  until  it  stood  a  little  above  the  middle  of  the 
slides.  Each  beaker  was  covered  with  a  glass  plate  to  prevent  too  rapid 
evaporation  (Fig.  1).  The  beakers  were  allowed  to  stand 
undisturbed  until  the  plasmodia  appeared.  During  the  time, 
however,  the  glass  plate  was  removed  for  an  hour  each 
morning,  for  fear  there  might  otherwise  be  too  great  an 
accumulation  of  CO 2  in  the  beaker.  But  no  further  pre- 
cautions were  taken.  Other  cultures  of  the  same  hay  were 
prepared  every  five  days,  in  that  way  the  various  stages  of 
development  could  be  studied  or  compared  at  any  desired 
time.  It  was  found  that  the  plasmodia  spread  as  readily 
upon  the  glass  slide,  on  the  side  turned  toward  the  surface  of  the  beaker,  as  on 
the  beaker  itself.  At  any  time,  therefore,  a  slide  could  be  taken  from  the  beaker 
and  studied  under  the  microscope  in  its  undisturbed  condition.  It  is  better  not 
to  cover  the  plasmodium  with  a  cover-glass,  as  it  does  not  live  under  water,  but 
just  at  the  surface,  and  either  from  the  pressure  of  the  cover-glass  or  the  excess 
of  water,  it  is  apt  to  go  all  to  pieces.  Very  little  of  the  hay  infusion  need  be 
added  from  time  to  time  to  keep  the  uncovered  plasmodium  moist. 

A  form  which  was  found  most  suitable  to  study  the  streaming  movements  of 
the  protoplasm  developed   in  cultures  made  from  hay  gathered  in  Aurora,  N.  Y., 
during  1S98.     This  same  hay  is  still  used  in  the  laboratory,  and  produces  plas- 
modia as  rapidly  as  it  did  the  first  year  it  was  gathered.     I  have  not  been  able 
to    identify    the     form 
illustrated       two-thirds 
its  natural  size  in  Fig.  2 
and  enlarged  in  Fig.  3. 
It     is    of     an    opaque 
r"'?:-  ■-■  cream  white  color  and 

always  forms  as  figured,  with  a  strong 
branch  just  at  the  surface  of  the  water, 
sending  down  secondary  branches 
somewhat  smaller  in  size,  which  con- 
nect by  still  smaller  branches  with  a 
very  vacuolated  network  just  within 
the  water.  Under  the  microscope  the 
clear  hyaloplasm  zone  and  the  granular  inner  zone  can  be  easily  distinguished ; 


Fig.  3. 


and  Laboratory  Methods.  11-1 

also  the  formation  of  new  branches  by  the  coalescence  of  the  pseudopodia  can 
be  observed.  It  will  not  be  necessary  to  go  into  further  details,  for  I  have 
nothing  new  to  add  to  the  general  description  given  in  the  English  edition  of 
DeBary's  Comparative  Morphology  and  Biology  of  Fungi,  Mycetozoa,  etc.  (p. 
425)  ;  all  that  is  spoken  of  there  can  be  readily  seen  and  followed.  In  fact, 
for  laboratory  demonstration  of  the  streaming  movements  of  protoplasm  to 
beginners,  this  plasmodium  has  proved  of  far  more  use  than  the  Amoeba  or  the 
other  objects  usually  used,  as  Chara  tips,  Tradescantia  hairs,  etc. ;  because, 
being  macroscopic  in  size,  you  waste  no  time  finding  it,  and  not  having  any 
very  dense  cell  walls,  at  the  ends  of  the  network  at  least,  the  motion  of  the  proto- 
plasm can  be  clearly  seen ;  besides,  being  able  to  put  one's  hands  on  it  any  time 
one  wishes  is  not  the  least  of  its  advantages. 

If  the  water  is  allowed  to  evaporate,  the  course  of  the  plasmodium,  following 
the  surface  of  the  water,  can  be  easily  traced  by  the  "  envelope  "  of  DeBary,  or 
the  "  hypothallus  "  of  Miller,  which  remains  behind,  and  the  outlines  of  which 
are  accentuated  by  the  refuse  gathered  around  it,  as  shown  in  Fig.  3. 

Permanent  mounts  of  plasmodia  may  be  made  by  plunging  the  slides  upon 
which  they  are  spread  into  strong  alcohol,  or  a  solution  of  picric  acid  in  strong 
alcohol.  The  alcohol  seems  to  be  necessary  to  coagulate  the  albuminous  sub- 
stances, and  so  fix  the  plasmodium  to  the  slide  at  the  same  time  that  it  is  killed. 
If  aqueous  killing  fluids  are  used,  such  as  picro-sulphuric  or  corrosive  acetic,  a 
considerable  evolution  of  gas  is  seen  to  arise  from  the  plasmodium  (probably 
CO 2  from  the  CaCOj),  and  sooner  or  later  it  floats  down  from  the  slide,  and 
it  is  difficult  to  successfully  remount  it.  With  the  alcohol  the  most  delicate 
threads  remain  intact.  The  slides  may  then  be  transferred  to  some  aqueous 
stain  (Griibler's  haematoxylin  gave  good  results),  and  if  desired  the  stain  may 
be  differentiated  with  acid  alcohol  without  harm.  The  vacuolated  structure  of 
the  protoplasm  is  very  beautifully  shown,  but  it  is  difficult  to  distinguish  between 
nuclei,  ingested  food  particles,  or  refuse  composed  of  unicellular  organisms, 
bacteria,  etc.,  which  sticks  all  over  the  plasmodium.  This  last  difficulty  might  be 
largely  obviated,  I  should  think,  if  Miller's  directions  on  the  Aseptic  Cultivation, 
etc.,  were  carefully  followed.  Bacteria,  according  to  Miller,  are  always  present, 
but  they  would  be  easily  distinguished.  Clara  Langenbeck. 

Wells  College. 


MICRO-CHEMICAL  ANALYSIS. 
X. 

POTASSIUM— Continued. 

VI.    IVit/i  Stannic  Chloride. 

The  hydrated  stannic  chloride  is  employed,  since  this  hydrated  salt  is  much 
more  easily  handled  than  the  anhydrous  liquid  compound  SnCl4.  In  the 
hydrated  salts  SnCl4;cH20,  x  may  be  either  3,  .5,  or  8.  All  three  of  these  salts 
are  crystalline,  and  are  to  be  referred  to  the  monoclinic  system. 

When  stannic  chloride  is  added  to  quite  concentrated  solutions  of  potassium 


1122 


Journal  of  Applied  Microscopy 


salts,  slightly  acidified  with  hydrochloric  acid,  beautiful,  large,  colorless,  octa- 
hedral crystals  of  the  compound  K2SnClg  sometimes  separate.  Generally  the 
conditions  which  obtain  are  such  that  owing  to  the  solubility  of  the  potassium 
stannic  salt,  nothing  is  seen  until  the  test  drop  has  evaporated  almost  to  dryness, 
or  until  alcohol  is  added. 

There  seems  to  be  some  doubt  as  to  whether  we  should  call  this  salt  a  true 
chlorstannate  or  a  double  salt  of  the  formula  "JKCl  •  SnCl4.  If  it  is  true  that 
we  have  chlorplatinic  acid  in  solutions  of  platinum  chloride,  by  analogy  we  can 
consider  that  in  the  case  of  the  potassium-tin  compound  we  have  to  do  with  a 
salt  of  chlorstannic  acid.  Moreover,  the  similarity  of  the  chlorstannates  of  K, 
Rb,  Cs,  and  NH^  to  the  chlorplatinates  of  these  elements  is  very  striking. 

Properly  speaking,  stannic  chloride  is  not  a  suitable  reagent  for  potassium, 
since  the  salt  formed  is  too  soluble.  This  reagent  is,  however,  one  of  our  most 
valuable  salts  for  the  detection  of  cesium  (q.  v.). 

The  chlorstannates  of  ammonium,  rubidium  and  cesium  (and  thallium)  are 
far  more  insoluble  than  the  potassium  salt. 

VII.    With  Cerous  Sulphate. 

Cerous  sulphate  added  to  solutions  of  salts  of  potassium  acidified  with  sul- 
phuric acid,  gives  rise  to  the  formation  of  potassium  cerous  sulphate. 

The  reagent  is  most  easily  obtained  in  the  form  of  the  eerie  oxide,  and  can 
be  kept  for  use  in  this  state.  It  can  be  brought  into  the  proper  condition  for 
use  by  being  treated  as  follows :  Place  a  small  drop  of  sulphuric  acid  on  platinum 
foil,  add  a  little  of  the  oxide,  and  heat  until  most  of  the  acid  has  been  driven  off. 
Add  more  acid  and  heat  again.  This  treatment  should  produce  a  product 
almost  completely  soluble  in  water.  Add  a  drop  or  two  of  hydrogen  peroxide, 
and  warm  the  preparation;  the  solution  being  acid,  the  HoOo  acts  as  a  reducing 
agent,  and  a  clear,  colorless  liquid  results.  Evaporate,  and  then  dissolve  in 
sufficient  water  to  make  a  dilute  solution. 

A  drop  of  a  solution  of 
the  reagent  is  allowed  to  jss 

flow  into  a  drop  of  that  of 
the  substance  to  be  tested. 
Both  drops  must  be  dilute. 
The  preparation  is  warmed 
very  gently  at  the  zone 
of  union.  The  salt 
K2SO4  .  Ce^CSOJg  . 
2H2O  rapidly  separates  as 
very  minute,  more  or  less 
spherical  masses.  When 
the  solutions  are  sufficient- 
ly dilute,  or  if  the  preparation  is  allowed  to  evaporate  spontaneously,  tiny  but 
well  formed  colorless  hexagonal  plates  are  obtained  (Fig.  34  B). 

Sodium  treated  in  the  same  way  gives,  as  has  already  been  stated  under  the 
head  of  this  element,  very  small  lenticular  and  fusiform  crystals,  and  dumb-bell- 


Fig.  34. 


and  Laboratory  Methods.  1123 

like  aggregates  (Fig.  34  A).  Rarely,  tiny  four-sided  prisms,  with  pyramidal 
ends,  are  formed. 

If  a  sufficiently  high  power  is  employed,  there  is  generally  no  difficulty  in 
distinguishing  the  double  salt  of  potassium  from  that  of  sodium. 

Owing  to  the  fact  that  higher  powers  must  be  employed  than  is  usual  in 
micro-chemical  work,  it  is  necessary  that  the  drop  be  spread  out  in  a  thin  layer, 
it  being  impossible  to  examine  a  well-rounded  deep  drop.  The  usefulness  of  the 
test  is  therefore  restricted. 

VIII.    With  Sodium-Cobalt  Nitrite. 

This  reagent  produces,  in  neutral  solutions  of  potassium  salts  or  solutions 
acidified  with  acetic  acid,  a  very  difficultly  soluble  double  nitrite  of  potassium  and 
cobalt  of  the  formula  3KNO2  •  Co(N02)3. 

It  is  greatly  to  be  regretted  that  this  interesting  and  delicate  reaction  can 
seldom  be  made  to  yield  more  than  very  minute  globular  grains.  While  it  is  a 
convenient  and  generally  reliable  reaction  for  potassium  in  ordinary  qualitative 
analysis  in  the  wet  way,  it  is  not  to  be  recommended  for  micro-chemical  testing. 

Either  the  standard  reagent,  all  prepared,  can  be  employed,  or  what  is  per- 
haps more  convenient  for  our  purposes,  the  sodium  nitrite,  is  added  to  the 
neutral  test  drop  and  a  solution  of  cobalt  acetate,  w^eakly  acidified  with  acetic 
acid,  is  allowed  to  flow  in.  The  formation  of  yellow  spheroids,  octahedra,  or  the 
skeletons  of  octahedra,  will  indicate  the  presence  of  potassium,  providing  that 
ammonium,  rubidium,  and  cesium  are  absent,  these  elements  giving  an  identical 
reaction. 

IX.    With  Sodium  Tarti-ate  or  Tartaric  Acid. 

The  reaction  taking  place  can  be  expressed  as  follows  :  KCl  +  HNaC4H406 
=  HKC4H40e  -]-  NaCl.  Since  the  product  of  the  reaction,  primary  potassium 
tartrate  (potassium  bi-tartrate),  requires  a  neutral  or  only  slightly  acid  solution, 
and  is,  moreover,  fairly  soluble,. it  is  convenient  to  proceed  as  follows  :  Evaporate 

the  test  drop  so  as  to  obtain  a  thin  uniform  film 
of  material.  Place,  near  by,  a  drop  of  water  into 
which  introduce  a  little  tartaric  acid  and  a  slightly 
greater  quantity  of  sodium  tartrate,  stir  until  all 
has  dissolved ;  then  draw  the  reagent  thus  pre- 
pared across  the  film  of  substance.  If  no  crys- 
tals appear  after  a  short  time,  add  a  drop  of  weak 
alcohol.  The  potassium  salt  separates  as  trans- 
parent, highly  refractive  prisms.  The  crystal 
forms  are  quite  varied,  those  most  frequently 
obtained  are  shown  in  Fig.  35.  Primary  potas- 
*"^-  35-  slum    tartrate    crystallizes  in    the    orthorhombic 

system,  and  exhibits  a  great  tendency  to  assume  hemihedral  and  skeleton  forms. 
Tartaric  acid  alone  can  be  employed  with  good  results,  but  primary  sodium 
tartrate  is  better.     The  addition  of  the  free  acid  suggested  above  is  for  the  pur- 
pose of  assuring  the  presence  of  the  primary  compound. 


1124 


Journal  of  Applied  Microscopy 


Rubidium  and  cesium  give  an  identical  reaction,  their  primary  tartrates  being 
even  more  insoluble  than  that  of  potassium,  hence  the  salts  of  these  two  elements 
will  separate  first.  In  fact,  this  last  property  can  be  utilized  for  detecting 
rubidium  or  cesium  in  the  presence  of  potassium,  if  a  sufficiently  dilute  solution 
be  employed. 

Ammonium  sometimes  gives  crystals  not  to  be  distinguished  from  those  of 
potassium  ;  at  other  times,  when  treated  as  above,  the  precipitate  is  distinctly 
different. 

Many  other  elements  yield  relatively  insoluble  tartrates,  which,  while  differing 
from  the  potassium  salt,  yet  resemble  it  sufificiently  to  lead  to  confusion  (see 
Calcium  and  Strontium). 

Double  tartrates  are  also  apt  to  be  formed.  This  is  far  more  liable  to  happen 
when  large  drops  are  employed  and  the  reagent  added  at  once  to  the  test  drop, 
than  when  evaporation  to  dryness  is  practiced,  and  the  reagent  then  drawn 
across. 

X.  Perchloric  acid  added  to  solutions  of  salts  of  potassium  precipitates  Potassium 
Perchlorate. 

K2S0^  +  2HC10^  =    -iKClO^  +  H.-,SO^. 

Method. — Next  to  the  dilute  solution  of  the  substance  to  be  tested  place  a 
tiny  drop  of  water,  and  to  the  latter  add  a  drop  of 
perchloric  acid  or  a  little  ammonium  perchlorate. 
Cause  the  drop  of  the  reagent  to  flow  into  the  drop 
to  be  tested.  In  a  few  seconds  colorless,  highly 
refractive,  clear  cut  crystals  of  potassium  perchlorate 
separate  (see  Fig.  36).  These  crystals  belong  to  the 
orthorhombic  system,  but  at  first  sight  those  first 
formed  seem  to  be  isometric,  while  later,  what  would 
be  mistaken  for  monoclinic  prisms  appear. 

Pemarks. — The  solutions  must  be  dilute,  other- 
wise the  potassium  perchlorate  is  precipitated  at 
once. 

If   the  solution   is  too   dilute,  crystals   may  not 
aopear  for  a  considerable  period.     The  addition  of  alcohol  will,  in  such  cases, 
greatly  hasten  matters. 

Rubidium  and  cesium  give  a  like  reaction  ;  their  perchlorates  are  more  insol- 
uble than  that  of  potassium.     Thallium  forms  a  still  more  insoluble  perchlorate. 

The  perchlorates  of  the  elements  of  the  other  groups  which  are  generally  met 
with  in  ordinary  work,  are  sufficiently  soluble  not  to  interfere. 

Behrens*  has  recently  shown  that  in  the  presence  of  potassium  permanganate, 
the  perchlorate  of  rubidium  is  colored  pink. 

Advantage  can  be  taken  of  a  similar  property  of  the  potassium  salt  to  obtain 
an  exceedingly  beautiful  test,  for  if  the  test  drop  contains  sodium  permanganate, 
the  potassium  perchlorate  separating  therefrom  will  be  colored.  To  obtain  this 
reaction,  add  to  the  test  drop  a  little   sodium  manganate,  so  as  to  impart  a  dis- 

*  van  Breukeleveen,  Rec.  trav.  chim.  Pays-Bas.  XVII,  i,  94. 


Fig.  3fci. 


and  Laboratory  Methods.  1125 

tinct  green,  then  add  a  tiny  drop  of  hydrochloric  acid,  thus  converting  the 
manganate  into  permanganate.  The  reagent  is  then  allowed  to  flow  in.  The 
crystals  of  potassium  perchlorate  which  separate  have  the  same  form  as  before, 
but  are  a  beautiful  deep  rose  color,  the  intensity  varying  with  the  amount  of  per- 
manganate present.  In  a  few  moments  the  liquid  is  completely  decolorized,  and 
the  precipitated  crystals  deeply  colored.  Performed  in  this  way  the  test  is  an 
elegant  and  very  striking  one. 

Exercises  for  Practice. 

Try  reaction  with  different  salts  of  potassium. 

Introduce  sodium  permanganate  into  the  test  drop,  and  test  as  above. 

Try  the  reaction  on  the  other  members  of  Group  I, 

Make  a  mixture  of  K  and  Na  salts.  Treat  a  drop  of  a  solution  of  this 
material  with  perchloric  acid,  evaporate,  treat  with  the  reagent  again,  and  again 
evaporate,  extract  the  dry  residue  with  alcohol,  and  test  the  alcoholic  extract  for 
sodium. 

Try  the  action  of  perchloric  acid  on  members  of  the  magnesium  group,  and 
the  calcium  group. 

XL  AmmoiiiuDi  Fluosilicate. 

As  performed  in  the  ordinary  manner,  with  solutions  of  moderate  concentra- 
tion, no  separation  of  crystals  results.  It  is  only  under  unusual  conditions,  or  by 
evaporation,  that  potassium  fluosilicate  can  be  made  to  appear.  From  a  practical 
standpoint,  therefore,  this  reagent  is  without  value  for  the  detection  of  potassium. 
The  salt  KgSiFg  crystallizes  as  cubes,  octahedra,  and  combinations  of  these 
forms. 

XII.   CofiTcrsion  into  Sulphate^  Double  Sulphates,  etc. 

The  remarks  made  under  Sodium,  with  reference  to  a  similar  method,  apply 
with  equal  force  to  potassium.  This  method  requires  too  much  care  and  great 
experience,  and  is  therefore  impracticable  save  for  the  expert  crystallographer. 

RUBIDIUM  AND  CESIUM. 

It  is  seldom,  indeed,  that  the  chemist  is  called  upon  to  make  tests  upon  a 
substance  containing  rubidium  or  cesium.  For  this  reason,  and  also  because  the 
present  series  of  articles  purports  to  give  merely  an  introduction  to  the  methods 
of  micro-chemical  analysis,  these  elements  can  be  discussed  together  and  dis- 
missed with  but  few  words. 

Among  the  reagents  which  can  be  employed  for  the  detection  of  these  two 
elements,  three  can  be  selected  as  being  the  most  satisfactory. 
I.  Potassium  Chlorplatinate. 
II.  Ammonium  Silicomolybdate. 
III.   Stannic  Chloride. 
Of  these,  I  and  II  serve  for  the  detection  of  both  rubidium  and  cesium,  and 
III  for  cesium  alone. 


1126  Journal  of  Applied  Microscopy 

/.  Potassium  Chiorplatifiate. 

Since  the  chlorplatinates  of  rubidium  and  cesium  are  so  much  more  insoluble 
than  that  of  potassium,  it  is  more  convenient  to  employ  a  saturated  solution  of 
the  potassium  salt  than  to  make  use  of  a  solution  of  chlorplatinic  acid.  The 
employment  of  this  salt  renders  it  possible  to  test  for  the  two  elements  under 
consideration  even  in  the  presence  of  salts  of  potassium  and  ammonium. 

Allow  a  drop  of  a  saturated  solution  of  potassium  chlorplatinate  to  flow  into 
a  drop  of  a  dilute  solution  of  the  material  to  be  tested.  The  test  drop  should  be 
neutral  or  only  slightly  acid  with  hydrochloric  acid  (see  Potassium,  Method  I). 
If  cesium  is  present  its  chlorplatinate  separates  immediately  as  exceedingly 
minute  crystals  of  the  same  form  as  those  of  the  potassium  salt.  The  crystals 
are  always  so  small  that  a  high  power  is  required  to  enable 
one  to  ascertain  that  the  precipitate  is  not  an  amorphous 
one.  Rubidium  chlorplatinate  being  a  trifle  more  soluble, 
separates  later  in  crystals  again  of  the  same  form,  but  at 
least  twice  as  large,  though  still  much  smaller  than  those 
^>  ^iL>     -^"^^       ^^  ^^^  corresponding  potassium  compound. 

If  the  solution  to  be  tested  is  not  exceedingly  dilute, 
skeleton    crystals    almost    invariably    result,    resembling 
ipiv=o.o>«„n,  crosses   or  5-  and  6-pointed  stars  ;  careful  focusing  will 

'*^'  ^^'  reveal  with  the  latter  the  fact  that  the  branches  of  the 

stars  do  not  lie  in  one  plane,  but  are  arranged  in  the  three  dimensions  of  space 
corresponding  to  the  axes  of  an  octahedron.  In  the  case  of  rubidium,  these  skele- 
ton forms  often  attain  a  considerable  size. 

The  usual  forms  assumed  by  rubidium  chlorplatinate  have  been  sketched  in 
Fig.  37.  The  crystal  forms  given  by  the  cesium  salt  are  the  same,  but  much 
smaller. 

Exercises  for  Practice. 

Try  the  reaction  of  chlorplatinic  acid  on  dilute,  and  on  concentrated  solutions 
of  K,  NH4,  Rb,  Cs. 

Repeat  the  experiments  in  the  presence  of  considerable  free  sulphuric  acid. 

Try,  as  directed  under  Rubidium  and  Cesium,  the  action  of  potassium  chlor- 
platinate on  these  two  elements.  Then  try  on  solutions  of  K  salts  and  of  NH^ 
salts. 

Make  a  mixture  of  Rb  and  Cs,  and  attempt  to  decide  upon  the  presence  of 
both  elements.     Then  try  a  mixture  of  NH4  and  Rb,  and  one  of  NH^  and  Cs. 

//.  Avimoiiium  Silicomolybdate. 

This  reagent,  which  can  be  prepared  according  to  the  method  given  in  a 
previous  article*,  forms  very  insoluble  compounds  with  rubidium  and  cesium. 
The  following  reaction  is  supposed  to  take  place : 

4RbCl+  [(NH^)4Si04.  I2M0O38H2O]  =:  [Rb4Si04.  12Mo03-  UHgO]  + 
4NH4CI. 


*  Jour.  App.  Micro.,  Vol.  Ill,  821. 


vp\v.  =  0.o 

Fig.  3S. 

and  Laboratory  Methods.  1127 

As  in  the  case  of  the  phosphomolybdates  (see  Potassium  IV),  the  composi- 
tion of  the  silicomolybdates  is  still  in  doubt. 

When  the  following  method  is  employed,  there  is  generally  no  difficulty  in 
distinguishing  between  rubidium  and  cesium.  A  drop  of  an  exceedingly  dilute 
solution  of  the  substance  is  spread  out  in  a  thin  layer,  and  evaporated  in  the 
usual  manner.  A  drop  of  a  dilute  solution  of  ammonium  silicomolybdate  con- 
taining a  trace  of  free  nitric  acid  is  drawn  across  the  dry  film  and  the  slide  held 
inclined  for  a  second  or  two,  placed  on  the  stage  of  the 
•*    '  microscope,  and  a  tiny  drop  of  a  saturated  solution  of  the 

reagent  added  to  the  reagent  drop  on  the  slide.  The 
rubidium  salt  separates  at  once  along  the  edges  of  the 
streak  of  reagent  in  the  form  of  lemon-yellow,  highly 
refractive  cubes,  octahedra,  dodecahedra,  and  the  usual 
combinations  of  these  forms,  often  rapidly  passing  into 
spheroidal  granules  (Fig.  38).  In  size  these  crystals 
approximate  those  of  rubidium  chlorplatinate. 
When  cesium  is  present,  and  the  test  is  thus  performed,  the  cesium  salt  is 
instantly  precipitated  in  the  form  of  grains  so  minute  that  even  a  high  power  fails 
to  reveal  any  definite  form  other  than  what  appear  to  be  minute  disks.  The 
solubility  of  the  cesium  salt  is  therefore  so  far  below  that  of  the  corresponding 
rubidium  compound,  that  there  is  little  difficulty  in  distinguishing  between  them 
even  when  both  are  present  in  the  same  substance.  So  delicate  is  the  reaction 
that  it  is  essential  that  there  be  only  the  smallest  possible  amount  of  these  two 
elements  present. 

It  is  advisable  to  have  no  salts  of  ammonium  present,  since  these  compounds 
seem  to  lower  the  solubility  of  the  reagent  sufficiently,  at  times,  to  cause  the 
appearance  of  octahedra  of  ammonium  silicomolybdate.  According  to  Laden- 
berg*  the  octahedra  of  ammonium  silicomolybdate  act  feebly  on  polarized  light, 
and  hence  are  not  to  be  referred  to  the  isometric  system. 

Potassium  salts  treated  in  the  above  manner  give  after  a  time,  near  the 
edges,  neat  prisms,  which  under  favorable  conditions  may  attain  a  considerable 
size.  These  crystals  are  far  too  soluble,  and  their  form  so  different  from  the 
rubidium  compound  that  it  is  impossible  for  them  to  be  mistaken  for  the  latter. 
Silver,  thallous,  and  mercurous  salts  are  also  precipitated  by  this  reagent. 
Sodium  and  lithium  yield  no  crystals  ;  the  same  is  true  of  the  magnesium  and 
the  calcium  groups. 

Better  crystals  of  rubidium  silicomolybdate  can  be  obtained  by  the  addition 
of  a  dilute  solution  of  the  reagent  to  a  dilute  solution  of  the  rubidium  compound 
than  by  the  method  suggested  above,  but  this  process  does  not  permit  of  so 
easily  distinguishing   between  rubidium  and  cesium. 

Exercises  for  Practice. 

Try  the  above  method  on  salts  of  Na,  K,  Rb,  Cs,  NH^,  Li. 
Test  mixtures  of  K  and  Rb ;  K  and  Cs  ;  NH^   and  Rb ;  NH^  and  Cs  ;  Rb 
and  Cs;  K,  Rb,  and  Cs. 

*Handworterbuch,  VII,  361. 


1128  Journal  of  Applied  Microscopy 

Make  a  mixture  of  Na,  K,  Ca,  and  a  trace  of  Rb,  and  test.  Repeat  the  last 
experiment,  after  having  introduced  Cs. 

///.  Stannic  Chloride. 

Solutions  of  stannic  chloride  added  to  hydrochloric  acid  solutions  of  cesium 
salts  give  rise,  even  in  dilute  solutions,  to  the  precipitation  of  cesium  chlorstan- 
nate  Cs2SnCl,;. 

In  order  to  avoid  the  possible  interference  of  other  salts,  it  is  advisable  to 
first  convert  the  substance  to  be  tested  into  the  form  of  a  chloride  by  repeatedly 
evaporating  with  hydrochloric  acid. 

Place  near  a  drop  of  the  moderately  dilute  solution  of  the  substance,  prev- 
iously acidified  with  hydrochloric  acid,  a  drop  of  a  concentrated  solution  of  the 
reagent,  also  acidified  with  hydrochloric  acid.  Cause  the  drops  to  unite.  Cesium 
chlorstannate  is  almost  immediately  precipitated  as  colorless  transparent  crystals. 
The  usual  forms  obtained  are  the  cube,  octahedron,  and  combinations  of  the  two. 
In  fact,  crystal  forms  identical  with  those  spoken  of  in  the  discussion  of  potassium 
chlorplatinate  (q.v.).  Fig.  29  will,  therefore,  also  represent  the  appearance  of 
these  crystals. 

Ammonium  salts  must  first  be  removed  by  gentle  ignition,  since  ammonium 
chlorstannate  is  a  salt  of  almost  as  low  solubility  as  that  of  the  corresponding 
cesium  compound. 

The  chlorstannates  of  potassium  and  rubidium  are  much  more  soluble  than 
that  of  cesium,  hence  there  is  little  danger  of  their  separating  from  dilute  solu- 
tions. If,  however,  the  solution  employed  has  not  been  of  sufficient  dilution,  the 
rubidium  salt  RbgSnCl^  will  first  separate  in  forms  identical  with  those  of  the 
cesium  salt,  but  of  slightly  larger  size,  then  after  a  time,  when  the  drop  has 
evaporated  sufficiently,  the  potassium  salt  K2SnClg  will  also  appear  in  yet  larger 
crystals. 

If  iron  is  present  in  any  considerable  amount  the  crystals  of  cesium  chlor- 
stannate which  separate  are  generally  colored  yellow. 

The  sodium  salt  Na2SnCl,5  •  SHgO  is  too  soluble  to  separate  under  the  con- 
ditions which  obtain  in  the  test.  The  same  is  true  of  the  chlorstannates  of  the 
calcium  group. 

In  cases  where  no  crystals  of  the  cesium  salt  appear  after  some  time,  a  little 
sodium  iodide  can  be  added,  thus  inducing  the  formation  of  cesium  iodostannate, 
which  is  considerably  less  soluble  than  the  chlorstannate.  Cesium  iodostannate 
appears  as  tiny  lemon  or  orange-yellow  octahedra. 

In  the  place  of  stannic  chloride,  the  chlorides  of  the  closely  related  elements, 
antimony  and  bismuth,  can  be  employed,  either  with  or  without  the  addition  of 
sodium  iodide.  Thus,  chlor-  or  iodo-  antimonates  or  bismuthates  are  obtained. 
The  iodo-  compounds  thus  produced  yield  very  beautiful  reactions. 

Exercises  for  Practice. 

To  chlorides,  in  HCl  solution,  of  Na,  K,  Rb,  Cs,  NH^,  add  stannic  chloride, 
first  trying  the  reaction  on  concentrated,  then  on  dilute  solutions. 

Make  a  mixture  of  K  and  Cs,  test.     Then  try  one  of  Rb  and  Cs. 

Try  the  reaction  of  chlorides  of  bismuth  and  of  antimony  on  cesium  chloride. 
Chemical  Laboratory,  Cornell  University.  E.  M.  ChamOT. 


and  Laboratory  Methods.  1129 


Easy  Method  of  Mounting  and  Preserving  Mosquitos. 

The  present  impetus  given  to  malarial  investigation  requires  the  collection 
and  identification  of  these  insects,  and  it  is  of  importance  that  scientists  and 
physicians  in  this  country  should  collect  and  identify  such  specimens  as  they 
can  obtain  in  their  immediate  vicinity,  and  more  especially  if  malarial  fever  is 
known  to  be  present  and  proved  microscopically.  The  calling  of  any  and  all  dis- 
eases malarial,  or  coupling  them  to  typhoid  or  pneumonia,  is  to  be  discouraged, 
and  all  physicians  should  have  a  positive  blood  test  before  treating  or  calling  a 
case  such. 

Papers  concerning  this  work  will  be  found  in  British  Medical  Journal  No. 
2054,  May  12,  1900,  pp.  1183-1188;  No.  2060,  June  23,  1900;  and  in  Prof. 
Adann's  Report  on  Tropical  and  Subtropical  Diseases  of  Canada,  p.  1544. 

A  pamphlet  on  "  How  to  Collect  Mosquitos  "  has  been  issued  by  the  Mon- 
treal Natural  History  Society,  and  edited  I  believe  from  the  British  Museum.  The 
insects  must  be  carefully  pinned  out  or  preserved,  or  they  are  injured  in  shipping. 
The  method  used  by  Dr.  D.  C.  Rees,  in  the  London  Tropical  School,  is  as  follows  : 

(1)  Kill  in  ordinary  killing  bottle,  or  chloroform,  or  tobacco. 

(2)  When  dead  turn  specimen  on  its  back,  separate  legs,  place  large  drop  of 
thick  xylol  balsam  on  slide,  invert  this  gently  on  to  the  mosquito  so  as  to  pick  it 
up  and  not  injure. 

(3)  With  fine  needle  spread  and  arrange  wings  and  legs,  and  if  necessary 
press  thorax  down  gently. 

(4)  Pour  on  thin  xylol  balsam  and  straighten  the  antennae  and  proboscis  as 
it  runs  out. 

(5)  Set  aside  to  harden  ;  chip  of?  excess. 

(6)  Place  a  glass  ring  about  yL  to  -^  inch  deep  over  specimen,  and  fill  up 
the  chamber  thus  formed  with  balsam,  the  upper  surface  of  which  should  be 
convex,  so  that  when  cover  is  applied  no  air  bubbles  are  included. 

(7)  Let  it  harden,  and  then  mail  if  desired. 

N.  B." — ^If  glass  rings  are  not  handy,  balsam  alone  will  do.  (I  use  zinc 
carpet  rings.)  If  desired  to  photograph  the  insect,  its  parts  must  lie  as  nearly 
in  one  plane  as  possible.  This  method  is  due  to  Dr.  G.  D.  Freer,  Colonial  Sur- 
geon, Penong  Hospital. — B.  M.  /.,  p.  1468. 

Dr.  John  Reid,  Redhill,  Surrey,  Eng.,  adds  a  note  in  regard  to  above.  In 
place  of  killing  aphides,  mosquitos,  etc.,  coax  them  to  get  entangled  in  a  drop  of 
glycerin,  and  with  fine  needles  put  in  best  position.  Chemistry  explains  how 
glycerol  menstrum  exhibits  structure  better  than  balsam,  which  refuses  abso- 
lutely to  mix  with  aqueous  media.  Care  and  patience  are  required  to  prevent 
injury  and  air  bubbles.  Glycerol  jelly  may  be  used  if  preferred,  and  the  insects 
die  in  more  natural  positions  than  in  balsam. — B.  M.J..,  p.  1592. 

In  reply  to  both,  I  prefer  the  carbolic  acid  method,  or  if  desired  carbolic  and 
glycerol,  which  I  published  some  years  ago.  Glycerin  is  not  always  to  be  relied 
on,  especially  if  any  chitinous  or  limy  deposits  are  present. 

V.  A.  Latham,  M.  D. 


11-^0  Journal  of  Applied  Microscopy 

With  the  beginning  of  our  fourth 

journal   or  volume  the  scope  of  the  Journal  is 

Annlicd     I^icrOSCOnV  broadened    so    as   to    include   general 

a„j  laboratory  methods  in  those  branches 

Laboratory   Methods.  ^^  science  and  industry  in  which  the 

microscope  is  used. 


Edited  by  L.  B.  ELLIOTT.  The  microscope  is  the  central  figure 


I       J  Mt    .ui    c       .u    o  Ki  „„,-,„  n»„ow,^»„.  around  which   a  host  of  contributory 

Issued  Monthly  from  the  Publication  Department  -' 

of  the  Bausch^ &^Lomb  Optical  Co.,  subjects  group  themselvcs,  and  a  record 

"  of   microscopical   progress    is,    to    the 


SUBSCRIPTIONS:  ,  .  ,   ^  j       r 

One  Dollar  per  Year.     To  Foreign  Countries.  $1.25       WOrker,     mcomplete     and     of    COmpara- 

per  Year,  in  Advance.     tivcly  little  valuc   without  a  record  of 

The  majority  of  our  subscribers  dislike  to  have  their     developments  in  the  acccssory  processcs 

riles  broken  in  case  they  fail  to  remit  at  the  expiration  ,   .    ,    ,  .  ,      ,  ,  ,_,       , 

of  their  paid  subscription.     We  therefore  assume  that  no       UpOn  whlCh  hlS  WOrk  depends.        i  0-day 
interruption   in   the   series  is  desired,  unless  notice  to        ^ ,  •        ^  e  t     r 

discontinue  is  sent.  the    muscum    IS    transformed    from    a 

curiosity  shop  to  a  substantial  aid  in 
demonstration,  just  as  the  microscope  has  been  elevated  from  the  function  of 
a  toy  to  that  of  the  biologist's  right  hand  assistant. 

The  camera,  popularly  employed  for  recreation,  now  supplies  one  of  the 
readiest,  most  useful  and  reliable  means  for  illustration  and  the  recording  of 
facts  and  conditions. 

The  stereopticon  has  advanced  from  the  companionship  of  children  to  the 
control  of  the  lecture  room. 

The  countryside  with  its  ponds  and  ditches,  once  the  exclusive  territory  of 
the  naturalist,  sneered  at  by  the  section  cutter,  is  again  sought  by  biologists,  and 
without  a  knowledge  of  the  life  of  its  denizens  his  work  is  balked. 

So  through  the  list  the  index  points  to  a  Journal  of  Applied  Microscopy 
AND  Laboratory  Methods  in  which  the  microscope  shall  be  the  principal  sub- 
ject and  the  related  methods  be  given  their  proper  share  of  consideration. 
In  this  decision  our  contributors  and  friends  to  whom  the  matter  was  first 
referred  have  unanimously  agreed.  It  is  not  proposed  to  lessen  the  amount  of 
material  devoted  to  the  microscope,  but  rather  to  give  an  additional  number  of 
pages  each  month  printed  on  a  finer  grade  of  paper  suitable  for  the  illustrations 

required. 

* 
*       * 

Beginning  with  this  number,  Mr.  Raymond  Pearl,  Zoological  Laboratory, 
University  of  Michigan,  will  conduct  a  department  of  General  Physiology,  which 
will  be  devoted  to  the  reviewing  of  current  literature  in  the  field  of  general 
physiology,  using  the  term  in  its  broadest  sense.  No  attempt  will  be  made  to 
keep  abreast  of  the  enormous  literature  of  medical  physiology  as  ordinarily 
defined,  but  we  shall  rather  have  to  do  with  that  physiology  which  treats  of  the 
life  phenomena  of  all  organisms.  A  special  feature  will  be  made  of  the  topics 
of  animal  reactions  and  behavior  which  are  now  exciting  such  general  interest. 
The  effort  will  be  made  to  give  practical  accounts  of  all  new  methods  of  work 
along  the  lines  indicated,  especially  such  as  can  be  used  by  teachers  in  secondary 
schools  or  colleges  in  demonstrating  to  classes. 


and  Laboratory  Methods.  1131 

CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Qoebel,  K.     Organographie  der    Pflanzen   ins-      -pj^g    f^^st    volume    of    this    work     has 
besondere   der  Archegoniaten  und  Samen- 
pflanzen,  Zweiter  Teil.    Specielle  Organo-      already  been  reviewed  in  the  Journal. 

graphie.    8vo,  pp.  xiii-xvi  4-385-648;    173      The    present    volume    deals    with   the 

illustrations.     Gustav  Fischer,  Jena.  M.    1. 

gametophyte  and  sporophyte  genera- 
tions of  the  Pteridophytes,  and  with  the  sporophyte  generation  of  the  Spermato- 
phytes.  The  gametophyte  of  the  Pteridophytes  is  discussed  under  the  headings 
(1)  Structure  and  Development  of  Sex  Organs  and  (2)  The  Form  of  the  Pro- 
thallia. 

In  treating  the  development  of  antheridia  the  author  advances  views  which 
are  at  variance  not  only  with  the  views  of  Belajeff  and  others,  but  also  with  his 
own  previous  accounts.  According  to  his  present  interpretation  we  have  within 
the  microspore  wall  of  Iscetes  a.  prothallium,  consisting  of  three  sterile  cells  and 
one  antheridium,  the  wall  being  represented  only  by  the  cover  cell.  In  treating 
the  development  of  antheridia  and  archegonia,  the  transition  from  free  to 
imbedded  forms  is  described  in  some  detail.  The  peculiar  prothallia  of  OpJiio- 
glossum,  Botrychiiim,  and  Lycopodium,  receive  particular  attention.  In  the 
second  part  of  the  book,  which  is  devoted  to  the  sporophyte  generation  of  the 
Pteridophytes  and  Spermatophytes,  the  various  organs  are  discussed  in  great 
detail.  Some  of  the  most  interesting  topics  are :  The  Comparative  Morphology 
of  the  Embryo ;  The  Transition  Between  Leaf  and  Shoot ;  Leaf  Formation  ; 
The  Relation  between  Leaf  Venation  and  Leaf  Development;  Transformed 
Leaves;  Branching,  etc.  The  treatment  throughout  is  dominated  by  experi- 
mental morphology,  and  cannot  fail  to  be  a  great  help  to  all  investigators,  and 
especially  to  those  who  are  too  rigid  in  their  morphology.  While  constantly 
calling  attention  to  the  variation  which  occurs  in  nature,  and  which  may  be 
induced  artificially,  the  author  also  recognizes  the  large  part  which  heredity 
plays  in  determining  the  plant  form.     An  English  translation  will  appear  soon. 

c.  J.  c. 

Byxbee,  Edith  S.    The  Development  of  the  ^^^^  Byxbee's  work  on  Lavatera  is  an 
Karyokinetic  Spindle  in  the  Pollen-mother-  •' 

Cells  of  Lavatera.    Proc.  Cal.  Acad.  Sci.,  addition  to  the  very  interesting  series 

Ser.  Ill,  Bot.  2:  63-82,  pis.  10-13,  1900.  of  contributions  on  spindle  formation 

recently  issued  from  the  laboratory  of  the  University  of  California.  While  differ- 
ing in  certain  minor  details,  the  writer's  conclusions  confirm  the  more  important 
points  previously  observed  in  Cobcea,  Passiflora,  Gladiolus^  etc.,  by  other  investi- 
gators. Her  observations  are  briefly  as  follows :  The  meshes  of  the  network, 
close  to  the  nuclear  wall,  form  a  felt  of  fibers  about  the  nucleus.  The  granular 
constituent  of  the  cytoplasm  collects  in  a  wide,  dense  zone  about  the  nucleus. 


113-  Journal  of  Applied  Microscopy 

The  linin  increases  in  quantity,  the  nuclear  wall  breaks  down,  and  the  fibers  out- 
side begin  to  grow  into  the  nuclear  cavity.  The  cytoplasmic  and  linin  fibers 
form  a  mass  in  which  the  chromosomes  lie.  The  mass  of  fibers  projects  out  at 
a  number  of  points,  forming  the  multipolar  spindle.  Two  of  the  cones  become 
more  prominent  than  the  others,  which  they  finally  absorb,  thereby  forming  the 
bipolar  spindle.  Just  how  this  absorption  of  the  cones  is  brought  about  is  not 
made  clear  either  in  the  description  or  in  the  figures.  Flemming's  strong  solu- 
tion of  chrom-osmo-acetic  acid  was  used  almost  exclusively  as  a  fixing  agent,  but 
fair  results  were  also  obtained  with  palladium  chloride  and  iridium  chloride  to 
which  a  small  amount  of  glacial  acetic  acid  had  been  added.  Of  the  stains  used 
the  safranin-gentian-violet-orange  G.  combination  gave  the  best  results.  The 
paper  is  well  illustrated  by  four  beautiful  lithographic  plates. 
Chicago.  A.  A.  Lawson. 

Timberlake,  H.  Q.     The  Development    and      This  work  was  undertaken  to  determine 
Function  of  the  Cell  Plate  in  Hieher  Plants.       •       i        •,     i  r 

Bot.  Gaz.  30:  73-99, 154-170,  pis.  8-9, 1900.      i"  detail  the  exact  sequence  of  events 

during  the  division  of  the  cell  body,  and 
to  correlate,  as  far  as  possible,  the  facts  thus  brought  out  from  the  point  of  view 
of  the  physiology  of  cell  reproduction.  The  formation  of  new  radiating  fibers 
around  the  daughter  nuclei  in  the  diaster  stage,  and  the  formation  of  a  spindle 
around  a  single  chromosome,  as  described  by  Juel  for  Hetnerocallis ,  are  taken  to 
indicate  that  the  chromatin  is  the  real  center  for  the  formation  of  kinoplasmic 
fibers.  Having  formed  as  fibers  around  the  nucleus  as  a  center,  the  kinoplasm 
takes  part  in  the  process  of  nuclear  division,  and  later  divides  the  cell  by  a  part 
of  the  fibers  being  transformed  into  a  membrane  which  becomes,  in  splitting,  the 
plasma  membranes  of  the  daughter  cells.  Prior  to  the  formation  of  the  cell 
plate  the  equatorial  zone  becomes  filled  with  a  substance  which  stains  strongly 
with  the  orange  of  the  triple  stain.  The  similarity  in  staining  of  this  substance, 
together  with  its  presence  in  the  region  of  the  spindle  in  which  the  cell  wall 
appears  later,  is  taken  to  signify  the  presence  of  a  carbohydrate  substance 
destined  for  the  formation  of  the  new  cell  wall.  The  relation  of  the  carbohydrate 
material  to  the  process  of  division  would  seem  to  show  that  the  substance  for  the 
formation  of  the  cell  wall  is  held  in  a  reserve  form  in  the  protoplasm  before  it  is 
actually  needed  for  the  process  of  wall  formation.  If  the  relation  of  the  carbo- 
hydrate material  to  the  spindle  fibers  be  taken  in  connection  with  the  facts  shown 
by  Klebs  and  Townsend,  that  the  presence  of  a  nucleus  is  necessary  for  the  forma- 
tion of  a  cell  wall,  there  would  be  some  evidence  for  the  hypothesis  that  the 
nucleus  forms  the  cell  wall  substance. 

The  material  used  for  investigation  was  Allium  cepa,  Lilium  longijlonim, 
Fritillaria  imperialis,  Hyacinthus  orie?italis,  Vicia  faba,  Phaseolus  vulgaris,  Fisuni 
sativum,  Larix  Americafia,  Larix  Eiiropcca,  Iris  versicolor,  z.x\di  Heinerocallis  fulva. 
Several  fixing  fluids  were  employed  :  Flemming's  chrom-osmo-acetic  acid  ;  Her- 
mann's platinum  chlorid-chrom-acetic  acid ;  Vom  Rath's  platinum  chlorid-picro- 
osmo-acetic  acid;  Reiser's  mercuric  chlorid-acetic-acid,  etc.  Of  these  methods 
the  material  fixed  in  Flemming's  stronger  solution  gave  the  best  results.  The 
triple  stain,  safranin-gentian-violet-orange,  was  used  to  stain  the  material  fixed  in 
fluids    containing  osmic    acid,   while     Zimmermann's   fuchsin   iodin  green   and 


and  Laboratory  Methods.  1133 

Heidenhain's  hsematoxylin,  preceded  by  Bordeau  red  as  a  ground  stain,  were 
used  to  stain  material  fixed  in  the  fluids  containing  mercuric  chlorid.  The  paper 
is  poorly  illustrated  by  a  series  of  reproductions  of  photographs.  While  photo- 
graphy is  a  convincing  method  of  illustrating  points  in  gross  histology,  it  has  so 
far  proved  a  failure  as  a  means  of  illustrating  protoplasmic  structures  within  the 
cell.  Lithographic  drawings,  or  even  diagrams,  are  much  more  satisfactory. 
The  work  is  a  valuable  contribution,  as  it  adds  much  to  our  knowledge  of  the 
origin  and  function  of  the  cell  plate.  A.  A.  Lawson. 

Chicago. 

Wager,  H.     The  Eye   Spot  of  Euglena  viridis.       ^j^jg  -^^    ^^^     ^^    interesting 

Jour.  Lmn.  Soc,  27 :  463-401,  1900.  ... 

account  of  investigations  on  the  struc- 
ture and  behavior  of  Euglena  viridis.  On  the  general  structure  of  Euglena,  Mr. 
Wager  gives  nothing  new,  merely  summarizing  what  is  already  known,  but  he 
reports  a  striking  feature  in  the  vacuole  system,  namely,  that  the  gullet  is  in 
permanent  connection  with  the  principal  vacuole  or  "  excretory  reservoir,"  as  he 
calls  it.  The  eye  spot  he  believes  to  be  derived  from  chlorophyll,  because  the 
action  of  its  granules  when  in  alcohol  shows  the  same  behavior  as  do  the  rusty, 
red  granules  of  Fucaceae,  which  are  known  to  be  derived  from  chlorophyll.  His 
most  interesting  discoveries  were  on  the  flagellum  and  its  relation  to  the  eye  spot. 
He  found,  by  the  use  of  osmic  acid,  that  the  flagellum  passed  into  and  was 
attached  to  the  excretory  reservoir  instead  of  terminating  in  the  gullet.  The 
base  of  the  flagellum  is  bifurcate,  and  on  one  of  the  limbs,  in  close  (but  not 
organic)  connection  with  the  concave  side  of  the  eye  spot,  is  a  large  oval  swelling 
or  enlargement.  Quoting  Englemann's  experiments  on  the  behavior  of  Euglena 
in  a  spectrum,  he  notes  that  the  greatest  gathering  of  the  Euglenae  is  in  the  blue 
end.  Since  the  red  pigment  of  the  eye  spot  allows  the  blue  rays  of  normal  light 
to  pass,  he  suggests,  tentatively,  that  there  is  possibly  a  definite  stimulus  exerted 
by  the  blue  rays  upon  the  swelling,  and  hence  on  the  flagellum.  The  other 
hypothesis  which  he  brings  forward  is  that  the  eye  spot  merely  causes  a  defi- 
nitely unequal  illumination  of  the  sensory  spot,  and  orientation  follows.  Further 
experiments  along  that  line  would  prove  interesting. 
Chicago.  P.  G.  WrightSON. 


CYTOLOGY,   EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Clajrpole,  Sage  College, 
Ithaca,  N.  Y. 

CURRENT  LITERATURE. 

Furst,  C.  M.     Haarzellen  und  Flimmerzellen.       This   investigation  was   carried   out  to 

Anat.    Anz.    18:   190-203,    iqoo.     (6  fia;s.  in         ,  1    ,-    •     ,        , 

text.)  "  show  definitely  the  resemblances  and 

differences      between      the      so-called 

"  hair-cell  "  and  the  ciliated  cell.     The  material  used  was  principally  embryonic 


1134  Journal  of  Applied  Microscopy 

salmon.  It  was  fixed  in  Perenyi's  fluid  and  stained  in  Heidenhain's  haematoxy- 
lin  and  Orange  G.  Especially  careful  study  was  made  of  hair-cells  from  the  crista 
and  macula  acoustica.  In  a  salmon  embryo  of  150  days  every  cell  from  this 
region  has  a  black  staining  layer  on  its  upper  border,  against  the  lymph  space. 
Next  inside  to  stain  is  a  similarly  staining  cone  with  a  clearly  marked  apex 
reaching  down  toward  the  nucleus.  The  hair-cell  complete,  possesses  an  in- 
ternal point  or  hair  which  can  clearly  be  shown  to  be  formed  of  cilia  fastened 
together  ;  a  basal  layer  shown  by  iron  haematoxylin  to  be  composed  of  deeply  stain- 
ing round  bodies  ;  and  a  cone  continued  into  the  cell  and  staining  deeply.  This 
whole  series  of  parts,  the  hair,  the  layer,  the  cone,  forms  a  definite  organ,  the 
"  hair  apparatus  or  organ."  The  different  parts  of  this  organ  correspond  to  the 
special  parts  of  the  ciliated  cell,  the  cilia,  the  basal  bodies,  the  ciliary  cone ;  as 
if  it  were  a  specialized  or  modified  ciliated  cell.  The  hair  cell  is,  due  to  the 
"  hair  organ,"  a  specific  form  of  cell  which  keeps  the  principal  morphological  fea- 
tures of  the  ciliated  cell.  Probably  this  "  hair  apparatus  "  is  the  sensory  organ 
of  the  cell.  The  opponents  of  the  Lenhossek-Henneguy  hypothesis,  concern- 
ing the  origin  of  the  basal  bodies  from  the  centrosome,  have  not  yet  deprived 
that  theory  of  its  probability.  a.  m.  c. 

Heidenhain,  M.    Ueber  die  erste  Entstehung      It    is    customary    to    see   the    surface 
der  Schleimpfropfe  beim  Oberflachenepithel  -^i     i-  r  .,         .  ,       -^i    -^         n 

des    Magens.     Anat.    Anz.    18:    417-425      epithelium  of  the  stomach  with  its  cells 
(4  abb.),  1900.  full  of  mucous,  a  condition  true  of  all 

animals  from  fishes  upward  throughout  the  vertebrate  series.  In  view  of  this 
fact  special  interest  was  aroused  by  finding  the  epithelial  cells  in  the  stomach  of 
a  full  grown  specimen  of  Triton  tceniatiis  contain  only  a  small  quantity  of 
mucous.  The  mucous  free  cells  showed  on  the  free  surface  a  striated  border 
similar  to  the  well  known  "  brush-border "  described  by  Tovnier  in  different 
glandular  epithelia.  In  all  these  preparations  the  brush-borders  show  clearly  in 
the  cells  of  the  stomach  glands,  but  in  spite  of  the  great  resemblance  of  the 
structures  it  is  doubtful  whether  the  border  of  fine  protoplasmic  hairs  of  the 
surface  epithelium  is  identical  with  the  "  brush-border  "  of  gland  cells.  How- 
ever, the  term  will  still  be  used  in  this  article. 

It  is  evident  that  the  width  of  the  border  varies  in  different  cells  and  the 
striated  border  shows  still  more  differences.  The  origin  of  the  mucous  plug 
commonly  seen  in  cells  is  by  the  pouring  out  of  the  mucous  substance  between 
the  rods  of  the  brush-border,  by  which  means  the  border  increases  in  width 
with  an  increase  in  the  amount  of  mucous  poured  out.  In  this  way  forms  the 
convex  mucous  "  goblet "  without  the  direct  participation  of  the  original  proto- 
plasmic cell  body.  These  preparations  were  stained  in  iron  haematoxylin  and 
rubin,  the  haematoxylin  being  bleached  until  it  remained  only  in  the  nucleus 
centrosome  and  granules  of  glandular  secretion ;  the  protoplasm  stained  solely 
by  the  rubin. 

Very  instructive  preparations  were  obtained  from  a  specimen  of  Trito)i 
tcaniatus  which  showed  a  larger  quantity  of  mucous.  The  brush-borders  stand 
in  a  close  relation  to  the  mucous  and  to  the  formation  of  the  well  known  mucous 
plug  of  the  surface  epithelium.  Black-tinted  protoplasmic  threads  rise  from  the 
surface  of  the  cell  and  are  here  thickened  into  root-like  processes ;  these  latter 


and  Laboratory  Methods.  1135 

show  no  particular  regularity  of  form.  These  "  roots  "  are  connected  at  the 
inner  limit  of  the  border  by  protoplasmic  fibres.  At  their  free  ends  the  little 
rods  swell  to  fine  irregular  knots,  a  condition  not  observed  for  the  brush-borders. 
Between  the  rods  lies  the  mucous  which  may  be  aided  in  its  separation  by  them. 
At  first  the  surface  of  the  cell  is  flat,  but  with  the  increasing  amount  of  mucous 
it  becomes  more  and  more  arched  until  the  familiar  beaker-cell  is  formed,  the 
fine  striations  still  remaining  visible  on  its  outer  border  and  sending  plasmic 
threads  downward.  When  this  form  has  been  obtained  the  cell-body,  hitherto 
free  from  mucous,  shows  a  large  drop  which  appears  below  the  striated  border. 
The  whole  presents  a  curious  appearance.  In  the  layer  of  mucous  coming  from 
the  cell-protoplasm  are  seen  many  fine  protoplasmic  columns.  These  support  on 
their  outer  ends  many  branches  looking  like  candelebra,  the  original  rods  of  the 
brush-border.  Continuing,  the  formation  of  mucous  causes  first  the  destruction 
of  the  protoplasmic  columns,  leaving  the  branches  only.  Then  these  too  disap- 
pear, leaving  the  mucous  clear  and  unlined.  These  thickened  "  roots  "  of  the 
rods  in  these  mucous  cells  are  not  to  be  compared  with  the  basal  pieces  of 
ciUated  epithelium.     The  controsome  lies  in  these  cells  within  the  mucous. 

The  special  reason  for  publishing  these  observations  in  such  detail  is  a 
recent  article  by  A.  Gurwitsch  (review  in  Journal  of  Applied  Microscopy, 
iii,  805,  1900),  on  the  development  of  ciliated  cells.  According  to  his  observa- 
tions, the  earliest  stages  show  a  border  of  purely  alveolar  structure  and  later  on 
the  full  surface  of  the  border  is  a  fine  protoplasmic  network.  These,  according 
to  the  present  writer,  are  both  impossible.  The  distal  thickenings  observed  by 
Heidenhain  account  for  the  distal  network.  a.  m.  c. 

Barrows,  A.  S.    Respiration  of  Desmognathus.      The    views    already    advanced    to    ac- 

Anat.  Anz.  18:461-464,  looo.  .    r  ■     ^-         •      ^         ^  ^ 

^      -t  t'    V  count  for  respiration  in  lungless  sala- 

manders, of  which  this  forms  a  type,  claim  extensive  "buccopharyngeal  "  respi- 
ration, excluding  the  skin  from  an  important  part  in  respiration.  The  discovery 
later  of  blood  vessels  in  certain  lunged  forms  that  reached  to  the  pharyngeal 
epithelium  supported  this  view.  It  has  been  shown  for  Spelerpes  fuscus,  a  lung- 
less  form,  that  there  is  a  similar  nearness  to  the  surface  of  the  pharyngeal  cap- 
illaries. In  the  work  done  on  Desmognathus  fusca  a  warm  carmin  injection 
mass  was  introduced  through  the  ventricle  of  the  heart  by  means  of  a  hypoder- 
mic syringe.  A  remarkable  net-work  of  capillaries  was  found  to  extend  through 
the  entire  wall  of  the  esophagus.  These  were  found  to  be  from  the  arteriae 
maxillares  externae  on  the  dorsal  wall  and  of  the  arteriae  pharygeae  on  the 
ventral  wall.  On  each  side  the  arteria;  pulmonales  anastomosing  with  the 
arteriae  gastrica  send  branches  to  the  esophagus.  The  blood  is  collected 
especially  in  the  venae  esophagae.  A  more  complete  consideration  will  follow 
this  preliminary  paper.  a.  m.  c. 

Stassano,  H.    Function  of  the  Nucleus.  Compt.      The  author  finds  cells  of  the  vascular 
Rend,  i,  30:    1780-1783,  igoo.     (Review  in  j    ^i     r  -r     1.  4.  cc.    -^ 

Jour.  Roy.  Micr.  Soc.  pt.  5,  1900.)  endothelium  manifest  a  strong  affinity 

for  mercury  and  other  poisons  intro- 
duced into  the  circulation,  and  believes  that  this  is  effected  by  the  nucleus  by 
virtue  of  its  nucleins,  which  form  compounds  with  metals  and  bases  analogous 


1136  Journal  of  Applied  Microscopy 

to  salts.  His  evidence  is  under  five  heads  :  1.  Leucocytes,  which  are  very  rich 
in  nuclein,  show  a  strong  affinity  for  metals.  '2.  In  young  dogs  the  endothelial 
cells  contain  granulations  shown  by  Kowalewsky  to  present  the  characters  of 
nuclear  granulations.  Organs  of  these  young  dogs  absorb  more  mercury  by 
weight  than  those  of  older  dogs  in  which  the  granules  are  absent.  3.  It  has 
been  shown  that  the  amount  of  nuclein  in  an  organ  depends  on  the  number  of 
cell  nuclei  present,  and  the  author's  experiments  show  that  the  amount  of  mer- 
cury absorbed  depends  on  the  amount  of  nuclein  present.  4.  The  non-nucle- 
ated red-blood  corpuscles  of  mammals  are  the  only  cellular  elements  that  do  not 
absorb  mercury.  5.  An  intravenous  injection  of  methyl-violet  reduces  the  absorp- 
tion of  mercury  by  the  cells  of  the  vascular  endothelium.  With  this  may  be 
compared  the  fact  that  cells  treated  with  such  substances  as  osmic  acid  do  not 
stain.  The  affinity  of  the  nucleus  for  basic  stains  is  itself  a  proof  of  the 
author's  view.  a.  m.  c. 

RECENT  LITERATURE. 

Aicbel,     Otto.      Vergleichende    Ent\\icklungs-  Doflein,  F.     Studien  zur  Naturgeschichte  der 

geschichte     und     Stammesgeschichte     der  Protozoon.  iv.  Zur  Morphologic  und  Physiol- 

Nebennieren.     Ueber    ein    neues  normales  ogie    der    Kern-  und    Zelhheilung.      Nach 

organ  des  Menschen  vind  der  Saugethiere.  Untersuchungen  an  Noctiluca  und  anderen 

3    Taf.    I  fig.    Arch.    f.    Mikrosk.    Anat.    u.  Organismen.    Zool.  Jahrb.  Abtheil  f.  Anat. 

Entwicklungsgesch.  56:   i-8o,  1900.  u.  Ontog.  v.  Thiere.  14:  1-60,  1900. 

Grosser,    0.     Mikroskopische  injectionen   mit  Phisalix-Picot.       Recherches    embryologiques, 

Eiweiss-Tusche.    Zeitschr.  f.  wiss.  Mikrosk.  histologiques    et     physiologiques     sur     les 

17:  178-181,  1900.  glandes  a  venin  de  la  salamandre  terrestre. 

Marcus,  H.      Zur    "  intravitalen  "    Neutralroth  These  de  doctorate  en.  med.  Paris,  1900. 

w^\"^    '^u'    Vra^'^^r"-      ^'^"^^    ^''""       Brunn,  Max  von.     Zur  Histologie  der  Epithe- 
Wochenschr.     13:  871-873,  1900.  jj^^   ^^^  3^,.^^^,^   Haute.  2  fig.  Centralbl.  f. 

Muhlmann,    M.     Atrophic   und    Entwicklung.  AUg.  Pathol,  u.  pathol.  Anat.,  II:   604-607, 

Deutsche   med.  Wochenschr.     26:  655-657,  1900. 

1900.  Reinke,     Johannes.      Die     Entwicklung     der 

Plato,  J.     Ueber  die  "  vitale  "  Farbbarkeit  der  Naturwissenschaften,  insbesondere  der  Biol- 

Phagocyten  des  Menschen  und  einiger  Saii-  ogie,  im  neunzehnten    Jahrhundert.    (Rede 

gethiere    mit   Neutralroth.     i  Taf.    Arch.  f.  zur   Feier  des    Jahrhundertmechsels   am  13 

Mikrosk.    Anat.   u.  Entwicklungsgesch.  .56:  Jan.,    1900,    Zu  Kiel.)    Kiel.   Univ.-Buchh. 

868-917,  1900.  1900  (215). 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Richard  M.  Pearce,  M.  D. 

University  of  Pa.,  Philadelphia,  Pa.,  to  whom  all  books  and  papers 
on  these  subjects  should  be  sent  for  review. 

Nichols,    E.    H.     On   the  Etiology  of   Cancer.       Nichols  investigated  this  subject  along 
First  Annual  Report  of  the  Cancer  Investi-       -         ,. 
gation  Committee  to  the    Surgical  Depart-       ^^^^  Imes  : 

ment    of    the     Harvard     Medical    School.  1.    A  histological  Study  of  tumors   in 

Journal  of  the    Boston  Society  of  Medical  1        ^      j    ^         •  u    ^i         ^u        u 

Sciences.    Vol.  V,  No.  2,  1900!  o'^der  to  determme  whether  the  char- 

acteristic  bodies    claimed    to   be    the 

cause  of  cancer  were  constantly  present. 

2.  The  inoculation  of  animals  with  portions  of  tissue  from  fresh  cancer. 

3.  The    inoculation    of    animals    with   the    blastomycetes   of    Sanfelice  and 
Plimmer. 


and  Laboratory  Methods.  1137 

4.  Attempts    to    isolate    parasitic    micro-organisms  from  malignant  tumors. 

First.  In  the  histological  study  tissues  were  hardened  in  alcohols  of  various 
strengths,  Hermann's  solution,  Flemming's  solution,  corrosive  sublimate,  and 
Zenker's  fluid.  Zenker's  fluid  gave  the  best  results.  Paraffin  imbedding  was 
used. 

For  staining,  the  methods  recommended  by  Sanfelice  and  PUmmer  were 
tried  at  first,  but  as  they  did  not  give  satisfactory  results,  the  following  method 
suggested  by  Mallory  was  used  : 

1.  Ten  per  cent.  aq.  sol.  ferric  chloride,  two  minutes. 

2.  Aq.  sol.  haematoxylin  (1-2  per  cent.),  freshly   made,  two  minutes. 

3.  Wash  in  water. 

4.  One  per  cent.  sol.  ferric  chloride  until  blue  color  is  removed 
from  protoplasm  and  nuclear  stain  is  distinct.  (Watch  under 
microscope.) 

5.  Wash  in  water. 

6.  In  the  following  solution  for  two  minutes. 

1  per  cent.  aq.  sol.  acid  fuchsin,  one  part. 
Sat.  aq.  sol.  picric  acid,  two  parts. 

7.  Wash  in  water. 

8.  Ninety-five  per  cent,  alcohol. 

9.  Xylol,  three  changes,  blotting  between  each  change. 
10.  Mount  in  Xylol  balsam. 

This  stain  colors  nuclei  black,  protoplasm  a  faint  greenish  pink,  and  con- 
nective tissue  a  brilliant  red.  The  peripheral  and  central  portions  of  inclusions 
stain  a  brilliant  red,  the  intermediate  portion  a  faint  pink. 

The  stain  is  simple  in  manipulation,  constant  and  even  in  its  results. 

Thirty-five  carcinomata  and  five  sarcomata  were  examined.  In  seventeen 
cases,  bodies  similar  to  those  described  by  Sanfelice  and  Plimmer  were  found. 
They  were  found  principally  in  cancer  of  the  breast,  in  thirteen  out  of  sixteen 
cases.  They  were  never  found  in  epidermoid  cancer  (thirteen  cases)  nor  in 
sarcoma  (five  cases). 

Although  the  writer  makes  no  definite  negative  statement,  he  apparently 
believes  that  these  bodies  have  nothing  to  do  with  the  causation  of  cancer. 

Second.  Inoculations  were  made  from  tumors  which  were  received  within  two 
hours  after  operation,  and  which  were  not  ulcerated.  Under  aseptic  precau- 
tions portions  of  tumor  were  removed  and  placed  in  the  peritoneal  cavity  of  a 
rabbit  and  a  guinea-pig.  In  all,  nine  rabbits  and  three  guinea-pigs  were  inoc- 
lated,  chiefly  with  tissues  from  cancer  of  the  breast.  All  inoculations  were 
negative. 

Third.  Subcutaneous  and  intraperitoneal  inoculations  and  injections  into  ear 
vein,  liver,  and  anterior  chamber  of  the  eye  of  rabbits  and  of  guinea-pigs,  of  the 
"  Saccharomyces  neoformans "  of  Sanfehce  and  the  micro-organism  of  Plim- 
mer produced  only  inflammatory  and  proliferative  changes.  No  tissue  resem- 
bling cancer  was  produced. 

Fourth.  Cultures  were  made  from  thirteen  cases.  In  three,  pyogenic  cocci 
grew,  the  other  ten  remained  sterile. 


1138  Journal  of  Applied  Microscopy 

The  writer  states  that  his  work  is  not  yet  completed  ;  and  he  therefore  can- 
not give  definite  conclusions.  The  results  so  far,  however,  have  been  definitely 
negative.  R.  m.  p. 

Qreenough,  R.  B.     On  the  Presence  of  the  So-  I"     this    work     Greenough     examined 

called  "  Plimmer's  Bodies  "  in  Carcinoma,  twenty-three  carcinomata  of  the  breast. 

Journal  of  the    Boston  Society  of  Medical  ™,      ..  j  •     '7      i       > 

Sciences.    Vol.  V,  No.  2,  1900  ^he  tissue  was  preserved  in  Zenker  s  or 

Hermann's    fluid,    the    former    giving 
the  best  results.     Sections  were  stained  according  to  Plimmer's  directions  with 
iron  haematoxylin  and  counterstained  with  either  Orange  G  and  fuchsin  or  with 
Bordeaux  red. 
Conclusions  : 

1.  The  appearances  known  as  "  Plimmer's  bodies  "  were  found  in  each  of 
twenty-three  cases  of  breast  cancer. 

2.  They  were  more  numerous  in  the  periphery  of  the  tumors  and  in  the 
metastases. 

3.  They  were  not  found  in  areas  which  had  undergone  even  slight  degenera- 
tion, whether  before  or  after  removal. 

4.  They  were  more  numerous  in  the  slow  growing  carcinomata,  and  less 
frequently  found  in  the  rapid  growing  ones. 

5.  They  were  more  numerous  in  scirrhous  than  in  medullary  or  adeno-carci- 
noma  types  of  cancer. 

6.  They  were  not  found  in  three  cases  of  epithelioma  (one  of  which  was  a 
typical  Paget's  disease  of  the  breast). 

7.  They  were  present  in  one  case  of  ovarian  cancer  and  absent  in  another 
case  of  general  peritoneal  cancer,  of  probable  ovarian  origin.  R.  m.  p. 


GENERAL   PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Parker,  0.  H.,  and  Burnett,  F.  L.    The  Reac-      ^^is  paper  gives  an  account  of  a  study 
tions  of  Planarians,  with  and  without  Eyes,  r-    r-       o 

to  Light.    Amer.  Jour.  Physiol.  4 :  373-385,      with  very  exact  methods  of  the  reactions 
'900-  to  light  of  normal  planarians  as  com- 

pared with  specimens  from  which  the  eyes  had  been  removed.  The  species  used 
was  Planaria  gonocephala  Duges.  The  method  employed  was  such  as  to  admit 
of  an  exact  statistical  treatment  of  the  problem,  and  is  on  that  account  very 
valuable.  In  detail  it  was  as  follows  :  A  planarian  was  placed  in  a  shallow  rec- 
tangular glass  dish  containing  water  to  a  depth  of  about  one  centimeter.  After 
the  animal  had  begun  to  creep  along  the  bottom,  the  dish  was  placed  on  a  black 
board  on  which  was  inscribed  a  circle  55  millimeters  in  diameter.  This  circle 
was  divided  into  quadrants  by  mutually  perpendicular  diameters,  and  the  arc  of 
each  quadrant  was  further  divided  by  short  cross  lines  into  intervals  of  ten 
degrees.     "  These  lines  were  designated  in  degrees,  the  one  at  the  end  of  one  of 


and  Laboratory  Methods.  1139 

the  diameters  being  taken  as  zero,  and  those  in  the  semicircles  to  the  right  and 
to  the  left  of  this  zero  being  numbered  in  corresponding  series  till  they  met  at 
180°."  The  dish  containing  the  animal  was  placed  over  this  circle  in  such  a 
way  as  to  bring  the  center  of  the  animal  over  the  center  of  the  circle,  and  its 
head  directed  towards  the  zero  point  on  the  circumference.  The  apparatus  was 
set  up  in  a  dark  chamber  in  order  to  exclude  extraneous  light,  and  the  illumina- 
tion for  the  experiment  was  obtained  from  a  Welsbach  burner  placed  at  a  con- 
stant distance  from  the  center  of  the  circle.  The  heat  rays  were  absorbed  by  an 
alum  solution  contained  in  a  parallel-sided  glass  vessel,  which  was  placed  between 
the  light  and  the  dish  containing  the  animal.  The  light. was  made  to  enter 
horizontally  only,  or,  by  the  use  of  a  screen  and  reflector,  vertically  from  above. 
The  anterior  end  of  the  animal  and  zero  point  of  the  circle  were  towards  the 
source  of  light  entering  horizontally  in  one  set  of  experiments,  and  away  from  it 
in  another  set,  while  in  a  third  the  light  entered  from  above.  For  one  series  the 
eyes  were  removed  by  cutting  off  the  head  with  a  sharp  scalpel.  In  each  experi- 
ment the  animal  was  observed  from  the  time  it  left  the  center  till  it  crossed  the 
circumference  of  the  circle.  Its  path  was  marked  free-hand  on  a  circle  which 
was  a  duplicate  of  the  one  on  the  black  board,  and  later  measured  in  millimeters. 
The  angle  on  the  circumference  at  which  the  animal  crossed  was  read  directly, 
and  the  time  taken  in  the  passage  from  center  to  circumference  was  obtained  by 
means  of  a  stop-watch. 

This  method,  with  modifications  to  suit  particular  cases,  will  undoubtedly 
prove  very  valuable  in  work  in  phototaxis.  Exact  records  can  be  obtained  of  the 
angle  of  deviation  in  the  path  caused  by  light  coming  from  one  direction ;  of  the 
form  of  the  path  taken  and  the  distance  travelled  ;  and  of  the  rate  of  travel  under 
constant  light  stimulation.  The  principal  result  of  the  investigation  was  to  show 
that  planarians,  with  or  without  eyes,  when  moving  horizontally  totuards  a  source 
of  light  are  more  deflected  from  an  ideal  course  (to  zero  on  the  scale)  than  when 
moving  under  a  vertical  light,  and  conversely,  when  moving  horizontally  away 
from  a  source  of  light  they  keep  nearer  to  an  ideal  course.  The  animals  without 
eyes  are  affected  by  light  in  the  same  way  as  those  with  eyes,  but  their  reaction 
is  less  precise.  The  rate  of  movement  of  the  decapitated  animals  is  slower  than 
that  of  the  normal.  The  reactions  are  believed  to  be  due  to  a  dermatoptic 
function.  R-  ^^ 

Matthews,  A.  P.    Some  Ways  of  Causing  Mi-      Several  new  methods  are  given  in  addi- 

totic  Division  in  Unfertilized  Arbacia  Eegs.         .        ^     ^i  u-    u  u  i        j      u^^^ 

Amer.  Jour.  Physiol.  4:  343-347,  1900.  ^lon  to  those  which  have  already  been 

described  for  producing  cell  division  in 
the  sea  urchin  egg.  Lack  of  oxygen  is  first  discussed.  Unfertilized  Arbacia 
eggs  were  placed  in  an  Engelmann  gas  chamber  in  sea  water,  and  hydrogen  gas, 
carefully  freed  from  acid,  was  passed  through  the  preparation.  After  twenty  to 
thirty  minutes'  exposure  to  the  hydrogen,  oxygen  was  admitted  for  ten  minutes, 
and  then  again  hydrogen  was  allowed  to  act  for  twenty  minutes.  The  eggs 
were  then  transferred  to  fresh  sea  water,  and  in  a  comparatively  short  time  clear 
areas  appeared  in  the  cytoplasm,  and  division  into  from  two  to  eight  cells  took 
place.  Continuous  exposure  to  hydrogen  kills  the  eggs  before  any  segmentation 
occurs.     Eggs  which  have  been  too  long  exposed  immediately  liquify  completely 


1140  Journal  of  Applied  Microscopy 

when  oxygen  is  admitted.  Warming  the  eggs  to  32°  or  33°  C.  for  from  two  to 
four  minutes  causes  the  clear  areas  to  appear,  and  segmentation  to  occur,  after 
the  return  to  sea  water  of  ordinary  temperature.  Segmentation  is  also  caused  by 
exposing  the  eggs  to  the  action  of  sea  water  in  which  either  ether,  or  alcohol,  or 
chloroform  has  been  dissolved.  In  all  cases  division  did  not  occur  until  the 
eggs  were  brought  back  into  ordinary  sea  water.  In  his  theoretical  conclusions, 
drawn  from  these  experiments,  the  author  is  inclined  to  abandon  his  earlier 
view  that  karyokinesis  is  allied  to  the  process  of  blood  clotting,  and  states  that 
he  believes  "  that  whatever  the  details  of  the  process  may  prove  to  be,  the  essen- 
tial basis  of  karyokinetic  cell  division  is  the  production  of  localized  areas  of 
liquefaction  in  the  protoplasm."  In  view  of  the  great  ease  with  which  Arbacia 
eggs  can  be  made  to  segment  by  a  variety  of  stimuli  of  different  physical  and 
chemical  character,  such  a  broad  generalization,  having  as  a  basis  the  phenomena 
shown  by  these  eggs  under  a  particular  set  of  conditions,  seems  to  be  of  uncer- 
tain value.  R.  P. 

Carlgren,  0.     Ueber  die   Einwirkung  des  con-       ^j^g  f^^g^  ^f  ^.^ese  papers  makes  a  note- 

stanten   galvanischen   btromes  auf   niedere  '^    '^ 

Organismen.    Arch.  Anat.  u.  Physiol.  Abth.,      worthy   advance  in   our  knowledge   of 

1899  pp^49-76,  I  Taf.  ^^^  gl=fect  of  the  constant  current   on 
Ueber  die  Einwirkung  u.  s.  w. :  Zweite  Mitth. 

Versuche  an  Verschiedenen   Entwicklungs-  organisms,    since    it   demonstrates    the 

stadien^emiger   Evertebraten.      Ibid,   1900,  importance    of    the    kataphoric,  or    SO- 

called  "  osmotic"  action  of  the  current. 
It  is  principally  given  to  an  account  of  the  electrotactic  response  of  Volvox. 
The  sense  of  the  reaction  is  at  first  kathodic,  but  later  changes  to  anodic.  Strik- 
ing changes  in  the  form  of  the  body  are  produced  by  the  current.  The  anode 
side  of  the  colony  becomes  crumpled,  while  the  kathode  side  is  correspondingly 
swollen  out.  The  parthenogonidia  move  to  the  anode  side  of  the  colony.  These 
changes  in  body  forms  and  movements  of  the  parthenogonidia  are  entirely  passive 
phenomena,  the  anode  crumpling  and  kathode  swelling  occurring  in  colonies 
which  have  been  killed  in  formalin  in  the  same  way  as  in  living  specimens. 
Various  Protozoa  killed  in  weak  formalin  or  ether  solution  show  the  same 
changes  in  form  under  the  action  of  the  current.  Carlgren  concludes  that  the 
purely  physical,  kataphoric  action  which  produces  these  results  is  of  very  great 
significance  as  a  factor  in  the  effect  of  the  current  on  organisms. 

In  the  second  paper  descriptions  are  given  of  the  electrotactic  responses  of  a 
number  of  marine  invertebrates.  The  point  of  most  general  interest  is  in  regard 
to  the  reactions  of  the  larva;  of  certain  echinoderms  {Strongylocentrotus  lividus, 
Spharechinus  granulans,  Ophiothrixfragilis,  ?iuA  Aster acanthion  glacialis).  Young, 
free-swimming  stages  of  these  forms  gave  no  response  whatever,  while  older 
larvae,  Plutei  and  Bipennaria;,  became  oriented  and  went  to  the  kathode.  Theor- 
etical discussion  of  the  results  is  left  for  a  later  paper.  No  new  methods  of 
work  are  described.  R.  p. 

Warren,    E.      On   the   Reaction   of    Daphnia       -pj^^     experiments     described     in     this 
magna  (Straus)  to  Certain    Changes  in   its  ' 

Environment.     Q.   J.    Mic.    Sci.    N.  S.  43 :       paper  have  to    do    with   the    effect   on 

'99-224,  1900-  Daphnia  of  certain  changes  in  the  con- 

ditions of  life.     It  was  found  that  the   time  of  killing   in  solutions   of  NaCl   of 


and  Laboratory  Methods.  1141 

different  strengths  (.8  per  cent,  to  G.O  per  cent.)  seems  to  depend  quite  exactly 
on  the  number  of  molecules  of  salt  which  strike  the  animal  in  a  unit  of  time. 
The  relation  of  time  of  killing  and  strength  of  solution  is  represented  by  the 
rectangular  hyperbola  y(x — 8) =277.  An  increase  in  temperature  causes  the 
molecules  to  move  faster  and  strike  with  greater  momentum,  and  hence  the  time 
of  killing  is  reduced  in  high  temperatures.  The  physiological  condition  of  the 
animal  is  a  most  important  factor  in  determining  its  power  of  resistance  to  NaCl. 
Perhaps  the  most  striking  result  of  the  investigation  is  that  animals  which  have 
become  acclimatised  to  a  .25  per  cent,  salt  solution  show  less  resistance  capacity 
than  do  normal,  unacclimatised  specimens,  to  solutions  of  greater  concentrations. 
The  author  thinks  that  this  is  probably  due  to  some  constitutional  weakness 
resulting  from  the  acclimatising  process.  Daphnia  living  in  a  confined  volume 
of  water  were  shown  to  have  shorter  caudal  spines,  and  to  reproduce  less  vigor- 
ously than  those  that  had  lived  in  an  unlimited  bulk.  Water  in  which  Daphnia 
has  lived  for  some  time  has  a  poisonous  effect  on  individuals  from  other  cultures. 
This  paper  is  of  interest  in  connection  with  the  recent  work  of  Miss  Enteman 
(Amer.  Nat.  34  :  879-890)  on  the  extreme  variability  of  a  related  species  Daphnia 
hyalina  under  differing  natural  environmental  conditions.  R.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Flexner.     The  Etiology  of  Tropical  Dysentery.       The    author    has    made     an     extensive 

Cent.  f.  Bac.  u.  Par.  I,  28 :  625,  1900.  •         r    i.    j-         r  j 

^  series  of  studies  of  dysentery  occurring 

in  the  Philippines,  and  comes  to  the  conclusion  that  dysentery  is  of  two  distinct 

types.     One,  the  chronic  form,  is  accompanied  by  the  presence  of  amoebae  in  the 

intestines  in  large  quantities,  and  is,  therefore,  what   has  been  called   amcebic 

dysentery.     The  other,  the   acute  form,  is  not   accompanied  by  these  protozoa, 

and  appears  to  be  produced  by  bacteria.     The  author   finds  universally  present 

in  these  cases,  a  bacillus  which  he  describes  and  with  which  he  experiments. 

This  bacillus  is  pathogenic  for  small  animals,  producing  symptoms  quite  similar 

to  those  of  dysentery,  and   is  believed   by  the  author  to  be  unquestionably  the 

cause  of  acute  dysentery  in  eastern  countries.     The  organism  is  identical  with 

that   isolated  by  Shiga  from   the   epidemic   of  dysentery   prevailing  in   Japan. 

Flexner  regards  this  cause  of  dysentery  as  widely  distributed  in  nature. 

H.  W.   C. 

Ritchie.     The    Bacteriology    of    Bronchitis.      The    author    makes    a   bacteriological 

Jour,  of  Path,  and  Bact.  7:   i,  looo.  ^     i         r  i.  r  r    l 

■'  ^  Study  of   a  number  of  cases   of  bron- 

chitis. In  most  instances  they  were  made  by  post  mortem  examinations,  and 
were  bacteriological  studies  of  the  lung  tissue.  Numerous  bacteria  are  found  in 
the  lungs  under  the  circumstances,  most  of  which,  as  would  be  expected,  have 
nothing  to  do   with  the  disease.     The  general  conclusion  of  the   author  is   as 


1142  Journal  of  Applied  Microscopy 

follows  :  Acute  bronchitis  is  an  infective  disease  but  not  due  to  any  specific 
organism.  Various  bacteria  are  found  in  the  secretions,  some  of  which  are  the 
exciting  causes  of  the  disease.  The  disease  is  more  often  due  to  a  mixed  infec- 
tion than  to  the  action  of  bacteria.  The  most  important  causal  bacteria  are  the 
Diptococcus  pneumonce  and  streptococci.  The  author  also  believes  that  the 
influenza  bacillus  is  not,  infectionally,  the  cause  of  bronchitis,  independent  of 
the  ordinary  form  of  influenza.  H.  w.  c. 

Moore.  Veranus  A.,  B.  S.,  M.  D.     Professor  of  This  excellent   manual,  originally  pub- 
Comparative   Pathology  and   Bacteriology,  Hshed  in   1898   (reviewed  in  the  JOUR- 
New  York  State  Veterinary  .College,  and  of  ,^    ,      ,       i.^        i^  -.-on     i 
Bacteriology,    Cornell    University    Medical  NAL,    Vol.     1,     No.    9,    page    1/2),   has 
College,  Ithaca,  N.  Y.     An  Introduction  to  already   gone    into    a    second    edition, 
Practical    Bacteriology    for    Students    and              ,  ,,             ,,          ,         ,    ,           ^, 
Practitioners  of   Comparative  and    Human  ''^"^  the    author    has  taken    the   oppor- 
Medicine.      Second    edition,   enlarged   and  tunity    to     revise     the     exercises     and 
revised.    Boston,  U.  vS.  A.,  Ginn  &  Co.,  Pub-  .            i  ,                    i  n 
lishers.    The  Athena:um  Press,  igoo.  ^o    add     several     more,    as    well    as 

an  appendix,  the  book  being  now 
twice  the  former  size.  The  new  chapters  are  on  the  morphology  of  the 
coccus  forms,  bacilli  and  spirilla,  a  study  of  Pseudovionas  pyocyafieiis,  of 
Bacillus  teta/ii,  and  of  the  bacteria  of  the  healthy  mouth.  The  appendix  con- 
tains a  reprint  of  the  method  of  determining  the  reaction  of  the  culture  media 
recommended  by  the  committee  of  the  American  Public  Health  Association  in 
1897,  together  with  brief  directions  for  inoculation  experiments  on  animals.  All 
of  the  exercises  are  exceedingly  practical  and  practicable,  the  directions  are 
concise  while  being  sufficiently  explicit,  and  references  to  standard  literature 
lead  the  student  to  extend  his  knowledge  by  consulting  the  authorities  in  the 
science.  The  selection  of  exercises  amply  justifies  the  title  of  the  book. 
University  of  Rochester.  Charles   WriGHT    Dodge. 

Hall,  H.  0.     The   Etiology  of   Scarlet    Fever.       The  feature  of  this  paper  consists  in   a 

New  York  Medical  Record,  56:  607,  i8gq.  ,        ^   ,,  1  •      1     i-       -i     ^• 

^       ^  Study  of  the  geographical   distribution 

of  scarlet  fever  and  its  relation  to  the  use  of  milk  as  a  food.     The  author  finds 

that  the  disease  occurs  in  all  countries  where  cow's  milk  is  an  important  article 

of  diet,  especially  for  children.     It  is  lacking,  however,  in  those  countries  where 

cow's   milk   is  not    used.       In    China    and    Japan,    where    cow's    milk   is  not 

used    as  food,    the    disease    is    unknown.      In    India,    where    cow's    milk    is 

used    for    adults    but    not    for    children,    the    disease    is    extremely    rare.       In 

countries  where  asses'  milk  or  goat's  milk  is  used,  scarlet  fever  is   unknown. 

The  author  is  of  the  opinion  that  this  is  a  disease  primarily  distributed  by  milk. 

H.  w.  c. 

Courmant.  ^'agglutination  der  bacille  de  Koch  The  author  has  .Studied  the  problem  of 
des  epauchements  tuberculeux.  Arch.de  the  agglutination  of  the  tubercle  bacillus 
Med.  Kxp.  12:    697,  1900.  °° 

by  the  various  exudations  from  tuber- 
culous animals.  He  finds  that  the  exudations  of  tuberculous  animals  always 
produce  an  agglutination  of  the  bacillus,  but  that  the  amount  of  agglutination  is 
not  proportioned  to  the  extent  of  the  disease.  Advanced  cases  of  the  disease 
produce  little  agglutination,  while  incipient  cases  produce  a  very  marked  effect. 
The  author  believes  that  the  phenomenon  may  be  of  a  decided  diagnostic  value. 


and  Laboratory  Methods.  1143 

If  the  serous  exudations  of  a  suspected  individual  produce  an  agglutination,  it 
indicates  the  presence  of  the  disease.  The  absence  of  the  reaction,  however, 
does  not  necessarily  indicate  the  absence  of  the  disease,  for  advanced  cases 
produce  no  reaction.  The  agglutinative  power  of  the  serai  exudations  is  greater 
than  that  of  the  blood  of  the  same  animal,  and,  hence,  the  author  concludes  that 
the  power  of  agglutination  is  developed  in  the  exudatio-ns,  and  are  not  simply  a 
phenomenon  transferred  from  the  blood.  h.  w.  c. 

Rogers.     Schutzimpfung   gegen    Rinderpest.      The      experiments     here      mentioned 

Zeit.  f.  Hyg.  u.  Infek.  21 :   SQ,  iQOO.  i  -i  i  •  r  •  ^-      i.- 

•"'  j^?'   ^'  describe  a  long  series  of  investigations 

upon  the  value  of  the  method  of  inoculating  against  rinderpest.  The  author 
experiments  not  only  with  the  method  of  Koch,  but  with  two  or  three  other 
methods  that  have  recently  been  devised.  His  general  conclusion  is  that  inocu- 
lation by  the  gall  method  produces  an  immunity  in  cattle  against  this  disease, 
but  that  the  immunity  is  quite  fleeting,  lasting  only  about  four  months.  He  finds, 
further,  that  different  classes  of  cattle  behave  quite  differently  towards  this 
inoculating  test.  Mountain  cattle  and  lowland  cattle  are  very  different  in  their 
sensitiveness  to  immunization  and  to  the  disease.  The  former  are  not  easily 
immunized  by  the  gall  method.  Of  the  several  methods  used  the  author  believes 
that  some  are  best  for  certain  breeds  of  cattle,  and  others  for  other  breeds  of 
cattle.     The  paper   hardly  admits  of  summary.  h.  w.  c. 

Saul.     Beitriige  zur  Morphologie  des  Staphy-      This    paper    consists    of    a    somewhat 
lococcus  albus.     Bed.  Klin.  Woch,  p.  1058.      unique  study  of  the  gelatin  colonies  of 
^    ■  this  well  known  organism.     The  author 

makes  his  studies  in  gelatin  plates  inoculated  in  such  a  way  that  each  plate  con- 
tained only  one  or  two  colonies,  and  preserved  under  conditions  to  retain  their 
moisture  so  that  the  colonies  could  continue  growing  for  months.  The  gelatin, 
with  the  contained  colony,  was  hardened  and  sectioned,  and  careful  studies  made 
of  the  sections.  Some  excellent  figures  are  given  of  the  colonies.  The  impor- 
tant conclusion  is  that  the  colonies  are  not  simply  irregular  aggregates  of  cells, 
but  appear  to  be  units,  and  should  be  regarded,  therefore,  rather  as  "  cell  states  " 
than  as  irregular  aggregations.  The  colonies,  though  varying  widely  in  form, 
are  always  reducible  to  a  fundamental  type  which  appears  to  be  based  upon  the 
principle  of  dichotomous  branching.  h.  w.  c. 

Trommsdorff.     Ueber  Gewohnung  von  Bak-      The  evidence  for  a  germicidal  action  of 
terien    an    Alexin.     Arch.    f.    Hyg.   39:   ^i,       ,       1  1      i  11        11         1 

jQQQ  -^  freshly  drawn  blood  has  been,  in  recent 

years,  subject  to  criticism.  It  has  been 
pointed  out  that  micro-organisms,  when  transferred  from  one  medium  to  another, 
commonly  suffer  some  injurious  influence,  and  for  a  time  fail  to  increase, — or 
may  even  decrease.  It  has  been  suggested,  therefore,  that  diminution  of  bacteria 
in  freshly  drawn  blood  is  due  to  the  change  from  bouillon  culture  to  blood,  and 
not  to  any  poisonous  alexin.  The  author  tests  this  theory  by  cultivating  the 
typhoid  cholera  bacteria  in  the  blood  and  serum  of  animals  whose  blood  is  inac- 
tive by  heat  at  56°  for  one  hour.  After  cultivation  in  this  inactive  blood  the 
bacteria  are  inoculated  in  active  blood,  and  are  found  to  be  just  as  rapidly  killed 


1144  Journal  of  Applied  Microscopy 

by  the  fresh  blood  as  they  are  in  a  control  test  when  they  are  taken  directly  from 
bouillon.  To  determine  whether  the  bacilli  could  adapt  themselves  to  the 
alexins  in  the  active  blood,  Trommsdorff  cultivated  the  organism  in  fresh  blood. 
After  being  cultivated  in  this  active  blood  for  a  time,  they  were  transferred  to 
fresh  active  blood,  and  were  found  to  suffer  no  diminution  in  numbers.  He 
found,  further,  that  organisms  thus  adapted  to  the  alexins  of  the  ordinary  blood 
are  checked  in  their  growth  if  transferred  to  a  pleuralexuadite  which  has  the 
alexins  present  in  larger  quantities  than  ordinary  blood.  He  concluded,  there- 
fore, that   bacilli  quickly  adapt  themselves  to  the  alexins  in  the  blood. 

H.  w.  c. 


NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Friedel,  0.     Nouveaux    essais  sur  les  Zeolites       JSfatroUte  (Mesofvpe).      Author  treats  of 
(Suite  i).     Bull.  Soc.  Min.  22:  84,  1899.  ^     .       ^-^ / 

the  manner  in  which  water  is  expelled 

during  heating,  and  gives  a  plate  showing  dehydration  curves.     Concludes  that 

all  the  water  of  natrolite  is  of  the  same  nature  ("  zeolitic  ").  l.  mcI.  l. 

Pratt,  J.  H.    On  the  Separation  of  Alumina      Author  treats  of  the  differentiation  of 
from  Molten  Magmas,  and  the  Formation  of       .  tit 

Corundum.    Am  Jour.  Sci.  iv,  8 :  227,  1899.      igneous  magmas  upon  cooling,  and  of 

the  genesis  of  minerals.  The  separa- 
tion of  corundum  from  molten  magmas  is  "  dependent  upon  the  composition  of 
the  chemical  compounds  that  are  the  basis  of  the  magma ;  upon  the  oxides  that 
are  dissolved  with  the  alumina  in  the  magma,  and  upon  the  amount  of  alumina 

itself."  L.  McI.  L. 

Ward,  H.  L.     New  Meteorite  from  Murphy,      Neuman  lines  noted,  also  the  presence 
Cherokee  Co.,  N.  C.    Am.  Jour.  Sci.  iv.  8:      ^f  troilite  and  daubreelite. 
22  s,  1009. 

^         ^^  L.  McI.  L. 

INDIVIDUAL  SPECIES. 

Calcites  (Siliceous)  from  the  Bad  Lands,  Wash-      The  specimens  resemble    in  character 
ington  Co.,  So.  Dakota.     S.  L.  Penfield  and         ,        t-.  •    i  1  i-         ^  j 

W  E.Ford.    Am.  Jour.  Sci. iv. 9:  352,  1900.      the   Fountainbleau   limestone,  and   are 

gray  in  color.  They  consist  of  about 
40  per  cent,  of  calcite,  enclosing  60  per  cent,  of  quartz  sand.  The  crystals  occur 
singly,  but  more  often  in  groups,  and  evidently  have  formed  in  a  stratified  deposit 
of  sand,  representing  a  phase  of  sand  cementation  with  the  crystalline  form  of 
the  calcite  preserved.  The  crystal  forms  are  steep  hexagonal  pyramids  of  the 
second  order  (rare  in  calcite),  which  are  somewhat  barrel-shaped,  with  rounded 
ends.     The  Fountainbleau  crystals  show  the  acute  rhombohedron  /(0221). 

L.  McI.  L. 


and  Laboratory  Methods.  1145 

Quartz.     Sur  un  groupe  de  cristaux  de  quartz       The   group    consists    of    three    crystals 
de    Striegan    (silesie).     F.   Gonnard.     Bull.  .  ,  ^.      ,  n   i  i  i- 

Soc  Min  22:  02  i8qq.  with  vertical  axes  parallel,  and  pecuhar 

arrangement  of  faces.     Eight  forms  are 

noticed,  of  which  three  [(13.0.  13.1),  (5051),  (2577)]  are  probably  new,  one  being 

a  new  plagiohedron.  l.  McI.  l. 

Quartz.     Etude  cristallographiquedu  quartz      Quartz    occurs    in    clear,    pellucid    bi- 
des    geodes     des    marnes    oxfordiennes    de  '■ 

Meylan   (Isere).      F.    Gonnard.     Bull.  Soc.       pyramids,  modified  by  very  small  prism 
Min.  22:  94,  1899.  faces,  and  also  showing  a  large   num- 

ber of  new  or  rare  forms.  Tabulation  of  these  forms  given.  Liquid  inclusions 
with  air  bubbles  also  noticed,  and  the  Assuring  of  the  crystals,  sometimes  ob- 
served, is  thought  to  be  due  to  the  expansion  of  this  liquid.  l.  McI.  l. 

Stokesite.    A.Hutchinson.    Phil.  Mag.,  Nov.,      Description  from  a  single  crystal  (10 
^  '  mm.  long)  in  Cambridge  Mineralogical 

Museum.  Orthorhombic,  with  forms  ^  (010)  and  z' (121).  a:/?:c=0.M7Q:l:- 
0.8117.  Cleavage  perfect,  parallel  to  l>,  and  also  good  parallel  to  prism,  taken 
as  unite;  fracture,  conchoidal ;  H.=6 — 6.5;  G.^3.185  ;  lustre,  vitreous,  pearly 
on  ^  ,•  colorless.  Partial  chemical  examination  determines  it  as  a  hydrated 
silicate  of  Na  and  Ca,  with  about  6  per  cent,  of  tin  oxide,  replacing  part  of  the 
SiOa- 

L.  McI.  L. 


Medical  Notes. 

Note  on  Examination  of  Blood. — A  microscopical  examination  of  the 
stained  specimen  of  pathological  blood  implies  a  comparison  with  the  appearance 
of  normal  blood  when  subjected  to  the  same  straining  process.  The  experienced 
observer  unconsciously  makes  use  of  his  mental  picture  of  the  normal  specimen 
in  doing  this  work,  and  to  him  it  is  sufficient.  In  fixing  and  staining  blood 
spreads,  however,  a  slight  variation  in  technique  may  produce  a  decided  differ- 
ence in  results,  consequently  those  who  have  had  comparatively  little  experience 
in  such  work  will  find  it  difficult  to  secure  uniform  results  without  a  considerable 
laboratory  equipment.  In  preparing  pathological  specimens  in  such  cases  a 
spread  of  normal  blood  may,  at  the  same  time,  be  subjected  to  the  same  technique 
and  mounted  on  the  slide  with  the  pathological  specimen,  making  exact  and 
reliable  comparison  a  very  easy  matter.  Dried  blood  spreads  can  be  kept 
indefinitely,  so  a  supply  of  normal  specimens  can  easily  be  kept  constantly  in 
readiness  for  use.  W.  A.  Fulton,  M.  D. 

Burlington,  Wis. 

Kober.     The  Presence  of  Diphtheria  Bacilli  in       Examinations  were  made   in  128  cases 
the  Mouths  of  Healthy   Individuals.     Zeit.  ,  .   ,  ,  ^      ,  , 

f.  Hyg.  31 :  4^3.  which  were   known  to   have   been  ex- 

posed to  diphtheria,  and  in  600  cases 
which  were  supposedly  not  so  exposed.  The  investigations  included  microscopical 
study,  cultivation  upon  serum,  and  inoculation  of  guinea  pigs.    Of  the  128  cases, 


1146  Journal  of  Applied  Microscopy 

only  10,  or  about  8  per  cent.,  gave  evidence  of  infection;  while  of  the  600 
cases,  but  15,  or  2.5  per  cent.,  showed  the  presence  of  bacilli,  and  10  of  these 
later  gave  evidence  of  having  been  previously  exposed,  thus  greatly  reducing  the 
percentage  in  the  last  experiment.  It  is  generally  estimated  that  diphtheria 
bacilli  are  found  in  the  mouths  of  about  18  per  cent,  of  all  healthy  individuals. 
The  results  above  cited  would  indicate  that  this  per  cent,  is  very  much  too  high. 

c.  w.  J. 

May,  Richard.   The  Use  of  Orcein  in  the  Dem-       The  sputum   is   thoroughly  mixed  with 
onstration  of  Elastic  Fibres  in  the  Sputum.  ,  ^     r  i  n 

Deut.  Archiv.  f.  Khn.  Med.  68:  427.  ^n  equal  amount  of  10  per  cent,  caustic 

potash  solution,  care  being  exercised 
that  no  more  heat  is  used  than  is  needed  to  dissolve  the  sputum.  When 
thoroughly  dissolved,  centrifugalize  and  pour  off  the  liquid  portion.  Add  to  the 
sediment  about  2  c.  c.  of  Unna-Tanzer's  orcein  solution,  the  composition  of  which 
is  as  follows  : 

Orcein, 1.0 

Alcohol,  absolute, 80.0 

Water,  dist., 40.0 

HCl,  cone, 40.0 

This  solution  has  a  red  color,  which  changes  to  violet  when  the  solution 
comes  in  contact  with  the  potash  of  the  sediment.  The  original  color  is  regained 
by  adding  three  to  five  drops  of  HCl. 

Place  the  centrifuge  tube  in  boiling  water  for  three  to  five  minutes,  as  heat  is 
necessary  to  hasten  the  staining  process.  Hydrochloric  acid  alcohol  is  then 
added,  and  after  gently  shaking  the  solution,  it  is  centrifugalized  by  a  few 
turns  of  the  machine ;  the  same  process  being  repeated  twice  with  fresh  acid 
alcohol.     The  formula  for  the  hydrochloric  acid  alcohol  decolorizing   solution 

is  as  follows : 

Hydrochloric  acid,  cone,  .     .  5.0 

Alcohol,  95  per  cent.,    .     .     .  1000.0 

Water,  dist., 250.0 

Malkes,  J.     Estimation  of  Mercury  in  Urine.       599  c.  c.  of  urine  are  mixed  with  5  c.  c. 
Chem.  Zeit.  35:  816,  1900. 

of  egg  albumm  and  15  drops  of  acetic 

acid,  and  heated  for  fifteen  minutes  on  the  water  bath.     The  mixture  is  poured 

into  a  beaker,  allowed  to  settle,  and  the  deposit  collected  in  a  filter.     The  paper 

and  its  contents  are  laid  on  a  porous  tile  for  a  few  minutes.     The  precipitate  is 

removed  to  a  small  cylinder  and  covered  with  50  c.  c.  strong  HCl,  a  spiral  of 

Cu  being  immersed  in  the  liquid.     After  about  fourteen  hours  all  the  Hg  has 

amalgamated  with  the  Cu,  and  the  acid  is  dark  in  color.     The  wire  is  washed 

with  water,  alcohol,  and  ether,  then  dried,  after  which  it  is  dropped  into  a  tube 

5  mm.  in  diameter  with  a  crystal  of  iodin,  and  heated   until  the  sublimate  of 

Hgl  2  appears  on  the  wall  of  the  tube.    The  amount  of  mercury  is  compared  with 

that  produced  by  a  ring  obtained  in  the  same  manner  from  a  urine  to  which  a 

known  quantity  of  HgClg  has  been  added.  c.  w.  j. 


and  Laboratory  Methods.  1147 

NEWS  AND  NOTES. 

Mr.  F.  B.  Kilmer  gives  the  following  report  of  the  December  meeting  of  the 
New  Brunswick  Society  : 

Under  the  reports  of  sections,  Dr.  Henry  R.  Baldwin  announced  that  experi- 
ments were  being  made  with  luminous  bacteria. 

Prof.  F.  C.  Van  Dyck  explained  a  new  application  of  the  microscope  to 
ascertain  the  tensile  strength  of  metals. 

The  President,  F.  B.  Kilmer,  delivered  a  paper  entitled  "  A  Study  of  Cotton," 
which  was  illustrated  by  lantern  slides  and  by  slides  under  the  microscope.  He 
showed  that  cotton  fibre  had  been  in  use  in  some  form  since  very  ancient  times ;  that 
while  the  principal  use  for  cotton  fibre  is  the  manufacture  of  thread  and  cloth,  in 
recent  years  many  new  and  important  uses  have  been  devised.  It  forms  a 
component  part  of  the  high  explosives  which  are  known  as  gun  cotton,  smokeless 
powder,  tonite,  blasting  gelatin,  etc.  In  the  form  of  nitrated  cotton,  which  is 
soluble  in  certain  liquids,  varnishes  and  lacquers  for  metal,  paper,  wood  and 
cloth,  imitation  leather  and  silk,  substitutes  for  India  rubber  and  gutta  percha 
are  manufactured.  Materials  of  this  character  made  of  cotton  were  exhibited  by 
the  speaker.  A  modern  application  for  cotton  is  its  use  as  a  dressing  for 
wounds. 

Cotton  for  surgical  purposes  is  known  as  absorbent  cotton,  which  means  that 
the  oil,  wax  and  varnish-like  coating  of  the  fibre  have  been  removed,  and  the  fibre 
thereupon  absorbs  water  and  other  liquids. 

The  speaker  explained  the  minute  structure  of  a  cotton  fibre,  and  while  this 
appeared  to  the  naked  eye  as  a  solid  cylindrical  hair,  under  the  microscope  it 
was  found  to  be  a  more  or  less  collapsed  tube  with  an  outer  sheath  and  an  inner 
opening  to  the  center  of  the  tube.  This  sheath  was  associated  with  a  varnish  or 
oily  substance  and  the  whole  permeated  with  wax  and  coloring  matter.  He 
stated  that  while  this  was  the  accepted  construction  of  the  fibre,  he  had  reason 
to  believe  there  was  much  yet  to  be  learned,  and  slides  were  exhibited  to  show 
that  the  structure  was  very  complex.  A  number  of  slides  showed  the  cotton 
plant  in  the  various  stages  of  growth ;  its  cultivation,  picking  and  preparation  for 
the  market  and  shipping.  Among  the  slides  were  those  which  gave  the  typical 
cotton  staples  of  the  world. 


The  second  annual  meeting  of  the  Society  of  American  Bacteriologists  was 
held  in  the  Pathological  Laboratory  of  Johns  Hopkins  University  on  December 
27  and  28.  A  full  programme  of  papers  was  presented  and  four  sessions  of  the 
society  were  held.  The  society  elected  Prof.  W.  H.  Welch  of  Johns  Hopkins 
University  as  its  president  for  the  year  1901.  Information  concerning  the 
society  may  be  obtained  by  writing  the  secretary,  H.  W.  Conn,  Middletown,  Ct. 


Methods  in  Germination  of  Spores.  —  The  hanging  drop  culture  is 
undoubtedly  the  most  convenient  method  of  observing  spore  germination.  It  is 
desirable  to  employ  large  rings,  which  should  be  cemented  to  the  glass  slips  by  a 


1148  Journal  of  Applied  Microscopy. 

mixture  of  refined  beeswax  and  pure  vaseline.  The  cover  should  be  cemented 
to  the  ring  with  vaseline.  The  same  character  of  liquid  should  be  used  at  the 
bottom  of  the  cell  as  employed  in  the  drop. 

This  form  of  cell  culture,  although  highly  accurate  for  culturable  forms  in 
nutrient  media,  will  not  give  best  results  when  a  careful  study  is  to  be  made  of 
particular  stimulants  in  water,  or  in  a  medium  not  ordinarily  causing  abundant 
germination.  Any  volatile  or  soluble  substance  besides  the  medium  employed  is 
apt  to  reduce  the  trustworthiness  of  the  results.  Even  the  purest  vaseline  may 
have  an  effect  on  sensitive  forms.  As  a  modification  of  the  above  method,  the 
cells  may  be  used  in  Petri  dishes,  on  the  bottom  of  which  is  placed  filter  paper 
with  holes  for  the  insertion  of  cells,  thus  securing  them  against  movement.  The 
covers  are  laid  on  without  vaseline  and  the  whole  kept  in  moist  chambers.  Petri 
dishes  with  ground  glass  tops  are  preferable. 

A  decoction  of  green  string  beans  or  of  sugar  beet  is  recommended  as  the 
best  culture  medium  for  must  readily  culturable  fungi ;  392  grams  of  green 
beans  per  liter  of  water,  or  about  50  grams  of  dry  matter  per  liter,  have  been 
found  satisfactory  proportions. 

As  a  standard  nutrient  salt  solution,  the  following  formula  is  well  known. 
It  may  be  used  without  the  sugar,  the  osmotic  influence  being  neglected  as  of 
little  consequence  in  comparison  with  the  desirability  of  having  equivalent  salt 
constituents  : 

Ammonium  nitrate     -     - 1.0  gram. 

Acid  potassic  phosphate      ------  0.5       " 

Magnesium    sulphate      -------  0.25    " 

Iron    -------------  trace. 

Cane   sugar      ----------  3  to  5  grams. 

Water      ---------.     -     -  100  c.  c. 


We  shall  be  very  glad  to  have  our  readers  avail  themselves  of  the  oppor- 
tunity which  we  have  previously  offered  in  the  Journal  and  as  herein  suggested 
by  Dr.  Bessey. 

Waterville,  Me.,  January  13,  1901. 

Journal  of  Applied  Microscopy  : 

Sirs  : — I  note  that  you  asked  your  patrons  to  suggest  anything  that  might 
occur  to  them  as  of  interest  in  making  the  Journal  more  interesting.  It  occurs 
to  me  that  if  you  should  set  apart  a  column  giving  the  subscribers  a  chance  to 
ask  questions  of  a  scientific  nature,  and  have  them  answered,  either  by  the 
editors  or  by  other  subscribers,  that  it  might  add  to  the  general  interest.  I 
think  almost  any  person  having  laboratory  work  would  like  to  ask  some  question 
that  is  not  in  publication,  and  could  be  answered  by  another  doing  such  work. 
I  merely  suggest  this.     Very  likely  you  have  already  considered  it. 

Most  sincerely  yours, 

M.  W.  Bessey,  M.  D., 
Colby  College.  Instructor  in  Zoology. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


Volume  IV. 


FEBRUARY,   1901. 


Number  2 


X'-. 


^  ►^  I*  ».   -- 


The  New  Biological  Laboratories  of  Ripoif 


College. 


BQT  Af^lO  A 


INGRAM    HALL  FROM    THE   NORTH. 


The  Biological  Laboratories  of  Ripon  College  are  located  in  the  first  story  and 
basement  of  Ingram  Hall,  which  was  occupied  for  the  first  time  at  the  beginning 
of  the  present  school  year. 

Ingram  Hall  contains,  besides  the  department  of  biology,  the  departments  of 
physics  and  chemistry,  physics  being  on  the  second  floor  and  chemistry  on  the 
third  floor.  The  building,  which  was  named  from  the  principal  donor,  Mr.  O. 
H.  Ingram  of  Eau  Claire,  is  73  by  121  feet  in  its  outside  dimensions,  and  is 
located  at  the  brow  of  the  hill  on  the  campus  in  such  a  way  that  the  south  side 

(1149) 


1150  Journal  of  Applied  Microscopy 

is  four  stories  in  height.  The  longer  dimension  of  the  building  is  east  and  west 
—the  entrance  to  the  first  floor  being  on  the  north  side,  and  to  the  basement  on 
the  south  side.  The  material  of  the  building  is  dark  red  vitrified  brick,  with 
rustications  of  Roman  brick  and  trimmings  of  Bedford  stone.  The  building  is 
in  form  a  rectangle,  with  only  such  projections  as  are  necessary  to  relieve  the 
monotony  of  its  exterior  surface.  The  architect  was  instructed  to  make  as  many 
windows  as  the  character  of  the  structure  would  permit,  and  the  result  is  that 
all  the  rooms  are  amply  lighted.  A  feature  of  the  construction  is  the  character 
of  the  partitions.  There  are  two  solid  brick  partitions  running  through  the 
whole  height  of  the  building.  The  other  partitions  are  only  two  inches  in 
thickness,  the  necessary  support  being  given  by  heavy  pillars.  These  partitions, 
a  device  of  the  architect,  Mr.  H.  K.  Holsman  of  Chicago,  have  a  core  of  wood, 
which  is  plastered  solid  on  both  sides.  At  first  thought,  they  would  seem  to  be 
very  unsubstantial,  but  as  a  matter  of  fact,  after  the  adamant  plaster  is  applied, 
not  only  are  the  walls  firm  and  substantial,  but  sound  is  not  carried  through  them 
to  any  disturbing  extent.  The  manifest  advantages  of  the  partitions  are  the 
economy  of  floor  space  and  the  fact  that  they  are  practically  fire-proof. 

The  building  is  finished  in  oak  throughout,  and  while  very  plain,  presents  an 
attractive  interior.  The  total  cost  was  about  ^33,000.  All  the  plun;bing  is 
"  open." 

The  laboratories  for  the  department  of  biology  are  so  located  as  to  use  north 
light  so  far  as  possible.  To  that  end,  the  room  for  museum  purposes  is  on  the 
south  side  of  the  building,  as  shown  in  the  annexed  floor  plan.  A  similar 
museum  room  on  the  second  floor  is  also  devoted  to  the  department  of  biology. 
These  museum  rooms  are  not  large,  and  were  not  intended  for  display  purposes, 
but  mainly  for  the  convenient  storage  of  materials  used  in  illustrating  the  lectures 
of  the  department.  A  lift  runs  through  the  entire  height  of  the  building,  and  by 
this  it  is  easy  to  convey  material  from  the  second  floor  to  the  first,  as  it  is 
required  for  use. 

Between  the  museum  and  the  lecture  room  is  an  apparatus  room,  or  prepara- 
tion room,  which  is  used  both  in  connection  with  the  work  of  the  museum  and 
in  preparing  material  for  lecture  work.  The  lecture  room,  in  the  southeast  cor- 
ner of  the  building,  has  the  floor  raised  in  four  steps,  so  that  all  students  can  see 
the  lecture  table  with  clearness.  The  blackboard  is  a  sliding  one,  and  back  of 
it  is  a  fixed  plate  of  ground  glass  which  can  be  used  for  illustrative  purposes  in 
the  lecture,  or  can  be  used  in  connection  with  the  lantern  work.  The  windows 
are  fitted  with  opaque  shades,  so  arranged  as  to  exclude  the  light,  in  order  that 
the  room  may  be  used  for  lantern  work. 

Adjoining  the  lecture  room,  on  the  north,  is  the  bacteriology  room.  This  is 
intended  as  a  place  where  the  various  forms  of  apparatus  connected  with  the 
work  in  bacteriology  can  be  permanently  set  up.  The  room  is  large  enough, 
also,  so  that  small  special  classes  in  bacteriology  can  do  their  work  there. 

The  two  rooms  at  the  west  end  of  the  building  are  the  general  laboratories, 
used  for  the  more  elementary  work,  and  for  the  vertebrate  work.  They  are  so 
arranged  that  by  a  movable  screen  the  two  rooms  can  be  used  separately  or  as 
one  large  room  for  especially  large  classes. 


and  Laboratory  Methods. 


1151 


11.") -J 


Journal  of  Applied  Microscopy 


At  the  right  of  the  north  entrance  is  the  private  laboratory  and  office  of  the 
head  of  the  department.  Adjoining  this  on  the  west  is  another  private  laboratory, 
from  which  a  door  opens  immediately  into  the  general  laboratory.  At  the  left  of 
the  north  entrance  is  a  room  used  for  the  more  advanced  classes  in  microscopical 
work.  This  opens  by  double  doors  into  the  laboratory  in  the  northeast  corner 
of  the  building,  which  is  also  used  for  microscopical  work.  By  means  of  the 
double  doors  these  two  rooms  can  be  thrown  together,  in  case  of  exceptionally 
large  classes.  Inasmuch  as  the  building  is  seldom  used  in  the  evening,  and  as 
it  was  necessary  to  have  gas  in  all  parts  of  the  building  in  any  case,  the  building 
has  not  been  wired  for  electric  light. 


PRIVATE   LABORATORY   AND  OFFICE. 


In  the  laboratories  for  microscopical  work,  each  table  is  supplied  with  gas 
jets,  and  when  it  is  necessary  to  use  artificial  light  for  class  work,  portable  gas 
lamps  with  Welsbach  burners  are  furnished  the  students.  In  the  author's 
experience,  the  Welsbach  burner  is  by  far  the  most  pleasant  light  for  microscopi- 
cal work.  The  plans  show  clearly  the  position  of  the  sinks  and  the  acid  closets, 
which  are  distributed  freely  in  the  various  laboratories.  In  the  arrangement  of 
the  tables  all  are  so  placed  as  to  face  the  windows.  The  author  has  a  personal 
objection  to  microscopical  work  with  a  side  light.  It  is  to  him  extremely  annoy- 
ing, and  it  seems  desirable  that  the  student  should  work  under  the  best  condi- 
tions ;  consequently,  in  the  various  laboratories,  the  tables  are  arranged  in  two 
lines,  one  on  the  wall  immediately  facing  the  windows,  and  the  other  a  few  feet 
back.  It  is  found  that  this  gives  much  better  working  light  than  to  arrange  the 
tables  in  alcove  fashion  from  the  walls,  and  it  is  also  fully  as  economical  of  floor 
space.     Our  room  is  so  ample  at  present  that  it  is  possible  in  all  our  classes  to 


and  Laboratory  Methods. 


1153 


furnish  students  with  tables  which  become  their  permanent  property  for  the 
term's  work.  This  has  a  great  many  advantages,  as  it  is  not  necessary  to  clear 
the  table  after  every  work  period,  and  it  is  possible,  in  the  case  of  enthusiastic 
students,  to  put  in  more  than  their  required  time  upon  their  laboratory  work. 
As  a  matter  of  fact,  in  many  cases,  more  than  double  the  amount  of  required 
time  is  spent  by  many  of  the  students  in  the  laboratory. 

For  laboratory  tables,  after  a  good  deal  of  thought,  we  have  used  hard  wood 
kitchen  tables.  The  essential  points  in  a  laboratory  table  for  an  under-graduate 
student  seems  to  be  a  sufficient  amount  of  room,  and  stability.  These  tables 
are  well  made — each  has  a  drawer — and  have  the  virtue  of  cheapness,  and  we 
prefer  to  spend  our  money  in  other  forms  of  apparatus  rather  than  in  elaborate 
laboratory  tables.     The  tops  of  the  tables  are  painted  a  dull  black. 


ELEMENTARY   LABORATORY. 


For  laboratory  chairs,  we  have  used  a  form  which  we  have  seen  in  no  other 
laboratory.  Many  of  our  best  laboratories  supply  stools  for  their  students.  It 
has  always  seemed  to  us  rather  hard  that  the  student,  who  is  working  two  hours 
or  more  continuously  at  the  laboratory  table,  should  have  nothing  more  com- 
fortable than  a  stool  without  a  back.  It  is  very  desirable  that  whatever  chairs 
are  used  should  be  adjustable  in  height,  because  of  the  varying  heights  of  the 
students  as  well  as  the  requirements  of  the  different  kinds  of  work.  We  have 
finally  adopted  in  all  of  our  biological  laboratories  a  chair  which  is  easily  under- 
stood from  the  illustrations.  The  base  is  a  swivel  stool,  upon  which  is  placed 
the  seat  of  an  ordinary  kitchen  chair.  These  chairs  are  very  comfortable,  and 
at  the  same  time  very  durable,  and  we  have  found  them  extremely  satisfactory 
for  the  laboratory  work. 


1154 


Journal  of  Applied  Microscopy 


In  both  the  general  laboratory  and  the  histological  laboratory,  is  a  large  drip 
tank  (one  of  them  is  shown  in  the  illustration  of  the  elementary  laboratory)  so 
arranged  that  jars  of  living  material  can  be  kept  alive  by  means  of  tubes  from 
the  overhanging  water  pipe,  or  the  tank  can  be  used  as  a  large  fish  tank  by 
plugging  the  outlet  with  a  hollow  plug  of  slightly  less  depth  than  the  tank  itself. 
In  the  general  laboratory  this  tank  is  used  for  keeping  such  animals  as  fresh 
water  mussels  at  the  times  when  the  classes  are  using  them. 

For  storage  of  the  thousand  and  one  things  that  are  in  constant  use  in  the 
laboratories,  as  well  as  the  working  collections  in  homeopathic  vials  and  similar 
small  receptacles,  we  have  a  series  of  tray  cases  (one  of  which  is  shown  in  the 
picture  of  the  private  laboratory).     The  trays  are  made  of  uniform  size,  and  are 


HISTOLOGICAL   LABORATORIES. 


interchangeable  in  position,  or  in  the  different  cases.  They  run  on  one-half 
inch  cleats  on  the  sides  of  the  cases,  and  each  one  has  a  label  holder  upon  the 
front.  In  the  trays  that  are  used  for  homeopathic  vials,  partitions  are  inserted 
parallel  with  the  front  and  back  of  the  trays.  For  such  collections  we  use  the 
ordinary  eight-drachm  homeopathic  vials.  Arranged  in  this  way,  these  trays 
will  hold  195  vials  each,  thus  being  extremely  economical  of  room.  Where 
the  trays  are  used  for  miscellaneous  material,  they  are  arranged  alphabetically, 
so  that  it  is  easy  to  place  one's  hand  upon  corks  of  various  sizes,  bottles,  etc. 
For  the  collections  in  homeopathic  vials  an  accession  catalog  is  kept,  and  the 
bottles  are  arranged  according  to  the  numbers  of  this  catalog.  By  means  of  a 
subject  catalog  upon  cards,  it  is  easy  to  find  any  desired  material.  The  trays 
are  of  such  a  size  that  they  hold  a  definite  number  of  Pillsbury  slide  boxes,  so 
that  they  can  be  used  also  for  packing  away  slide  collections.     The  collection 


and  Laboratory  Methods. 


1155 


of  slides  is  also  arranged  numerically,  the  accession  catalog  being  the  one 
devised  by  Dr.  Ward.  The  subject  card  catalog  makes  it  comparatively  easy  to 
locate  the  slides  of  any  particular  subject. 

Between  the  museum  and  the  general  laboratory  is  a  room  in  which  is  placed 
the  department  library,  where  are  kept  all  the  working  books  on  biology.  To 
this  the  students  of  the  department  have  free  access  at  all  times. 

The  storerooms  of  the  department  are  located  upon  the  north  side  of  the 
basement.  Upon  the  south  side  of  the  basement  are  two  rooms  for  botanical 
laboratories.  On  the  south  side  of  the  basement,  also,  is  located  the  vivarium, 
in  which  are  kept  the  animals  which  must  be  kept  in  stock  for  the  biological 
laboratory.  This  room  is  floored  with  cement,  inclining  towards  an  opening  in 
the  center  which  is  connected  with  the  sewer,  so  that  by  means  of  a  hose  the 
room  can  be  thoroughly  flushed  and  cleansed.  Connected  with  this  room  is  the 
injection  room,  which  has  a  cement  floor  arranged  in  the  same  manner.  In  this 
room  all  the  dirty  work  of  killing  large  animals  and  of  injection,  in  fact  all  work 
which  would  be  liable  to  foul  the  laboratories,  is  taken  care  of.  In  this  way  a 
large  part  of  the  disagreeable  work  is  kept  out  of  the  laboratory  rooms. 
Ripon  College.  C.  DwiGHT  MarSH. 


A  Combined  Condenser  and  Polarizer  for  Petrographical 

Microscopes. 

The  attachment  consists  of  a  double  lens  condensing  system,  and   a   Nicol 
prism  mounted  as  shown  in  the  following  illustration. 

The  upper  condensing  lens  is  mounted 
on  a  revolving  arm  so  that  it  may,  at  the 
will  of  the  operator,  be  instantly  thrown  in  or 
out  of  the  optical  axis  by  a  lever ;  a  suitable 
stop  bp'ng  provided  to  bring  it  to  a  central 
positi  .ii. 

The  lower  lens    is    mounted   at  the   proper 
distance  below  the  upper  surface  of  the  appa- 
ratus so  that  when  the  upper  lens  is  moved 
out  of  optical  axis,  the  lower  lens  focuses  upon 
the  slide,  thus  avoiding  the  necessity  of   re- 
focusing  the  condenser  system  when  changing 
from    the    double    to    single    combination.       The    Nicol    prism   is    mounted  in 
revolving  sleeve  with  graduated  collar  and  a  stop  to  indicate  zero  or  the  position 
of  Nicol  prisms  when  crossed. 

The  advantages  of  this  arrangement  over  others  for  accomplishing  the  same 
results  are,  briefly,  as  follows : 

First — It  is  not  necessary  to  increase  the  size  or  thickness  of  the  microscope 
stage. 

Second — The  attachment  is  always  in  focus  whether  one  or  both  lenses  are  used. 
Third — Compactness  and  freedom  from  liability  of  disturbance  while  operat- 
ing stage  or  slide.  W.  L.  Patterson. 


1156 


Journal  of  Applied  Microscopy 


7 


r\ 


M 


A  Plan  for  a  Ureometer. 

The  plan  for  a  ureometer  (or  modification  of  Doremus'  apparatus)  here  pre- 
sented would,  it  is  believed,  offer  some  advantages  as  to  accuracy  and  facility  of 
manipulation  over  other  methods  of  estimating  urea. 

C  is  a  tube  of  about  '^5  cubic  centimeters  capacity,  closed  at  the  upper  end, 
and  accurately  graduated  from  the  upper  end  downward  for  IG  c.  c,  to  tenths  of 
a  cubic  centimeter.     The  graduation  also  gives  urea  percentages  to  tenths  per 

cent.,  at  20°C,  and  barometric  pressure  of  760  mm.  of 
mercury.  At  the  bottom  of  tube  r  is  a  curved  neck 
communicating  with  a  bulb  />,  which  opens  into  a  tube 
(7 ;  this  has  a  funnel  top,  and  is  of  the  same  length  as 
tube  c.  The  capacity  of  bulb  ^  is  about  30  c.  c,  which 
quantity  is  indicated  by  a  mark  on  tube  a. 

Tube  c  communicates  near  its  lower  end  with  a 
small  tube  ^/,  joined  at  such  an  angle  and  position  that 
gas  generated  in  ^/  will  rise  easily  into  tube  c  and 
none  pass  into  bulb  />.  A  removable  accurately  fitted 
glass  stopper  and  stop-cock  e  separates  tube  ^  from 
the  cavity  of  tube  r,  and  the  capacity  of  tube  t/  below 
the  stopper  is  exactly  1  cubic  centimeter  when  the 
stopper  is  inserted.  A  transverse  perforation  «,  rather 
large,  passes  through  the  stopper,  so  that  when  turned 
in  the  right  direction  it  opens  free  communication 
between  tubes  t/  and  c.  When  the  stopper  is  so  turned 
that  it  shuts  off  tube  //,  the  perforation  n  should  open 
into   the    cavity    of    tube    r.      A    glass    cross-bar    // 

strengthens  the  appara- 
tus, which  is  entirely  of 
glass.  A  separate  base 
or  stand  for  the  support 
of  the  instrument  can  be 
provided. 

Method  of  Use.  — 
The  stopper,  being  re- 
moved, the  tube  t/  is  by 
means  of  a  dropper  filled 
with  urine.  The  stopper 
e  is  then  inserted  with 
opening  /i  so  placed  as 
to  cut  off  tube  t/  from  tube  r.  Tube  //  then  contains  exactly  1  c.  c.  of 
urine.  The  usual  sodium-hypobromite  solution  is  then  poured  into  tube  a, 
up  to  the  30  c.  c.  mark.  The  apparatus  is  then  tilted  so  that  the  solution  runs 
into  tube  r,  entirely  filling  it ;  the  perforation  ;/  should  also  be  filled  with  the 
solution.     The  apparatus  being  held  upright,  the   stopper  e  is  turned  so  as  to 


and  Laboratory  Methods. 


1157 


make  communication  between  tubes  d  and  c.  Tube  d  and  perforation  //  should 
be  large  enough  to  enable  the  fluids  to  mix  readily.  The  rapidity  with  which 
the  fluids  mix  can  be  controlled  by  the  stop-cock.  The  urine  mixes  with  the 
test  solution,  the  urea  is  decomposed,  and  the  nitrogen  evolved  rises  into  the 
upper  part  of  tube  c.  When  the  reaction  is  complete  and  the  temperature  has 
subsided  to  that  of  the  room,  water  is  added  to  (or  removed  from)  tube  a  until 
the  level  of  the  fluid  in  the  two  arms  is  the  same.  The  amount  of  gas  in  tube  c 
is  then  read  ofi^,  and  from  it  the  amount  of  urea  can  be  calculated ;  or  the  per- 
centage of  urea  can  be  read  directly  from  the  graduation. 

The  advantages  of  this  method  would  be  as  follows :  The  measured  amount 
of  urine  ( 1  c.  c.)  is  obtained  accurately  and  easily,  as  it  were  automatically. 
None  of  the  gas  generated  is  lost,  but  all  is  saved  in  tube  c  for  measurement. 
The  equalization  of  the  level  of  the  fluid  in  the  two  arms  equalizes  the  hydro- 
static pressure  and  thus  gives  an  accurate  reading  of  the  amount  of  gas  free 
from  that  source  of  error.  The  graduation  in  cubic  centimeters  enables  the 
urea  to  be  calculated  with  the  greatest  nicety,  applying  any  corrections  for  tem- 
perature and  barometric  pressure ;  while  for  localities  near  the  sea  level  and 
ordinary  room  temperatures  the  percentage  graduation  on  the  tube  (in  the  sketch 
given  on  the  theoretical  basis)  gives  a  direct  reading  of  sufficient  accuracy. 
The  apparatus  is  compact,  not  cumbersome,  easily  kept  clean,  and  always  in 
working  order.     It  combines  accuracy  with  facility  of  manipulation. 

J.  B.  Nichols,  M.  D. 
Washington,  D.  C. 


A  Device  for  Supporting  Pasteur  Flasks. 

Pasteur  flasks  are  difficult  to'handle  on  account  of  their  peculiar  shape.     A 
collar  of  asbestos,  cork,  or  straw  is  ordinarily  used,  but  has  to  be  fitted  closely 

to  the  base  in  order  to  keep  the  flask  erect. 

The  photograph  shows  a  device  for  supporting 
these  flasks,  which  permits  greater  freedom  and 
safety  in  manipulation  than  is  obtained  with  the 
ordinary  collar  support.  The  device  consists  of  a 
solid  disk  of  wood  about  5^  inches  in  diameter 
and  2  inches  in  thickness.  This  is  hollowed  out  in 
the  center,  leaving  a  concavity  into  which  the  base 
of  the  flask  fits.  One  end  of  a  piece  of  heavy  brass 
wire  is  fastened  into  the  margin  of  the  base,  the 
other  end  of  the  wire  is  bent  so  that  the  bend  of  the 
tube  of  the  flask  fits  into  it  loosely.  The  wire  sup- 
ports the  flask  in  the  erect  position,  so  that  the  base 
of  the  flask  need  not  fit  closely  into  the  hollowed 
wooden  base.  Katherine  E.  Golden. 


1158  Journal  of  Applied  Microscopy 

LABORATORY  PHOTOGRAPHY. 

HIGH-POWER  PHOTO-MICROGRAPHY. 

There  is  a  fascination  about  the  use  of  the  microscope  and  camera  together 
that  can  hardly  be  experienced  when  either  instrument  is  operated  alone.  In 
its  simpler  aspects,  moreover,  photo-micrography  may  be  enjoyed  by  any  one  who 
possesses  the  ordinary  microscopical  and  photographic  apparatus.  By  make- 
shift adjustments  and  adaptations,  it  is  possible  to  arrange  the  separate  parts 
into  a  workable  series  so  that  the  amateur  photographer  may  add  the  making  of 
enlarged  pictures  of  small  objects  to  his  other  accomplishments  and  the  micros- 
copist  may  secure  permanent  records  of  the  transitory  images  that  have  so  often 
delighted  him. 

It  is  otherwise,  however,  with  those  who  attack  the  problem  of  producing 
photo-micrographs  which  represent  a  high  amplification  of  the  object — 1000 
diameters  or  over.  This  branch  of  the  work  should  not  be  undertaken  without 
serious  purpose  and  the  best  of  apparatus. 

Here  makeshifts  are  out  of  place.  The  great  degree  of  accuracy  and  the 
nicety  of  adjustment  demanded  of  each  part  of  the  apparatus  makes  it  necessary 
to  employ  an  installation  that  is  especially  designed  for  its  own  particular 
purpose.  With  such  assistance,  only,  can  the  scientist  achieve  any  valuable 
results,  for  it  is  only  the  scientist  who  would  have  the  time  and  patience 
requisite  for  work  of  this  character.  There  must  be  some  end  in  view  aside 
from  the  mere  gratification  of  an  idle  curiosity  to  see  how  big  a  picture  of  a  small 
object  can  be  made.  This  purpose  finds  itself  in  the  desire  of  investigators  to 
present  to  their  fellows  as  accurate  and  as  complete  a  conception  of  their  material 
as  it  is  possible  to  give.  The  value  of  photography  as  an  aid  in  this  direction  is 
being  more  and  more  appreciated  and  nowhere  more  than  among  those  who 
have  to  deal  with  the  almost  ultra-microscopic  structure  of  the  organic  cell. 

Not  that  photographs  are  designed  to  supercede  the  customary  drawings. 
Both  sun  image  and  pencil  image  have  their  places  as  aids  in  the  elucidation  of 
the  text.  The  former  exhibits,  often  in  a  bewilderment  of  detail,  the  whole  field 
of  the  study ;  the  latter  presents  concretely  the  investigator's  interpretation  of 
the  essential  facts.  A  comparison  of  the  two  by  one  acquainted  with  the  subject 
will  enable  him  to  reach  an  opinion  as  to  the  validity  of  the  writer's  conclusions 
such  as  would  be  impossible  if  only  one  method  of  delineation  had  been  used. 

In  recognition  of  this  fact,  many  writers  upon  histological  and  cytological 
subjects  now  enrich  their  papers  by  supplementary  plates  of  photographs  and 
drawings,  which,  with  the  text,  enable  a  reader  to  obtain  as  complete  a  mental 
image  of  the  subject  as  can  be  acquired  without  a  personal  examination  of  the 
specimens.  Such  a  work  as  Wilson's  "  Fertilization  and  Karyokinesis  of  the 
Ovum,"  wherein  the  author's  skill  in  observation  is  supplemented  by  the 
beautiful  photographs  of  Dr.  Leaming,  is  an  excellent  example  of  what  may  be 
done  in  this  direction. 

As  a  realization  of  the  importance   of  this  class  of    illustration  grows,  the 


and  Laboratory  Methods. 


1159 


demand  for  information  concerning  the  methods  employed  increases  until  micro- 
scopical journals  find  it  advantageous  to  assign  a  separate  department  to  the 
discussion  of  methods  and  apparatus  involved  in  the  production  of  scientific 
photographs.  It  is  a  matter  of  congratulation  that  workers  in  this  country  are 
now  afforded  such  a  means  of  communication  through  the  columns  of  the 
Journal  of  Applied  Microscopy  and  Laboratory  Methods. 

The  department  here  being  new,  it  is  but  natural  that  matters  of  an  elementary 
nature  should  be  discussed.  With  this  in  mind,  I  have  thought  to  describe  an 
actual  installation  of  apparatus  for  high-power  photo-micrographic  work  and  to 
exhibit  some  of  the  results  attained  by  its  use.  In  Vol.  Ill,  No.  5,  of  the 
Journal  appeared  an  account  of  the  outfit  employed  in  the  Johns  Hopkins  labora- 
tory.    The  one  at  the   University  of   Kansas  is  very  similar,  but   it   has   been 


1  • 


Figure  1. — Spermatogonia!  mitoses  of   the  grasshopper,  j¥z>/«c?^r  p/iaiiho/'/erus,  in  the  metaphase  and 

anaphase.     These  divisions  take  place  very   rapidly  and  the  archoplasmic  threads  of  previous 

spindles  may  still  be  seen  between  the  centrosomes  of  different  cells,     1000  diameters. 

further  modified  from  the  original  Zeiss  arrangement  than  has  the  one  at 
Baltimore.  Upon  the  optical  bench  are  placed  the  illuminating  apparatus,  two 
iris  diaphragm  supports,  and  the  microscope.  The  other  accessories  furnished 
with  the  complete  outfit  are  not  employed.  The  camera  itself  has  not  been 
altered. 

Following  is  the  arrangement  of  the  apparatus :  The  microscope — I  use  a 
Van  Heurck — Watson  stand — is  firmly  clamped  on  the  end  of  the  bench  nearest 
the  camera.  Next,  the  carbons  of  the  arc  light  are  roughly  adjusted  so  as  to  lie 
approximately  within  the  optical  axis  of  the  microscope.  With  a  low  power 
objective  focused  upon  the  object,  the  arc  is  projected  upon  a  small  screen 
suspended  upon  the  front  of  the  camera  which  is  pushed  back  on  its  sliding  bed 
to  a  distance.  By  means  of  the  adjustment  screws,  the  arc  is  then  brought  into 
such  a  position  that  the  glowing  crater  occupies  the  center  of  the  field.     Prelim- 


1160 


Journal  of  Applied  Microscopy 


inary  to  this,  however,  the  substage  condenser  has  been  racked  up  with  the 
coarse  adjustment  until  it  brings  the  image  of  the  crater  into  focus  at  the  level 
of  the  object.  If  the  camera  has  not  been  adjusted  with  reference  to  the  optical 
bench,  it  is  now  arranged  so  that  the  image  of  the  crater  falls  in  the  center  of 
the  ground  glass.  Provided  the  substage  condenser  is  properly  centered,  the 
linear  adjustment  of  the  combination  is  complete. 

The  next  step  is  to  arrange  the  object  with  reference  to  the  condenser  and 
the  objective  which  is  to  be  used  in  making  the  negative.  I  have  found  it 
advantageous  to  connect  these  three  elements  with  homogeneous  immersion 
fluid  and  for  a  condenser  employ  the  "  parachromatic  "  oil  immersion  form  made 
by  Watson  &  Sons.     The  objective  is  an  apochromatic  'I  mm.  of  Zeiss. 


Figure  2. — Transformation  stages  between  tlie  telophase  of   the  spermatogonia  and  the  ]irophase  of  the  first 

spermatocytes.     The    gradual   accimuilation    of   the   chromatin  into   a    thread   may    he    noted. 

Successive   stages  shown  at  "  a  ",  "  b  ",  and  "  c  ".     Same  object.     1000  diameters. 


For  a  number  of  reasons,  it  is  convenient  to  interpose  temporarily  an 
incandescent  gas  lamp  in  the  substage  series  while  getting  the  proper  focus  and 
adjustment  of  the  object  with  the  eye.  When  this  adjustment  has  been  accom- 
plished, it  remains  only  to  get  the  final  projection  of  the  image  upon  the  ground 
glass  of  the  camera  before  the  exposure  is  made. 

The  projection  eyepiece  is  now  substituted  for  the  observation  ocular,  which 
has  been  used  up  to  this  time,  and  an  image  thrown  upon  the  small  screen  which 
still  hangs  upon  the  front  of  the  camera.  Here  an  approximate  focus  of  both 
object  and  source  of  illumination  is  obtained  and  the  composition  of  the  picture 
studied.  If  this  is  satisfactory,  the  screen  is  removed,  the  camera  pulled  forward 
and  joined  to  the  microscope,  and  connections  made  between  the  fine  adjust- 
ments of  the  tube  and  of  the  substage  condenser  with  rods  that  lead  back  to  the 


and  Laboratory  Methods. 


1161 


end  of  the  camera  carriage.     Here  these  may  be  manipulated  while  the  image  is 
being  examined  and  focused  on  the  screen. 

At  this  stage  of  the  proceedings,  there  are  visible  upon  the  ground  glass 
indistinct  images  of  the  incandescent  carbon  and  of  the  object.  By  means  of 
the  fine  adjustments,  these  are  brought  into  a  sharp  focus  upon  the  glass. 
Owing  to  the  nature  of  the  crater,  the  illumination  is  not  uniform  over  the  whole 
field  and  it  is  necessary  to  place  a  piece  of  ground  glass  between  the  source  of 
illumination  and  the  object.  This  should  not  be  more  than  dense  enough  to 
properly  diffuse  the  light,  otherwise   it  unduly  lengthens  the  time  of   exposure. 


Figure  o. — Prophase  of  the  first  spermatocyte.     Tlie  chromatin  tliread  has  broken  into  segments.     Between 

these  run  delicate  )inin  fibers  as  at  "  a  '',     At  "  b  "an  element  distinguishable  from  the  others 

by  its  greater  transparency  and  sharper  outline.     In  these  cells  it  is  known  as 

the  accessory  chromosome.     Same  object.     1000  diameters. 


There  should  now  be  visible  a  sharply  defined,  evenly  illuminated  image  of  the 
object  wherein  the  details  are  neither  obliterated  by  excessive  illumination,  nor 
rendered  granular  and  obscure  by  deficient  light. 

In  obtaining  a  good  negative,  the  manipulation  of  the  substage  iris  diaphragm 
is  as  important  as  the  proper  adjustment  of  the  objective.  No  pains  are  spared, 
accordingly,  to  bring  about  the  appropriate  arrangement  of  the  cone  of  light, 
and  the  lever  of  the  diaphragm  is  swung  back  and  forth  until  the  very  best 
possible  result  is  obtained.  The  Watson  condenser  above  mentioned  has  a 
graduation  upon  the  mounting  indicating  the  numerical  aperture  afforded  by  the 
opening  of  the  diaphragm.  This  is  a  convenience  which  should  find  a  place 
upon  the  condensers  of  other  manufacturers. 

The  further  stages  of  the  process  are  those  which  pertain  to  photographic 
manipulations  in  general,  and  the  limits  of  this  article  will  not  permit  their  con- 
sideration.    Summing  up  the  steps  to  be  followed,  we  have  : 


1162 


Journal  of  Applied  Microscopy 


1.  Linear  adjustment  of  substage  condenser,  crater,  and  center  of  camera- 
back  to  obtain  their  coincidence  with  the  optical  axis  of  the  microscope. 

2.  Focus    of   source    of    illumination    upon    object    by    means    of    substage 
condenser. 

3.  Focus  of  objective  upon  object. 


%    A 


m\  m 


■  V 


-^A*  ^  ^4 


Figure  4. — Metaphase   of   the   first   spermatocyte.     The    chromosomes    of   the    same  cell    do    not    divide 

simultaneously,  as  may  be  seen  at  "  a  ".     Sometimes  they  form  rings  as  seen  at  "  b  ". 

In  the  cell  marked  "  c  "  the  archoplasmic  fibers  are  sharply  in 

focus.     Same  object.     1000  diameters. 

4.  Final  simultaneous  projection  of  crater  and  object  images   upon  ground 
glass  of  camera. 

5.  Diffusion  of  light  by  ground  glass  between  source  of  illumination  and  object. 

6.  Adjustment  of  substage  iris  diaphragm. 

7.  Exposure  of  plate.  C.  E.  McClung. 
University  of  Kansas. 


An  Improvised  Microtome. 

On  a  recent  visit  to  the  College  of  Physicians  and  Surgeons  in  Boston,  the 
writer  saw,  among  other  ingenious  contrivances,  a  microtome  devised  by  Dr. 
Shurtleff,  of  the  above  named  institution,  and  made  by  him  at  a  cost  of  fifty 
cents,  for  cutting  the  micrometer  screw.  Since  my  visit  I  have  myself  made 
a  similar  microtome  at  the  small  cost  of  two  hours'  work,  since  a  common  screw 
was  employed  instead  of  a  micrometer.  Thinking  that  possibly  the  idea  may  be 
of  use  to  some  other  investigator,  I  will  venture  to  offer  the  following  description  : 

The  first  essential  is,  of  course,  a  knife;  and,  while  a  regular  section  razor  is 
preferable,  an  ordinary  razor  will  answer.     In  the  absence  of  either,  however,  I 


and  Laboratory  Methods. 


1163 


have  seen  a  shoe  knife  successfully  used ;  but  in  this  case  the  back  was  strength- 
ened by  soldering  a  knitting  needle  on  one  side  and  a  rod  cut  from  a  stove  poker 
on  the  other.  Assuming,  then,  that  a  knife  is  at  hand,  the  next  requisite  is  the 
holder,  which  consists  of  a  piece  of  wood  about  four  by  seven  inches,  having  a 
U-shaped  cut-out  at  the  top,  two  inches  wide  and  three  inches  deep.  This  leaves 
two  prongs  each  an  inch  wide  into  which  small  wire  nails  are  driven  so  that  the 
razor  (R)  may  rest  upon  them  when  it  is  in  position.  Spring  clips  of  some  sort  hold 
the  razor  firmly  in  place.  For  the  clips,  stout  wire  an  eighth  of  an  inch  thick  is 
good.  Each  wire  may  be  fastened  to  the  board  by  double-pointed  tacks.  Near 
the  bottom  of  the  board  and  in  the  center  is  the  screw  (S).  A  common  screw  will 
answer,  but  a  fine  threaded  screw  passing  through  a  nut  is  better.  In  either  case, 
however,  a  large  disc  may  be  soldered  to  the  screw  head  for  increased  delicacy 
in  operation.  The  "holder"  complete  is  now,  by  means  of  a  pair  of  hooks  and 
eyes,   to  be    made  attachable  to  the  end  of  a  box   so   that  turning  the  screw 


'l  '  ' 

|Ur 

1 

1 

1     W 

BASE. 

Fig.  I. 


Fig.  2. 


gives  a  delicate  movement  to  the  razor.  The  screw  point  should  work  against  a 
small  metal  plate  on  the  box.  Tension  is  secured  with  a  rubber  band  or  spiral 
spring.  (Reference  to  the  diagram  will  make  the  idea  clear.)  The  "  holder " 
should  be  so  placed  that  the  razor  edge  will  be  two  or  more  inches  higher  than 
the  top  of  the  box.  Now,  when  an  adjustable  object-holder  is  provided,  the 
microtome  is  completed.  To  make  the  object-holder,  a  board  somewhat  shorter 
than  the  box,  a  block,  and  a  straight-grained  stick  about  one-half  an  inch  in 
cross  section  are  necessary.  Fasten  the  block  near  one  end  of  the  board,  nail 
the  stick  to  the  block  (as  indicated  in  the  diagram),  and  the  microtome  is  ready 
for  service. 

In  use,  the  paraffin  block  (O)  is  fastened  to  the  end  of  the  stick  with  melted 
paraffin,  and  proper  adjustments  are  made  with  reference  to  the  razor.  Then, 
downward  pressure  on  the  stick  cuts  the  section,  while  clockwise  movement  of 
the    screw    regulates    the  thickness.     Serial    sections   are   readily  made,  if  the 


1164  Journal  of  Applied  Microscopy 

paraffin  block  is  carefully  squared ;  but,  for  this  work,  the  object-holder  should 
be  steadied  by  a  weight  of  five  or  six  pounds  (W). 

While  the  microtome  can  by  no  means  take  the  place  of  such  a  splendid 
instrument  as  the  Bausch  ^:  Lomb  "  Student,"  yet  it  is  a  practical  and  serviceable 
apparatus,  and  its  usefulness  has  been  demonstrated  in  everyday  histological 
work.  Irwin  LaVerne  Powers. 

Randolph,  Mass. 


The  Study  of  Bacteria  in  the  Public  Schools. 

The  highest  aims  in  "  municipal  housekeeping  "  can  never  be  attained  by 
Boards  of  Health  or  by  Departments  of  Street  Cleaning  alone,  however  efficient 
these  organizations  may  be.  Unless  these  city  departments  are  backed  by  a 
strong,  intelligent  public  sentiment  we  shall  experience  nothing  better  than  spor- 
adic reform  in  the  cleaning  of  our  streets,  in  the  construction  of  tenement  houses, 
and  in  the  general  care  for  the  public  health.  When  conditions  get  sufficiently 
bad  in  a  community,  it  is  comparatively  easy  to  arouse  the  voters  and  roll  in  a 
reform  administration  by  big  majorities.  But  alas  !  we  soon  tire  of  our  attempts 
at  public  virtue,  we  reverse  our  votes  at  the  next  election,  and  sink  back  into 
easy  toleration  of  filth  and  its  resulting  disease.  One  might  indeed  become 
pessimistic  with  reference  to  the  future  of  our  cities  were  it  not  true  that  democ- 
racy possesses  a  most  powerful  means  of  developing  a  public  sentiment  which  may 
be  at  once  intelligent  and  lasting.  Gathered  in  our  schools  of  to-day  are  the  boys 
and  girls  who  will  be  the  voters  and  the  home-makers  of  to-morrow.  Hence  to  the 
teacher,  especially  in  the  public  schools,  is  given  the  opportunity  to  exert  a 
telling  influence  in  developing  the  better  city  of  the  future. 

The  discoveries  in  bacteriology  within  a  few  years  have  made  new  sciences 
of  surgery,  medicine,  and  sanitation.  Epidemics  of  typhoid  fever  have  ceased 
to  be  regarded  as  "  a  dispensation  of  an  all-wise  Providence,"  for  we  have  come 
to  know  that  the  presence  of  this  disease  usually  means  a  contaminated  water 
supply  or  imperfect  sewerage.  Scientific  men  have  learned,  too,  how  to  check 
the  ravages  of  yellow  fever  and  cholera,  and  even  consumption  is  found  to  be  a 
preventable  disease.  To  make  these  discoveries  of  practical  use,  however, 
this  knowledge  must  be  possessed  by  a  large  majority  of  the  citizens  in  a  com- 
munity, and  the  most  effective  means  of  attaining  this  end  is  by  educating  the 
pupils  in  our  public  schools.  With  this  object  in  view,  in  the  Peter  Cooper  High 
School,  New  York  City,  we  devote  considerable  time  in  the  course  in  biology  to 
the  study  of  bacteria,  yeast,  and  moulds. 

In  this  study,  it  is  necessary  at  the  very  first  to  impress  the  pupil  with  some 
idea  of  the  omnipresence  of  these  micro-organisms  in  everyday  life ;  and  for 
this  purpose  an  experiment  performed  by  the  boy  or  the  girl  is  always  more  tell- 
ing than  a  talk  by  the  teacher  or  a  dozen  pages  of  description.  We  begin  with 
the  study  of  a  hay  infusion.  The  work  is  done  by  each  pupil  at  home,  and  the 
report  presented  at  the  next  recitation.  The  following  account  is  selected  from 
the  one  hundred  and  fifty  papers  received  from  the  first  year  pupils  : 


and  Laboratory  Methods.  1165 

"  Straw  Infusion.  I  procured  about  a  handful  of  straw  at  a  livery  stable 
and  put  it  in  a  Mason  jar  three-quarters  full  of  water,  and  put  it  in  a  warm  place 
where  the  temperature  was  on  an  average  of  75°,  on  Thursday,  March  22.  Its 
color  was  tan  and  the  mixture  smelt  like  musty  straw. 

"  The  23d,  temperature  73°,  mixture  getting  darker  in  color,  and  smell 
becoming  more  noticeable.  Saturday,  temperature  74°.  A  thin  scum  is  forming 
and  small  things  are  coming  up  from  the  bottom  and  straw.  The  smell  is  get- 
ting very  strong. 

"  Sunday,  temperature  74°,  scum  becoming  thicker  and  bubbles  appearing  in  it." 

Discussion  and  microscopical  examination  in  the  class  room  brought  out  the 
fact  that  the  scum  was  composed  of  countless  bacteria  and  other  micro-organ- 
isms which  had  grown  from  the  germs  on  the  dried  hay.  The  inference  was 
drawn  from  the  experiment  that  bacteria  grow  rapidly  in  a  warm  temperature, 
when  water  and  organic  matter  are  present,  and  that  decay  is  one  of  the  results 
of  their  activity. 

The  cultivation  of  bacteria  in  the  laboratory  was  the  topic  next  considered. 
Nutrient  gelatin,  the  most  useful  medium  in  which  to  grow  all  kinds  of  bacteria, 
may  be  readily  prepared  in  the  laboratory  or  in  the  home  kitchen.  The  ingre- 
dients necessary  are  the  following :  one  pound  of  lean  beef  chopped  fine  (or 
better  run  through  a  meat  cutter) ;  60  grams  (2  oz.)  of  the  best  French 
gelatin  ;  6  grams  (1-5  oz.)  of  peptone,  which  can  be  bought  for  10  cents  at  any 
drug  store ;  a  teaspoonful  of  salt,  and  a  little  baking  soda.  Put  the  beef  in  a 
porcelain  or  agate  dish,  add  a  pint  of  cold  water,  and  allow  the  mixture  to  boil 
slowly  for  a  half  hour.  Strain  the  broth  through  muslin  and  then  allow  the 
liquid  to  run  through  filter  paper.  Pour  in  enough  water  to  make  the  quantity 
of  broth  equal  to  about  a  pint  and  a  half.*  The  gelatin,  cut  into  small  pieces,  is 
then  added  to  the  broth,  together  with  the  peptone  and  salt.  The  mixture  should 
be  heated  sufficiently  to  cause  the  gelatin  to  dissolve,  but  should  not  be  allowed 
to  boil.  Just  enough  cooking  soda  is  added  to  cause  red  litmus  paper  dipped 
in  the  mixture  to  turn  blue,  that  is,  the  liquid  should  be  faintly  alkaline.  Filter- 
ing the  hot  gelatin  sometimes  involves  more  or  less  difficulty.  The  process  can 
be  easily  carried  on,  however,  within  a  steam  cooker.  A  glass  funnel  should  be 
put  in  the  mouth  of  a  Florence  flask  (used  commonly  in  a  chemical  laboratory) 
and  one  or  two  layers  of  absorbent  cotton  placed  within  the  funnel.  If  the  gela- 
tin, flask,  and  funnel  are  kept  hot  within  the  cooker  the  liquid  will  readily  pass 
through  the  cotton.  After  filtering,  close  the  mouth  of  the  flask  with  a  plug 
of  absorbent  cotton,  and  boil  for  a  few  moments.  The  flask  may  be  set  aside 
as  stock  gelatin  until  needed  for  use.  (If  the  gelatin  mixture  is  not  clear,  it 
should  be  filtered  through  the  same  cotton  a  second  time.) 

Some  of  the  liquid  gelatin  was  poured  into  clean  Petri  dishes  (Fig.  1), 
or  test  tubes  plugged  with  cotton  may  be  used.  After  the  gelatin  had 
solidified  some  of  the  dishes  were  opened  to  the  air.  Several  days  after 
this  exposure  the  cultures  were  placed  upon  the  desks  of  the  pupils,  and  they 
were  asked  to  make  drawings  of  the  bacteria  colonies,  and  to  answer  certain 

*This  broth  may  be  prepared  more  easily  from  Liebig's  beef  extract.  Four  grams  should 
be  dissolved  in  the  pint-and-a-half  (750  c.  c. )  of  water,  and  the  solution  should  be  filtered  through 
filter  paper. 


,„■„;  Jourmil  of  Applica  Microscopy 


-  •■ :-  K-#<^»v  "tf-' 


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cd^wt^  ff^JL^.^^^^M*-^*^:^^j^  ti^^ 


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f«^««#o 


Fig-^i 


and  Laboratory  Methods.  1107 

questions  stated  in  tlie  Laboratory  M;inual.  On<;  of  tin:  |)ap(;rs  prepared  rlurini^ 
a  recitation  period  is  reproduced  in  J'ig.  2.  I'i^iircs  .'5  and  4  arc  drawin^yi  ol 
l^etri  dish  cultures  made  by  two  other  pupils. 

One  of  the  boys,  not  satisfied  with  the  anrifjunt  <<f  lai^fjratory  work  ^ivcn  in 
school,  prepared  nutrient  gelatin  at  home.      He  writes  thus  of  his  experiences: 

"  I  took  about  a  half  pound  of  lean  beef  and  after  cutting  into  pieces  placed 
in  a  [jot  and  covered  with  water,  then  brought  to  a  boil.  I  should  also  mention 
that  I  used  a  moderate  fire  so  that  the  process  occupied  about  twet)ty  minutes. 
After  obtaining  my  broth  I  added  gelatin  and  brought  again  tf;  a  boil.  Here  I 
added  some  salt  and  carbonate  of  soda,  after  which  I  strainerl  the  broth  through 
cotton  into  a  sterilized  bottle  and  corked. 

"  I  experienced  such  trouble  in  clearing  the  gelatin  of  colonies  that  I  hnally 
melted  the  gelatin  and  poured  it  into  test  tubes  and  in  them  brougfit  if  to  a  boil 
with  the  result  of  one  tube  burnt  and  five  cleared.  In  three  of  the  tubes  Mr.  I'ea- 
body  inoculated  pure  cultures;  one  of  the  tubes  has  proflueed  a  very  large  red 
colony,  the  others  have  not  grown." 

This  laboratory  work  on  the  growth  of  bacteria  was  followed  by  an  «;xperi- 
ment  performed  at  home  by  the  pupils.  On(;  of  tlu;  girls  gives  the  following 
report  of  her  work  : 

"Tjik  Hiiiiiv  Ol  iiACTKkiA  IN  Mn.K.  I  procured  three  bottles  of  about  th*; 
.same  size.  1  then  thoroughly  cleansed  each  bottle  before  I  used  it.  'I'wo  ol 
the  bottles  had  stoppers  ;  the  other  had  none.  One  of  the  bottlf;s  I  lialf  filled 
with  good  fresh  milk,  put  the  sto|;per  on,  and  set  it  outside  the  window.  I 
labeled  this  bottle  'No.  1.' 

"Into  the  second  bottle  1  poured  about  the  same  amount,  oi  rnilk,  and  set  it 
aside  in  a  warm  temperature  of  about  70'\      I  labehrd  it  '  No.  2.' 

"The  third  bottle  I  cleaned  in  very  hot  water.  I  then  boiled  the  same 
amount  of  milk  that  1  put  in  each  of  the  other  bottles.  1  allowed  it  to  boil  for 
about  three  minutes.  After  the  milk  had  cooled  a  little  J  poured  it  into  the  thirri 
bottle.     I  placed  it  beside  bottle  No.  2,  and  labeled  it  'Sterilized  Milk.' 

"  At  the  end  of  fifteen  hours  1  examined  each  of  the  bottles.  I  noticed  that 
No.  1  had  very  little  smell  at  all.  No.  2  had  a  .sour  like  smell.  It  smelled  as  if 
the  milk  were  turning.  No.  8  had  hardly  any  smell  at  all,  Jf  there  was  any  smell 
at  all,  it  was  a  sweet  one.  I  now  boiled  the  milk  in  \o.  '•',  again.  1  first 
thoroughly  cleansed  the  bottle  and  cork  before  I  put  the  rnilk  in.  I  then  placed 
it  Vjeside  No.  2,  and  put  No.  1  again  outside  the  window. 

"At  the  end  of  twenty-four  hours  I  again  examined  my  bottles.  I  found 
that  No.  1  had  not  any  smell  at  all.  .No.  2  had  a  very  decidedly  .sour  smell,  and 
No,  8  had  a  sweet  smell. 

"  The  changes  in  the  milk  are  due  to  the  growth  of  the  bacteria  from  the  air, 
or  on  the  bottles,  or  the  stoppers.  As  far  as  my  experiment  has  worked  I  do 
not  think  a  cold  temperature  kills  the  bacteria,  but  I  think  it  numbs  them.  I 
think  a  boiling  temperature  kills  the  bacteria,  and  J  think  a  mr^derate  tempera- 
ture increases  the  growth  of  the  bacteria." 

.Successful  microscopical  work  was  done  with  magnifying  powers  of  about  .000 
diameters.  Pure  cultures  of  spherical-,  rod-,  and  spiral-shaped  bacteria  grow- 
ing in  test  tubes  of  gelatin  were  supplied  us  by  Dr.  T.  Mitchell  Prudden  of  the 


llfiS  Journal  of  Applied  Microscopy 

College  of  Physicians  and  Surgeons,  to  whom  I  am  much  indebted  for  help  in 
this  bacteriological  work.  Microscopical  slides  are  easily  prepared  thus  :  Hold 
upside  down  the  test  tube  in  which  bacteria  are  growing,  and  carefully  remove 
the  cotton  from  the  mouth.  Touch  one  of  the  colonies  of  bacteria  with  the  point 
of  a  needle,  and  then  rub  the  needle  point  on  a  clean  glass  slide ;  add  a  drop  of 
water  to  the  spot  touched  by  the  needle,  cover  with  a  cover-glass.  Stains  (Loef- 
fler's  methylen  blue  and  Ziehl's  carbol  fuchsin)  bring  out  more  clearly  the  struc- 
ture of  the  bacteria.  Each  of  the  thirty-five  pupils  in  a  division  examined  the 
stained  bacteria,  and  watched  under  another  microscope  the  motion  of  the  living 
forms.     One  pupil's  written  account  of  this  study  is  here  given  : 

"Microscopic  Study  of  Bacteria.  1.  The  bacteria  which  I  saw  under 
the  microscope  last  Wednesday  were  of  red  and  blue  colors.  This  was  caused 
by  the  coloring  matter  (stains). 

"  2.  They  were  of  three  different  shapes,  round,  pencil-shaped,  and  corkscrew. 

"  3.  The  bacteria  which  I  saw  to-day  under  the  microscope  are  moving  around. 

"  4.  There  were  also  under  the  microscope  egg-shaped  animals  which  were 
moving  around."  (This  slide  was  prepared  from  the  hay  infusion  and  contained 
infusoria.) 

A  little  mathematical  problem  worked  out  by  each  student  helped  to  make 
real  the  rapidity  of  multiplication  among  these  micro-organisms.  The  pupils 
were  told  that  a  rod-shaped  bacterium,  when  conditions  are  favorable,  divides 
in  about  an  hour  to  form  two  bacteria.  The  problem  was  stated  something  like 
this:  Suppose  we  start  with  a  single  bacterium  this  morning  at  10  o'clock;  if 
conditions  are  favorable,  how  many  cells  would  be  seen  at  11  o'clock?  The 
answer  was  "two."  Between  11  and  12  o'clock  each  of  the  two  would  divide  to 
form  two ;  hence  at  12  o'clock  it  was  evident  that  there  would  be  four  bacteria 
in  place  of  the  single  cell  at  10  o'clock.  The  pupils,  continuing  the  calculation, 
found  that  if  the  process  were  to  go  on  until  10  o'clock  the  next  morning,  the  origi- 
nal bacterium  would  give  rise  to  16,776,216.  The  completion  of  this  calcula- 
tion for  a  second  day's  crop  of  bacteria  was  not  attempted  for  obvious  reasons. 

Thus  far  the  experiments  and  discussions  had  made  real  to  the  pupils  the 
existence  of  countless  millions  of  micro-organisms.  They  had  learned  something 
of  the  form,  size,  and  motions  of  the  individual  bacteria ;  and  they  had  become 
acquainted  with  some  of  the  results  of  their  activity  in  causing  decay,  in  souring 
milk,  and  in  producing  colors. 

Some  of  the  conditions  which  tend  to  check  the  growth  of  bacteria  were 
learned  from  the  milk  experiment  performed  at  home.  A  laboratory  demonstra- 
tion developed  this  subject  still  further.  One  of  the  boys  described  the  experi- 
ment thus : 

"  Sterilization.  Mr.  Peabody  took  three  test  tubes  and  inoculated  some 
of  the  bacteria  from  the  hay  infusion.  The  first  test  tube  contained  nourishment 
in  a  solid  form  (nutrient  gelatin),  and  after  the  bacteria  had  been  inoculated  it 
was  set  aside.  The  second  test  tube  was  prepared  the  same  way,  but  Mr.  Pea- 
body  poured  some  corrosive  sublimate  over  the  surface  of  the  gelatin.  The 
third  test  tube  was  prepared  in  the  same  way  as  the  first,  but  was  put  in  the 
(steam)  sterilizer  for  five  minutes,  and  then  set  aside. 


and  Laboratory  Methods. 


11()9 


/^?ri      /• 


''^ — ■■ f      ■    ,' 

u     u 


-^^^T-i-^n.^/^  ^-^^/y^-^-ti^. 


■<^'<^2^'C'C&<^. 


-'i^-ti'^^Z^^^ 


Fig.  .S. 


Q^Xi^i^Vf        Of-  fo  (MlAX/l'CCL/  ■ 


Bsicier/^.    Yellow. 
Aoii</.   White. 


Fig.  4. 


1170  Journal  of  Applied  Microscopy 

"  At  the  end  of  five  days  we  examined  the  tubes  and  found  that  the  two  tubes, 
one  sterilized  by  heat  and  the  other  by  poison,  were  perfectly  clean,  while  the 
other  had  a  large  colony  growing.  From  this  I  infer  that  corrosive  sublimate 
and  the  heat  killed  the  bacteria." 

We  are  fortunate  in  possessing  a  hundred  copies  of  "  The  Story  of  Bacteria  " 
and  a  like  number  of  "  Dust  and  its  Dangers  "  by  Dr.  T.  M.  Prudden.  These 
books  were  loaned  to  the  192  pupils  who  were  studying  the  subject,  and  about 
one-fourth  of  the  chapters  w^ere  assigned  for  text-book  lessons.  One  may  judge 
of  the  interest  in  this  study  by  the  following  figures  :  When  the  books  were 
returned  it  was  found  that  103  pupils  had  read  the  whole  book;  that  the  books 
had  been  read  by  197  parents  or  friends  of  the  pupils;  and  that  various  topics 
in  bacteriology  had  been  discussed  in  over  half  of  the  homes. 

The  practical  applications  of  the  subject  were  brought  out  in  discussion  of  a 
list  of  questions  from  which  the  following  are  selected  : 

1.  From  all  your  experiments  state — 

a.  What  conditions  seem  to  favor  the  growth  of  bacteria  ? 
/>.  What  conditions  seem  to  hinder  the  growth  of  bacteria  ? 

2.  Why  are  fruits  cooked  before  canning? 

3.  Why  should  fruit  jars  be  filled  completely  before  screwing  on  the  cover  ? 

4.  Why  is  grass  dried  before  putting  it  in  the  barn  ? 

5.  Why  are  milk,  meat,  etc.,  put  in  the  refrigerator  in  summer  time? 

6.  Why  should  the  prohibition  against  spitting  in  public  places  be  rigidly 

enforced  ? 

7.  Why  should  sweeping  be  done  as  far  as  possible  without  raising  a  dust  ? 

8.  Why  are  hard  wood  floors  more  healthful  than  carpets  ? 

9.  Why  should  the  teeth  be  brushed  often  ? 

10.  Why  should  the  refuse  be  removed  from  the  streets  every  morning  early, 

especially  in  summer  time  ? 

11.  Why  should  sink  drains  be  carefully  inspected  ? 

12.  Why  should  wounds  be  carefully  cleansed  and  dressed  at  once  ? 

13.  Why  are  typhoid  fever,  diphtheria,  and  other  infectious  diseases  often 

best  treated  in  hospitals  ? 
The  tables  of  the  New  York  Board  of  Health  give  figures  and  charts  which 
serve  to  clinch  the  argument  in  favor  of  good  city  housekeeping.     The  pupils 
copied  into  their  note-books  the  following  figures  giving  the  annual  death-rate 
per  thousand  of  the  population  in  New  York  City,  188G  to  1896  inclusive: 

1886,  25.99  1891,  26.31 

1887,  26.32  1892,  25.95 

1888,  26.39  1893,  25.30 

1889,  25.32  1894,  22.76 

1890,  24.87  1895,  23.11 
1896,  21.52  (first  part  of  year). 

There  was  little  need  to  suggest  that  the  sudden  decrease  in  death-rate  in  1894 
and  in  succeeding  years  was  doubtless  due  in  no  small  measure  to  the  efficiency 
of  the  Street  Cleaning  Department  organized  and  directed  by  the  late  Col. 
Waring. 


and  Laboratory  Methods.  ^^"1 

After  reading  Dr.  Prudden's  books,  and  after  class-room  discussions,  each 
pupil  was  asked  to  outline  at  home  the  arguments  in  favor  of  and  against  the  bac- 
teria.    The  case  is  stated  thus  in  one  of  the  papers : 

"  Benefits  of  Bacteria  to  Mankind.  They  construct  food-stuffs  for  plants 
out  of  the  nitrogen  gas  and  the  solutions  absorbed  from  the  soil. 

"  They  ripen  the  cream  before  churning  and  thus  form  butter. 

"  They  give  flavor  to  butter. 

"  They  are  an  absolute  necessity  in  making  cheese. 

"  In  making  vinegar  from  cider,  yeast  and  bacteria  work  together, 

"  Bacteria  perform  a  very  necessary  work  in  the  process  of  '  retting '  flax  in 
the  linen  industry,  without  which  we  would  not  have  our  fine  linen  and  delicate 
laces. 

"  Bacteria  play  a  prominent  part  in  the  curing  of  tobacco. 

"  Sprouting  of  seeds  is  promoted  by  bacteria. 

"  Streams  and  lakes  are  cleared  by  bacteria. 

"They  decompose  dead  animals  into  the  dust  from  whence  they  came. 

"The  Ways  Bacteria  Prove  to  be  '  Man's  Invisible  Foes.'  Bacteria  cause 
the  diseases,  consumption,  typhoid  fever,  scarlet  fever,  pneumonia,  leprosy,  lock- 
jaw, influenza,  cholera. 

"  They  cause  blood  poisoning. 

"  They  destroy  foods." 

The  primary  aim  of  these  eight  lessons  in  bacteriology,  as  already  stated, 
was  a  practical  one,  namely,  to  present  to  the  boys  and  girls  of  our  city  a  most 
telling  argument  for  cleanliness  in  the  care  of  the  home  and  in  the  care  of  the 
city.  The  colored  charts  portraying  the  cases  of  consumption  in  the  region  of 
Mott  street  and  of  diphtheria  in  the  Tenth  and  Twelfth  wards  will  not  soon  be 
forgotten.  Hence  the  New  York  of  to-morrow  will  doubtless  number  among 
its  citizens  at  least  a  few  more  staunch  supporters  of  an  efficient  Board  of  Health  ; 
a  few  more  homes  will  probably  be  free  from  the  danger  of  disease  contagion, 
and  a  few  more  house-wives  will  exercise  greater  care  to  secure  abundance  of 
light  and  of  fresh  air  in  their   homes  and  to  select  and  prepare  nutritious  foods. 

The  treatment  of  the  subject,  however,  was  not  allowed  to  leave  in  the  minds 
of  the  pupils  the  lasting  impression  that  we  have  discovered  in  bacteria  an  omni- 
present and  well-nigh  omnipotent  enemy.  They  were  led  to  see  that  consump- 
tion, cholera,  typhoid,  and  all  the  other  diseases  charged  to  these  micro-organ- 
isms are  due  to  the  ignorance  or  carelessness  of  man,  and  that  these  diseases 
can  be  prevented.  While,  on  the  other  hand,  they  learned  that  the  bacteria  are 
toiling  incessantly  to  clear  our  earth  from  the  debris  of  decay,  and  to  prepare  the 
soil  and  the  air  for  the  growth  of  the  higher  plants.  Thus  this  study  becomes  a 
part  of  the  great  study  of  biology,  and  in  this  fact  lies  the  deeper  interest  of  the 
subject.  In  the  hay  infusion  all  the  functions  of  living  nature  are  in  full  oper- 
ation. There  one  may  study  assimilation,  oxidation,  respiration,  excretion,  the 
life  and  death  struggle  for  food,  reproduction,  and  even  something  akin  to  sen- 
sation ;  for  who  of  us,  after  an  hour  at  the  microscope,  watching  the  varying 
movements  in  this  world  of  micro-organisms,  is  prepared  to  deny  absolutely  all 
sentient  impressions  even  among  bacteria  ?     Biological  study  of  this  sort  should 


in2  Journal  of  Applied  Microscopy 

not  only  result  in  more  healthy  bodies  for  our  pupils  and  in  a  more  healthful 
community,  but  it  should  contribute  largely  to  broaden  and  deepen  the  mental 
life  of  the  student.  James  E.  Peabody. 

The  Peter  Cooper  High  School,  New  York  City. 


Biology  Wall  Charts. 

"A  Method  of  Making  Biology  Wall-Charts,"  by  F.  D.  Heald  of  Parsons 
College,  Fairfield,  Iowa,  published  in  the  Journal  of  Applied  Microscopy 
for  November,  1900,  induces  me  to  speak  of  a  method  of  chart  making  which  I 
have  adopted  with  considerable  satisfaction,  to  myself  at  least. 

The  method  is  not  original  with  me,  but  was  suggested  by  Prof.  H.  P.  Johnson 
of  the  University  of  California.  My  charts  are  made  of  material  such  as. is  used 
by  "  millers  "  for  the  manufacture  of  flour  sacks.  It  is  well  known  that  this  sack 
muslin  has  incorporated  in  the  meshes  of  the  cloth  a  filling  of  paste  material 
which  renders  the  surface  smooth  and  very  suitable  to  draw  upon.  Instead  of 
a  pen  I  use  an  ordinary  paint  brush  of  suitable  size  and  shape.  My  pigments 
are  such  as  painters  use  for  the  ordinary  canvas  advertising  streamers  and  are 
procured  ready  mixed  at  the  paint  shop  at  a  cost  of  a  few  cents.  With  these 
materials,  charts  of  all  sizes,  colors,  and  kinds  may  be  readily  made.  I  find 
these  "  home-made  "  charts  more  satisfactory  in  my  classes  than  any  others  that 
I  have  heretofore  used,  as  there  can  be  represented  upon  them  exactly  what  it 
is  wished  to  illustrate.  These  charts  are  so  inexpensive  and  so  easily  made  that 
any  school  may  provide  itself  with  a  sufficient  number  for  illustration  in 
Physiology,  Zoology,  Botany,  and  other  subjects.  As  they  are  made  in  water- 
colors,  when  not  in  use  they  should  be  kept  in  a  dry  place. 

Orson  Howard. 
University  of  Utah,  Salt  Lake  City. 


Staining  in  Toto  with  Delafield's  Haematoxylin. 

The  stain  is  prepared  according  to  the  method  found  in  Ruber's  "  Directions 
for  Work  in  Histological  Laboratory,"  p.  153;  except  that,  before  using,  it  is 
diluted  with  an  equal  amount  of  distilled  water,  instead  of  five  to  ten  times  with 
water,  as  according  to  the  directions. 

The  specimens,  which  should  not  be  of  too  great  size — not  more  than 
one-fourth  inch  in  thickness — are  left  in  the  stain  five  days.  After  rinsing  in 
water  they  are  decolorized  for  two  hours  in  acid  alcohol  made  as  follows : 

Hydrochloric  Acid,  .  .  .  .  .1  part 

Alcohol,  06  per  cent.       .  .  .  .70  parts 

Water,  .  .  .  .  .  .30  parts 

They  are  then  washed  in  running  water  for  at  least  two  hours  to  remove  all 
the  acid.     Dehydrate,  and  imbed  in  paraffin.  Newton  Evans,  M.  D. 

American  Medical  Missionary  College. 


and  Laboratory  Methods.  1173 

Journal   of  "Men  of  science  form,  as  it  were,  an 

A          !•     J     IV/I*  organized  army,  laboring  on  behalf  of 

Applied     Microscopy  the  whole  nation,  and  generally  under 

""'^  its  direction  and  at  its  expense,  to  aug- 

Laboratory   Methods.  ^ent  the  stock  of  knowledge  as  may 

Edited  by  L.  B.  ELLIOTT.  ^^'""^  ^°  promote  industrial  enterprise, 

to  increase  wealth,  to  adorn  life,  to  im- 


issued  Month^y^fron.  H,e^p_ubjicj,tio„^  Department  p^oye  political  and  social  relations,  and 

Rochester,  N.  Y.  ^o  further  the   moral   development   of 

SUBSCRIPTIONS:  individual  citizens."     The  full  signifi- 

Orie  Dollar  per^Year.^Jo^Fordgn^Countries,$^  CanCC      of      these      WOrds,      written      by 

=^-^===-==^=^-==-=^=^=^^=-^=^=  Helmholz  nearly  half  a  century  ago,  is 

The  majority  of  our  subscribers  dislike  to  have  their  i      u       •        •                i_       r    11 

files  broken  in  case  they  fail  to  remit  at  the  expiration  nOW    Only   beginning  tO  be   fuUy    appre- 

of  their  paid  subscription.     We  therefore  assume  that  no  •    4.    j         o    •         -n 

interruption   in   the   series  is   desired,  unless   notice   to  Ciateu.        hCientlnC    men,   in   SpitC   Of  the 
discontinue  is  sent. 


—  — :     old  popular   idea  to  the  contrary,  have 

demonstrated  indisputably  their  ability  to  cope  with  problems  of  the  greatest 
practical  and  economic  value.  Industrial  progress  is  more  and  more  dependent 
upon  the  results  of  their  labors. 

The  policy  of  our  government  has  always  been  to  support  liberally  men  and 
institutions  which  undertake  to  promote  the  welfare  of  the  people ;  and  yet  we 
must  admit  that  we  have  not  met  our  highest  possibilities,  for  we  have  but  to 
look  to  certain  other  progressive  nations  to  see  points  wherein  we  can  make 
decided  improvement.  It  is  no  longer  a  theory  that  governmental  support  of 
scientific  work  pays  in  every  sense  of  the  word,  for  Germany  has  long  since 
demonstrated  it  to  be  a  fact.  Her  scientific  instruments  have  been  brought  to 
the  highest  degree  of  perfection,  by  cooperation  with  individuals  capable  of  im- 
proving them,  and  through  them  her  industrial  progress  has  been  most  advanced. 
"  Made  in  Germany "  has  been  a  key  to  every  market  in  the  world.  This 
development  must  be  attributed  to  the  cooperation  of  science  and  government ; 
a  condition  of  mutual  support,  toward  which  our  own  country  is  rapidly  trending. 

The  work  accomplished  by  our  science  departments  and  bureaux  is  only  the 
preface  of  what  may  be  expected  in  the  near  future.  Nation,  state,  university, 
and  individual  are  forming  one  great  combination  for  the  pursuit  of  pure  and 
applied  science.  In  this  cooperation  the  needs  of  science  will  be  largely  brought 
to  light  by  individuals  who  are  actually  engaged  in  the  work.  These  needs  must 
be  met  by  the  institution,  state,  or  nation  in  whose  interest  the  individual  pursues 
his  investigations. 

There  are  now  needs  which  handicap  our  progress  and  place  us  at  a  dis- 
advantage in  the  competition  with  other  nations.  Governmental  cooperation  in 
the  development  of  the  methods  of  chemical  glass,  and  many  other  manufactures, 
and  in  the  standardizing  and  control  of  apparatus  used  for  weighing  and  measur- 
ing, together  with  the  adoption  of  the  metric  system  of  weights  and  measures, 
would  produce  practical  results. 

Scientific  men  have  adopted  the  metric  system  in  their  work.  The  industries 
recognizing  its  advantages,  do  not  wait  for  the  adoption  of  the  system  by  the 
government,  but  are  rapidly  introducing  it  into  their  various  calculations. 


1174  Journal  of  Applied  Microscopy 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  sliould  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Juel,  H.  0.     Beitrage  zur  Kentniss  der  Tetra-      This     important     contribution     really 
denbildung.    Jahrb.  f.  wiss.  Bot.  35:  626-      consists  of  three  distinct  papers,  which 

659,  pis.  15-16,  1900.  r    r        > 

can  be  considered  separately. 
I.   Tetrad  formation  in  the  ovule  of  Larix. 

The  homologies  between  the  reproductive  organs  of  the  vascular  cryptogams 
and  the  phanerogams  have  long  been  known  in  their  grosser  features.  The 
pollen  chambers  in  the  anther  are  microsporangia  and  the  pollen  grains  are 
microspores.  The  ovule  is  a  modified  megasporangium  and  the  embryo-sac  is  a 
megaspore ;  but  while  it  is  accepted  that  the  pollen  grain,  like  the  spore  of  a 
vascular  cryptogam,  arises  by  a  tetrad  division,  it  is  generally  believed  that  the 
embryo-sac  is  formed  without  a  tetrad  division. 

Dr.  Juel  investigated  the  ovule  of  Larix  sibirica  from  an  early  stage  in  the 
development  of  the  mother  cell  of  the  megaspore  up  to  the  beginning  of  endo- 
sperm formation.  The  paper  is  of  particular  interest  because  it  is  the  first  to 
treat  this  portion  of  the  life  history  of  a  Gymnosperm  from  the  standpoint  of 
modern  cytology. 

In  material  collected  about  the  middle  of  April,  before  the  snow  had  disap- 
peared, the  mother  cell  of  the  megaspore  was  easily  distinguished  by  its  large 
size  and  by  the  abundance  of  starch  which  it  contained.  The  first  division  is 
heterotypic  and  shows  the  reduced  number  of  chromosomes  (12).  At  the  poles 
of  the  spindle  are  granular  masses  which  may  possibly  represent  centrosomes. 
During  the  anaplase  the  starch  disappears,  a  cell  wall  is  formed  and  each  of  the 
daughter  nuclei  divides  again  by  a  homotypic  division  and  thus  gives  rise  to  a  row 
of  four  megaspores,  the  lowest  of  which  germinates  and  produces  the  prothallium. 

By  comparing  these  series  with  the  development  of  the  microspore  from  the 
mother  cell,  which  has  already  been  thoroughly  studied  in  Larix,  Prof.  Juel  comes 
to  the  conclusion  that  the  two  series  are  homologous,  the  megaspore  arising  like 
the  microspore  by  a  tetrad  division.  While  this  conclusion  is  not  new,  the  evi- 
dence supporting  it  is  a  real  contribution. 

//.    The  tetrad  division  in  a  hybrid  plant. 

It  has  long  been  known  that  hybrids  are  generally  sterile,  and  it  has  also 
been  known  that  the  pollen  of  hybrid  plants  is  commonly  imperfect.  The  pres- 
ent writer  investigated  the  formation  of  the  tetrad  in  Syringa  rothomagensis,  a 
hybrid  between  S.  persica  and  S.  vulgaris.  The  form  did  not  prove  to  be  a  favor- 
able one  for  such  a  problem,  because  the  pollen  of  both  parents  is  poor,  in  S. 
vulgaris  about  50  per  cent,  of  the  pollen  grains  appearing  to  be  incapable  of 


and  Laboratory  Methods.  1175 

functioning,  and  in  6".  persica  normal  pollen  grains  being  quite  rare.  The  latter 
form  is  almost  as  sterile  as  the  hybrid. 

In  all  three  forms  the  development  is  normal  up  to  the  formation  of  the  pollen 
mother  cells.  In  the  hybrid  S.  rothomagensis  it  was  found  that  while  most  of  the 
divisions  in  the  pollen  mother  cells  were  mitotic,  there  were  also  numerous  cases 
of  amitotic  division,  and  abnormalities  in  the  chromatin  and  in  the  achromatic 
figure  were  frequent. 

///.   The  development  of  the  pollen  grain  of  Carex. 

As  a  rule  the  pollen  mother  cell  of  a  flowering  plant  gives  rise  to  four  pollen 
grains,  but  it  has  been  reported  that  in  the  Asclepiadaceae  and  Cyperaceae  the 
mother  cell  gives  rise  to  but  one  pollen  grain. 

A  careful  examination  of  Carex  acuta  gave  the  following  results :  The  wall 
of  the  pollen  mother  cell  becomes  the  wall  of  the  pollen  grain.  The  tetrad  divis- 
ions take  place,  but  the  walls  separating  the  four  cells  are  imperfect  and  only  one 
cell  of  the  tetrad  develops  into  a  pollen  grain,  the  other  three  being  crowded  out, 
just  as  in  the  megaspore  series  three  potential  megaspores  are  crowded  out  by 
the  one  functioning  megaspore.  c.  j.  c. 

Dixon,  H.  H.    On  the  first  mitosis  of  the  spore      ^^^    ^^^^or    States    very    clearly    the 
mother  cells    of    Libtun.     Notes   from   the  •'  ■' 

Botanical  School  of  Trinity  College,  Dublin,      points    in    regard    to    which    there    is 
No.  4,  pp.  129-140,  pis.  7-8,  1 90 1.  essential  agreement  among  cytologists, 

and  outlines  the  debated  questions.  While  admitting  that  there  is  still  ample 
room  for  dispute,  he  concludes  that  in  both  the  first  and  second  nucler  divisions 
by  which  the  spores  are  formed  from  the  mother  cell,  the  splitting  of  the  chro- 
mosomes is  longitudinal  and  that,  consequently,  there  is  no  reducing  division. 

c.  J.  c. 

Holm,  Theodore.    Erigenia  bulbosa,  Nutt.    A      Mr.    Holm,   who  has  done  more  than 
Morphological  and  Anatomical  Study.    Am.  ^^^    gigg    j^    this    country    on    the 

Jour.  Sci.  IV.     II:  63-72.     6  figs.  ^  ■' 

minute  anatomy  of  plants,  presents  in 

his  latest  paper  some  interesting  morphological  and  anatomical  facts  on  this 
unique  plant.  The  Erigenia  possesses  a  single  cotyledon.  The  blade  of  the 
cotyledon  is  held  in  a  horizontal  position  and  raised  above  the  ground  by  a  long 
slender  petiole.  In  the  second  year  after  germination  the  first  proper  leaf 
appears  and  has  a  ternately  decompound  leaf,  with  divisions  of  the  same  shape 
as  those  of  the  mature  leaf.  The  third  year's  growth  is  not  much  advanced  as 
only  a  single  green  leaf  is  developed  with  a  few  additional  divisions.  The  Eri- 
genia germinates  then  with  only  one  cotyledon. 

The  tuber  as  it  appears  during  the  seedling  stage,  as  Holm  has  shown  from 
anatomical  consideratiohs,  is  a  swollen  part  of  the  primary  root. 

The  structure  in  fully  matured  specimens  is  very  different,  and  here  is  what 
the  writer  says  concerning  this : 

"  The  mature  tuberous  root  possesses  a  number  of  cork-layers,  a  secondary 
bark  of  very  considerable  width,  filled  with  starch,  and  inside  the  bark  is  a  band 
of  collateral  mestome-bundles  with  cambium  between  the  leptome  and  hadrome 
and  besides  well  defined  strata  of  interfascicular  cambium,  while  a  broad  pith 


liTf)  Journal  of  Applied  Microscopy 

occupies  the  central  portion  of  the  root,  of  which,  however,  the  innermost  part  is 
broken  down  into  a  cavity ;  thus  the  principal  features  of  the  primary  root  are 
almost  totally  obliterated.  Oil-ducts  are  quite  numerous  in  the  mature  root ; 
they  are  located  in  the  same  radii  as  the  mestome  bundles  and  occur  in  four  or 
five  concentric  bands.  The  innermost  oil-ducts  are  to  be  seen  in  the  leptome 
itself,  the  others  some  distance  apart,  the  outermost  being  very  near  the  peri- 
phery, though  not  in  contact  with  the  cork.  It  appears  as  if  the  ducts  of  the 
outermost  two  bands  are  mostly  pentagonal  in  transverse  sections  ;  while  those 
of  the  inner  are  rhombic  and  somewhat  narrower  in  circumference." 

There  are  thirty-eight  oil-ducts  in  each  mericarp  in  each  fruit,  twelve  on  the 
commisural  side,  one  outside  each  of  the  five  mestome  bundles,  and  from  five  to 
six  in  the  intervals  between  these.  The  mericarps  are  not  glabrous,  but  hairy, 
consisting  of  short  unicellular  pointed  hairs  which  cover  the  entire  dorsal  face. 
Agri.  College,  Ames,  la.  L.  H.  Pammel. 


CYTOLOGY,   EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  Sage  College, 
Ithaca,  N.  Y. 

CURRENT  LITERATURE. 

Prenant,  A.     Cellules  Tracheales  des  Oestres.      Material  used  for  this  investigation  was 
Archiv.     D'Anat.     Microscop.    3:    201-3-16       ^   ,  ...         .  ^,        ^  1       r  ^i 

(Planch.  15-16),  1900.  taken,  livmg,  from  the  stomach  of  the 

horse,  and  consisted  in  larvae  of   the 

bot-fly  {Gastrophilus  eqi/i).     Examining  these  larvae  an  area  of  red  coloration 

is  always  seen  at  the  posterior  extremity  of  the  animal.     On  dissection  two  bodies 

are  found,  one  on  each  side  of  the  digestive  tube,  which  are  white  and  opaque  in 

their  anterior  three-quarters,  and   lobulated  in  structure.     In  the  posterior  part 

they   are  more  granular,  and  of  a  reddish   color,  varying  to  a  purple-red.     This 

part  of  the  organ   is  provisionally  called  the  "  organe  rouge."     This  organ  is 

found  to  be  composed  of  a  number  of  large  oval  cells,  surrounded  thickly  by 

tracheae,  which  branch  and  finally  enter  into  the  interior  of  the  cell.     Such  cells 

are  called  "  tracheal  cells  "  to  distinguish  them  from  the  fat  cells  of  the  anterior 

part  of  the  organ.     Besides  using  fresh  material,  organs  were  fixed  in  Flemming's 

stronger  solution;  Mann's  fluid  (picric  acid,  sat.  sol.,  10  pts.;  sublimate  sat.  sol., 

10  pts.;  formol,  5  pts.)  ;  formo-picric  sol.  of  Bouin  (sat.  sol.  picric  acid,  75  vols.; 

formol   "25 ;    glacial   acetic  acid,   5  pts.) ;    platinum  mixture  of  Bouin  (platinic 

chloride  of  1  per  cent,  sol.,  10  pts. ;  sat.  sol.  picric  acid,  20  pts. ;  formol,  10  pts.;  or 

platinic  chloride  of  1  per  cent,  sol.,  20  pts.;  sat.  sol  of  sublimate,  20  pts.;  formol, 

10  pts.;  acetic  or  formic  acid,  2  to  5  pts.)  ;  Weigert's  neuroglia  fluid,  consisting  of 

T)  per  cent.  sol.  of  acetate  of  copper,  5  per  cent,  acetic  acid,  chrome  alum  5  per 

cent.,  and  10  per  cent,  formol;  saline  saturated  sublimate  solution  ;  Golgi's  fluid. 


and  Laboratory  Methods.  1177 

Of  these  the  formo-picric  of  Weigert,  and  Flemming's,  gave  the  best  results.  The 
stains  used  were  Benda's  safranin  and  Hght  green,  Flemming's  triple  safranin- 
gentian-orange,  Mann's  blue  of  toluidin-eosin ;  especially  good  results  were 
given  by  Heidenhain's  iron  haematoxylin  after  the  picric-formol  fixative.  All 
sections  were  cut  in  paraffin.  The  author  summarizes  his  results  as  follows  : 
As  before  stated,  there  is  present  in  the  larvae  of  Gastrophilus  equi,  Fabr,  or 
pecorum,  Fabr.  but  not  in  those  of  Hypodenna  bovis  L.  and  Cephaloinyia  ovis  L., 
an  organ  occupying  the  posterior  fourth  of  the  animal,  having  a  characteristic 
red  color.  This  organ  has  anatomical  continuity  with  the  fat  body.  It  is  com- 
posed of  large  cells,  between  which  the  tracheae  branch  and  subdivide.  The 
smallest  branches  penetrate  to  the  interior  of  these  "  tracheal  cells."  Nothing 
further  could  be  observed  as  to  the  ends  of  the  branches  beyond  fine  subdivision. 
The  cytoplasm  of  the  cells  is  distinguished  from  the  tracheal  branches  by  the 
different  form  and  stain  affinity  of  its  filaments,  which  come  into  close  relation 
with  the  walls  of  the  tracheae,  but  do  not  represent  the  fine  continuations  of  these 
tubes.  These  tracheal  cells  pass  gradually  over  into  adipose  cells  in  the  transi- 
tion region  of  the  organ.  This  transition  is  effected  by  filling  the  tracheal  cell 
with  fat  globules  and  the  reduction  of  the  intracellular  trachea.  Independent  of 
this  tracheal  organ,  certain  irregular  subcutaneous  tracheal  cells  are  found  in 
certain  regions  of  the  body.  The  reason  for  this  specialization  is  considered  to 
lie  in  the  peculiar  habitat  of  the  larvae,  since  closely  related  forms  living  under 
different  conditions  show  no  such  structures.  It  is  an  example  of  limited  adapta- 
tion. Physiologically,  the  function  of  these  cells  is  respiratory,  and  hence  the 
cells  are  really  "  cenocytes,"  differing  however  from  the  latter  in  a  red  instead 
of  a  yellow  coloration.  The  transformation  of  these  tracheal  cells  into  fat  cells 
argues  for  their  "  cenocytic  "  nature,  and  they  represent  the  first  step  in  respira- 
tory differentiation.  They  are  abundantly  supplied  with  oxygen,  and  in  conse- 
quence easily  elaborate  fatty  granules  ;  hence  the  functions  of  the  two  parts  of 
this  organ  are  not  distinct,  but  successive.  a.  m.  c. 

Zollikofer,  R.     Kammerfiirbung   der  Leucocy-       In  the  Study  of  leucocytes  two   objects 
ten.     Zeit.  f.  wiss.  Mikros.  u.  f.  Mik.  Techn.  .        .  ,        ^        .  ^     .  ,     . 

17:  m-l2i   iqoo.  ^''^  ^'^  view,  the  fixation  of  the  whole 

mass  of  them  and  the  differentiation  of 
this  mass  into  its  different  kinds.  In  order  that  the  study  may  be  done  in  a 
counting  chamber,  it  is  necessary  to  mix  the  blood  and  the  staining  fluid,  and  to 
have  this  mixture  take  place  in  one  pipette.  To  determine  the  numerical  rela- 
tions of  the  different  kinds  of  leucocytes  on  a  cover-glass  preparation  is  impos- 
sible, since  there  is  an  unequal  distribution  of  the  kinds.  Lymphocytes  are 
found  in  thick  places  of  the  film,  and  are  rare  or  crushed  in  thin  places.  No 
such  objection  can  be  made  to  a  film  of  blood  in  a  counting  cell.  The  thing 
needed  was  a  diluting  fluid  for  this  "  staining-chamber  "  which  would  render 
the  red  corpuscles  invisible  and  stain  the  white  differentially.  Thin  aqueous 
formalin  solution  answers  the  first  requirement,  and  for  a  stain  a  mixture  of 
eosin  and  methylin  blue  (eosin  W.  G.  and  methylin  blue  B.  x.  of  Griibler,  Leip- 
zig) was  most  satisfactory  in  the  following  composition  :  Eosin  W.  G.  0.05,  con- 
centrated formalin  1.0,  distilled  water  100.00;  methylin  blue  0.05,  concentrated 
formalin  1.0,   distilled  water   100.00.      These  solutions  must  be  filtered;  the 


1178  Journal  of  Applied  Microscopy 

formalin  mixture  needs  to  be  kept  in  the  dark,  and  a  dark  glass  dropper  was 
used.  About  equal  parts  of  the  two  liquids  were  taken.  A  Thoma-Zeiss  pipette 
was  filled  with  blood  to  0.5  mark,  and  filled  to  1.20  with  the  mixture  ;  1.10  does 
not  completely  destroy  the  red  cells.  After  five  minutes,  the  chamber  is  filled 
from  the  pipette,  and  the  white  cells  are  allowed  to  settle.  Then  the  blood  plates 
are  found  to  be  arranged  in  characteristic  masses,  and  stain  a  light  gray-blue  ;  the 
erythrocytes  are  destroyed  ;  nucleated  red  cells  are  sometimes  recognizable  by 
their  greenish  "  discoplasm."  Malaria  plasmodia  stain  blue,  but  their  recognition 
is  uncertain.  Leucocytes  are  stained  as  to  both  nuclei  and  granules.  Eosino- 
phil granules  are  clearly  outlined,  and  the  nuclei  remain  bright.  Neutrophil 
granules  are  gray-violet.  Most  cell  granules  are  unstained.  The  mononuclear 
or  ungranulated,  leucocytes  of  normal  blood  are  homogenous,  with  faintly  blue 
cytoplasm  and  varying  nuclei.  Nuclei  of  the  larger  lymphocytes  are  clearer,  and 
light  violet,  the  others  bright  blue  and  oval.  The  nuclei  of  granulated  leucocytes 
stain  lightly.  The  granulated  mononuclear,  or  "Mark"  cells,  are  conspicuous 
by  their  size  and  varying  form  of  nuclei.  These  are  principally  recognized  by 
their  granules  and  the  size  of  the  nucleus.  For  a  counting  chamber,  Elzholz's 
(Reichert)  was  used.  This  has  a  capacity  of  0.9  cubic  miUimeter.  The  blood 
is  diluted  twenty  times,  and  the  whole  field  is  counted.  The  contained  number 
is  multiplied  by  ^^^  or  22.222.  Many  cover-glass  stained  preparations  were  also 
studied.  Preparations  were  stained  for  a  few  minutes  in  eosin,  and  for  half  a 
minute  in  methylen  blue  diluted  five  times  with  water.  Careful  fixation  is  neces- 
sary for  good  results  in  staining;  heating  in  a  hot  chamber  at  115°  for  an  hour, 
or  for  a  few  minutes  at  120 — 125°,  gives  good  results  for  both  red  and  white 
cells.  The  triacid  stain  was  used  when  it  was  desired  to  stain  the  neutrophil 
granules.  The  mixtures  given  above  afford  excellent  results  not  only  on  blood 
but  also  on  sputum,  pus,  and  other  secretions.  a.  m.  c. 

Lewinson,  J.    Zur  Methode  der  Fettfiirbung      Osmic  acid   is  the  usual   fixation  and 
Zeitschr.  f.  wiss.    Mikros.  u.  f.    Mikr.  Tech.         ^   .    .        n    •  ^  r       r  ^     ^     ^  ■    ,  ^ 

17:  321-326   iQoo.  stammg  fluid  for  fats,  but  it  has  several 

disadvantages.  It  is  expensive,  it  fixes 
but  a  small  part  of  the  tissue  put  into  it  or  any  of  the  liquids  in  which  it  is  an 
active  agent.  Any  fat  near  the  middle  of  the  tissue  remains  unfixed  and  un- 
stained. The  stain  of  osmic  acid  is  very  often  of  short  duration,  and  it  is  almost 
impossible  to  use  other  stains  after  this  fixative.  Experimenting  on  myelinic 
fibers,  a  haematoxylin  method  of  Wolters'  was  used.  By  modifications  it  was 
found  that  other  tissues  than  myelinic  fibers  took  this  stain.  The  author 
tried  concentrated  nuclear  stain,  warmed,  after  definite  fixation  methods, 
to  see  if  any  result  in  staining  fat  could  be  obtained.  A  concentrated  solution 
of  methylen  blue  in  2  to  5  per  cent,  salt  solution,  and  haematoxylin  in  acetic  acid, 
were  first  tried  on  objects  fixed  in  different  fluids.  Celloidin  sections  of  the 
ovary  of  a  rabbit  fixed  in  picric  acid  were  stained  in  such  a  warmed  solution  of 
methylen  blue  as  described  above  for  10  to  15  minutes.  After  decolorizing  with 
weak  aqueous  hydrochloric  acid  and  counterstaining  with  alcoholic  picric  acid, 
the  following  results  :  Nuclei  of  the  cells  are  blue,  protoplasm  yellow-green,  and 
the  connective  tissue  is  violet.  The  fat  in  the  follicles  takes  the  form  of  small, 
dark,  almost  black  fat-corpuscles.     For  a  modification  of  Wolters'  method  the 


and  Laboratory  Methods.  11T9 

tissues  are  fixed  in  Miiller's  fluid  ;  the  object  can  have  a  large  surface,  but  should 
not  be  thick.  From  the  fixation  fluid  the  tissue  is  put  into  70  per  cent,  alcohol, 
as  treatment  with  weak  alcohol  and  water  renders  the  fat  unstainable.  The 
celloidin  sections  are  put  in  the  stain  for  twelve  hours  at  a  temperature  of  40°C. 
A  2  per  cent,  solution  of  Kultschitzki's  haematoxylin  (hematoxylin  2  gms.,  dis- 
solved in  a  little  absolute  alcohol  added  to  100  c.  c.  of  2  per  cent,  acetic  acid). 
When  the  mixture,  which  is  at  first  yellow,  becomes  red,  it  is  ready  for  use.  The 
principal  point  is  good  decolorization,  only  well  bleached  preparations  show  the 
fat  clearly.  The  whole  process  is  as  follows  :  (1)  Fix  in  Miiller's  fluid  2  to  6 
weeks,  depending  on  the  size  of  the  object ;  wash  out  in  70  to  85  per  cent, 
alcohol,  etc.,  imbed  in  celloidin.  (2)  Cut  sections  10  to  15  mikrons  thick,  and 
put  them  directly  from  alcohol  into  the  stain  for  twelve  hours  at  a  temperature  of 
40 °C.  (3)  Wash  out  with  water.  (4)  Wash  in  a  1  per  cent,  solution  of  potas- 
sium permanganate  10  to  15  minutes.  (5)  Wash  in  water.  (6)  Treat  with  a  2 
per  cent,  solution  of  oxalic  acid,  or  a  mixture  of  two  parts  of  2  per  cent,  oxalic 
acid  to  one  of  2  per  cent,  solution  of  potassium  sulphate,  for  five  minutes.  Should 
the  preparation  show  a  yellow  or  gray-black  color,  return  it  to  the  potassium  per- 
manganate, then  pass  to  the  oxalic  acid.  If  no  fat  is  present  the  sections  lose 
their  color  entirely ;  if  fat  is  there  the  sections  are  light  ash-gray  to  an  intense 
gray-violet,  depending  on  the  amount  present.  In  this  way  fat  is  shown  on  a 
colorless  background  in  gray-violet  fat  globules.  If  it  is  desired  to  stain  the 
nuclei  and  protoplasm  of  the  cells,  a  counterstain  of  concentrated  carmin  solu- 
tion may  be  used  as  follows  :  (1)  The  sections  decolorized  in  oxalic  acid  are 
washed  in  water  and  left  in  an  ammoniacal  solution  of  borax  carmin  for  twenty- 
four  hours.  (2)  Treated  in  acid  alcohol  (1  per  cent,  in  70  per  cent,  alcohol)  for 
two  minutes,  {o)  Sat.  alcoholic  sol.  of  picric  acid  for  one  minute.  (4)  85  per 
cent,  alcohol,  absolute,  xylol  or  origanum  oil,  balsam.  The  fat  is  now  dark  blue, 
almost  black  ;  nuclei,  red  ;  protoplasm,  yellow.  This  method  is  valuable  for  four 
reasons  :  (1)  Fat  is  clearly  differentiated  to  the  smallest  particle.  (2)  This  fat- 
stain  is  very  lasting;  preparations  remain  good  for  several  months.  (3)  Miiller's 
fluid  is  an  easily  available  fixation  fluid.  (4)  The  method  is  both  inexpensive 
and  simple,  requiring  no  complicated  technique.  a.  m.  c. 

RECENT    LITERATURE. 

Lavdowsky,    M.     Ueber  eine  Chromsublimat-  Nicolas,  A.     Recherches  sur  I'embryologie  des 

verbinclung  und  ihre  histologische  Anwend-  Reptiles.       Contribution    a    I'etude    de    la 

ung,  unter  anderem  audi  zur    Restauration  Fecondation  chez  I'Orvet.     Archiv.  D'Anat. 

alterer  Objecte.     Zeit.  f.  wiss.  Mikros.   u.  f.  Micros.  3:  456-489,  i  pL,  1900. 

Mikros.  Technik.  17:  301-31 1,  1900.  Goodrich,    E.    S.       Nephridia    of    Polycliasta. 

«;  u        i       /--     ..  •!-   .•        '  1.'.    J     J      1         '.  Quart.  Jour.  Micr.   Sci.  43:    609-748,  6  pis., 

Weber,  A.     Contribution   a  1  etude  de    la  met-  ^  ■'  .^  /t  >      r 

amerie  du  cerveau   anterieur  chez  quelques  "     '  •  /-. 

Oiseaux.     Archiv.  D'Anat.   Micros.  3 :  369-       Yasuda,  A.     Adaptution  of  Infusorians  to  Con- 

424   2  pis.    igoo.  centrated  Solutions.    Jour.  Coll.  Sci.  Tokyo. 

13:   101-140,  3  pis.,  1900. 


1180  Journal  of  Applied  Microscopy 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.;  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Bielschowsky  and   Pllen.      Zur  Technik  der      Ehrlich    and    Lazarus    introduced   the 
Nervenzellenfarbung.    Neurologisches  Cent-  ,  i     •    i    ,      r  •    •  . 

ralblatt,  19:  1441,  1900.  "se    of    cresyl-violet    for    staining  the 

basophilic  granules  of  mast-cells.  Lit- 
ten  has  used  the  same  anilin  dye  for  coloring  the  basophilic  granules  which  are 
found  in  the  red  blood  corpuscles  in  cases  of  anaemia.  The  writers  of  this  article 
have  found  cresyl-violet  a  good  stain  for  the  chromophilic  substance  of  the  nerve- 
cells.  They  used  chiefly  the  preparation  which  bears  the  trade  name  "  cresyl- 
violet  R.  R." 

In  composition  this  new  stain  is  probably  related  to  methylen  blue ;  in 
staining  properties  it  resembles  thionin  or  toluidin  blue,  but  is  superior  to  them, 
chiefly  because  the  preparations  are  more  permanent,  and  as  dilute  solutions 
are  employed  it  is  more  economical.  Another  advantage,  which  the  writers 
claim,  is  that  the  sections  are  never  lost  in  the  dilute  transparent  solution  of 
cresyl-violet. 

The  best  results  were  obtained  with  the  following  method  : 

1.  Harden  in  alcohol  or  formalin. 

2.  Imbed  in  celloidin. 

3.  Stain  in  a  thin  aqueous  solution  of  cresyl-violet  for  i!4  hours.  It  is  suffi- 
cient to  add  six  to  ten  drops  of  a  concentrated  aqueous  solution  to  50  c.  c.  of 
water. 

4.  Wash  quickly  in  water. 

5.  Dehydrate  in  a  series  of  alcohols  of  increasing  strength.  The  alcohol,  by 
removing  excess  of  color  from  the  diffusely  stained  sections,  differentiates  the 
gray  and  white  matter  of  the  central  nervous  system. 

6.  Clear  in  oil  of  cajeput. 

7.  Xylol. 

8.  Mount  in  Canada  balsam. 

Equally  good  results  are  obtained  after  imbedding  in  paraffin.  When  a  quick 
method  is  desirable,  a  concentrated  solution  of  the  stain  may  be  used. 

Cresyl-violet  gives  a  metachromatic  effect  with  amyloid,  coloring  amyloid 
substance  bright  blue,  the  remainder  of  the  section  violet.  j.  h.  p. 

Krompecher.     Glandhke  Carcinoma  of  Epi-      Krompecher  describes  a  peculiar  type 
dermic    Origin.     Ziegler's    Beitrage,   28:    i,  ,  ,  ,  ,,         ,  .       ,         1  •   1    i 

j„QQ  00  o  Qf  tumor  of  the  skin,  to  which  he  gives 

the  name    "  carcinoma  epithelialeade- 

noides."     He  believes  that  the  gross  and  histological  appearances  are  sufficiently 

characteristic  to  establish  it  as  a  distinct  group. 

Braun,  in  1892,  studied  this  class  of  tumors,  and  regarded  them  as  endothel- 

iomata.     Krompecher  asserts  that  the  diagnosis  of  endothelioma  can  be  made 

only  when  the  origin  of  the  tumor-masses  from  the  endothelium  of  the  larger 


and  Laboratory  Methods.  1181 

lymph-spaces  can  be  directly  traced,  or  when  the  tumor  is  found  in  places,  such 
as  bones  and  lymph-nodes,  where  epithelium  is  lacking,  and  the  structure  of  the 
tumor  corresponds  to  that  of  undoubted  endotheliomata.  Braun  did  not  fulfill 
these  requirements  of  Krompecher.  He  based  his  diagnosis  on  the  difference  of 
structure  as  compared  with  ordinary  epidermoid  carcinomata  ;  on  the  absence  of 
epithelial  pearls ;  and  especially  on  the  lack  of  any  connection  between  the  tumor 
and  the  skin. 

Krompecher  studied  thirty-three  cases.  The  tumors  occurred  on  various 
parts  of  the  body.  By  means  of  serial  sections,  he  demonstrated  the  connection 
of  these  tumors  with  the  surface  epithelium,  thus  proving  their  epithelial  origin. 
The  striking  feature  of  these  tumors  is  their  microscopic  structure.  While  the 
epidermoid  cancer  is  composed  of  the  cylindrical  cells  of  the  stratum  Malpighii, 
and  of  polygonal  prickle  cells,  which  by  cornification  give  rise  to  epithelial  pearls, 
the  group  of  tumors  under  consideration  is  distinguished  by  the  fact  that  only 
the  cylindrical  layer  of  the  stratum  Malpighii  proHferates.  The  cells  retain  their 
embryonic  character.  The  tumor  consists  of  nests  of  high  cylindrical  cells,  which 
stain  intensely.     There  is  no  formation  of  epithelial  pearls.  j.  h.  p. 

Wright,  J.  H.    A  Case  of  Multiple  Myeloma.      ^^.j  ^^  defines  multiple  myeloma  as  a 
Irans.  Assoc.  Am.  Phys.  15:   137,  1900.  ,  °  f  j 

primary  neoplasm  of  the  bone  marrow, 

affecting  chiefly  the  sternum,  the  ribs,  the  vertebrae,  and  the  skull ;  the  substance 
of  the  bone  being  more  or  less  extensively  replaced  by  the  tumor  tissue.  The 
affection  was  first  recognized  by  von  Rustizky  in  1873.  It  is  a  rare  condition. 
Less  than  twenty  cases  have  been  reported.  The  association  of  albumosuria 
with  multiple  myeloma  is  an  interesting  feature,  and  an  aid  in  diagnosis.  In  the 
case  studied  by  the  writer,  the  tissues  were  hardened  in  Zenker's  fluid  and  in 
Flemming's  solution.  The  sections  were  stained  in  various  ways,  but  eosin  and 
Unna's  alkaline  methylen  blue  solution,  and  fuchsin,  either  alone  or  in  combina- 
tion with  aurantia,  were  found  most  satisfactory. 

The  tumor  is  chiefly  made  up  of  small  cells  closely  crowded  together.  Most 
of  the  cells  have  all  the  appearances  of  plasma  cells,  except  that  the  cytoplasm 
does  not  in  all  cases  show  a  marked  affinity  for  methylen  blue,  as  does  the 
typical  plasma  cell.  Wright  holds  that  these  cells  are  plasma  cells,  and  their 
deviations  from  the  type  of  the  parent  cell  are  quite  analagous  to  those  seen  in 
the  cells  of  other  neoplasms. 

The  author  found  that  plasma  cells  are  a  normal  constituent  of  the  red  mar- 
row, and  he  concludes  that  the  tumor  arose  from  an  abnormal  proliferation  of 
these  cells.  Hence  his  case  of  multiple  myeloma  is  to  be  regarded  as  a  neoplasm 
originating  not  in  the  red  marrow  cells  collectively,  but  in  only  one  of  the  varie- 
ties of  the  cells  of  the  red  marrow,  namely,  the  plasma  cells.  j.  h.  p. 


1182  Journal  of  Applied  Microscopy 

GENERAL   PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Gamble,  F.  W.,  and  Keeble,  F.  W,    Hippolyte      This  paper  deals  with  the  color  changes 

Scfsd.  N%"% i^S-e,"  p'r/^e.  P,i      ^h°«"  by  'he  prawn  Hippolyte  varians, 

which  lives  in  shallow  water  clinging  to 
seaweeds  and  zoophytes,  in  relation  to  different  environmental  conditions  and 
to  stimulation  by  light.  It  has  been  often  noted  that  Hippolyte  shows  a 
very  remarkable  similarity  in  its  coloration  to  its  surroundings,  and  it  was  the 
purpose  of  the  authors^to  determine  under  exact  experimental  conditions 
what  the  nature  of  these  adaptive  color  changes  was,  and  how  they  were  brought 
about.  As  an  introduction  a  description  is  given  of  the  different  natural  varieties 
of  this  prawn  and  of  the  condition  of  the  chromatophores  associated  with  these 
varieties.  Uniform  brown,  pink,  red,  and  green  adult  specimens  were  collected, 
while  among  immature  individuals  "red  liners,"  "black  liners,"  "green  liners  " 
and  jellow  barred  specimens  were  common.  The  "  liners  "  are  animals  which 
are  transversely  striped  with  the  color  indicated.  The  pigments  are  contained  in 
chromatophores  which  lie  under  the  epidermis  in  the  connective  tissue  and 
muscles  and  about  the  alimentary  canal  and  blood  vessels.  The  chromatophore 
itself  consists  of  a  central  body  from  which  diverge  a  number  of  line,  ramifying, 
hollow  tubes.  In  these  tubes  and  the  central  body  are  contained  the  pigments 
which  give  the  color  to  the  animal.  There  are  three  pigments,  red,  yellow  and 
blue,  and  color  changes  are  caused  by  the  movement  of  these  pigments  in  the 
tubes.  There  is  a  layer  of  chromatophores  in  the  connective  tissue  just  beneath 
the  epidermis,  the  processes  of  which  form  a  close  meshwork  about  the  clear 
transparent  cells  of  the  epidermis  itself.  To  the  combination  of  the  three 
pigments  in  this  epidermal  meshwork  is  due  the  color  of  the  animal  as  a  whole. 
For  example,  in  a  green  prawn  the  tubes  of  the  meshwork  are  found  to  contain 
both  yellow  and  blue  pigment  side  by  side. 

The  animal  exhibits  three  sorts  of  color  changes:  (a)  slow,  sympathetic 
changes  of  color  accompanying  changes  in  the  color  of  the  weed  to  which  the 
individual  is  attached ;  (b)  rapid  color  changes  caused  by  changes  in  light 
intensity  ;  (c)  periodic  nocturnal  color  changes,  (a)  In  regard  to  adaptive  color 
changes  in  response  to  changes  of  the  weeds,  it  was  found  that  the  animals  were 
capable  of  only  very  slow  sympathetic  changes.  The  adaptations  observed  in 
nature  are  the  result  of  the  selection  by  the  animal  of  those  weeds  whose  color 
most  closely  matches  their  own.  (b)  The  intensity  of  illumination  has  a  pro- 
nounced effect  on  the  color  of  the  animals  and  this  effect  is  produced  in  a  very 
short  time.  In  high  light,  or  in  low  light  scattered  evenly  from  the  surface  of 
the  containing  dish,  there  is  a  retraction  of  the  red  pigment  and  an  evolution  of 
the  blue  and  the  yellow,  producing  a  green  coloration  of  the  animal ;  while  in  low 
light  absorbed  by  the  walls  of  the  vessel  the  red  remains  expanded.  The  color 
quality  of    the    light  has   no   effect  on  the  color  of   the    animal  provided   the 


and  Laboratory  Methods.  1183 

intensity  remains  the  same,  (c.)  It  was  found  that  at  evening  the  prawns  change 
regularly  and  uniformly  from  their  diurnal  color  to  a  deep,  transparent  blue. 
This  blue  color  passes  away  in  the  morning  and  the  diurnal  color  of  the  previous 
day  reappears.  This  nocturnal  change  with  its  associated  diurnal  recovery  is  a 
periodic  phenomenon.  The  nocturnal  blue  appears  at  certain  intervals  even  if 
the  illumination  is  kept  constant  for  several  days  at  a  time,  and  on  the  other 
hand  the  recovery  of  the  diurnal  color  occurs  regularly  in  specimens  kept  in  the 
dark  during  similar  long  periods.  Blinded  prawns  exhibit  the  same  periodicity. 
The  general  physiological  condition  of  the  animals  is  very  different  during  the 
night  and  the  day. 

Other  experiments  showed  that  the  condition  of  the  chromatophores  at  any 
time  is  the  result  of  impulses  passing  to  them  from  the  central  nervous  system. 
Color  changes  can  be  induced  by  a  variety  of  stimuli  such  as  temperature, 
chemicals,  electricity,  etc.,  which  affect  the  nervous  system.  Removal  of  the 
eyes  causes  a  change  in  the  impulses  going  to  the  central  nervous  system 
and  hence  a  change  in  the  color.  The  nocturnal  condition  is  a  result  of  a 
periodicity  in  the  action  of  the  nervous  system.  The  relation  of  the  nocturnal 
color  of  these  prawns  to  the  color  of  deep  sea  animals  is  discussed. 

This  paper  is  an  important  contribution  to  the  physiology  of  coloration. 
Unfortunately  in  the  space  of  a  brief  review  it  has  been  impossible  to  do  more 
than  mention  some  of  the  most  significant  points  in  the  great  amount  of  interesting 
detail  presented  in  the  work.  The  admirably  executed  plates  are  a  feature  of 
the  paper.     The  experimental  methods  described  are  numerous  and  valuable. 

R.  p. 

Yerkes,  R.  M.     Reactions  of  Entomostraca  to       -j^j^g  purposes  of  the  investigation  were 
Stimulation    by    Light.     II.  Reactions     of  *^      "^  ° 

Daphnia  and  Cypris.  Amer.  Jour.  Physiol.  4  :       to  determine  the    relation   between   the 
405-422, 1900.  j-^^g  q£  movement  and  the  intensity  of 

light ;  to  determine  whether  there  is  a  reversal  of  the  phototactic  reaction  of 
Daphnia  and  Cypris  under  certain  conditions  ;  and  finally  to  study  the  effect  of 
other  stimuli  on  the  light  reactions.  In  regard  to  the  first  point  it  was  found 
that  Daphnia  moved  only  slightly  faster  as  the  intensity  of  the  light  increased, 
while  Cypris  showed  a  still  less  definite  relation  between  rate  and  intensity  than 
Daphnia.  No  very  conclusive  results  came  from  the  experiments  devised  to  test 
the  question  of  reversal  of  phototaxis  from  positive  to  negative  and  vice  versa. 
Daphnia  usually  gives  a  positive  reponse  which  could  be  maintained  indefinitely 
by  changing  the  direction  of  the  light.  In  some  cases  it  was  possible  to  change 
this  positive  reaction  into  a  weak  negative  one  by  taking  the  anirhals  up  in  a 
pipette  as  described  by  Towle  (Amer.  /our.  Physiol.  3:  345-365),  but  this 
result  was  not  constant.  The  reverse  change  from  negative  to  positive  was  not 
obtained,  owing  apparently  to  lack  of  negatively  phototactic  animals  on  which  to 
experiment.  In  the  case  of  Cypris,  negative  animals  were  made  positive  by 
contact  with  the  sides  of  the  pipette.  Raising  the  temperature  does  not  affect 
the  phototaxis  of  these  crustaceans.  Sudden  illumination  of  the  animals  from 
above  in  such  a  way  that  the  directive  influence  of  the  light  is  excluded  does  not 
cause  any  change  in  the  direction  of  swimming.  In  one  set  of  experiments  some 
strong  HCl  was  put  into  the  trough  at  the  end  nearest  the  source  of  light.     The 


1184 


Journal  of  Applied  Microscopy 


animals  swam  towards  the  light  until  they  encountered  the  acid  solution,  and 
then  instead  of  turning  back  stayed  there  till  they  were  killed.  In  conclusion 
the  author  gives  a  rather  unconvincing  answer  to  certain  criticisms  of  his 
earlier  work. 

In  the  paper  several  useful  pieces  of  apparatus  for  phototaxis  work  are 
described.  The  method  of  changing  the  direction  of  the  light  rays  impinging  on 
the  animal  without  disturbing  any  of  the  other  conditions  seems  especially 
valuable  and  will  be  described  in  detail.  "  A  tin  trough  8  x  >4  x  ^  inches 
(T)  mounted  on  a  wooden  base  was  painted  dead  black ;  at  either  end  of  this 
trough  a  glass  box,  A,  A',  containing  alum  solution  was  placed.  Screens,  S,  S', 
were  arranged  so  that  side  rays  and  reflected  light  were  cut  off,  and  the  trough 
was  illuminated  exclusively  by  rays  parallel  with  its  long  axis  coming  through 
holes  six  inches  high  and  two  inches  wide   cut  in  the  screens,  S,  S'.     At  either 

end,  ten  inches  from  S  and 
S'  respectively,  was  a 
Welsbach  burner,  L,  L'. 
For  observations  this  appa- 
ratus was  set  up  in  a  dark 
room.  After  the  trough  had 
been  filled  with  water  and 
the  screens  s,  s',  which 
shut  off  all  light,  had  been 


S' 


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placed  in  position,  an  animal  was  carefully  dropped  into  the  middle  of  T.  One 
of  the  screens  (s)  was  then  removed  and  the  animal  responded  usually  with  a 
-f  reaction, — it  moved  toward  the  end  from  which  the  screen  had  been  removed, 
that  is,  toward  the  light.  As  soon  as  the  animal  came  within  two  centimeters  of 
the  +  end  of  the  trough,  s  was  quickly  replaced  and  s'  removed,  thus  giving  light 
from  the  opposite  direction  without  the  inconvenience  of  moving  the  burner. 
By  this  means  it  could  easily  be  observed  whether  the  response  was  continued 
as  before  or  reversed."  R-  ?• 

Piitter,  A.  Studien  iiber  Thigmotaxis  bei  The  author  deals  in  a  thorough  and 
Protisten.  Arch.  Anat.  u.  Physiol.  Physiol.  exhaustive  way  with  the  effect  of  COn- 
Abth.  Suppl.  Bd.   IQOO:  pp.  243-302.  .,,.,,,. 

tact  with  solid  bodies  on  the  reactions 

of  the  Protozoa.  After  a  brief  historical  introduction  and  description  of  methods 
employed,  the  reactions  of  a  large  number  of  Protozoa,  including  nearly  all  the 
main  groups  from  the  rhizopods  to  the  hypotrichous  ciliates,  are  described  in 
detail.  Positive  and  negative  forms  of  thigmotaxis  are  distinguished  according 
as  the  animal  remains  in  contact  with  a  solid  body  which  it  encounters,  or  moves 
away  from  it.  The  positive  reaction  displays  two  forms  or  factors.  The  first 
factor  is  the  one  which  affects  the  locomotor  organs  (pseudopodia,  fiagella  or 
cilia)  and  results  in  a  lessening  or  inhibition  of  their  movement.  The  second 
factor  in  the  thigmotaxis  is  the  secretion  of  a  sticky  slime  which  helps  to  fasten 
the  animal  to  solid  bodies.  This  secretion  factor  is  very  evident  among  the 
rhizopods,  less  apparent  among  the  flagellates  and  ciliates  where  the  first  factor 
is  most  important,  and  finally  it  is  the  most  essential  phenomenon  in  the  thigmo- 


and  Laboratory  Methods.  1185 

taxis  of  Oscillaria,  diatoms  and  desmids,  and  the  Gregarinidse.  It  is  evident 
that  these  phenomena  of  thigmotaxis  are  very  important  in  the  life  of  the 
Protozoa. 

This  importance  is  well  shown  by  the  effect  of  the  thigmotaxis  on  the  reac- 
tions to  other  stimuli.  The  other  reactions  studied  were  the  electrotactic  and 
the  thermotactic.  In  regard  to  the  electrotaxis  it  was  found  that  among  some 
of  the  ciliates,  individuals  which  were  kathodically  electrotactic  when  swimming 
freely  through  the  water,  when  in  contact  with  a  solid  body  (i.  e.,  thigmotactic) 
oriented  themselves  more  or  less  transversely  to  the  direction  of  the  current  with 
the  oral  side  of  the  body  towards  the  kathode.  This  transverse  orientation  was 
investigated  in  a  number  of  forms.  It  is  evidently  the  same  reaction  as  that 
which  has  been  described  by  the  reviewer  (Amer.  Jour.  Physiol.  Z\  96-123) 
and  explained  as  due  to  the  conflict  between  two  sets  of  ciliary  activities. 
It  is  now  shown  to  be  also  in  part  the  result  of  the  thigmotaxis  of  the  animal. 
The  author  confirms  previous  investigators  as  to  the  reversal  of  the  cilia  on  the 
kathode  side  of  the  body  during  the  action  of  the  current.  The  permanent 
transverse  electrotaxis  of  Spirostomum  is  thought  to  be  a  result  of  the  thigmo- 
taxis of  the  posterior  end  of  the  body.  Professor  Loeb's  theory  of  the  action  of 
the  external  electrolytes  in  electrotaxis  is  thoroughly  examined  and  strong 
evidence  against  it  is  presented.  The  effect  of  heat  or  cold  is  different  accord- 
ing as  the  animal  is,  or  is  not,  in  contact  with  a  solid  body.  Many  forms 
(Euglena,  Chilodon,  Stylonychia,  Spirostomum  and  others)  cannot  be  made  to 
leave  the  bottom  by  heating.     They  die  while  still  thigmotactic. 

The  reactions  of  Stylonychia  mytilus  are  described  in  more  detail  than  those 
of  any  other  form  and  some  interesting  curves  are  given  showing  the  relative 
activities  of  the  different  groups  of  cilia  at  different  temperatures.  All  the  cilia 
show  maximal  activity  at  two  widely  separated  temperatures  (5-10°  and 
25-35°C.)  while  the  minimal  activity  of  all  is  between  15°  and  20°C. 

This  excellent  piece  of  work  puts  our  knowledge  of  another  of  the  reactions 
of  the  Protozoa  on  a  firm  basis.  R.  p. 

Delage,  Y.,  and  Delage,  M.    Sur  les  relations      In  this  note  are  presented  the  results  of 
entre  le  constitution  chimique  des  produits  rhemiral  analvses   of  the    sexual 

sexuels  et  celle  des  solutions  capables  de      ^ome  ciiemicai  analyses  or  tne  sexual 

determiner  la  parthenogenese.    C.  R.  Ac.      productsof  the  male  and  female  in  the  sea 
8ci.  Paris,  131 :  1227-1229,  1900.  ^^^^^.^^  Strongylocentrotus  lividus.  The 

starting  point  of  the  work  is  the  idea  that  if,  as  has  been  stated,  it  is  the  Mg-ion 
which  causes  the  artificial  parthenogenetic  development  of  the  egg,  and  if 
normal  development  is  the  result  of  the  same  sort  of  a  process,  analysis  ought 
to  show  a  greater  proportionate  amount  of  this  salt  in  the  sperms  than  in  the 
eggs.  This  was  not  found  to  be  the  case.  The  magnesium  content  of  the 
products  of  both  sexes  is  essentially  the  same,  so  that  normal  fertilization  cannot 
depend  merely  on  the  bringing  of  more  of  this  salt  into  the  egg  by  the  sperm. 
This  result  in  no  way  affects  Professor  Loeb's  later  views,  which  point  to  osmotic 
pressure  as  the  essential  factor  in  the  production  of  artificial  parthenogenesis. 


1186  Journal  of  Applied  Microscopy. 

CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Jensen.    Studien  uber  die  Enzyme  im  Kase.  -pj^g  question  whether  various  fermen- 

Cent.  f.  Bac.  II.  6:  734,  1900.  .     ^                         . 

„     „  ,    .       r  ,    T-  tative    processes    in   nature  are  to   be 
Babcock  and  Russell.     Relation  of  the  Enzymes 

of  Rennet  to  Ripening  of  Cheddar  Cheese.  ascribed    properly    tO     the      action     of 

Cent.f.  Bac.  II.  6:  817,1900.    (See  also  sev-  micro-organisms    or    to    the    action    of 

enteenth  annual  report  of  Agri.  Expt.  bta.  of  '^             . 

Wis.)  enzymes,  has,  in  late  years,  become  a 

Babcock  and  Russell.  Causes  Operative  in  the  somewhat  burning  one  with  our  bacteri- 
Formation  of  Silage.  Seventeenth  An.  Rep.  ologists  and  chemists.  In  large  degree 
of  Agri.  Expt.  Sta.  of  Wis.,   1900.  ^  .  ,  .       ,,      .        , 

,.        ,       ^  ,       the  question   reduces   itself  simply  to 

Behrens.     Ueber   die  oxydierenden    Bestand- 

theile  und  die  Fermentation  des  Deutschen       determining     whether      the      enzymes, 
Tabaks.    Cent.  f.  Bac.  II.  7 :  i,  1901.  which  are  the  direct  cause  of  the  action, 

are  produced  by  bacteria  or  from  some  other  source.  The  four  papers  here 
referred  to  discuss  different  aspects  of  this  problem.  It  has  been  shown  by 
Babcock  and  Russell  that  fresh  milk  contains  an  enzyme  which  they  have  named 
galactase.  They  believe  that  this  enzyme,  rather  than  micro-organisms,  plays 
the  important  part  in  the  ripening  of  cheese.  The  first  of  the  articles  here 
referred  to  contains  an  especially  careful  series  of  experiments  to  test  this  con- 
clusion. As  the  result  of  a  long  series  of  most  careful  experiments,  Jensen 
concludes,  in  brief,  that  in  the  ripening  of  soft  cheeses  the  effect  is  produced, 
(1)  by  enzymes  which  are  produced  by  yeasts  and  bacteria  growing  on  the 
surface  of  the  cheese  ;  and  (2)  by  enzymes  in  the  center  of  the  cheese  which  are 
not  derived  from  bacteria  growth,  but  rather  from  the  rennet  which  was  added 
to  curdle  the  casein,  the  enzyme  in  this  case  being  pepsin.  The  ripening  of 
hard  cheeses  depends  partly  upon  the  action  of  an  enzyme  produced  throughout 
the  mass  as  the  result  of  bacteria,  and  partly,  especially  in  the  early  part  of  the 
ripening,  upon  the  galactase,  which,  as  Babcock  and  Russell  have  shown,  is 
present  in  the  fresh  milk. 

In  the  second  article  Babcock  and  Russell  test,  by  an  entirely  different  line 
of  experiments,  the  question  whether  the  pepsin  present  in  the  rennet  has  an 
important  agency  in  the  ripening  of  cheese.  The  conclusion  they  reached  is 
essentially  identical  with  that  of  Jensen,  namely,  that  the  ripening  of  cheese  is 
dependent  in  considerable  degree  upon  the  pepsin  present  in  the  rennet.  The 
agency  of  bacteria  in  the  ripening  of  cheeses  is  not  especially  studied  by 
these  authors. 

The  third  paper  records  a  series  of  experiments  to  determine  whether  the 
production  of  silage  is,  as  has  previously  been  believed,  the  result  of  the  growth 
of  micro-organisms.  The  authors  reached  the  conclusion  that  micro-organisms 
have  nothing  whatsoever  to  do  with  the  production  of  normal  silage.  Both  the 
initial  heating  and  the  subsequent  ripening  of  silage  are  due  to  entirely  different 
agents.     The    production  of   silage,  the    authors   believe,    is    due,    (1)    to   the 


and  Laboratory  Methods.  1187 

respiratory  processes  of  plant  tissues  which  continue  for  some  time  in  the  silage 
after  the  silo  is  packed,  thus  producing  the  initial  heating ;  (2)  to  the  presence  of 
enzymes  which  are  liberated  from  the  plant  cells  after  the  death  of  the  plant  tissue. 

Micro-organisms,  the  authors  believe,  only  injure  the  silage  and  are  of  no 
significance  in  a  properly  constructed  silo. 

The  paper  by  Behrens  deals  with  the  question  of  the  fermentation  of  tobacco, 
which  has  been  regarded  as  due  to  micro-organisms,  but  which  Loew  has  some- 
what recently  insisted  is  the  result  of  enzymes  formed  in  the  tobacco  leaves. 
Behrens  has  tested  Loew's  conclusion  and  was  able  to  isolate  from  the  leaves  of 
German  tobacco  the  same  chemical  products  referred  to  by  Loew.  After  making 
a  somewhat  careful  study  of  them  and  their  action,  he  reaches,  however,  quite 
different  conclusions  from  those  of  Loew.  His  conclusions  are,  briefly,  that 
these  bodies  (oxydase,  peroxydase)  are  formed  in  tobacco  leaves.  He  is 
doubtful  as  to  whether  they  are  properly  to  be  called  enzymes,  and  is  convinced 
from  his  experiments  that  they  cannot  be  the  cause  of  the  tobacco  fermentation, 
inasmuch  as  they  disappear  from  the  leaves  before  the  important  fermentation 
takes  place.  His  experiments  further  show  that  these  oxydases  will  not  produce 
ammonia  from  nicotine,  a  phenomenon  of  tobacco  fermentation  which  he 
attributes  to  bacteria.  He  finds,  also,  that  micro-organisms  will  grow  in  tobacco 
when  the  amount  of  water  is  not  over  25  per  cent.,  contrary  to  Loew's  claims, 
and  is,  therefore,  convinced  that  the  chief  factor  in  the  proper  tobacco  fermenta- 
tion is  due  to  bacteria  growth  rather  than  to  these  chemical  bodies  produced  in 
the  tobacco  leaves.  H.  w.  c. 

Harrison.    Die    Lebensdauer    des  Tuberkel-      Harrison  has   experimented  upon  the 
Bacillus    im     Kase.     Landw.     Jahrb.     der       ,  ,       ^    •         ■         .  •   ,        i         i     i       mi- 

Schweiz.  looo.  length  of  time  m  which  tubercle  bacilli 

remain  alive  in  cheese.     His  method  of 

experiment  has  been  to  inoculate  milk  with  a  considerable  quantity  of  tubercle 

culture  and  then  to  make  the  milk  into  cheese  in   the  ordinary  fashion.     At 

varying  intervals  the  cheese  was  tested  by  inoculation  into  guinea  pigs.     These 

animals   were   studied  both   clinically  and  microscopically.     He  found  that,  in 

Emmenthaler  cheese,  the  tubercle  bacilli  were  dead  at  the  end  of  33-40  days, 

while  in  Cheddar  cheese  they  might  remain  alive  for  104  days.     The  conclusion 

is  that  neither  of  these  cheeses  is  a  source   of  danger  to   man,  since  they  are 

seldom  eaten  until  they  are  four  months  old,  or  even  older.  h.  w.  c. 

Lameris  and  Harrevelt.    Bakterienbefund   in      The    authors    made    a   study    of    the 
Kuhnmilch  nach  algeheilter  Mastitis.     Zeit.       ,  .  .  ,        .,, 

f.  Fl.  u.  Milch  Hyg.  11 :  114,1901.  bacteria    content  of    some  cows    milk 

which  had  produced  cases  of  diarrhoea. 

Inspection  of  the  source  of  the  milk  showed  that  some  of  the  cows  had  formerly 

suffered  from  mastitis,   but  had  apparently   recovered.     In   the   milk,  however, 

there  was  present  a  species  of  streptococcus  which  is  uniformly  found  and  which 

is  really  the  cause  of  the  intestinal  disturbance  produced  by  the  use  of  the  milk. 

Inasmuch,  however,  as  the  milk  produced  these  disturbances,  even  after  boiling, 

and  the  streptococci  were  shown  to  be  killed  by  this  temperature,  the  authors 

conclude  that  the  trouble  arose  from  the  toxines  developed  in  the  milk  by  the 

streptococcus  rather  than  by  the  direct  action  of  the  organisms.  h.  w.  c. 


1188  Journal  of  Applied  Microscopy. 


Medical  Notes. 

The  Epilepsy  Parasite. — The  short  paper  published  in  the  December  num- 
ber of  the  Journal  does  not  represent  the  present  status  of  this  parasite,  and  in 
justice  to  myself  and  the  cause  of  science  a  little  more  should  be  said  about  it. 
That  a  parasite,  represented  in  the  cut  accompanying  the  December  article,  is 
the  cause  of  some  forms  of  reflex  epilepsy,  is  an  undisputed  fact.  In  the  first 
case  at  Chester,  Illinois,  the  boy  was  cured  by  the  permanent  removal  of  the 
parasite,  and  has  remained  cured.  Three  other  cases  are  known  to  the  writer 
where  the  parasite  was  found.  Only  one  of  these  three  was  known  to  the  writer 
in  detail,  and  the  removal  of  the  parasites  cured  this  case  as  in  the  first. 

The  parasite  was  found  to  be  new  to  science,  and  a  description  of  it  was  pub- 
lished in  the  September  number,  1900,  of  the  Canadian  Entomologist,  under  the 
name  of  Gastrophilus  cpilcpsalis.  In  the  September  number  of  the  Alkaloidal 
Clinic  of  Chicago  was  published  a  more  extended  account  of  what  was 
known  of  the  three  cases  that  had  been  found  then.  This  article  treated  the 
subject  more  from  a  pathological  standpoint  than  the  article  in  the  Canadian 
Entomologist.  Since  that,  one  other  case  has  been  found  in  Kentucky,  where  the 
parasite  was  connected  with  epilepsy. 

The  parasite,  instead  of  being  a  Nematoid  worm,  is  the  larva  of  a  fly,  related 
to  the  horse  bot-ily,  Gastrophilus  equi.  The  adult  fly  has  not  yet  been  recog- 
nized, nor  has  it  been  ascertained  definitely  how  it  first  gains  entrance  to  the 
system.  In  the  investigation  of  this  parasite,  two  other  fly  parasites  infesting  the 
human  intestines  have  been  found  by  the  writer  that  the  books  do  not  tell  us 
about,  one  a  species  of  Eristalis,  of  the  family  Syrphidae,  and  the  other  a  species 
of  Sarcop/iaga,  of  the  family  Sarcophagidae. 

I  should  like  to  get  specimens  of  intestinal  parasites,  and  have  correspond- 
ence relative  to  the  effect  of  such  parasites  on  the  system  of  the  host. 
Carbondale,  111.  G.  H.  FRENCH. 


Dr.  Klett  of  Wiirtemberg  has  recently  made  important  researches  upon  the 
phenomena  of  anaerobic  life.  In  his  investigations  on  the  production  of  spore- 
less  anthrax  outside  the  living  body,  Dr.  Klett  found  that  if  nitrogen  is  substituted 
for  air  in  the  anaerobic  conditions,  the  growth  of  the  organism  is  not  impaired,  and 
spores  develop  as  freely  as  under  ordinary  conditions.  If,  however,  hydrogen 
is  substituted  in  place  of  air,  no  spores  develop  providing  the  medium  is  such  as 
to  permit  intimate  contact  of  gas  with  the  culture.  These  results  would  indi- 
cate that  absence  of  oxygen  is  responsible  for  non-production  of  spores  in  anaero- 
bic cultivation  of  anthrax. 


At  a  meeting  of  Pathologists  and  Bacteriologists  in  New  York  on  January 
26th,  an  American  association  was  organized.  The  officers  elected  were  :  Dr.  W. 
T.  Councilman,  president ;  Dr.  H.  C.  Ernst,  secretary  ;  Dr.  Eugene  Hodenpyl, 
treasurer.     The  first  regular  meeting  will  be  held  in  Boston  on  April  5th. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 

VOLUME   IV.  MARCH,    1901.  NUMBER  3 

MICRO-CHEMICAL  ANALYSIS. 
XL 

AMMONIUM. 

The  salts  of  the  radical  NH^  resemble  so  closely,  in  their  behavior,  those 
of  the  alkalies  that  it  is  more  convenient  to  discuss  ammonium  in  connection  with 
the  elements  of  Group  I  than  under  the  head  of  Nitrogen. 

As  has  already  been  seen  from  the  preceding  work,  ammonium  reacts  with 
most  of  the  reagents  used  for  the  detection  of  potassium,  rubidium,  and  cesium. 
Hence  it  is  usually  not  practical  to  test  for  ammonium  directly  in  the  substance. 
It  follows,  therefore,  that  it  is  generally  necessary  to  first  volatilize  the  ammonia 
and  test  for  this  substance  after  its  separation. 

Although  the  salts  of  ammonium  are  easily  driven  of¥  by  heat,  any  attfempt  to 
sublime  them  and  then  to  test  the  sublimate  will  be  found  unsatisfactory.  A  far 
better  plan  is  to  expel  the  NH,  by  the  action  of  an  alkali  and  heat  and  to  absorb 
the  evolved  gas  in  dilute  acid.     The  method  of  procedure  is  as  follows  : 

Place  in  a  deep  25  mm.  watch  glass  a  tiny  bunch  of  fibrous  asbestos  which 
has  just  been  ignited  to  redness  by  being  held,  with  the  forceps,  in  the  flame  of 
the  Bunsen  burner.-  In  the  absence  of  asbestos,  a  tiny  piece  of  thick  filter  paper 
can  be  employed,  but  in  this  case  the  paper  must  be  tested  for  ammonia. 

On  the  absorbent  place  a  small  amount  of  the  substance  to  be  tested  and 
sufficient  water  to  just  thoroughly  moisten  the  mass,  but  no  more.  Now  add  a 
fragment  or  two  of  sodium  hydroxide  so  as  to  obtain  an  alkaline  reaction.  Invert 
over  the  watch  glass  thus  prepared,  a  glass  slide  bearing  at  its  center  a  minute 
drop  of  water  acidified  with  hydrochloric  acid. 

Hold  the  watch  glass  thus  covered  (by  grasping 
its  edges  between  the  thumb  and  fore-finger)  over  a 
small  flame  (see  diagram,  Fig.  39)  so  as  to  expel  the 
ammonia.  The  heating  is  kept  up  until  the  slide 
becomes  bedewed  with  moisture.  Heating  to  boil- 
ing should  be  avoided,  since  in  such  cases  there  is 
danger  of  some  of  the  contents  of  the  watch  glass 
spirting  upon  the  slide.  Pm  ^g 

(1189) 


1190  Journal  of  Applied  Microscopy 

Remove  the  slide  from  the  watch  glass  and  turn  it  over.  Cause  the  con- 
densed moisture  to  unite  in  one  drop  by  stirring  with  a  glass  rod.  This  drop 
will  contain  the  ammonia  which  has  been  expelled  from  the  substance.  The 
danger  of  a  possible  loss  has  been  guarded  against  by  the  drop  of  dilute  hydro- 
chloric acid  employed. 

In  order  to  test  for  ammonium  in  the  drop  thus  obtained,  it  is  only  necessary 

to  add  a  suitable  reagent.     Since  it  is  extremely  unlikely  that  any  compound 

other  than  ammonium  chloride  is  present,  a  number  of  methods  are  available. 

There  are  two,  however,  which  will  be  found  more  satisfactory  than  the  others. 

I.  Chlorplatinic  Acid  (Platinum  Chloride). 

II.  Magnesium   Acetate   and    Sodium    Phosphate  in  alkaline 

solution. 

In  practice  Method  I  is  the  most  convenient,  simple,  and  satisfactory. 
It  is  essential  that  a  blank  experiment  be   always  performed  to  ascertain 
whether  the  reagents  employed  are  free  from  ammonium  salts. 

/.  Chlorplatinic  acid  added  to  solutions  of  Animonimn  salts  precipitates  Ammo- 
nium Chlorplatinate. 

2NH4CI  +  HgPtCle  =  (NH4)2PtCl6  +  2HC1. 

Method.  Cause  a  drop  of  platinum  chloride  to  flow  into  the  drop  obtained 
after  the  manner  described  above.  In  a  few  moments  yellow  octahedral  crystals 
of  (NH4)2  PtClg  are  obtained.  These  crystals  resemble  those  of  the  corres- 
ponding potassium  compound  in  size,  form,  and  color.  The  reader  is  therefore 
referred  to  Potassium  Method  I,  for  a  discussion  of  the  appearance  of  the  crys- 
tals and  to  Fig.  29  for  a  representation  of  their  form. 

Remarks.  When  much  ammonium  is  present  there  is  apt  to  be  an  immediate 
precipitation  of  the  chlorplatinate  in  the  form  of  very  minute  or  skeleton  crystals. 
It  is  then  advisable  to  add  a  drop  of  water  and  recrystallize  by  heating.  If,  on 
the  other  hand,  the  amount  of  ammonium  is  small,  no  crystals  will  appear  until 
the  liquid  has  been  concentrated  by  gentle  heat. 

//.  The  addition  of  Magnesium  Acetate  and  Sodium  Phosphate  to  alkaline  solu- 
tions of  Ammonium  salts  gives  rise  to  the  formation  of  Magnesium  Ammotiiutn 
Phosphate. 

NH4CI  +  Mg(C2H302)2  +  HNa2P04  +  NaOH  = 

MgNH^PO^  .  6H2O  +  NaCl  +  2Na(C2H302)  +  H2O. 

Method.  To  the  test  drop  add  a  fragment  of  sodium  phosphate  and  a  very 
little  magnesium  acetate  ;  stir  thoroughly.  Beside  the  drop,  place  a  drop  of  a 
moderately  concentrated  solution  of  primary  sodium  carbonate  (HNaCOg)  or  of 
sodium  hydroxide.  Cause  the  drops  to  flow  together.  There  is  generally  an 
amorphous  precipitate  immediately  produced,  which  soon  partly  changes  into 
star-  and  X-like  crystallites,  see  Fig.  40.  A  little  farther  away,  roof-  and  envelope- 
like crystals  are  obtained. 

Remarks.  In  dilute  solutions  the  Xs  and  stars  are  generally  absent,  being 
replaced  by  prismatic  forms. 


and  Laboratory  Methods. 


1191 


Fig.  40. 


The  crystals  of  magnesium  ammonium  phos- 
phate belong  to  the  orthorhombic  system  and 
have  a  great  tendency  to  assume  hemihedral  and 
hemimorphic  and  skeleton  forms. 

Only  very  little  magnesium  acetate  should  be 
used  since  either  a  dense  amorphous  precipitate 
of  magnesium  phosphate  will  result,  or  if  the  con- 
ditions are  favorable  this  salt  will  itself  crystal- 
lize in  star-like  prism  aggregates. 

This  method  can  be  applied  directly  to  the 
solution  of  the  substance  without  the  necessity  of 
having  recourse  to  the  separation  of  the  am- 
monia. When  applied  directly  it  is  advisable  to 
substitute  sodium  hydroxide  for  the  carbonate. 
The  objection  to  this  procedure  is  that  many  elements  are  precipitated  as 
phosphates  in  alkaline  solution,  and  that  magnesium  hydroxide  almost  invariably 
separates  as  a  flocculent  mass. 

Exercises  for  Practice. 

Expel  the  ammonia  from  an  ammonium  salt  by  the  method  above  described, 
and  test  by  Method  I. 

Repeat,  and  test  the  drop  by  Method  II.  First  employing  primary  sodium 
carbonate,  then  using  sodium  hydroxide. 

Make  a  mixture  of  various  compounds  introducing  a  salt  of  ammonium. 
Test  directly  by  II.     Expel  the  ammonia  and  test  by  either  I  or  II. 

LITHIUM. 

The  element  lithium  can  be  considered  as  marking  the  transition  between 
the  alkalies  on  the  one  hand  and  the  alkaline  earths  on  the  other,  and  is  there- 
fore a  link  between  Groups  I  and  II.  Because  of  this — -its  peculiar  behavior — 
lithium  is  worthy  of  a  brief  consideration,  although  it  is  so  seldom  that  the 
chemist  is  required  to  test  for  its  presence  that  it  should  properly  not  be  con- 
sidered in  these  articles. 

The  solubility  of  its  sulphate  and  oxalate  excludes  its  appearance  in  testing 
for  calcium,  strontium  and  barium ;  while  its  precipitation  with  ammonium  (or 
potassium)  carbonate  and  sodium  phosphate  brings  it  into  close  relationship  with 
these  elements. 

With  almost  all  the  reagents  used  for  Group  I,  as  for  example,  chlorplatinic 
acid,  potassium  antimonate,  tartaric  acid,  ammonium  silicomolybdate,  bismuth 
thiosulphate,  etc.,  lithium  resembles  sodium  in  its  behavior;  yet  on  the  other 
hand  the  fact  that  in  rare  cases  phosphomolybdic  acid  may  cause  a  precipitate 
and  that  hexagons  are  obtained  with  bismuth  sulphate  brings  this  element  in 
close  analogy  to  potassium. 

Lastly,  like  magnesium,  lithium  forms  a  double  ammonium  phosphate  of  low- 
solubility.  Moreover,  this  salt  is  isomorphous  with  the  magnesium  ammonium 
phosphate.     In  this  respect  lithium  resembles  the  magnesium  group. 


1192  Journal  of  Applied  Microscopy 

The  microchemical  detection  of  lithium  is  satisfactory  only  when  this  element 
is  present  in  considerable  amount  and  in  quite  simple  mixtures. 

At  their  best  the  methods  are  apt  to  prove  unsatisfactory  and  require  not  a 
little  care  and  experience. 

Practically  only  three  reagents  are  available ;  these  are  : 
I.  Sodium  Phosphate. 
II.  Ammonium  Carbonate. 
III.  Ammonium  Fluoride. 
Only  I  and  II  will  be  described,  since  it  is  not  advisable  for  the  beginner  to 
make  use  of  fluorides,  because  of  the  great  danger  of  corrosion  of  the  objectives 
of  the  microscope. 

/.  Sodium  Phosphate  added  to  solutions  of  Lithium  salts  precipitates  Lithium 
Phosphate. 

SLiaSO^  +  2HNa2P04  =  ^^LigPO^    H2O  +  2Na2SO^  +  H2SO4. 

Method.  Allow  a  drop  of  a  solution  of  sodium  phosphate  to  flow  into  a  drop 

of  a  moderately  concentrated  solution  of  the  substance  to  be  tested.     Heat  the 

preparation  almost  to  boiling.     An  exceedingly  fine  crystalline  precipitate  is  at 

once  formed.     Examined  with  a  high  power,  this  precipitate  is  seen  to  consist  of 

strongly  refractive  lenticular  and  fusiform  bodies  either  singly  or  arranged  in 

cross-,  star-,  or  dagger-like  groups  which  are  very  characteristic  of   lithium  (see 

t^y^  Fig.  41).     These  crystals  extinguish  when  their  length 

J.    Vi^      xaS    '^S  lies  parallel  to  the  cross-hairs  of  the  polarizing  micro- 

C3      ^    ^  scope.     Globulites  are  also  formed  in  abundance,  which 

^  /}    '^<U    ~^_     '"y        when  examined  between  crossed  nicols  show  the  black 

n    i^^  '^A  cross  of  "  sphero-crystals." 

ri  rx    00    V  Reinarks.  The   reaction   should    be    performed    in 

\j  \j   60  neutral  (or  slightly  alkaline)  solution.     If  acid,  neutral- 

*^^^3*    //'    V^  ize  with  sodium  carbonate  or  sodium  hydroxide.    Since 

'  \-p■^'^,  --o.oim.^     i^  ^^  reaction  an  uncombined  acid  probably  results. 

Fig.  41.  it  is  advisable  to  have  a  small  amount  of  free  alkali 

present,  and  in  practice  it  is  found  that  faintly  alkaline  solutions  yield  the  best 

results.     An  excess  of  alkali  is  to  be  avoided. 

If  much  sulphuric  acid  is  present  it  is  advisable  to  heat  the  material  on  plati- 
num until  most  of  the  acid  has  been  driven  off,  after  which  the  residue  is  dis- 
solved in  water  and  the  solution  neutralized. 

Elements  of  the  calcium  group  can  be  advantageously  removed  by  treatment 
with  sulphuric  acid  or  ammonium  oxalate. 

If  much  magnesium  is  present,  globulites  seem  to  predominate. 
In  the  presence  of  ammonium  there  arises  the  possibility  of  the  formation  of 
a  double  phosphate  LiNH4P04«  GHgO  :  hence  if  any  ammonium  salts  have  been 
employed  in  preceding  operations,  they  should  be  removed  by  ignition  before 
testing  for  lithium. 

In  testing  mixtures,  if  in  doubt  as  to  the  nature  of  the  phosphate  obtained,^ 
draw  off  the  supernatant  liquor,  wash  the  precipitate,  dissolve  in  dilute  acid  and 


and  Laboratory  Methods. 


1193 


'vv.  iO.O\*v\*i\ 


add  ammonium  hydroxide.     If  the  precipitate  was  lithium  phosphate  no  turbidity 
should  result,  while  the  alkaline  earths  are  again  precipitated. 

//.  Addition  of  Atnmonium  Carbonate  to  neutral  solutions  of  salts  of  Lithium 
causes  the  separation  of  Lithium  Carbonate. 

Li2S04  +  (NH4)2C03  =  Li2C03  +  (NH  J2SO4. 

Method.  To  the  drop  of  the  neutral  solution,  which  should  not  be  too  dilute 
with  respect  to  lithium,  add  a  fragment  of  solid  ammonium  carbonate.  After  a 
short  time  there  will  appear  near  the  circumference  of  the  drop,  globulites, 
bunches  of  needles  and  thin,  more  or  less  irregular  plates  and  bristly  masses 
(Fig.  42).  The  general  appearance  of  these 
aggregates  will  vary  somewhat  with  the  con- 
centration of  the  test  drop. 

Remarks.  All  substances  forming  difficultly 
soluble  carbonates  interfere. 

The  reaction  requires  care  and  experience 
in  order  that  it  can  be  made  to  always  yield 
acceptable  results. 

Since  the  lithium  carbonate  separates  only 
after  some  little  time,  there  are  apt  to  appear, 
almost  simultaneously,  crystals  of  other  salts, 
particularly  if  the  test  drop  contains  sulphates. 
In  such  an  event  add  to  the  drop  a  little  dilute  alcohol ;  lithium  carbonate  will 
remain  undissolved  for  some  time,  while  the  sulphates  or  chlorides  of  the  other 
members  of  the  first  group  will  pass  into  solution. 

Exercises  for  Practice. 

Try  the  above  methods  for  lithium  first  on  a  simple  salt  of  this  element,  then 
on  mixtures  of  lithium  and  other  members  of  its  group,  and  lastly  try  mixture 
containing  ammonium  and  others  containing  calcium,  magnesium,  etc. 

EXAMINATION    OF   SUBSTANCES   CONTAINING   THE    ELEMENTS    OF 

GROUP  I. 

Ammonium  is  tested  for  in  a  portion  of  the  material  by  expelling  it  in  the 
manner  previously  given. 

The  material  to  be  tested  is  brought  into  solution  by  any  suitable  means,  not 
introducing  alkalies. 

If  soluble  in  HCl,  the  solution  of  the  chlorides  is  evaporated  to  dryness,  and, 
if  ammonium  salts  are  present,  the  residue  is  ignited  until  all  these  salts  have 
been  driven  off.  The  residue  is  extracted  with  absolute  alcohol  or  with  carefully 
purified  amyl  alcohol.  The  alcoholic  extract  is  evaporated  to  dryness  and  the 
residue  tested  for  the  members  of  Group  I. 

When  sulphuric  acid  has  been  employed,  or  if  the  substance  contains  sul- 
phates, it  is  first  necessary  to  convert  the  material  into  chlorides ;  this  is  accom- 
plished by  treating  with  barium  chloride  in  sufficient  amount  to  precipitate  all 
the  sulphuric  acid  and  removing  any  excess  of  barium  by  means  of  ammonium 


1194  Journal  of  Applied  Microscopy 

carbonate,  carefully  added.  The  turbid  liquid  is  then  either  filtered,  or,  what  is 
better,  whirled  in  the  centrifuge.  The  clear  liquor  is  evaporated  to  dryness,  the 
ammonium  salts  driven  off  and  the  residue  extracted  with  alcohol  or  amyl  alcohol. 

Or,  in  the  absence  of  sulphates,  another  method  is  open  which  avoids  the 
\ise  of  alcohol.  Treat  the  aqueous  solution  with  ammonium  hydroxide  and 
ammonium  oxalate,  then  add  sufficient  ammonium  carbonate  to  precipitate  the 
magnesium  ;  separate  the  precipitate  by  filtration  or  by  means  of  the  centrifuge. 
Evaporate  the  clear  liquid  and  ignite  the  residue.  Treat  with  HCl,  evaporate 
to  dryness,  and  test  the  residue  as  given  below.  This  method  is  open  to  a 
number  of  objections  and  is  somewhat  limited  in  its  application. 

Test  a  portion  of  the  material  with  potassium  chlorplatinate  or  ammonium 
silicomolybdate  for  Rb  and  Cs. 

Another  portion  is  treated  with  platinum  chloride  for  K. 

Sodium  is  tested  for  with  uranyl  acetate  or,  if  thought  to  be  in  only  small 
amount,  with  uranyl  acetate  and  magnesium  acetate.  If  much  K  has  previously 
been  found,  either  separate  this  element  by  the  perchloric  acid  method,  or  test 
for  Na  at  once  with  potassium  antimonate  or  bismuth  sulphate. 

There  now  remains  Li  to  be  searched  for.     This  can  be  done  by  any  one  of 
the  three  methods  mentioned  under  the  head  of  this  element. 
Cornell  University,  Chemical  Laboratory.  E.  M.  ChamoT. 


Staining  Sections  for  Class  Work. 

Being  under  the  necessity  of  staining  large  numbers  of  sections  of  tissues 
sectioned  in  both  paraffin  and  in  celloidin,  for  class  work,  we  have  adopted  the 
following  methods  for  shortening  the  time  and  facilitating  the  technique. 

The  paraffin  sections  being  mounted  on  cover-slips,  and  the  paraffin  removed 
by  the  usual  method,  they  are  placed  in  rows  on  a  corrugated  glass  disc  of  a 
staining  dish,  the  disc  being  provided  with  handles  of  wire,  and  resting  in  a 
dish  of  xylol  deep  enough  to  cover  the  sections.  Upon  this  disc  or  tray  the 
sections  can  be  quickly  transferred  from  one  reagent  to  another,  if  necessary, 
allowing  the  disc  to  drain  upon  pieces  of  filter  paper  between  the  different 
reagents.  In  this  way  thirty  or  thirty-five  sections  can  be  stained  in  a  very  few 
minutes  on  one  staining  disc. 

For  celloidin  sections,  the  reagents  being  in  small,  deep  dishes,  the  sections 
are  placed  on  a  piece  of  copper  or  brass  wire  gauze,  folded  into  the  form  of  a 
cup,  and  resting  in  a  dish  of  alcohol  or  water.  By  means  of  this  porous  cup  the 
sections  can  be  quickly  transferred  from  one  dish  of  reagent  to  another  and 
easily  drained  ;  the  number  of  sections  that  can  be  stained  at  once  being 
indefinitely  large.  Newton  Evans,  M.  D. 

American  Medical  Missionary  College. 


and  Laboratory  Methods.  1195 


Home  Made  Wall  Charts. 

I  can  vouch  for  the  value  of  Professor  Heald's  method  of  making  wall  charts^ 
described  in  the  November  Journal,  as  many  years  ago,  when  connected  with 
the  State  College  of  Iowa,  I  made  some  very  serviceable  charts  in  this  way.  I 
soon  found  that  I  could  use  a  camel's  hair  brush  for  inking  the  pencil  lines. 
After  a  little  practice  one  learns  just  how  rapidly  the  brush  must  be  drawn  over 
the  surface  to  produce  the  right  kind  of  a  line,  and  to  avoid  spreading  and 
blotting.  I  still  have  a  few  of  these  old  muslin  charts,  which  are  as  good  as 
ever,  after  at  least  twenty  years  of  service.  One  great  advantage  which  such 
charts  have  over  all  others  is  that  they  may  be  folded  into  small,  fiat  parcels,^ 
and  tucked  away  in  one's  traveling  bag,  and  not  be  any  the  worse  for  it,  after  an 
extended  lecture  trip. 

Another  method  of  making  charts  is  one  which  grew  out  of  the  foregoing, 
and  which  I  prefer  for  charts  for  hanging  on  the  walls  of  the  lecture  room  or 
laboratory,  although  less  convenient  for  carrying  about  the  country.  I  buy  a 
roll  of  "opaque  "  curtain  cloth,  white  or  of  a  light  shade,  and  about  100  cm,  in 
width.  This  is  cut  into  sections  of  the  desired  length,  say  1}4  to  2  meters, 
and  on  these  the  desired  figures  are  drawn.  I  buy  one  pound  boxes  of  paints, 
ground  and  mixed,  ready  for  use.  In  order  to  hasten  the  drying  of  the  oily 
paint,  I  take  out  a  little  from  the  box,  allowing  the  surplus  oil  to  drain  off,  and 
then  mix  it  with  the  proper  amount  of  spirits  of  turpentine  to  make  it  flow 
readily  from  the  brush.  The  figures  having  been  traced  in  lead  pencil,  a  good 
camel's  hair  brush  is  used  in  applying  the  paint.  Since  the  curtain  stuff  is  a 
kind  of  "  filled  "  canvas,  its  surface  takes  the  paint  very  easily,  and  there  is  no 
danger  of  its  spreading.  When  the  material  is  white,  colored  paints  may  be 
used  to  good  advantage.  I  have  been  able  to  get  good  effects  from  the  use  of 
green,  yellow,  and  brown  paints  of  the  quality  found  in  the  pound  boxes  men- 
tioned above.  Other  colors,  especially  the  reds,  and  the  delicate  shades  of  pink, 
lavender,  gray,  etc.,  are  not  as  satisfactory  with  these  coarser  paints  as  with  the 
"  tube  "  paints,  which  I  have  used  for  finer  work,  as  in  cytological  charts.  For 
the  charts  made  in  black  throughout,  any  good  lampblack  paint  will  prove 
satisfactory. 

In  mounting  the  charts,  I  have  found  that  the  best  way  is  to  use  pairs  of  pine 
or  whitewood  "  half  round  "  strips  of  the  proper  length,  clapping  the  end  of  the 
chart  between  the  two,  and  fastening  them  together  with  small  wire  nails.  They 
thus  form  a  cylindrical  roller  at  each  end,  and  the  cloth  is  fastened  much  more 
securely  than  when  a  solid  roller  is  used. 

I  have  a  hundred  or  more  charts  in   the  Botanical  department  of  the  Uni- 
versity of  Nebraska,  made  in  this  way,  and  they  have  been  found  very  satisfactory, 
while  the  cost  for  material  has  been  little. 
University  of  Nebraska.  CHARLES  E.  BeSSEY. 


1196  Journal  of  Applied  Microscopy 


Flattening  and  Fixing  Paraffin  Sections  on  Slide. 

One  of  the  difficulties  in  mounting  paraffin  sections  in  series  is  the  loss  due 
to  imperfect  fixing  on  the  slide. 

The  following  methods  are  generally  recommended  :  Water  (Lee's  Vade- 
Mecum,  sec.  182,  5th  ed.);  alcohol  (70  per  cent,  alcohol  is  used  instead  of  water. 
Method  described  in  above  reference);  Mayer's  albiimi?i  (Lee's  Vade-Mecum, 
sec.  183,  5th  ed.).  Each  of  these  methods  is  open  to  some  objections,  either  on 
account  of  extreme  care  necessary  for  good  results  or  clouding  of  sections  in 
staining. 

Of  these,  the  alcohol  method  seems  to  be  the  most  satisfactory,  as  it  does  not 
require  that  the  slides  be  absolutely  clean,  nor  are  the  sections  clouded  in  stain- 
ing as  sometimes  occurs  in  the  albumin  method.  The  improvement  on  the 
alcohol  method  suggested  by  Eisen  (Zeit.  f.  wiss.  Micros.  Bd.  xvi)  makes  it  as 
certain  as  the  albumin  method,  without  its  objectionable  features. 

The  essential  steps  in  the  process  are  as  follows  : 

a.  Flood  the  slide  with  70  per  cent,  to  85  per  cent,  alcohol.     Arrange  sec- 

tions in  order.  Hold  slide  a  few  inches  above  small  flame  until  sections 
are  flattened. 

b.  Drain    off    surplus    alcohol     (use    filter-paper    or    cloth).       Rearrange 

sections  in  desired  positions. 

c.  Cut  out  two  pieces  of  smooth  blotting  paper  same  size  as  slide.     Wet  one 

in  same  strength  alcohol  as  used  in  {a^,  and  place  over  sections. 
Over  this  put  the  other  piece  dry.  Pass  small  rubber  roller  (such  as 
used  by  photographers),  firmly  over  the  dry  blotting  paper  two  or 
three  times.  Instead  of  using  the  roller,  any  weight  with  smooth  sur- 
face may  be  pressed  against  the  blotting  paper.  The  object  of  this 
step  is  to  flatten  the  sections  completely,  so  that  every  part  of  the 
section  will  come  in  contact  with  the  slide. 

d.  Remove  any  lint  which  adheres  to  the  slide  and  dry  in  a  place  protected 

from   dust.     At   the  ordinary  temperature   of  the  room,  two   or  three 
hours   are   necessary  for  complete   drying.     The   process  may  be  has- 
tened by  keeping  the  sections   at  a  temperature  a  few  degrees  below 
the  melting  point  of  the  paraffin  (below  40°  C). 
If  this  method  has  been   carried   out   carefully,   the   sections  may  be  taken 
through  as  many  stains  or  reagents  as  desired,  or  left   indefinitely  in   any  solu- 
tion which  will  not  act  chemically  on  them.  B.  M.  Davis. 
Biological  Laboratory,  State  Normal,  Los  Angeles,  Cal. 


A  new  building  is  being  erected,  at  a  cost  of  $125,000,  for  the  Medical 
Department  of  Cornell  University.  It  will  be  the  finest  building  on  the  Cornell 
campus,  and  will  offer  facilities  for  scientific  and  practical  study  which  are  not 
excelled  by  any  institution  in  the  world. 


and  Laboratory  Methods.  119*^ 


A  Ventilated  Dish  for  Bacteria  Cultures. 

When  Koch's  original  form  of   plate  for  bacteria  culture  was  abandoned  for 

the  more  convenient   Petri  dish,  a  step  was   undoubtedly    taken  in    the    right 

direction  ;  yet  in  one  respect  it  was  a  step  backward.     Koch's  plates  were  placed 

for  incubation  in  a  large   air-tight  receptacle,  as  a  bell-jar,  which  contained  wet 

filter  paper.     The  object  of  this  jar  was  to  prevent  the  gelatin   from  drying  up 

as  it  would  do  if  exposed  to  the  atmosphere  of  the  ordinary  incubator.     In  the 

Petri  dish  drying  was  prevented  by  making  the  cover  fit  the  bottom  plate  tightly 

— at  least  such  was  the  intention.     As  a  matter  of  fact,  however,  the  dishes  are 

seldom  air-tight  because  the  bottom  plates  arid  covers  become  mismated  in  the 

laboratory.     Consequently  when  Petri  dishes  are  used  there  is  almost  always  a 

slight  drying  of  the  gelatin.     The  loss   of  water  by  evaporation   in  the  ordinary 

Petri  dish  may  be  as  great  as  15  per  cent,  in  72  hours  at  "20°,  but  it  is  ordinarily 

about  5    per    cent.     Although   this    evaporation    is    comparatively  small,  it    is 

sufficient  to  cause  a  thickening  of  the  gelatin  at  the  surface,  and  this  thin  film 

tends  to  exclude  oxygen  from  the  medium  and  thus  retard  or  prevent  the  growth 

of  aerobic  bacteria.     In  a  former  publication*   the   writer   has   shown  how  the 

amount  of  moisture  in  the  atmosphere  of  the   incubator  affects  the  number  of 

bacteria  that  will  develop  from  a  sample  of  water.     The  results  there  set  forth 

were  summarized  in  the  following  table  : 

Per  Cent,  which  the  Number 

of  Bacteria  that  Developed 

Relative    Humidity    of    the  in  the  Incubator  was  of  the 

Atmosphere  of  the  Incuba-  Number  that  Developed  in 

tor  in  per  cent,  of  Saturation.  a  Moist  Chamber. 

GO  75 

75  82 

95  98 

98  97 

100  100 

It  was  also  shown  that  air-tight  Petri  dishes  are  unfavorable  for  the  growth 
of  aerobic  bacteria,  because  of  the  partial  exhaustion  of  the  oxygen  from  the 
somewhat  limited  air  space  and  the  collection  there  of  gaseous  products  of 
growth.  For  example,  the  air  in  five  sealed  Petri  dishes  was  collected  and 
analyzed  after  bacteria  had  been  allowed  to  develop  in  them  for  72  hours,  and 
was  found  to  contain  only  5  per  cent,  of  oxygen  and  5  per  cent,  of  CO.^  ; 
whereas  in  ordinary  Petri  dishes  with  ill-fitting  covers  the  percentage  of  oxygen 
was  15  per  cent,  and  of  carbonic  acid  2  per  cent. 

From  these  two  facts,  that  an  air-tight  Petri  dish  gave  too  low  results  on 
account  of  exhaustion  of  oxygen  and  that  an  ordinary  Petri  dish  gave  too  low 
results  on  account  of  evaporation  of  moisture,  it  was  argued  that  the  best  condi- 
tions would  be  obtained  by  cultivating  bacteria  in  a  ventilated  dish  placed  in  an 


*On  the  Necessity  of  Cultivating  Water  Bacteria  in  an  Atmosphere  Saturated  with  Moisture. 
Technology  Quarterly,  Dec.   1899. 


1198  Journal  of  Applied  Microscopy 

incubator  of  which  the  atmosphere  was  saturated  with  moisture.  Thus  drying 
of  the  gelatin  was  prevented  and  a  sufficient  amount  of  oxygen  was  provided. 
Experiments  showed  that  the  results  obtained  from  these  conditions  warranted 
the  trouble  necessary  to  provide  them.  The  atmosphere  of  the  incubator  may 
be  easily  kept  nearly  saturated  by  shallow  pans  of  water  placed  beneath  the 
shelves,  and  ventilation  of  the  dishes  may  be  accomplished  in  a  number  of  ways. 
I  have  recently  had  made  a  very  convenient  form  of  ventilated  dish,  which  is 
shown  in  the  accompanying  diagram.  The  cover  is  supported  about  'J  mm. 
above  the   lower  plate  by  means  of  three  projections  of  glass,  which  are  merely 


^£ 


•y 


indentations  in  the  cover,  obtained  by  heating  the  edge  and  pressing  in  the 
softened  glass  with  a  sharp  point.  The  sides  of  the  cover  are  made  deeper 
than  in  the  Petri  dish  by  an  amount  about  equal  to  that  which  the  cover  is 
raised  above  the  dish.  With  the  cover  thus  elevated  there  is  abundant  op- 
portunity for  a  free  circulation  of  air,  as  indicated  by  the  arrows.  Ordinary 
Petri  dishes  may  be  thus  ventilated,  but  unless  the  work  is  done  by  a  skilled 
glass-blower  the  breakage  is  liable  to  be  great.  Furthermore,  the  cover  of  the 
ordinary  Petri  dish  is  too  shallow. 

If  the  ventilated  dish  is  desirable  for  the  cultivation  of  aerobic  bacteria,  it  is 
even  more  necessary  for  the  cultivation  of  anaerobic  forms.  When  these  ventilated 
dishes  are  placed  in  a  jar,  like  the  Novy  jar,  for  example,  the  air  in  them  may 
be  easily  replaced  with  hydrogen,  while  with  the  ordinary  Petri  dishes  this  is 
sometimes  a  difficult  matter.  George  C.  Whipple. 

Mt.  Prospect  Laboratory,  Brooklyn,  N.  Y. 


As  a  result  of  the  investigations  on  malaria  carried  on  in  Italy  by  Professors 
Celli  and  Grassi,  the  Italian  government  will  soon  consider  the  appropriation  of 
a  sum  of  money  to  continue  the  work  already  begun.  Employers  of  labor  in 
malarial  districts  will  be  compelled  to  provide  the  proper  precautions  against 
infection,  and  also  supply  medical  aid  to  laborers  who  contract  the  disease,  at  the 
same  time  providing  a  fixed  amount  for  the  support  of  their  families  when  the 
employer  fails  to  comply  with  the  requirements  of  the  bill. 


and  Laboratory  Methods.  1199 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 
THE  PROCESSES  OF  PHOTO-MICROGRAPHY. 

In  a  previous  paper,  I  described  the  apparatus  employed  in  high-power  photo- 
micrography and  outlined  the  method  used  for  the  projection  of  the  luminous 
image  upon  the  focusing  screen.  To  complete  this  description  and  to  reduce  the 
whole  matter  to  a  concrete  example,  I  will  here  explain  the  further  steps  involved 
in  the  production  of  a  negative,  giving  actual  details  of  exposure  time,  speed  of 
plate,  etc. 

Before  proceeding  to  a  discussion  of  processes,  however,  it  will  be  well  to 
consider  the  materials  which  are  to  be  employed.  Chief  among  these  is  the  sen- 
sitive plate  designed  to  receive  the  light  impressions  and  to  make  permanent 
record  of  them.  Formerly  the  old  collodion  wet  plate  was  used  for  this  purpose, 
and  many  of  the  best  photo-micrographs  ever  made  are  the  productions  of  the 
earlier  workers  who  employed  this  process.  Workers  now-a-days,  though,  do  not 
resort  to  the  tedious  methods  the  use  of  the  wet  plate  involves,  but  choose  the 
more  convenient  and  rapid  gelatin  dry  plate. 

Of  dry  plates,  there  are  any  number  on  the  market,  and  most  of  them  are 
good  for  ordinary  photography ;  but  in  photo-micrographic  work  the  conditions 
are  different  from  those  which  prevail  where  the  image  is  smaller  than  the  object 
and  the  light  is  plentiful.  The  strong  diffusion  of  light  involved  in  the  produc- 
tion of  an  image  1000  diameters  larger  than  the  object  itself  renders  it  expedient 
to  use  plates  that  will  record  strongly  all  dififerences  of  lighting  and  thereby  pro- 
duce negatives  with  good  "contrast."  In  this  respect  the  slower  plates,  those 
of  lower  sensitometer  numbers,  are  the  best ;  and  if  the  operator  has  any  one 
with  which  he  is  familiar,  he  will  do  well  to  make  use  of  it  when  undertaking  the 
unfamiliar  work  of  photo-micrography. 

To  such  as  are  not  adept  in  the  manipulation  of  any  particular  plate,  I  would 
strongly  recommend  the  orthochromatic  "  Carbutt  Process  Plate."  After  an 
extended  trial  of  many  plates,  I  find  this  one  in  a  large  degree  satisfactory.  It 
produces  negatives  with  clear,  sharp  details  and  abundant  contrast.  The 
film  is  hard  and  firm,  washes  readily,  and  dries  quickly.  Beginners  will  make  no 
mistake,  I  am  sure,  in  starting  with  this  plate.  Since  so  many  sections  are 
stained  by  some  one  or  other  of  the  blue  dyes,  it  is  best  to  make  use  of  the  ortho- 
chromatic  plates,  as  they  give  better  balanced  negatives.  The  "  Process  "  plate 
may  be  obtained  in  this  form  and  I  prefer  it  to  the  plain  form. 

Next  in  importance  to  the  plate  is  the  developer  with  which  the  latent  image 
upon  it  is  made  manifest.  Here,  as  in  the  former  case,  I  have  nothing  to  rec- 
ommend to  those  who  are  well  acquainted  with  the  action  of  any  good  develop- 
ing fluid.  The  agent  itself  is  really  not  of  so  much  importance  as  is  the  knowl- 
edge of  how  to  use  it.  By  this,  I  do  not  mean  that  there  is  no  choice,  but  it  is 
better   to    manipulate  a  poor  developer  well  than  a  good  one  poorly. 

It  is  recognized  that  certain  reducing  agents,  e.  g.,  eikonogen,  produce  "  thin  " 


1200  Journal  of  Applied  Microscopy 

negatives  with  much  detail ;  while  others,  such  as  hydrochinone,  afford  density. 
Contrast  being  desirable,  it  is  well  to  choose  a  developer  containing  a  reducing 
agent  that  will  best  produce  it.  In  doing  this,  however,  it  is  often  advantageous 
to  combine  reducing  agents  with  opposing  characters  in  order  to  secure  a  nega- 
tive well  balanced  in  detail  and  density. 

If  asked  to  recommend  developers  of  this  character  I  should  suggest  three 
that  do  the  work  well  and,  at  the  same  time,  differ  otherwise  so  as  to  make  them 
applicable  under  different  conditions. 

The  simplest  of  these  is  known  by  the  trade  name  "  Rodinal."  To  prepare 
it  for  use,  it  is  only  necessary  to  dilute  it  with  twenty  parts  of  water.  This,  with 
its  good  keeping  qualities,  makes  it  convenient  for  those  who  have  only  an  occa- 
sional negative  to  develop.  Persons  unfamiliar  with  its  use  will  perhaps  be 
startled  at  the  suddeness  with  which  the  image  flashes  into  view  under  its  action, 
and  will  consider  a  normally  exposed  negative  overtimed.  It  is,  further,  some- 
what deceptive  in  the  relative  amount  of  density  apparent  before  and  after  fixing, 
since  the  negatives  produced  by  its  use  lose  more  under  the  action  of  the  hypo 
than  do  almost  any  others.  As  it  does  not  produce  chemical  fog,  even  after  long 
action,  it  is  well  to  let  it  operate  until  an  apparent  excessive  density  is  produced. 
Among  the  class  of  one-solution  developers  that  may  be  prepared  by  the 
operator  and  kept  for  some  time,  I  like  the  metol-hydrochinone  mixture  prepared 
after  the  following  formula  : 

Water,  distilled  ....         500    c.  c. 

Metol  3    gm. 

Hydrochinone     .....  .5   " 

Sodium  Sulphite     .  .         .         .  .        18      " 

Sodium  Carbonate       .  .  .  .  14      " 

Mix  in  the  order  given,  and  when  wanted  for  use  dilute  with  an  equal  quantity 
of  water. 

As  a  type  of  the  two-solution  developers,  I  would  recommend  the  one  given 
by  Carbutt  for  use  with  his  plates.     This  is  prepared  as  follows  : 
Solution  1  : 

Water,  distilled  ....         600  c.  c. 

Sulphite  of  soda     .....     120  gm. 

Eikonogen         .....  '2'2    " 

Hydrochinone  .  .  .  .  .       10    " 

Add  water  to  make    ....  960  c.  c. 

Solution  2 : 

Water,  distilled         ....  600  c.  c. 

Carbonate  of  potash     ....  60  gm. 

Carbonate  of  soda     .         .         .         .         .     60    " 
Add  water  to  make       ....         060  c.  c. 
To  use,  mix  one  part  each  of  1  and  "2  with  four  parts  of  water. 
Greater  contrast  can   be  obtained  from   any   developer  by  decreasing  the 
amount  of  alkaline  solution  or  by  the  addition  of  a  few  drops  of  10  per  cent, 
potassium  bromide  solution. 


and  Laboratory  Methods.  1201 

The  fixing  bath,  while  by  no  means  as  important  as  the  developing  solution, 
may  have  its  value  underestimated.  It  is  not  uncommon  to  find  operators  who 
mix  up  water  and  hypo  in  almost  any  proportion  and,  without  filtering  the  solu- 
tion, use  it  until  it  becomes  so  discolored  as  to  afi^ect  the  film  of  the  plate  inju- 
riously. A  little  care  will  greatly  economise  the  use  of  the  hypo  and  at  the  same 
time  produce  much  better  negatives.  I  find  the  following  bath  entirely  satisfac- 
tory, cheap  and  convenient:  Prepare  and  filter  saturated  aqueous  solutions  of 
hypo  and  boric  acid.  Mix  one  part  of  the  hypo  with  three  of  the  boric  acid 
solution.  This  bath  will  keep  until  the  hypo  is  exhausted  without  discoloring 
and,  being  acid,  hardens  the  film. 

Plate  and  developer  are  important  agents  in  the  production  of  a  good  nega- 
tive, but,  without  the  proper  adjustment  of  light  effect  to  the  speed  of  the  plate, 
they  are  worthless.  The  exposure  must  be  judiciously  regulated  so  that  the 
darkest  parts  of  the  object  will  not  be  allowed  to  produce  any  effect  upon  the 
sensitive  film,  while,  at  the  same  time,  the  light  must  be  allowed  to  act  long 
enough  to  be  effective,  in  various  degrees,  over  the  lighter  portions  of  the  object. 
General  instructions  regarding  this  part  of  the  work  are  of  little  value,  so  I  will 
outline  the  actual  conditions  under  which  negatives  have  been  produced. 

Source  of  illumination- — the  crater  of  an  arc  light  placed  at  a  distance  of 
two  feet  from  the  object  and  having  interposed  between  it  and  the  condenser  a 
ground  glass  disc.  This  disc  stands  about  six  inches  from  the  crater  and  its 
matt  surface  is  made  somewhat  transparent  by  rubbing  with  glycerin. 

Condenser — a  parachromatic  of  1.30  N.  A.  in  homogeneous  contact  with 
the  lower  portion  of  the  slide.  The  diaphragm  registers  a  numerical  aperture 
of  .5. 

Object — a  section  of  embryonic  tissue  6^^  micra  thick,  stained  with  iron- 
haematoxylin  and  mounted  in  balsam. 

Objective — a  '2  mm.  homogeneous  immersion  apochromatic  of  1.30  N.  A. 

Ocular — a  No.  2  projection. 

Magnification — 1000  diameters. 

Plate — a  "  Carbutt  Ortho.  Process." 

Under  these  circumstances,  an  exposure  of  20-30  seconds  is  sufficient. 

Two  things  are  to  be  guarded  against  during  the  time  of  exposure  ;  viz., 
flickering  of  the  light  and  vibration  of  the  microscope  or  camera.  Either  of 
these  untoward  circumstances  will  ruin  what  might  otherwise  prove  to  be  a  good 
negative. 

With  the  proper  exposure  and  by  the  use  of  the  metol-hydrochinone  devel- 
oper, the  image  will  begin  to  appear  upon  the  plate  in  about  30  seconds,  and  develop- 
ment will  be  complete  in  about  five  minutes.  In  the  "  Process  "  plate,  very  little 
of  the  image  will  show  upon  the  reverse,  or  glass,  side  of  the  plate.  The  prog- 
ress of  the  development  is  best  observed  by  examining  the  image  under  trans- 
mitted light.  Somewhat  greater  density  than  is  finally  desired  should  appear, 
since  some  of  it  is  lost  in  the  hypo. 

Fix  until  all  the  unreduced  silver  salts  are  removed,  and  the  shadows  are 
clear.  To  be  sure  of  this,  allow  the  hypo  to  act  some  minutes  after  the  last  trace 
of  milkiness  has  disappeared  from  the  film. 


1-0'J  Journal  of  Applied  Microscopy 

Wash  for  an  hour  in  running  water  and  dry.  With  this  plate,  heat  may  be 
used  to  hasten  the  evaporation  of  the  water  from  the  gelatin. 

The  time  limits  here  set  are.  of  course,  operative  only  under  the  conditions 
given.  Increased  light,  either  from  greater  transparencv  of  the  ground  glass. 
enlarged  aperture  of  substage  diaphragm,  or  decreased  magnification  will  mate- 
rially shorten  the  time  of  exposure.  It  is  possible,  also  to  decrease  considerably 
the  exposure  by  employing  a  rapid  plate  instead  of  the  slow  one  recommended. 
Some  circumstances  may  justify  this  use,  but  ordinarily,  the  slow  emulsion  will 
give  the  better  results. 

Finally,  one  other  suggestion  may  prove  of  advantage.  Excessive  contrasts 
may  exist  in  the  object  itself,  and  it  is  desirable  to  reduce  these.  This  may  be 
accomplished  by  an  over  exposure  producing  a  flat  negative  which  may  be  made 
printable  by  subsequent  intensification.  This  will  give  general  detail  even  when 
the  object  is  dark.  Variations  in  density  and  detail,  within  more  or  less  narrow 
limits,  may  also  be  secured  by  choosing  printing  papers  of  different  kinds ;  this 
choice  is  particularly  important  when  prints  are  made  for  micro-mechanical  repro- 
duction, since  the  balance  of  light  and  shade  is  not  equally  preserved  by  the 
various  papers  under  these  circumstances.  C.  E.  McClung. 

University  of  Kansas. 

A  LABORATORY  CAMERA  STAND. 

Photographic  reproductions  ot  material  for  illustrating  Experiment  Station 
and  other  literature  have  become  important  aids  in  technical  work  and  have  been 
used  with  more  or  less  success — frequently  the  latter.  The  ditticulty  does  not  lie 
in  the  photographic  processes,  but  rather  in  carrying  them  out.  There  are  cer- 
tain lines  of  work  in  which  the  photographic  processes  are  not  easily  employed, 
such  as  illustrating  microscopic  insects  and  fungi.  Even  this  field  mav  be  occu- 
pied in  time.  As  long  as  botanists  and  entomologists  depended  upon  the  por- 
trait photographer  to  prepare  the  negatives,  the  work  was  usuallv  most  disap- 
pointing :  but  with  the  advent  of  plant  and  insect  photographers,  some  most 
excellent  and  pleasing  results  have  been  obtained. 

In  his  little  booklet  on  photographing  trees  and  rlowers.  Mr.  j.  Horace 
McFarland  has  shown  some  things  that  mav  be  done  with  simple  apparatus. 
Before  seeing  this  pamphlet  an  order  was  let  for  a  laboratorv  stand  that  differs 
greatly  from  the  one  illustrated  by  Mr.  McFarland.  and  also  from  the  one  used 
in  the  botanical  laborator}-  of  the  Florida  Agricultural  College.  The  one  at 
Clemson  College  is  used  for  photographing  diseased  plants,  individual  plants, 
and  similar  m.aterial.  with  no  idea  of  using  it  for  illustrating  bouquets  or  pot 
plants. 

The  source  of  light  is  from  a  high  window  to  the  north,  making  the  illumi- 
nation like  a  skylight. 

The  ST.\Nn. — The  frame  is  made  of  one-inch  angle-iron  and  holds  the 
camera  post  and  a  ."iO  x  oO-inch  glass  plate.  At  the  lower  end  of  the  post  is  a 
mirror,  attached  by  mechanical  contrivances  in  such  manner  as  to  allow  it  to  be 
raised  or  low^ered ;  tilted  forward,  backward,  or  sidewise;  brought  nearer  to  the 
object  or  drawn   back  from  it ;  or   so  adjusted   as  to  throw  the  reflection  oft" 


and   Laboratory  Methods. 


20:; 


entirely.  With  this  arrangement  any  portion 
of  the  field  rnay  be  illuminated,  or  the  illumi- 
nation rnay  be  dispensed   with  entirely. 

Above  the  mirror  attachment  is  the  camera 
attachment,  which  allows  the  camera  to  be 
raised  and  lowered  to  any  point  on  the  po.st 
and  securely  clamped.  'J'he  front  ?.»oard  hav- 
ing been  brought  into  position  and  the  ground 
glass  adjusted,  the  whole  camera  may  be 
lowered  or  raised,  or  racked  entirely  out  of  the 
way  until  wanted. 

Accessories.  Besides  the  camera  and 
stand  proper  there  are  several  accessories  that 
are  excellent  time  savers. 

(\)  A  Four-Foot  Rule,  seen  in  the  figure 
leaning  against  the  camera  frame.  This  rule 
has  marked  upon  it  the  exact  distance  from 
the  glass  plate  to  the  front  board  and  to  the 
ground  glass  for  all  combinations  needed, 
'i'hus,  by  using  a  Zeiss  8x10  series  V  lens, 
to  enlarge  the  object  two  diameters  the  front 
board  should  be  lOj^ 
inches  from  the  glass 
plate  and  the  ground 
glass  30 j^  inches  ; 
for  making  a  'yi',  nat- 
ural size  negative  the 
front  board  should 
be  18  inches  and  the 
ground  glass  29  inches,  and  so  on,  for  other  enlargements  and  reductions.  The 
advantage  of  this  rule  is  that  the  camera  is  adjusted  quickly  and  accurately  with- 
out experimenting.  When  the  specimen  is  in  place  the  camera  may  be  racked 
to  such  position  as  to  bring  the  highest  part  of  the  object  Tthat  nearest  the  lens) 
into  sharpest  focus.  Those  who  do  not  use  a  rule  of  this  kind  will  find  it  a  sur- 
prising convenience.  If  the  stand  is  of  a  different  design  it  is  sometimes  prac- 
ticable to  mark  these  distances  upon  the  post  to  serve  the  same  purpose. 

(2)  A  Ruled  Card  is  prepared  from  a  piece  of  heavy  cardboard  30  x  30 
inches,  the  size  of  the  glass  plate,  and  ruled  so  as  to  have  areas  corresponding 
to  multiples  of  different  sized  plates.  Where  a  large  number  of  plates  are  used, 
the  cost  becomes  an  item  worth  considering,  and  there  is  no  occasion  for  using 
an  8x  10  plate  if  G^/^  x  8j^  or  .5  x7  will  answer.  The  areas  for  the  4x5  and 
8  X  10  plates  are  30  x  24,  2.5  x  20,  20  x  16,  15  xl2,  10  x  8,  5  x  4,  and  2i^  x  2.  The 
reverse  side  of  the  card  is  ruled  for  the  5x7  and  6|4  x  8^  plates.  These  two 
sizes  do  not  coincide  as  is  the  case  with  the  4x5  and  8  x  10,  so  a  dotted  line  is 
used  for  the  6^/^  x  8^  fields  and  a  line  for  the  5  x  7. 

This  ruled  card  serves  two  purposes  :  (1)  The  object  is  placed  upon  it  to  ascer- 


1204  Journal  of  Applied  Microscopy 

tain  what  sized  plate  will  cover  it  with  the  least  waste.  It  also  shows  at  a  glance 
how  much  the  object  will  be  reduced  or  enlarged  for  that  particular  plate,  and 
by  reading  the  rule  the  camera  may  be  adjusted  at  once.  (2)  The  card  being 
placed  imder  the  glass  plate  shows  the  exact  field  that  the  object  should  occupy 
to  be  included  on  the  ground  glass. 

(3)  A  Glass  Plate.  A  method  for  posing  insects,  and  one  equally  serviceable 
for  arranging  flowers,  is  to  secure  a  clean  glass  plate,  such  as  the  glass  from  a 
photographic  plate  or  other  equally  good  sheet  of  glass  of  the  desired  size.  The 
object  is  arranged  upon  this  glass  and  when  properly  posed  is  slipped  into  posi- 
tion under  the  lens.  The  glass  being  clean,  the  plate  of  the  stand  likewise,  and 
both  free  from  defects,  no  image  of  either  will  be  formed  on  the  sensitive  plate. 
This  method  was  developed  by  Prof.  A.  L.  Quaintance  while  associated  with  the 
writer. 

(4)  A  Paper  Rule  such  as  is  sold  by  the  Cambridge  Botanical  Supply  Company, 
with  sharp  lines  upon  clear  white  paper,  a  little  heavier  than  heavy  herbarium 
paper,  makes  a  convenient  object  to  focus  upon.  Such  a  rule  is  so  light  that  it 
may  be  placed  upon  the  object  to  be  photographed  for  the  purpose  of  verifying 
the  focus  before  inserting  the  plate-holder.  In  many  cases  the  rule  may  be  left 
in  an  appropriate  portion  of  the  field  to  serve  as  an  index  of  the  enlargement  or 
reduction.  In  the  absence  of  a  light  paper  rule,  a  visiting  card,  as  Mr.  McFar- 
land  suggests,  makes  a  convenient  object  to  prove  the  focus.  A  wooden  rule  is 
anything  but  a  desirable  substitute. 

In  conclusion  I  would  recommend  heavier  angle-iron  for  the  frame,  say  about 
2^  or  3  inches.  The  one-inch  makes  a  frame  appear  light  and  the  post  not  so 
firm  as  might  be  desired.     In  practice  it  has  given  no  trouble. 

A  suitable  background  is  supplied  in  the  same  way  as  suggested  in  the  book- 
let referred  to  before.  P.  H.  Rolfs,  Botanist. 
Clemson  Agricultural  College,  Clemson  College,  S.  C. 


Received  for  Notice. 

Modern  Photography  in  Theory  and   Practice.      '^'^is  book  is,  as   its   title  page  states, 

H.  S.  Abbott,  Chicago;  Geo.  K.  Hazlett  &      "A   Hand  Book  for  Amateurs,"  con- 

'  taining  chapters  on  the  principal  forms 

of  cameras  and  apparatus  likely  to  be  used  by  the  amateur,  methods  of  loading 

plate-holders,   recording  exposures,   focusing,   exposing,  development,   and   the 

various  processes  in  the  manipulation  of  the  negative,  paper,  etc.,  to  produce  a 

satisfactory  print.     Standard  formulae  for  the  various  solutions  are  freely  quoted, 

and  numerous  illustrations  show  the  principal  kinds  of  paper  for  making  prints. 

The  book  is  intended  for  the  studious  amateur,  and  as  a  repository  of  useful 

formula;  and  hints  for  the  beginner  it  serves  its  purpose  admirably. 

L.   B.  E. 


and  Laboratory  Methods.  1205 

Journal    of  Owing  to  extreme  pressure  of  work 

AnnlieH     Miorn^^Onnv  '"^  connection  with  his  new  courses,  at 

APPliea    microscopy  ^^e  university  of  Pennsylvania,  Dr.  R. 

Laboratory'  Methods.  ^-  ^'^'''  ^^"  "°'  ^"  ^^'"  ^°  '^'''"^  '^^ 

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^     hterature  of  "  Normal  and  Pathological 
Edited  by  L.  B.  ELLIOTT.  Histology  "  as  heretofore.     The  work 

which    Dr.    Pierce    has    done    for  the 


Issued  Monthly  from  the  Publication  Department 

of  the  Bausch  &  Lomb  Optical  Co.,  TouRNAL  in  this  department  has  been 

Rochester,  N.  Y.  ^ 


-  greatly    appreciated    by    our    readers. 

SUBSCRIPTIONS:  \Ye    are    fortunate    in     securing    the 

One  Dollar  per  Year.     To  Foreign  Countries,  $1.25  . 

per  Year,  in  Advance.  cooperation  of  Dr.  J.  H.  Pratt,  Harvard 


The  majority  of  our  subscribers  dislike  to  have  their     University  M^dical  School,  to  Continue 

files  broken  in  case  they  fail  to  remit  at  the  expiration        i^Uf,    ^nrW         Dr     Pratt  w;i  11    rnnrlin-t    tViA 
of  their  paid  subscription.     We  therefore  assume  that  no       ^'^^    WOrK.       ur.   rrau  Will    COnOUCt    tnc 

discontki'uTis  sent"  ''""'  ''  ^''''"'^'  ""'"''  "°""  '°     department  along  the  lines  heretofore 

followed. 


The  interest  expressed  by  our  readers  in  Zoological  methods,  both  in  the 
laboratory  and  in  the  field,  has  made  it  necessary  to  add  to  our  reviews  a  depart- 
ment of  "  Current  Zoological  Literature."  In  this  department  will  be  included 
reviews  of  important  zoological  investigations,  especially  those  which  deal  with 
types  most  frequently  used  in  laboratory  work;  methods  in  use  in  zoological 
laboratories  and  by  zoological  investigators  in  the  preservation  and  preparation 
of  animal  forms  for  microscopical  examination,  for  dissection  and  for  exhibition 
purposes ;  methods  in  field  work  in  zoology,  apparatus  for  collecting  aquatic  and 
marine  life,  and  suggestions  for  maintaining  aquaria  and  vivaria  in  the  labora- 
tory ;  notes  on  methods  in  vogue  at  fresh  water  and  marine  biological  stations. 
The  fact  that  Mr.  C.  A.  Kofoid,  University  of  California,  will  conduct  this 
department,  is  in  itself  a  guarantee  of  the  practical  nature  of  the  matter,  which 
will  be  selected  from  the  mass  of  American  and  foreign  literature  on  the  subject, 
and  of  the  faithful  rendering  of  the  author's  meaning.  Separates  of  papers  or 
books  for  review  should  be  sent  addressed  to  C.  A.  Kofoid,  University  of  Cali- 
fornia, Berkeley,  Cal.  Authors  will  confer  a  favor  by  sending  separates  as 
soon  as  issued,  in  order  that  our  reviews  may  be  as  little  delayed  as  possible. 

Now  that  we  are  on  time  in  publication  once  more,  notes  and  news  items 
from  the  various  laboratories  will  be  welcomed,  and  we  ask  those  in  need  of 
assistance  to  make  use  of  the  question-box.  Inquiries  will  be  answered  through 
the  Journal,  or,  if  in  pressing  need  of  information,  at  once  by  letter. 

* 
*  * 

Numerous  requests  for  an  exchange  department  for  the  exchange  or  sale  of 
books,  material,  and  apparatus,  have  reached  the  editor  from  time  to  time.  We 
do  not  consider  the  conduct  of  such  a  department  advisable,  as  we  are  not  in 
position  to  know  the  responsibility  of  every  sender  of  an  exchange  notice,  or  of 
the  merits  of  the  article  offered. 


1206  Journal  of  Applied  Microscopy 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

..  „     _     ,     ^         .     .     ,  ,  In  reviewing  the  literature  of  this  sub- 
Dlxon,  H.  H.     On  the  first  mitosis  of  the  spore-  ° 

mother-cells  of  Lilium.    Notes   from   the  ject  Prof.  Dixon  finds  that,  while  there 

Botanical  School  of  Trinity  College,  Dublin,  j^  ^  ^j^g  divergence  of  opinion  in  regard 
No.  4,  pp.  129-139,  pis.  7-8,  Jan.,  1901.  °  .  ... 

to  the  phenomena  involved  in  this 
mitosis,  there  are,  nevertheless,  certain  stages  which  are  admitted  and  which 
have  been  constantly  observed.  How  these  stages  are  derived  from  one  another 
is  the  most  debated  question.  The  writer  figures  and  describes  six  well  ascer- 
tained stages  and  then  proceeds  into  the  debated  territory.  Nearly  all  observ- 
ers describe  a  longitudinal  splitting  of  the  entire  thread  just  previous  to  the  seg- 
mentation into  chromosomes,  but  Prof.  Dixon  believes  that  the  stage  so  con- 
stantly observed  arises  from  the  looping  on  each  other  and  approximation  of  two 
portions  of  the  thread.  Several  very  suggestive  objections  are  urged  against  the 
commonly  accepted  interpretation.  While  believing  that  each  of  the  two  twisted 
portions  undergoes  a  longitudinal  splitting  while  still  in  the  spirem  stage  or  imme- 
diately after  differentiation  into  chromosomes,  regarded  as  a  seco7id  longitudinal 
splitting  by  Guignard  and  others,  the  author  believes  that  this  is  the  first  and 
only  longitudinal  splitting. 

A  series  of  very  clear  diagrams  illustrates  the  writer's  interpretation  of  the 
composition  of  the  chromosomes  and  their  behavior  during  the  later  phases  of 
mitosis.  According  to  this  interpretation  there  is  no  differential  or  "  reducing  " 
division  during  the  first  mitosis  of  the  spore-mother-cell.  c.  j.  c. 

Britton,  Elizabeth  0.,  and  Taylor,  Alexandria.  This  is  the  first  fairly  complete  account 
Life  History  of  Schizeapusilla  Bull.  Torrey  ^f  j^e  life  history  of  this  interesting 
Bot.  Club,  28:  1-19,  pis.  1-6,  1901.  \  *» 

fern.     The  material  was  collected  at 

Forked  River,  New  Jersey,  in  July,  1900.  Sections  do  not  seem  to  have  b?en 
made  except  in  the  study  of  the  root,  stem  and  leaf.  While  the  peculiar  game- 
tophyte  and  general  aspect  of  the  young  sporophyte  is  shown  more  clearly  with- 
out sections,  we  cannot  help  feeling  that  the  development  of  antheridia  and  arche- 
gonia  and  also  the  very  young  sporophyte  would  have  been  more  satisfactory  if 
the  study  had  been  made  from  microtome  sections. 

A  part  of  the  description,  which  could  hardly  be  abbreviated,  reads  as  follows  : 
"  The  gametophyte  is  composed  of  numerous,  erect,  branching,  dark  green  pro- 
tonemal  filaments ;  monoecious,  bearing  5-12  archegonia,  usually  on  a  slightly 
thickened  and  expanded  series  of  cells  in  the  nature  of  an  archegoniophore  ( ? ) 
or  directly  on  the  filaments ;  antheridia  more  numerous,  often  on  separate 
branches  and  nearer  the  extremities  of  the  filaments ;  radicles  seldom  borne  on 
the  filament,  but  produced  from  specially  modified,  large  spherical  cells,  appar- 


and  Laboratory  Methods.  1207 

€ntly  in  symbiotic  relation  with  a  fungus."  The  filamentous  prothallium  persists 
until  the  young  sporophytes  have  attained  considerable  size. 

In  the  development  of  the  antheridium,  one  figure  shows  a  filament  of  three 
cells.  The  outermost  cell  "  becomes  large  and  globular  and  cuts  off  a  cap  cell 
at  the  summit,  with  the  wall  oblique  ;  the  large  cell  divides  into  the  mother  cells 
of  the.  antherozoids  and  one  ring  cell." 

The  archegonia  are  not  at  all  imbedded,  but  are  entirely  free,  and,  at  first 
glance,  bear  a  striking  resemblance  to  the  archegonia  of  certain  liverworts. 
Each  archegonium  is  derived  from  a  single  superficial  cell  which  divides  into 
three  cells.  The  basal  cell  forms  the  venter  and  from  the  middle  cell  arises  the 
central  cell  and  the  canal  cell.     The  other  cell  forms  the  neck. 

The  anatomy  of  the  root,  stem  and  leaf  is  described  in  considerable  detail. 
The  six  plates  of  careful  drawings  form  no  small  part  of  the  contribution. 

c.  J.  c. 

Macallum,  A.  B.  On  the  Cytology  of  Non-  This  work  was  undertaken  with  the 
nucleated  organisms  University  of  Toronto  j^^pg  ^f  throwing  some  light  on  the 
Studies.     Physiological  Series,  No.  2,  1900.  ^  1  i_      • 

origin  of  the  cell  nucleus  and  to  obtam 

data  to  determine  the  morphological  character  of  the  primal  life  organism.  The 
work  is  divided  into  three  parts,  each  dealing  with  a  separate  group  of  low  organ- 
isms, namely,  the  Cyanophyceae,  Beggiatoa,  and  the  yeast  cell. 

His  results  on  the  Cyanophyceae  are  briefly  as  follows :  The  cell  consists  of 
two  portions,  the  central  body  and  the  peripheral  zone  holding  the  pigment. 
There  is  no  evidence  of  the  presence  of  a  special  chromatophore.  There  are 
two  types  of  granules  present  in  the  cell.  The  one  stains  with  haematoxylin, 
contains  "  marked  "  iron  and  organic  phosphorus,  and  therefore  resembles  chro- 
matin. The  other  type  is  found  in  the  peripheral  layer,  and  chiefly  adjacent  to 
the  cell  membrane.  It  stains  with  picro-carmin,  and  is  free  from  organic  phosphor- 
us and  "  marked  "  iron.  It  is  probably  a  proteid.  There  is  no  nucleus,  nor 
any  structure  which  resembles  a  nucleus  in  the  Cyanophyceae. 

In  Beggiatoa  there  is  no  differentiation  of  the  cytoplasm  into  a  central  body 
and  a  peripheral  layer  such  as  Biitschli  describes.  The  compound  of  "  marked  " 
iron  and  organic  phosphorus  are  uniformly  diffused  throughout  the  cytoplasm  in 
the  threads.  In  the  "spirilla,"  "comma"  and  "cocci"  forms  the  cytoplasm 
shows  characters  like  those  of  the  threads,  but  there  are  also  granules  present 
which  give  slight  reaction  for  "  marked  "  iron  and  organic  phosphorus  and  there- 
fore is  considered  analogous  to  chromatin.  No  specialized  chromatin-holding 
structure  in  the  shape  of  a  nucleus  was  found  in  any  of  the  forms  of  Beggiatoa 
studied. 

In  his  studies  on  the  yeast  cell  Macallum  finds  the  cytoplasm  takes  a  diffuse 
stain  with  haematoxylin  and  gives  a  diffuse  reaction  for  "  marked "  iron  and 
organic  phosphorus.  In  addition  to  the  chromatin-like  substance  diffused 
throughout  the  cell,  there  is  usually  present  a  homogeneous  corpuscle.  This  is 
not  considered  to  be  a  nucleus  although  held  as  such  by  other  investigators. 
The  chromatin-like  substance  in  Saccharomyces  is  soluble  in  artificial  gastric 
juice,  thus  differing  from  the  chromatin  of  the  higher  animal  and  plant  cells. 

In  his  investigations  Macallum  used  the  ordinary  cytological  methods  and 


1208  Journal  of  Applied  Microscopy 

also  micro-chemical  reactions.  Many  fixing  fluids  were  employed,  but  the  best 
results  were  obtained  with  picric  acid  and  corrosive  sublimate.  The  staining 
reagents  employed  were  Ehrlich's  and  Delafield's  haematoxylin,  Czokor's  alum, 
cochineal,  safranin,  eosin,  picro-carmin  and  methylen  blue.  Picro-carmin  was 
employed  to  stain  the  cyanophycin  granules.  A  strong  solution  of  hydrogen 
peroxide  containing  traces  of  sulphuric  acid  was  used  to  liberate  the  "  marked  " 
iron. 

The  paper  is  a  most  valuable  addition  to  the  literature  of  this  important 
problem.  A.  A.  Lawson. 

Chicago. 

Pierce,  0.  J.  The  nature  of  the  association  of  Both  cultures  and  microtome  sections 
Alga  and  Fungus  in  Lichens.  Proc.  Calif.  ^gj.g  ^gg^  j^  ^j^jg  ^Q^k  Various  fixing 
Acad.  Sci.  Ser.  Ill,  I:  207-240,  pi.  41,  1899.  ° 

agents  were  used,  but  a  saturated  solu- 
tion of  corrosive  sublimate  in  35  per  cent,  alcohol  just  below  the  boiling  point 
proved  most  satisfactory.  Dehydration  must  be  thorough,  but,  on  account  of 
the  gelatinous  coating  of  the  lichen,  must  not  be  too  rapid. 

The  results  show  that  the  hyphae  and  gonidia  are  in  the  most  intimate  con- 
nection, the  hyphae  developing  branches  which  clasp  the  algal  cell  or  even  enter 
it  as  haustoria.  This  relation  stimulates  the  algal  cell  to  internal  cell  divisions. 
The  haustoria  drain  the  living  contents  of  the  algal  cells,  leaving  only  the  empty 
cell  walls.  The  fungus  is  fed  by  the  alga  and  it  is  doubtful  whether  the  alga 
receives  any  benefit,  since  it  is  known  that  in  their  resting  forms  free  algse  with- 
stand extremes  of  heat,  dryness,  etc.,  as  successfully  as  do  the  algae  which  are 
associated  with  fungi  in  lichens.  c.  j.  c. 


CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  Sage  College, 

Ithaca,  N.  Y. 

CURRENT  LITERATURE. 

Beard,  J.  The  Source  of  Leucocytes  and  the  The  function  of  this  obscure  gland  has 
True  Function  of  the  Thymus.  Anat.  Anz.  ^^g^  hitherto  but  little  known.  Its 
lo:  550-573,  1900. 

origin  from  the  epithelium  of  the  gill 

pouch  by  Koelliker  for  mammals,  and  his  subsequent  statement  that  the  original 
epithelial  cells  gave  rise  to  leucocytes,  has  been  followed  by  two  views  :  one  that 
these  leucocytes  have  migrated  into  the  gland  from  outside,  the  other  that  they 
originated  within  the  gland.  A  complete  developmental  study  of  ScylUnm 
canicula  made  by  the  author  disclosed  the  fact  that  for  a  relatively  long  period 
the  blood  contains  only  nucleated  red  corpuscles,  no  leucocytes  of  any  kind 
being  present,  as  had  been  noted  by  Koelliker  some  years  before.  In  these 
studies  investigation  was  carried  on   in   two  lines.     First,  a  careful  search  was 


and  Laboratory  Methods.  1209 

made  for  the  stages  in  which  leucocytes  first  appeared  in  the  blood  and  meso- 
blast.  Second,  the  development  of  the  thymus  was  followed  from  its  earliest 
stages  till  the  permanent  characters  appeared.  The  best  material  was  found  to 
be  that  fixed  in  Rabl's  picro-platino-chloride,  or  in  corrosive  sublimate.  The 
most  satisfactory  stain  is  picro-carmin.  In  this  stain  the  red  blood  corpuscles 
are  yellow  and  the  nuclei  often  unstained  (in  picric  acid  preparations) ;  the 
leucocytes  are  differentially  stained,  nuclei  a  brilliant  red  and  their  scanty  proto- 
plasm a  yellowish-brown.  All  the  sections  were  of  embryos  less  than  30  mm. 
in  length  and  have  been  studied  with  the  Zeiss  2  mm.  apochromatic  and  a  Leitz 
^  oil  immersion. 

The  origin  of  the  thymus  element  is  from  a  small  area  of  modified  epithelimn. 
The  term  "placode"  will  be  used  for  this  thymus  element,  a  name  introduced 
by  von  Kupper  for  such  a  small  piece  of  modified  epithelium.  The  thymus 
elements  arise  in  the  skate  as  specialized  portions  of  the  epithelium  of  the  gill 
pouches  before  these  open  to  the  outside,  and  hence  the  thymus  is  of  hypoblastic 
origin.  These  placodes  are  five  in  number  on  each  side.  In  most  cases  the 
histogenesis  of  the  thymus  does  not  take  place  until  the  embryo  is  17-18  mm. 
long,  leucocytes  appearing  at  the  same  time.  It  is  evident  that  the  first  of  these 
originate  in  the  thymus  epithelium,  and  until  some  are  found  there  none  are 
present  elsewhere  in  the  mesoderm  or  blood.  The  first  changes  seen  in  the 
transition  of  the  epithelial  cell  into  a  leucocyte  is  an  increase  in  the  refractive 
power  of  the  cytoplasm  and  under  favorable  conditions  it  assumes  a  somewhat 
brownish  color.  Gradually  the  nucleus  changes  from  its  oval  shape  and  becomes 
rounder,  a  form  later  assumed  by  the  whole  cell.  Finally  the  nucleus  comes  to 
its  eccentric  position.  Many  of  these  newly  formed  leucocytes  wander  out 
immediately  into  the  tissue,  while  others  remain  in  the  gland  and  increase  by 
division,  as  is  shown  by  their  being  in  groups  of  2  and  4.  Emigration  of  leuco- 
cytes begins  at  first  singly,  but  later  the  even  contour  of  the  gland  is  broken,  one 
break  occupies  nearly  the  whole  lower  surface,  and  here  the  leucocytes  are 
wandering  out  en  masse.  Many  are  here  in  the  blood  itself.  These  breaks  in 
the  placode  walls  are  very  characteristic,  all  the  thymus  elements  of  all  the 
embryos  from  this  stage  up  to  those  42  mm.  in  length. 

The  thymus  of  an  embryo  of  71  mm.  has  practically  reached  its  adult 
condition.  The  corpuscles  of  Hassall  have  never  been  seen,  in  the  embryo, 
young  skate  or  adult.  Their  presence  in  fishes  is  uncertain,  only  one  author 
mentions  finding  them.  The  transition  from  the  epithelial  structure  is  as 
follows  :  In  an  embryo  of  33  mm.  the  epithelial  cells  are  restricted  to  the  basal 
portion  of  each  placode ;  the  emigration  of  leucocytes  is  in  active  progress ;  no 
blood  vessels  are  as  yet  within  the  thymus  and  it  is  without  a  connective  tissue 
capsule.  No  spleen  has  yet  been  formed.  In  an  embryo  of  43  mm.  a  capsule 
is  in  process  of  formation,  but  no  blood  vessels  have  formed.  Connective  tissues 
strands  are  forcing  their  way  in  and  lobulating  the  organ.  In  an  embryo  of  71 
mm.  the  thymus  elements  are  free  from  the  epithelium  of  the  clefts,  separated  by 
the  capsule  growth ;  this  latter  still  permits  the  emigration  of  leucocytes,  and 
there  are  many  such  within  the  organ.  Blood  capillaries  are  now  within  the  organ, 
brought  there  by  the  connective  tissue,  and  afford  easy  transport  for  the  leucocytes. 


I'^IO  Journal  of  Applied  Microscopy 

The  author  considers  that  there  is  no  other  source  of  leucocytes  in  the 
vertebrate  body  for  several  reasons.  1.  The  first  leucocytes  clearly  rise  from 
the  thymus,  as  there  are  no  others  present  in  the  body  when  this  organ  first  forms 
such  structures.  These  first  or  parent  leucocytes  quickly  infiltrate  the  blood,  and 
other  lymphoid  tissues  rise  in  all  probability  from  such  migrating  cells.  2.  No 
other  lymph  organ  is  known  which  resembles  the  thymus  in  origin  and  develop- 
mental history.  3.  The  thymus  alone  is  sufficient  to  account  for  all  the  leucocytes 
of  the  body  and  it  is  an  organ  characteristic  of  all  true  vertebrates.  4.  Except  in 
the  case  of  paired  or  metameric  organs  it  is  not  usual  to  find  the  same  function 
in  any  two  organs  of  the  body.  The  thymus  is  a  paired  metameric  structure  of 
the  branchial  region  only.  That  the  thymus  should  be  the  parent  source  of 
leucocytes  explains  its  functional  activity  in  young  animals  and  its  later  atrophy. 

A.   isi.   c. 

Folsom     J.    W.     The    Development    of     the       -pj^g  q^^-     ^  ^f   ^^e    paper    was    twofold, 
Mouth    Parts  of    Amirnia    marjti7na,  Guer.  -^  *^    ' 

Bull,  of  the  Museum  of  Comparat.  Zool.,  to  supplement  a  previous  account  of  the 
Harvard  College,  36:  87-157,8  pis.,  1900.  anatomy  and  functions  of  the  mouth- 
parts  of  a  representative  collembolan  and  to  discuss  the  morphology  of  mandibu- 
late  mouth-parts  of  insects  and  their  nearest  allies  upon  anatomical  and 
embryological  evidence  derived  from  the  most  primitive  insects,  the  Apterygota. 
Material  was  killed  in  hot  water  and  carried  through  successive  stages  of  alcohol 
to  be  preserved  in  absolute  alcohol.  Material  was  imbedded  in  hard  paraffin 
and  sections  cut  from  5-10  }x  in  thickness.  Delafield's  or  Kleinenburg's 
haematoxylin  followed  by  safranin,  Grenacher's  alcoholic  borax-carmin,  and 
Heidenhain's  iron-haematoxylin  were  used  for  staining. 

Nine  consecutive  stages  were  taken  for  representing  the  development  stages, 
and  the  following  parts  are  considered  :  The  procephalic  lobes,  labrum  and 
clypeus,  antennae,  premandibular  appendages  (intercalary),  mandibles,  lingua  and 
superlinguae,  maxillae,  labium,  skull,  tentorium,  segmentation  of  the  head. 

The  proto  cerebrum  of  the  Apterygota  agrees  with  that  of  other  insects  in 
development  and  structure.  The  ocular  segments  of  the  Hexapoda  and  decapod 
Crustacea,  as  well  as  the  compound  eyes  of  the  two  groups,  are  homologous. 
The  labrum  and  clypeus  of  insects  develop  from  a  single  median  evagination 
between  the  procephalic  lobes,  and  do  not  represent  a  pair  of  appendages.  The 
labrum  of  Apterygota  is  homologous  with  that  of  other  insects,  and  of  the 
Symphyla,  Diplopoda,  Chilopoda,  and  higher  Crustacea.  The  antennae  of  the 
Apterygota  evaginate  from  the  posterior  boundaries  of  the  procephalic  lobes, 
and  agree  with  those  of  the  Pterygota.  In  both  groups  the  antennae  are  first 
post-  and  later  pre-oral  in  position.  The  dentocerebrum  of  insects  is  homologous 
with  that  of  Crustacea,  and  the  antennae  of  Hexapoda  are  equivalent  to  the 
antennules  of  Crustacea,  and  the  embryonic  pre-antenncX'  of  Chilopoda.  Pre- 
mandibular or  intercellary  appendages  exist  in  the  embryo  of  Anurida,  and 
appear  to  be  represented  in  the  adults  of  several  Apterygota  genera.  The  trito- 
cerebrum  of  Apterygota  is  homologous  with  that  of  Orthoptera  and  decapod 
Crustacea,  and  the  rudimentary  premandibular  appendages  of  Collembola  and 
Thysanura  represent  the  second  antennae  of  decapod  Crustacea,  and  probably 
the  antennae  of  Diplopoda  and  Chilopoda.     A  distinct  ganglion  for  the  intercalary 


and  Laboratory  Methods.  1211 

segment  shows  it  to  be  a  primary  head  segment.  The  mandibles  develop  from 
simple  papillae,  and  are  only  lobed  in  Campodea  ;  they  are  homologous  with  the 
mandibles  of  Pter)-gota,  Scolopendrella,  Crustacea,  and  probably  Diplopoda  and 
Chilopoda.  The  hypopharynx  consists  of  two  dorsal  "  superlinguae,"  develop- 
ing from  a  pair  of  papillae  between  the  mandibular  and  first  maxillary  segments, 
also  a  ventral  lingua.  First  maxillae  develop  as  in  Orthoptera.  A  palpus 
appears  in  the  embryo,  which  disappears  before  hatching.  The  labrum  of 
Anurida  develops  from  a  pair  of  papillae  from  which  the  entire  gular  region  is 
derived.  A  palpus  appears,  but  is  resolved.  It  is  homologous  with  the  Pterygota 
structure,  agrees  in  detail  with  the  first  maxillipeds  of  decapod  Crustacea.  The 
sides  of  the  face  develope  from  a  lateral  evagination  near  the  mandibular  seg- 
ment, which  eventually  involve  the  labral  and  labial  fundaments.  These  folds 
are  of  Collembola,  Campodea,  and  Japj'x,  are  homologous  with  the  genae  of 
Pterygota.  The  dorsal  region  of  the  skull  in  Anurida  does  not  differentiate 
into  sclerites  comparable  with  those  of  the  Pterygota.  The  evidence  is  for  seven 
segments  in  the  head,  as  is  probably  true  for  all  Hexapods.  Ocular,  antennal, 
intercalary,  mandibular,  superlingual,  maxillary,  labial,  with  ganglia,  and  a  pair 
of  appendages  for  each.  The  Collembola  resemble  Campodea  and  Japyx  in 
structure,  their  entognathous  characteristics  separating  these  groups  from  the 
rest  of  these  insects.  The  Collembola  are  somewhat  more  specialized  than  the 
Thysanura  in  general  structure.  The  Aphoruridae,  including  Anurida,  are  the 
more  generalized  and  probably  degenerate  forms.  The  resemblance  in  most 
parts  indicates  that  the  primitive  collembolan  descended  from  the  stem  form  of 
Campodea,  the  affinities  of  Campodea,  and  in  two  directions,  towards  Machilis  and 
Lepisma,  and  towards  Scolopendrella.  a.  m.  c. 

Kizer,  E.  J.     Formalin  as  a  Reagent  in  Blood      This  has  been  found  a  useful  reagent 
Studies      Proceed.  Indiana  Acad,  of  Sci.,  p.       j^    bringing    out   blood    Structures.      It 

222-2,    1898.  .     . 

produces  no  visible  distortion,  does  not 
interfere  with  staining,  and  is  an  excellent  preservative.  One  volume  of  fresh 
blood  is  mixed  with  three  volumes  of  two  per  cent,  formalin,  and  after  standing 
for  an  hour  a  drop  is  pipetted  from  the  sediment  to  a  cover  slip,  and  allowed  to 
dry  by  evaporation  after  being  spread  evenly.  Slips  are  fixed  in  a  flame,  and 
dipped  once  or  twice  in  a  five  per  cent,  solution  of  acetic  acid.  The  acid  is 
removed  by  water,  and  two  per  cent,  gentian-violet  is  used,  or  methyl-blue  and 
gentian-violet,  or  haematoxylin  and  eosin,  methyl-green  and  safranin,  or  Ehrlich's 
triple  stain.  Excess  of  stain  is  removed  by  water  or  alcohol,  as  the  fluid  requires. 
Mounted  in  balsam.  a.  m.  c. 

Baum,  J.    Beitrage  zur  Kenntniss  der  Muskel-       The  author   used  the    muscles    of   man 

spindeln.    Anat.  Hefte.  H.  42, 43.  249-306,      ^nd   Other    mammals     (especially   the 
4  Tafler,  1900.  ^     ^  ^ 

hedgehog,  guinea  pig,   dog,  cat,  rabbit 

sheep,  pig,  mole)  also  of  Pristiurus  melanostomus,  Syngtiathus  phlegon,  Petro- 
myzon,  and  the  frog.  The  muscle  was  studied  fresh;  it  was  isolated  in  con- 
centrated caustic  potash,  which  does  not  affect  the  nuclei  an-d  fibres,  but  loosens 
the  connective  tissue,  so  that  in  fifteen  minutes  separation  is  easy,  but  great 
pressure  on  the  cover-glass  must  be  avoided.     Acetic  acid  is  used  for  isolation 


1212  Journal  of  Applied  Microscopy 

since  the  nerve  fibres  are  rendered  very  easily  distinguishable.  For  nerve  endings 
the  gold  stain  of  Lowit  was  used.  Miiller's  fluid,  and  occasionally  sublimate, 
were  the  best  fixatives.  Embryonic  and  small  animals  were  decalcified  with 
picric  or  hydrochloric  acid.  Imbedding  was  sometimes  in  collodion  and  some- 
times in  paraffin.  Staining  was  mostly  in  haematoxylin  and  eosin.  Both  bulk 
and  section  staining  were  used.  a.  m.  c. 

Smith,  S.  Note  on  Staining  of  Sections  while  The  author  leaves  the  sections  stretched 
Imbedded  in  Paraffin.  Jour,  of  Anat.  a.  ^^  ^^^^  ^^^^^.  ^^  ^j^j^j^  ^^^  staining 
Physiol.  31:  151-152,  1900.  ° 

solution  has  been  added.     Subsequent 

washing  in  clear  water  was  followed  by  treatment  in  the  usual  manner. 

A.  M.  c. 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Schonichen,  W.,  und  Kalberlah,  A.   B.  Eyferth's      A    third    fully    revised    and    enlarged 
Einfachste     Lebensformen    des    Tier-  und  ,.•  CT^rii  •  iri 

Pflanzenreiches.  Naturgeschichte  der  Mi-  edition  of  Eyferth  s  treatise  on  the  fresh 
kroskopischen  Siisswasserbewohner.  516  water  micro-fauna  and  flora  has  been 
pp.,   16    Taf.,  Braunschweig,   igoo.     Verlag  ,  ,       „         ^    ,  ..    .   ,  ,  x^    1 

von  Benno  Goeritz.  prepared  by  Drs.  Schonichen  and  Kal- 

berlah, assistants  in  the  Royal  Botan- 
ical Gardens  at  Halle,  Germany.  The  present  work  is  a  very  decided  advance 
upon  previous  editions,  the  revision  having  been  most  thorough  and  painstaking. 
The  authors  have  endeavored  to  include  only  those  forms  which  are  most  com- 
mon and  most  widely  distributed.  Many  genera  and  species  described  in  recent 
years  have  been  added  in  this  edition.  The  cosmopolitan  character  of  the 
organisms  found  in  fresh  water  makes  a  treatise  of  this  nature  useful  everywhere, 
quite  as  much  in  America  as  in  Europe.  The  scope  of  the  book  is  indicated  in 
the  title.  The  groups  included  are  the  bacteria,  alga;,  desmids  and  diatoms, 
the  protozoa  and  the  rotifers.  The  main  body  of  the  text  is  made  up  of  brief 
diagnostic  descriptions  with  synoptic  keys  to  the  various  divisions  down  to 
species,  over  OOO  of  which  are  figured  on  the  plates.  The  specific  descriptions 
are  necessarily  very  brief.  The  book  is  thus  not  for  the  use  of  the  specialist, 
but  is  intended  for  the  general  student,  and  the  amateur  microscopist.  It  is  a 
very  convenient  manual  for  the  biological  laboratory.  c.  a.  k. 

Keibel,  F.,  und  Abraham,  K.    Normentafel  zur  I'^e     second     number     of     Keibel's 

Entwicklungsgeschichte  des  Huhnes  (Gal-  "  Normentafeln  "  of  the  development 
lus  domesticus).     132  pp.,  3pls.,4to.     Jena,  ^    1  .1  11  •  , 

1900.    Verlag  von  Gustav  Fischer.  o^  the  vertebrates  has  been  written  by 

the  editor-in-chief  of  the  series,  with 
the  assistance  of  one  of  his  students.  The  growth  of  the  chick  embryo  has  been 
systematically  traced  from  the  earliest  stages,  through  the  first  ten  days  of  incu- 
bation.    Carefully  drawn  figures  are  given  of  a  series  of  embryos  viewed,  how- 


and  Laboratory  Methods.  1213 

ever,  only  as  opaque  objects.  A  tabular  view  is  given '  of  the  stage  of  develop- 
ment of  the  various  organs  of  the  body  in  132  embryos  of  successive  ages  up  to 
ten  days.  The  authors  call  attention  to  those  features  of  the  development  which 
are  subject  to  individual  variation  in  the  chick.  The  care  with  which  this  work 
has  been  done  makes  this  book  a  valuable  work  of  reference  in  establishing  the 
age  of  embryos  as  well  as  in  the  selection  of  embryos  for  the  study  of  organology. 
The  book  is  an  indispensable  aid  in  every  embryological  laboratory.  A  very 
full  bibliography  of  the  subject  occupies  fifty  quarto  pages  of  the  book. 

The  fixing  agents  used  were  sublimate-acetic  and  chrom-acetic,  and  the 
stains  borax-carmin  (followed  in  some  cases  by  bleu  de  Lyon),  para-carmin, 
and  haematein.  c.  a.  k. 

Linko,  Alex.     Ueber  den   Bau  der  Augen  bei       Material     was     prepared     with     aceto- 

den  Hydromedusen.     Mem  del' Acad  imp.      sublimate,    Perenyi's    fluid,    etc.,    and 

des  Sci.  St.  Petersbourg.     CI.   Phys.    Math.  \  ^  '  ' 

10:  No.  3,  1-23,  pis.  1-2,  1900.  Stained   with    Delafield's    haematoxylin 

or  alum  or  borax-carmin.  All  attempts 
to  use  methylen-blue  or  the  Golgi  method  in  any  of  their  modifications  were 
futile.  The  depigmentation  of  the  eyes  was  effected  neither  by  Grenacher's 
method,  by  chlorin,  nor  by  eau  de  Javelle.  In  some  species  the  pigment  was  par- 
tially removed  by  exposure  to  Perenyi's  fluid  for  3-4  hours,  though  this  induced 
some  maceration  of  the  tissvies.  Eight  genera  were  examined,  exhibiting  a  wide 
range  in  structure.  In  Catahlevia  the  eye  is  a  simple  pigment  fleck,  composed 
of  pigmented  and  of  visual  cells.  In  Oceania  a  pigmented  area  of  similar  struc- 
ture is  found  in  a  shallow  pit.  In  Staurostotna  the  eyes  are  numerous  (400)  and 
vary  from  a  simple  pigment  spot  to  the  beaker-form  eye  with  vitreous  body.  In 
Codoniiim  the  sensory  cells  are  somewhat  retracted  and  their  outer  ends  exhibit 
thickenings  which  terminate  in  sensory  "hairs."  In  Sarsia  a  vitreous  body 
occurs  and  the  sensory  cells  terminate  in  conical  end  organs.  Sarsia  is  quite 
sensitive  to  the  stimulus  of  light.  The  eyes  of  Itaropsis  are  of  the  inverted 
type  with  pigment  cells  of  entodermal  origin.  c.  a.  k. 

Bergh,  R.  S.    Beitrage  zur  Vergleichende  His-  Various   writers   have    stated  that  the 

tologie  II.    Ueber  den  Bau  der  Gefiisse  bei  blood  vessels  of  Annelids  are  provided 

den  Anneliden.     Anat.  Hefte  IS:   1500-623,  -,11               r  1         •.     i-      1          1               r 

pis   48-i;i    iQoo  with  a  layer  of  longitudmal,  and  one  of 

circular  muscle-fibers,  with  a  lining  of 
connective  tissue  intima,  folds  of  which  form  the  valves.  Others  have  reported 
that  the  blood  vessels  have  an  endothelial  lining.  Bergh  has  found  a  number  of 
errors  in  these  statements.  Lumbriciis  was  cut  open  and  pinned  out  with  porcu- 
pine spines  in  silver  nitrate.  The  silver  was  reduced  by  exposure  to  sunlight  or 
in  alcohol  slightly  acidulated  with  formic  acid.  The  mixture  of  formic  acid  with 
the  silver  solution  directly  produced  too  excessive  blackening  and  precipitation. 
Silver  preparations  were  stained  in  haematoxylin.  Blood  vessels  for  sectioning 
were  freed  of  their  blood  by  slight  pressure  before  fixing  in  aceto-sublimate. 
Sections  were  stained  in  haematoxylin  or  by  van  Gieson-Hansen's  haematoxylin- 
acid  fuchsin-picric  method,  which  leaves  the  muscle  fibers  yellow,  and  the 
connective  tissue  ground  substance  a  bright  red.  Bergh  was  not  able  to  find  an 
endothelial  lining  in  any  of  the  blood  vessels,  neither  could  he  detect  any  longi- 


1214  Journal  of  Applied  Microscopy 

tudinal  muscle  fibers.  The  valves  are  not  folds  of  the  intima,  but  are  composed 
of  masses  of  cells.  The  blood  vessels,  contractile  and  non-contractile  alike,  are 
lined  throughout  by  a  homogeneous  non-cellular  connective  tissue  membrane 
(Leydig's  intima),  which  is  sharply  limited  internally  and  externally.  Outside  of 
the  intima  is  a  layer  of  connective  tissue  cells  which,  in  the  non-contractile 
vessels  contains  fibrous  or  band-like  elements  in  circular  arrangement.  In  con- 
tractile vessels  this  connective  tissue  layer  contains  strong  circular  muscle  fibers 
with  characteristic  nuclei.  Free  blood  vessels  are  covered  by  the  peritoneal 
cells,  which  have  various  forms.  The  formed  elements  of  the  connective  tissue 
layer  in  silver  preparations  exhibit  endothelial-like  boundaries,  and  adherent 
blood  cells  in  the  vessels  resemble  endothelial  nuclei,  hence  the  endothelium 
reported  by  previous  authors.  c.  a.  k. 

Ritter,  W.  E.,  and  Crocker,  G.  R.    Multiplication  Young  stars  of  this  species  were  found 

of  the  Rays  and  Bilateral  Symmetry  in  the  r       •         r  •  •  ■         n 

2o-Rayed  Starfish,  Pycnopodia  helianthoides  havmg  from  SIX  to  Sixteen  arms   m   all 

(Stimpson).     Proc.    Wash.  Acad.   Sci.  2:  stages  of  growth.     The  six  arms  are 

247-274,  pis.   13,   14,    1900.  J  •        1 

arranged  m  a  group  of  five  and  a  smgle 
one,  the  budding  zones  being  placed  between  the  two  and  the  younger  arms 
coming  in  simultaneously  on  each  side  of  the  group  of  five.  The  stars  are  thus 
bilateral,  but  the  madreporite  is  not  a  median  organ.  The  arms  arise  as  inter- 
radial  outgrowths  of  the  water-vascular  ring-canal  and  the  perihsemal  canals, 
forming  ambulacra  and  receiving  radial  nerves,  which  at  first  project  into  an 
ectodermal  pocket  from  the  outer  edge  of  the  nerve  ring.  A  comparison  is 
made  of  the  position  of  the  sixth  arm  and  that  of  the  larval  organ  (preoral  lobe) 
of  Asterina.  c.  A.  K. 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Melnikow-Raswedenkow.  Pachymeningitis  Hae-  In  his  study  of   the  normal  structure 

morrhagica  Interna.     Ziegler's  Beitrage,  28:       ^f  ^j^^  ^  ^j^g  ^^^^j^^^  f^^^^  Weigert's 

217,  1900.  ... 

elastic  tissue  stain  of  great  value.     In 

the  inner  portion  of  the  dura  the  following  layers  can  be  distinguished:  (1)  A 
single  layer  of  epithelium  which  covers  the  inner  surface  ;  (2)  a  hyaloid,  fenes- 
trated, elastic  membrane,  which  varies  with  age  and  with  the  individual ;  (3) 
the  inner  capillary  network ;  (4)  a  layer  of  connective  tissue,  mixed  with  elastic 
fibers.  The  dura  mater  is  a  peculiar  formation  and  has  nothing  in  common  with 
the  plural  and  peritoneal  serosae. 

Internal  pachymeningitis  is  regarded  as  an  inflammation.  A  fibrinous  exuda- 
tion occurs  upon  the  surface  of  the  internal  elastic  membrane.  Organization  of 
the  exudate  follows  ;  thin-walled  capillaries  grow  out  from  the  capillary  layer  and 
pass  through  spaces  in  the  internal  elastic  membrane.  Rupture  of  the  newly 
formed  blood  vessels  is  common  and  haemorrhage  into  the  delicate  connective 
tissue  results.  j,  h.  p. 


and  Laboratory  Methods.  1215- 

„      ,    ,     TT  .         u  TVT        1  J      Hauck     found,     in      agreement     with 

nauck,  L.    Untersuchungen  zur  Normalen  una  " 

Pathologischen  Histologic  der  Quergestreif-       Other    observers,    that    in    infants     the 

ten  Musculatur.     Deutsche  Zeitschr.  f.  Ner-       individual    fibers    of    different    Striated 
venheilk.,  17  :    57,  1900. 

muscles    have    practically    the     same 

diameter,  while  in  adults  fibers  from  different  muscles  vary  greatly  in  diameter. 
The  thickness  of  the  fiber  is  dependent  upon  the  general  nutrition  of  the  indi- 
vidual.    Rigor  mortis  causes  a  decrease  in  the  diameter. 

In  a  series  of  experiments  upon  young  dogs  the  author  studied  the  influence 
of  rest,  work  and  enervation  upon  the  size  of  the  muscular  fibers.  Cutting  the 
sciatic  nerve  produces  simple  atrophy  of  the  muscle  supplied  by  it.  The  width 
of  the  muscular  fibers  is  diminished  about  one-half.  Simple  muscular  inactivity 
due  to  ankylosis  gives  the  same  result.  j.  h.  p. 

Moser,  A.  Tuberculosis  of  the  Heart.  Med.  Moser  reports  a  case  of  tuberculosis 
and  Surgical  Reports  of  the  Boston  City  ^f  ^j^g  myocardium,  and  presents  an 
Hospital,  11:  194,  1900.  ■'  '  r- 

analysis  of  forty-five  other  cases  col- 
lected from  the  literature.  In  the  case  studied  by  the  author,  a  firm  yellow 
thrombus,  two  cm.  in  size,  was  found  attached  to  the  wall  of  the  left  ventricle. 
The  heart  muscle  was  yellow  and  fibrous.  Histological  examination  showed 
that  the  muscle  underlying  the  thrombus  was  tuberculous,  and  that  tuberculous 
tissue  was  growing  into  the  thrombus.  The  process  apparently  began  with  the 
formation  of  subendocardial  tubercles,  which  later  fused  together.  Tubercle 
bacilli  were  found  in  over  half  of  the  sections  examined. 

Moser  states  that  the  following  method  of  staining  tubercle  bacilli  in  sections, 
devised  by  Mallory  and  Wright,  is  superior  to  the  common  method  known  as  the 
Ziehl-Neelsen :  Stain  lightly  in  alum  haematoxylin ;  then  in  steaming  carbol- 
fuchsin  two  to  three  minutes  ;  decolorize  in  one  per  cent,  acid  alcohol  one-half 
minute ;  wash  thoroughly  in  water ;  dehydrate  in  alcohol ;  clear  in  xylol  and 
mount. 

Birch-Hirschfeld,  in  reporting  a  similar  case  of  tuberculous  mural  thrombus 
of  the  heart,  gave  two  possible  modes  of  origin  :  (1)  Bacilli  wandered  into  a 
mural  thrombus,  or  ("2)  bacilli  clung  to  the  heart  wall,  grew,  and  formed  a 
thrombus.  The  latter  view  was  regarded  as  the  more  probable,  as  Ribbert  pro- 
duced endocarditis  by  injecting  into  the  circulation  particles  of  potato  laden  with 
micrococci.  j.  h.  p. 

Fujinami.    Ueber  das  Histologische  Verhalten  Fujinami    studied  a  large    number  of 

des  Quergestreiften  Muskels  an  der  Grenze  cases  and  found  that  both  cancers  and 
bosartiger  Geschwiilste.     Virchow's  Archiv.,  .  ,  ,       .  ,       , 

161:  III  1900.  sarcomas  mvade   muscle   m  much  the 

same  way.  They  may  infiltrate  be- 
tween the  separate  muscle  fibers  ;  they  may  press  against  the  muscle  fibers  as  a 
mass  ;  or  they  may  be  separated  from  the  muscle  fibers  by  bands  of  connective 
tissue,  thus  only  affecting  them  indirectly. 

The  infiltration  by  the  tumor  takes  place  through  the  sarcolemma  sacs,  as 
well  as  through  the  tissue  spaces,  and  through  the  lymph  and  blood  vessels. 
The  invasion  of  the  sarcolemma  sac  is  especially  marked  when  the  infiltration  of 
the  muscle  is  parallel  to  the  muscle  fibers,  and  is  much  more  common  in  cancers^ 


1216  Journal  of  Applied  Microscopy 

than  in  sarcomas.  In  fact,  the  round-celled  type  is  the  only  form  of  sarcoma  in 
which  the  invasion  of  the  sarcolemma  sac  has  been  observed. 

A  variety  of  changes  occurs  in  the  muscle  fibers  as  a  result  of  the  presence 
of  the  neoplasm.  Simple  atrophy  is  the  most  frequent.  Usually  the  muscle 
nuclei  disappear  as  the  muscle  fibers  atrophy.  Sometimes,  however,  the  nuclei 
increase  greatly  in  number.  Multiplication  occurs  chiefly,  if  not  entirely,  by 
direct  division.  The  nuclei  may  be  found  in  masses,  which  may  be  mistaken 
for  giant  cells. 

The  tumor  cells  may  compress  muscle  fibers,  giving  rise  to  an  irregular 
atrophy,  which  causes  the  fibers  to  assume  a  beaded  appearance. 

All  the  changes  which  occur  in  regenerating  muscle  and  which  have  been 
regarded  as  regenerative  processes  are  found  in  the  degenerating  muscle. 
Hence,  it  is  impossible  to  tell  by  histological  examination  alone  whether  regen- 
eration or  degeneration  is  in  progress.  j.  h.  p. 

Benedict,  Dr.  A.  L.,  Buffalo.  Clinical  Quanti-  In  the  examination  of  stomach  con- 
tative  Analysis  of  Proteids  in  Stomach  ^^^^  ^^^^  f^  jj^g  ^.g^j  function  of  the 
Contents. 

Stomach,  and  how  well   or  how  poorly 

that  function  is  performed,  has  not  been  ascertained.  It  has  been  learned  how 
much  hydrochloric  acid  remained  in  excess  of  that  taken  up  by  food ;  whether  a 
similar  excess  of  ferments  was  present ;  how  much  the  stomach  had  interfered 
with  starch  digestion,  and  when  the  stomach  passed  its  contents  into  the  small 
intestine  ;  but  the  direct  issue  of  the  amount  of  albumin  transformed  into  albu- 
moses  and  true  peptones  has  been  ignored. 

The  method  consists  in  the  successive  precipitation  of  the  proteids  in  solu- 
tion in  the  stomach  contents  and  their  approximate  measurement  by  centrifugal- 
izing  the  three  precipitates,  acid  albumin;  albumoses  and  peptones.  At  first 
thought,  this  would  seem  to  be  a  very  simple  matter,  but  I  assure  you  that  to 
place  it  on  a  practical  clinical  basis  required  a  large  amount  of  research  and 
laboratory  experiment,  as  well  as  interviews  and  correspondence  with  chemists. 
Strangely  enough,  no  analytic  chemist  seems  to  have  undertaken  the  problem 
before.  Any  physiological  chemistry  contains  directions  regarding  the  reactions 
of  the  various  forms  of  nitrogenous  matter,  but,  in  practically  every  case,  it  was 
assumed  that  an  unlimited  supply,  usually  prepared  artificially,  was  available. 
In  all  instances,  the  tests  were  given  with  the  understanding  that  the  investi- 
gator would  perform  them  as  a  matter  of  scientific  curiosity  and  not  with  the 
practical,  analytic  object  of  separating  and  quantitating  the  ingredients  of  a 
mixed  mass  of  proteids.  For  several  years,  it  has  been  my  custom  to  take  up 
some  special  problem,  either  in  physical  diagnosis,  applied  chemistry  of  diges- 
tion, microscopical  technic,  or  some  other  theoretic  topic  that  seemed  likely  to 
yield  practical  results  if  properly  applied,  and  to  make  a  winter's  study  of  it. 
But  the  problem  of  proteid  digestion  in  the  stomach  has  occupied  two  winters, 
simply  because  my  ignorance  of  certain  scientific  details  of  chemistry  compelled 
me  to  grope  in  the  dark;  while,  on  the  other  hand,  lack  of  familiarity  with  the 
conditions  of  medical  practice  prevented  '  chemists  from  giving  me  exactly  the 
information  which  would  have  been  of  the  greatest   use.     I  mention  this  point 


and  Laboratory  Methods.  1217 

only  to  urge  a  more  general  co-operation  between  the  medical  scientist  and  the 
medical  practitioner,  in  attacking  the  many  problems  that  lie  before  us,  the  solu- 
tion of  which  will  make  medicine  more  and  more  an  applied  science,  as  well  as 
an  art. 

One  of  the  difficulties  in  the  way  of  careful  analysis  of  chyme  is  the  small 
amount  obtainable  —  not  usually  over  eighty  and  often  less  than  thirty  cubic 
centimeters,  after  the  ordinary  test  meals.  Free  HCl  and  total  acidity  can  be 
estimated  at  one  titration,  if  we  are  careful  and  meet  with  no  mishap.  Com- 
bined acidity  requires  another  titration,  proteids  another,  and  at  least  a  small 
quantity  must  be  reserved  for  various  qualitative  tests.  While  the  tests  for 
acidity  are  best  applied  to  unfiltered  chyme,  proteolysis  requires  a  clear  filtrate, 
and  a  considerable  loss  occurs  on  account  of  the  mass  left  on  the  filter.  As  a 
matter  of  practical  experience,  I  have  found  that  the  tests  for  proteids  must  be  made 
with  ten  or  sometimes  only  five  cubic  centimeters.  Filtration,  especially  if  much 
mucus  is  present  in  the  stomach  contents,  is  a  tedious  process.  I  have  tried  all 
sorts  of  expedients,  such  as  the  use  of  absorbent  cotton,  separation  by  a  colan- 
der, etc.,  but  have  not  succeeded  as  yet  in  obtaining  rapid  filtration.  By  centri- 
fugalizing  the  stomach  contents,  they  can  readily  be  separated  into  three  layers, 
the  lower  one  consisting  of  undigested  food,  the  upper  one  of  butter  and  mucus, 
the  middle  one  of  comparatively  clear  liquid.  By  removing  the  upper  layer,  the 
middle  one  can  be  decanted  and  filtered  in  the  usual  way,  without  the  delay 
required  if  the  stomach  contents  are  simply  poured  into  the  filter. 

There  is  no  natural  separation  of  the  various  steps  of  the  peptonizing  pro- 
cess, but,  by  common  consent,  chemists  consider  that  every  proteid  not  precipi- 
tated by  ammonium  sulphate  in  saturated  solution  is  a  peptone,  and  that  every- 
thing between  albumin  and  peptone  may  be  called  albumose.  Of  course,  the 
process  of  peptonization  could  be  further  subdivided  by  using  different  reagents. 
To  precipitate  albumose — or  rather  albumoses — I  add  one  gram  of  ammonium 
sulphate  to  ten  cubic  centimeters  of  decantate,  dissolve  the  salt  by  heat,  and 
cool.  As  the  mixture  cools,  a  turbidity  forms,  due  to  albumose.  This  is  very 
light  and  is  precipitated  only  with  the  greatest  difficulty;  in  fact,  I  do  not  usually 
try  to  clear  it  absolutely  by  the  centrifuge,  but  simply  estimate  what  is  thrown 
down  by  10,000  revolutions  ;   ]  per  cent,  may  be  taken  as  the  normal   maximum. 

To  precipitate  peptones,  I  employ  phospho-molybidic  acid,  which  makes  a 
very  bulky  precipitate.  I  should  prefer  tannic  acid,  the  precipitate  from  which 
is  only  about  a  sixth  as  bulky,  and  tannic  acid  is  the  reagent  usually  recom- 
mended by  chemists,  even  in  experimenting  with  stomach  contents  ;  but  they 
forget  that  tannic  acid  also  precipitates  starch,  which  is  almost  invariably  present 
in  chyme.  This  little  oversight  alone  cost  me  several  months'  time,  as  it  neces- 
sitated throwing  out  quite  a  series  of  observations.  Normally,  the  precipitate 
with  phospho-molybdic  acid  is  from  ten  to  nearly  thirty  per  cent,  of  the  filtrated 
chyme.  This  relatively  enormous  bulk  suggests  that  something  else  than  pep- 
tones is  precipitated,  and  it  was  only  after  careful  search  of  chemic  literature  and 
consultation  with  chemists  that  I  became  convinced  that  we  could  rely  on  this 
reagent.  Phospho-molybdic  acid  precipitates  alkaloids  and  certain  biliary  con- 
stituents, but  it  is  impossible   that  there   should  be   anything  of   a  non-proteid 


1218  Journal  of  Applied  Microscopy 

nature  in  the  filtered  chyme,  in  sufficient  quantity  to  interfere  with  the  result 
desired.  For  instance,  to  show  that  ordinary  saline  matters  and  waste  that 
might  be  present  in  the  stomach  could  not  cause  a  precipitate,  we  need  only 
add  phospho-molybdic  acid  to  urine,  when  we  find  a  reassuring  absence  of  any 
precipitation. 

You  will  ask  what  practical  result  we  can  derive  from  such  an  examination. 
The  method  is  comparatively  simple,  and  by  it  we  can  tell  exactly  how  much 
digestive  work  the  stomach  is  accomplishing.  In  general,  we  shall  find  an 
excess  of  lower  forms  of  proteids  in  cases  of  subacidity  and  deficient  formation 
of  ferments.  For  instance,  in  cancer,  we  should  expect  at  least  2  per  cent,  of 
acid  albumin,  probably  as  much  albumose,  and  only  5  or  10  percent,  of  peptone, 
by  bulk.  We  must  also  bear  in  mind  that  an  excess  of  an  end-product  may 
mean  either  unusually  good  digestion  or  poor  absorption.  a.  l.  b. 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Driesch,  H.    Studien  liber  das  Regulationsver-  I"  this  paper  are  collected  the  results 

mogen    der    Organismen.    5.    Ergiinzende  of  several  sets  of  experiments  on  the 

Beobachtungen   an  Tubularia.     Arch.   Ent-  1      1      ■  ,    t-.   ,     7      •  ,  , 

wick.-Mech.  11 :  i8i;-2o6   igoi.  hydroid  lubiilana  inesembryant/iemum, 

all  having  as  a  general  problem  the 
regulatory  processes  of  the  organism.  The  first  point  considered  is  the  number 
of  tentacles  formed  in  successive  oral  reparations.  In  the  experiments  the  polyps 
were  cut  off  and  formed  again  five  times  in  succession.  In  each  successive 
reparation  the  average  number  of  tentacles  is  less  than  in  the  preceding  one. 
The  difference  in  the  number  of  tentacles  between  the  original  polyp  and  the 
individual  resulting  from  the  first  reparation  is  greater  than  that  between  the 
individuals  of  any  of  the  succeeding  reparations.  It  is  believed  that  this  differ- 
ence between  the  original  hydranth  and  the  subsequent  formations  is  due  to 
differences  in  nutritive  conditions  in  the  two  cases.  The  second  point  deter- 
mined was  that  fewer  tentacles  are  formed  on  the  polyp  at  the  oral  end  of  an 
aboral  piece  of  stem  than  on  the  oral  polyp  of  an  oral  piece,  and  that  the  number 
of  tentacles  of  a  polyp  formed  at  the  aboral  end  of  an  oral  piece  was  less  than  in 
the  case  of  oral  polyps  of  either  piece.  From  experiments  with  dififerent  lengths 
of  stems  it  appears  that,  the  longer  the  piece  of  stem  is,  the  more  tentacles  the 
polyp  formed  on  it  has.  The  author  repeats  his  former  conclusion  that  the  "  red 
substance  "  is  the  "  means  "  by  which  the  regulatory  processes  are  effected  in 
Tubularia.  All  the  facts  of  tentacle  formation  are  explained  as  due  to  the  greater 
aggregation  of  this  formative  "  red  substance  "  at  the  oral  end  of  any  piece  of 
the  hydroid,  whether  original  or  secondary  (resulting  from  operation).  The  next 
main  topic  of  the  paper  is  the  healing  of  wounds  in  the  perisarc.  If  the  perisarc 
is  removed  over  a  certain  area  of  the  coenosarc  the  wound  heals  very  quickly. 


and  Laboratory  Methods.  1219 

This  quick  closing  of  the  wound  is  thought  to  be  due,  in  the  first  instance,  to  the 
elasticity  of  the  healing  tissue.  It  was  found  that  small  pieces  of  stem  without 
perisarc  were  capable  of  regeneration.  In  some  cases  pieces  about  1  mm.  in 
diameter  developed  into  complete  polyps  bearing  tentacles.  The  last  general 
subject  considered  was  the  reparation  of  pieces  of  the  stem  split  longitudinally. 
The  method  of  carrying  out  the  experiments  was  to  divide  the  aboral  two-thirds 
of  the  animal  into  two  cross  pieces,  and  then  to  split  longitudinally  the  more 
aboral  of  these  two.  The  polyps  produced  by  each  of  the  split  pieces  are  sym- 
metrical. The  number  of  the  tentacles  formed  by  each  of  the  three  pieces  was 
determined,  and  it  was  found  that  the  sum  of  the  tentacles  produced  on  the  split 
pieces  was  always  greater  than  the  number  produced  by  the  intact,  oral  pieces  of 
the  same  animal.  The  reason  for  this  appears  to  be  found  in  the  relations  of 
the  surfaces  of  the  pieces.  The  sum  of  the  volumes  of  the  split  pieces  is  evi- 
dently equal  to  the  volume  of  the  unsplit  piece,  but  the  sum  of  their  surfaces 
stand  to  the  surface  of  the  uncut  piece  in  the  relation  of  \^,  as  they  are  approx- 
imately cylindrical  and  of  equal  length.  On  the  other  hand,  the  relation  of  the 
number  of  tentacles  in  split  and  unsplit  pieces  is  -^-Jw.  Reducing  these  two  frac- 
tions to  a  common  denominator,  we  have  ^-|^  and  i|^f .  The  close  similarity  of 
these  fractions  indicates  the  validity  of  the  conclusion  that  the  number  of  tenta- 
cles formed  is  directly  related  to  the  extent  of  surface  of  the  formative  basis. 
The  paper  is  one  of  great  interest  and  importance.  R.  p. 

Holmes,  S.  J.  Observations  on  the  Habits  and  This  paper  describes  quite  fully  the 
Natural  History  of  Amphithoe  longimana  behavior  and  general  "  natural  history  " 
Smith.     Biol.  Bull.  2:  165-193,1901.  °  ■'  , 

of  the  amphipod  crustacean,  Amphi- 
thoe. The  scope  of  the  work  is  well  indicated  by  the  titles  of  the  sections, 
which  are  as  follows :  "  Specitic  Description,  Habitat,  Enemies,  Food,  Move- 
ments, Nests  and  Nest-Building,  Moulting,  The  Seat  of  Smell,  Color  and  Color 
Changes,  Sexual  Habits,  The  Disposal  of  Excrement,  Timidity  and  Pugnacity. 
Phototaxis,  Thigmotaxis,  The  Instincts  of  the  Young,  Regeneration,  The  Effect 
of  Cutting  the  Animal  in  Two."  Some  points  of  particular  interest  are:  1. 
Amount  of  food  eaten.  It  was  found  that  the  animals  eat  in  twenty-four  hours 
an  amount  of  food,  as  estimated  from  the  excrement  voided,  equal  to  approxi- 
mately one-tenth  of  their  own  bulk.  2.  Method  of  keeping  a  straight  course 
while  swimming.  The  constant  state  of  partial  flexion  of  the  abdomen,  together 
with  the  beating  of  the  pleopods,  tends  to  make  the  animal  move  in  a  curved 
path  while  swimming.  This  tendency  is  counteracted  by  the  rotation  of  the 
body  on  the  long  axis  through  180°,  at  frequent  intervals.  The  result  of  the 
frequent  repetition  of  this  rotation  on  the  long  axis  is  a  fairly  straight  path, 
having  for  component  parts  arcs  of  circles.  This  method  of  keeping  a  straight 
course  resembles  that  shown  by  some  of  the  Protozoa.  3.  Nest-building.  The 
nests,  which  are  tubular  structures  open  at  both  ends  and  attached  to  water 
plants,  etc.,  are  constructed  from  a  secretion  which  is  poured  out  from  glands  in 
the  first  two  pereopods.  This  secretion  hardens  as  it  comes  out,  and  is  fastened 
at  different  points  by  the  pereopods  touching  their  ends  to  the  object  on  which 
the  nest  is  being  constructed.     New  nests  are  built  in  a  very  short  time,  "  often 


1220  Journal  of  Applied  Microscopy 

in  less  than  a  half  hour."  4.  Sense  of  smell.  The  most  important  olfactory 
organs  are  the  first  antennse,  but  from  the  fact  that  there  is  some  reaction  to 
olfactory  stimulation  after  the  removal  of  the  antennae,  it  is  thought  that  there  is 
a  second  organ  for  this  sense.  The  author,  however,  did  not  succeed  in  pre- 
cisely localising  this  second  seat  of  "  chemo-reception."  5.  Color.  Descrip- 
tions are  given  of  several  color  varieties  of  Amphithoe  which  exist  in  nature  and 
of  the  relation  of  the  pigments  in  these  varieties.  The  color  changes  adapting 
the  animal  to  its  surroundings  are  less  perfect  than  those  shown  by  the  prawn, 
Hippolyte  varians,  as  described  by  Gamble  and  Ashworth  (Q.  J.  Mic.  Sci.  N.  S. 
43:  589-698,  and  this  Jour.  4:  1182-1183).  6.  Thigmotaxis.  Amphithoe  is 
very  strongly  thigmotactic  over  all  parts  of  the  body.  The  author  believes  that 
this  thigmotaxis  forms  the  basis  of  many  of  the  animal's  instincts.  7.  The  young 
animals  soon  after  hatching  show  most,  if  not  all,  of  the  instincts  and  peculiari- 
ties of  behavior  exhibited  by  the  adults. 

The  paper  is  a  good  example  of  the  tendency,  which  is  becoming  strongly 
manifest,  to  return  to  the  old  "  Natural  History  "  view  point,  and,  by  the  appli- 
cation of  modern  methods  of  thought  and  investigation,  to  attempt  to  solve  the 
same  sort  of  problems  as  those  at  which  the  "  naturalists  "  of  the  early  part  of 
the  century  worked.  R.  p. 

Yasuda,  A.    Studien  uber  die  Anpassungsfa-      This  paper  deals  with  the  results  of  a 
higkeit    einiger   Infusorien    an    concentrirte       study  of  the   power   of   acclimatisation 

A  considerable  amount  of  chemical 
acclimatisation  work  has  been  done  on  the  lower  Algae  and  Fungi,  but  hitherto 
there  have  been  no  extensive  results  from  correspondingly  low  animal  forms 
available  for  comparison.  Some  of  these  needed  results  this  paper  presents. 
As  objects  of  experimentation  the  following  species  of  infusoria  were  employed : 
Eiiglena  viridis,  Chilomonas  paramoeciiim,  Mallotnonas  Plosslii,  Colpidimn  colpoda, 
and  Faramoscium  cmidatiwi.  Cultures  of  these  infusoria  were  put  into  solutions 
of  milk  sugar,  cane  sugar,  grape  sugar,  glycerin,  magnesium  sulphate,  potassium 
nitrate,  sodium  nitrate,  potassium  chloride,  sodium  chloride,  and  ammonium 
chloride.  The  solutions  of  these  substances  were  of  different  strengths,  begin- 
ning with  very  low  concentrations  and  going  up  to  those  in  which  death  occurred 
immediately.  In  all  cases  observations  were  made  on  the  length  of  time  the 
animal  lived  in  the  solution,  the  changes  in  structure,  the  effect  on  multiplication 
and  movement,  etc.  Detailed  accounts  are  given  of  all  experiments,  but  only 
the  most  important  results  will  be  mentioned  here.  It  was  found  that  in  isotonic 
solutions  all  the  different  substances  have  nearly  the  same  effect  on  the  same 
organism,  but  this  relation  is  only  an  approximate  one.  The  maximal  limit  of 
concentration  to  which  infusoria  can  become  acclimatised  is  considerably  lower 
than  in  the  cases  of  the  Algae  and  Fungi.  It  is  noteworthy  in  this  connection 
that  Euglena  showed  the  highest  resistance  capacity  of  any  of  the  forms  studied, 
while  it  is  of  course,  structurally,  more  closely  related  to  the  Algae  than  any  of 
the  others.  Increase  in  concentration  is  accompanied  by  a  checking  of  the  mul- 
tiplication of  the  organisms  ;  by  a  retardation  of  the  movement ;  by  an  increase 
in  the  size  of  the  vacuoles,  and  chromatophores.     In  strong  solutions  the  body 


and  Laboratory  Methods.  1221 

of  the  infusorial!  becomes  rounded  and  uneven  in  contour,  and  there  is  a  ten- 
dency, as  the  concentration  approaches  the  maximal  point,  for  the  chromatophores 
or  amylum  bodies  to  join  together  and  form  large  masses. 

A  method  is  described  for  making  pure  cultures  of  infusoria.  A  culture  fluid 
is  prepared  according  to  the  following  formula : 

Meat  extract 1  gram. 

Cane  sugar 20  grams. 

"  Concentrated,  cooked  infusion  of  Porphyi'a  vulgaris  " .  250  c.  cm. 

Distilled  water 729  c.  cm. 

This  culture  fluid  is  sterilized  and  the  desired  infusoria  are  introduced  into 
it  by  means  of  a  capillary  tube.  The  capillary  tube  can  be  examined  under  the 
microscope,  and  only  that  part  of  it  which  contains  the  species  wanted  is  emptied 
into  the  culture  fluid.  The  medium  is  thus  inoculated  with  a  single  species  and 
by  multiplication  a  pure  culture  results.  This  method  is  stated  to  be  very  suc- 
cessful in  practice.  R.  p. 

Moore,  A.  Further  Evidence  of  the  Poisonous  The  purpose  of  this  work  is  to  deter- 
Eff ects  of  a  Pure  NaCl  Solution.  Amer.  jj^j^e  whether  pure  solutions  of  various 
Jour.  Physiol.  4  :    386-396,1900. 

electrolytes  have  the  same  poisonous 

effects  on  fresh  water  animals  as  they  have  been  shown  to  have  on  those  living 
in  sea  water.  The  organisms  used  were  young  trout,  and  frog  tadpoles.  The 
trout  were  taken  just  after  hatching  and  immersed  in  solutions  of  known  concen- 
trations. The  time  of  death  as  indicated  by  the  cessation  of  respiration  was 
noted  and  the  results  from  different  combinations  of  salts  were  tabulated.  The 
results  are  entirely  confirmatory  of  Loeb's  work  on  other  forms.  It  was  found 
that  pure  solutions  of  the  chlorides  of  Na,  Ca,  K,  Mg  and  Li  were  poisonous. 
The  poisonous  effects  of  NaCl  were  antagonized  by  Ca,  but  the  latter  was  not 
found,  however,  to  be  in  itself  necessary,  since  it  made  a  sugar  solution  more 
harmful.  K  did  not  counteract  the  effects  of  Na,  but  was  antagonistic  to  Ca 
used  in  small  quantities.  Sugar  in  weak  solutions  was  as  poisonous  as  NaCl  in 
solutions  of  equal  osmotic  pressure,  while  in  stronger  solutions  it  was  less  pois- 
onous. The  solutions  in  which  the  animals  lived  longest  were  combinations  of 
NaCl  and  CaCl2,  or  of  these  two  salts  with  the  addition  of  KCl.  The  young 
trout  lived  indefinitely  in  distilled  water,  showing  that  no  salts  are  directly  neces- 
sary for  the  preservation  of  life.  A  point  of  interest  was  that  in  case  of  the  trout 
the  heart  beat  continued  for  some  time  after  respiration  had  ceased.  Many  of 
the  solutions  caused  a  remarkable  shrinkage  in  the  volume  of  the  frog  tadpoles 
kept  in  them.  R.  p. 

Galloway,  T.  W.  Studies  on  the  Cause  of  the  It  is  known  that  an  increase  of  temper- 
Accelerating  Effect  of  Heat  upon  Growth.  ^^^^^  causes  an  increased  rate  of  growth 
Amer.  Nat.  34  :  949-957,  1900.  .  .  ° 

in  many  organisms.     The  purpose  of 

this  paper  is  to  determine  whether  in  accelerated  growth  due  to  increase  of  tem- 
perature, the  imbibition  of  water  and  the  anabolic  metabolism  are  equally  accel- 
erated, and,  if  not,  which  of  the  two  processes  is  more  accelerated.  Larvae  of 
Rana  sylvestris,  Amblystoma  puiictatmn  and  Bufo  america?ia  were  used  in  the 
experiments.     Fertilized  eggs  were  subjected  to  three  different  temperature  con- 


1222  Journal  of  Applied  Microscopy 

ditions:  (1)  6°-8°  C,  (2)  12°-18°  C.  (12°-15°  C.  in  Rana),  and  22°-25°  C. 
(20°-24°  C.  in  Amblystoma).  Measurements  were  made  of  tlie  length,  of  the 
total  weight  when  freed  of  superficial  water,  and  of  the  dry  weight,  and  the  results 
were  tabulated.  It  was  found  that  the  dry  weight  is  practically  unaffected  by 
temperature,  and  that,  therefore,  the  acceleration  of  growth  accompanying  a  rise 
of  temperature  is  almost  entirely  due  to  "  the  changed  rate  of  imbibition  of  water." 
The  maximum  percentage  of  water  in  tadpoles  reared  in  high  temperatures  is 
slightly  greater  than  in  those  which  have  lived  in  lower  temperatures.  The 
maximum  total  weight  of  the  animals  reared  in  low  temperatures  is  greater  than 
that  of  those  in  higher  temperature  conditions.  Animals  kept  for  seven  days  in 
a  temperature  of  12°-15°  and  then  placed  in  a  warm  chamber  show  a  greater 
rate  of  increase  of  imbibition  than  those  which  have  been  in  the  high  tempera- 
ture from  the  beginning.  r.  p. 

Jennings,  H.  S.  Demonstrations  of  the  Reac-  In  a  report  of  a  recent  meeting  of  the 
N°l  iz  Un^^^"^^^^^^  Organisms.  Science,  Zoological  Journal  Club  of  the  Univer- 
sity of  Michigan,  the  author  gives  an 
account  of  a  series  of  demonstrations  by  means  of  the  projection  apparatus,  of 
some  of  the  more  striking  facts  in  the  reactions  of  the  unicellular  organisms. 
Among  many  matters  demonstrated,  the  most  important  were :  (1)  The  collec- 
ting ("positive  chemotaxis  ")  of  Paramcecia  about  a  bubble  of  CO 2  and  in  min- 
eral acids.  (2)  The  spontaneous  collections  of  the  organisms,  due  to  CO  2 
excreted  by  themselves.  (3)  Negative  chemotaxis  to  salt  solutions.  (4)  The 
absence  of  orientation  in  chemotaxis.  (5)  The  "  motor  reaction  "  of  Oxytricha. 
(6)  The  essential  identity  of  "positive  chemotaxis  "  and  "negative  chemotaxis." 
In  view  of  certain  recent  criticisms  of  the  author's  brilliant  and  fundamentally 
important  results,  this  record  of  demojistratioiis  of  the  facts  in  the  case  is  espe- 
cially welcome.  r.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Eckles.    An  Abnormal  Fermentation  of  Bread.      Several  instances  of  a  slimy  fermenta- 
Pmceedings   of  the    Iowa  Acad,  of  Sc.  7:      ^^^^  ^f  ^read  appearing   a   day  or  two 

Juckenack,  A.    Beitragzur  Kenntnis  desfaden-      after  the  baking  have  been  recorded 
ziehenden  Brotes.     Zeit.  f.  Analyt.  Chemie.       ^^^   studied    in    recent   years.      Eckles 
Pp.  73-01, 1900.  -' 

has  found  the  trouble  quite  common  in 

a  number  of  localities.     The  sliminess  appears  only  in  bread  that  is  kept  warm 

for  some  hours  after  baking,  and  makes  its  appearance  on  the  third  or  fourth 

day.     The  bread  is  disagreeable  in  odor,  becoming  quite  musty  and  stale,  and 

extremely  slimy.     Eckles  finds  a  number  of  bacteria  present  in  such  bread,  but 

concludes  that  the  trouble  is  due  to  two  species  :     B.  mesentericus  vulgatus  and 

B.  liodermos,  both  of  which  organisms  are  found  capable  of  producing  such  a 


and  Laboratory  Methods.  1223 

sliminess  under  proper  conditions.  The  last  organism  produces  a  greater  slimi- 
ness,  but  the  first  one  a  yellow  color,  which  commonly  accompanies  this  fermen- 
tation. They  frequently  act  together.  After  studying  the  various  sources  which 
may  serve  as  the  cause  of  this  infection,  the  author  concludes  that  the  trouble  is 
probably  due  to  impure  yeasts.  As  a  remedy  he  suggests  either  the  use  of  purer 
yeasts,  or  the  simple  practice  of  cooling  the  bread  directly  after  baking. 

The  second  article  here  referred  to  describes  a  similar  fermentation  of  bread, 
developing  a  very  unpleasant  odor  and  producing  sickness  among  children  when 
used  as  food.  The  cause  of  this  slimy  fermentation  the  author  found  to  be 
neither  of  the  bacilli  mentioned  by  Eckles,  but  the  well  known  species  B.  mesen- 
tericus  fuscus.  The  author  traced  the  trouble  to  the  flour  and  attributed  the 
infection  to  the  fact  that  this  flour  had  been  allowed  to  stand  after  the  milling  in 
a  damp,  mouldy  cellar,  where  it  became  impregnated  with  the  bacilli,     h.  w.  c. 

Newfeld.    Beitrag  zur  Kenntnis  der  Smegma  The  very  great  interest  which  has  devel- 
bacillus.   Arch.  f.  Hyg.   39:    184,1000.  ,  .  ^  .  i    ^      ^1 

Fraenkel.    Zur  Kenntnis  der  Smegma  bacillus,  oped  in  recent  years  m  regard  to  the 

Cent.  f.  Bak.  u.  Par.  I,  29 :  1,1901.  tubercle  bacillus  and  all  other  bacilli 

Russell   and   Hastings.     The  Thermal    Death  11,  ^,  ^   •    •  i-^- 

Point  of  Tubercle  Bacilli.     27  An.  Rep.  of  which  have  the  same  staining  qualities, 

the  Agr.  Exp.  Sta.  of  Wis.  led  the   author  to   institute   a    careful 

Rabinowitsch,  L.     Befund  von  sauref esten  Tub-         .     i        r  ,,  n  .  1       -i 

erkelbacillen  ahnhchen  Bakterien  bei  Lun-  Study  of  the  well  known  smegma  bacil- 

gengangran.     Deutsche    med.  Wochenschr.  lus,  which   has   many  points   of   resem- 

Korn,'ottJ.^°Weitere  Beitrage  zur  Kenntnis  Glance  to  the  tubercle  bacillus.  The 
der  siiurefesten  Bakterien.  Cent.  f.  Bak.  u.  smegma  bacillus  has  shown  COnsider- 
•    7.  P-  4    .9    •  a.h\e  variation  as  studied  by  different 

observers,  and  Newfeld  attempts  to  determine  whether  this  indicates  a  number  of 
species,  or  simply  variations  under  different  conditions.  He  concludes  that 
among  the  smegma  bacilli  that  there  are  at  least  two  types,  one  resembling  the 
tubercle  bacillus,  which  holds  its  color  in  spite  of  the  action  of  acids,  and  the 
other  having  a  similarity  to  the  diphtheria  bacillus,  whose  power  of  holding  the 
stain  is  less.  In  addition,  there  are  numerous  varieties  which  are  probably 
simply  polymorphic  forms  of  these  two  types.  These  two  types  remain  distinct 
in  spite  of  changes  in  the  medium  in  which  they  grow,  but,  nevertheless,  a  change 
in  the  sub-stratum  produces  very  noticeable  differences  in  the  character  of  the 
different  bacilli,  affecting  their  power  of  holding  stains  in  a  considerable  degree. 
The  smegma  bacillus,  in  short,  represents  two  distinct  types,  capable  of  wide 
variations  under  different  conditions. 

Fraenkel  has  made  a  study  of  the  same  problem.  His  methods  of  study  have 
•differed  from  those  of  Newfeld,  but  he  has  reached  practically  the  same  conclu- 
sion. He  finds  that  there  are  two  types  of  the  so-called  smegma  bacillus,  one 
resembling  the  diphtheria  bacillus,  and  the  other  adhering  more  closely  to  the 
characteristics  of  the  tubercle  bacillus.  The  latter  only  he  regards  as  the  smegma 
bacillus.  He  is  inclined  to  believe  that  the  former  represents  the  pseudo-diph- 
theria bacillus,  which  has  acquired  the  power  of  resisting  discoloration  by  acids. 

The  question  of  pasteurization  of  milk  for  the  purpose  of  destroying  patho- 
genic bacteria  is  one  of  great  practical  interest  to  the  dairy  industry.  Pasteur- 
ization at  a  temperature  of  75°  to  85°  C,  temperatures  which  have  been  commonly 


1224  Journal  of  Applied  Microscopy 

employed,  unquestionably  produce  certain  changes  in  the  milk  and  cream  which 
detract  somewhat  from  their  value.  The  question  whether  pasteurization  at  a 
lower  temperature  of  G0°  C  (140°  F)  is  not  sufficient  to  kill  the  tubercle  bacilli 
has  been  investigated  by  several  observers.  The  authors  of  this  paper  have 
tested  this  subject  more  carefully  than  others  up  to  the  present  time,  and  they 
reach  the  extremely  important  conclusion  that  an  exposure  of  tuberculous  milk 
in  a  tightly  closed  pasteurizer  for  ten  minutes  to  a  temperature  of  60°  C  destroys 
the  pathogenic  character  of  the  tubercle  bacilli  that  are  present.  When,  how- 
ever, the  milk  is  exposed  under  conditions  that  enable  a  scum  to  form  on  the  sur- 
face, the  organism  resists  this  temperature  for  a  longer  time.  The  authors,  how- 
ever, regard  a  pasteurization  of  milk  at  60°  C,  for  not  less  than  20  minutes,  under 
conditions  that  prevent  formation  of  a  scum,  entirely  sufficient  to  destroy  the 
pathogenic  character  of  the  tubercle  bacilli  present. 

The  author  finds  in  a  case  of  chronic  pulmonary  gangrene  a  species  of  bacil- 
lus which,  in  its  microscopic  appearance  and  in  its  staining  properties,  agrees 
with  the  tubercle  bacillus.  Its  culture  relations  on  various  media  are,  however, 
different  from  those  of  the  tubercle  bacillus.  For  example,  in  glycerin  agar  it 
produces  an  intense  orange  yellow  pigment.  It  is  not  pathogenic  for  guinea  pigs 
and  seems  to  be  identical  with  one  previously  isolated  by  the  author  from  butter. 

This  last  article  describes  the  characters  of  a  tubercle-like  bacillus,  found  in 
butter.  The  chief  characteristic  of  this  organism  is  that  it  will  not  grow  in  gela- 
tin stab  cultures  at  an  ordinary  room  temperature.  In  this  respect,  as  well  as  in 
the  fact  that  it  cannot  be  adapted  to  room  temperatures,  it  agrees  with  the  tuber- 
cle bacillus.  For  mice  and  birds  the  organism  is  not  pathogenic,  but  for  guinea 
pigs  and  rats  it  produces  an  infection  which  cannot  be  distinguished  from  true 
tuberculosis.  The  author  believes  that  this  organism,  though  not  the  typical 
tubercle  bacillus,  is  a  closely  related  variety.  h.  w.  c. 


NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Ternler,  P.  Nouvelle  contribution  a  I'etude  The  crystals  were  obtained  by  distilla- 
cristallographique  du  cadmium  et  du  zinc  ^[q^  of  ^heir  metals  in  a  vacuum  at  low 
metalliques.     Bull.  Soc.  Min.     23:   18,1900. 

temperatures. 

Zinc  crystals  were  very  small  (less  than  1  mm.  diam.),  quite  clear  and  in  hexa- 
gonal tablets,  with  periphery  formed  of  rhombohedral  facettes.  c  =  1.356. 
Nine  forms  noted. 

Cadmium  crystals  showed  a  marked  similarity  to  those  of  zinc,  with  c  = 
1.335;  the  zinc  crystals,  however,  sometimes  showed  two  prisms.  Seven  forms 
noted. 


and  Laboratory  Methods. 


1225 


Both  metals  also  yielded  spherolitic  aggregates  with  polyhedral  facettes  ;  and 
when  the  cooling  was  rapid  showed  confused,  interlaced  aggregates,  with  the 
free  surfaces  of  the  globules  remaining  spheroidal. 

The  "  slipping  figures  "  obtained  with  a  needle  point  upon  these  facettes, 
were  also  discussed,  their  hexagonal  nature  being  undoubtedly  proved. 


L.  MCI.   L. 


Goldschtnidt,  V.     Ueber  Erkennung  eines  Zwil 
lings.    Zeit.  f.  Kryst.     30:346-351,1898. 


In  previous  numbers*  reference  has 
been  made  to  the  two  circle  method  of 
measuring  crystals  and  the  gnomonic  projection  used  in  conjunction  therewith. 
It  may  therefore  be  of  interest  to  see  how  these  can  be  applied  to  the  recogni- 
tion of  twinned  crystals. 

The  crystal  is  measured  with 
one  individual  in  normal  position, 
with  its  prism  zone  normal  to  the 
vertical  circle.  Symbols  are  ob- 
tained for  this  individual  in  the 
usual  manner,  and  then  for  the 
other  either  by  comparison  with  the 
first  or  by  separate  setting  up  and 
measurement.  The  gnomonic  pro- 
jection is  then  made  and  the  cor- 
responding faces  distinguished  for 
instance  by  a  a,  b  b,  c  £,  etc.  If 
the  grouping  is  a  twinning  there 
will  be  found  a  symmetry  point  j 
at  the  intersection  of  the  zones 
connecting  corresponding  faces  of 
the  two  individuals,  and  this  point 
s  will  bisect  the  angle  between  any 
two  corresponding  faces.  Furthermore  it  will  be  the  pole  of  an  important  face 
or  zone  or,  rarely,  at  90°  to  such  a  face  or  zone. 

If  the  poles  of  two  corresponding  faces  are  superposed,  then  s  is  either  coin- 
cident with  these  or  at  90°  thereto. 

If  the  two  poles,  a  />,  of  one  individual  coincide  with  two,  a'  b' ,  of  the  other, 
then  s  is  the  pole  of  the  zone  a  b. 

When  it  is  not  known  ivhich  points  are  eqiiivaletit  several  poles  of  the  one  are 
connected  by  straight  lines  with  one  pole  of  the  other,  then  all  of  the  first  to  a  second 
point  of  second,  then  to  a  third  point,  and  so  on.  If  several  lines  go  through  a 
common  point,  especially  if  it  is  the  pole  of  an  important  face  or  zone,  then  the 
test  is  made  whether  this  point  bisects  the  angle  between  the  two  poles  on  any 
line. 

The  graphic  determitiation  of  this  equality  in  angle,  and  indeed  the  graphic 
measurement  of  the  angle  between  any  two  faces  in  gnomonic  projection,  consists 
in  finding  the  stereographic  projection  of  the  pole  of  the  zone  of  the  two  faces 


Vol.  3,  Nos.  2  and  7. 


1226  Journal  of  Applied  Microscopy 

(this  Goldscmidt  calls  the  angle  point)  as  follows,  see  Fig.:  Let  a  a'  be  the  zone. 
Draw  the  diameters  ^/^/ parallel  to  <7  a'  and/  c  perpendicular  to  a  a'.  From  / 
with  radius/^/ describe  an  arc.  The  intersection  w  with/r  is  the  desired  pole 
of  the  zone  a  a'  and  the  angle  a  7v  a'  is  the  true  angle  between  a  and  a',  and  if 
J-  is  a  point  of  symmetry,  then  a  7v  s  must  equal  a'  iv  s. 

In  most  cases  the  opposite  faces  are  equivalent  faces  ;  sometimes,  however,  as 
in  case  of  positive  and  negative  tetrahedra,  they  are  not  equivalent.  If  the  poles 
equidistant  from  the  point  s  are  non-equivalent,  s  is  the  pole  of  a  plane  of  sym- 
metry, but  if  the  equidistant  poles  are  equivalent  s  is  the  rotation  point. 

If  no  point  of  symmetry  is  found  the  group  is  not  a  twinning. 

If  s  is  known  it  may  be  made  the  pole  face  and  the  measurements  so  obtained 
will  give  corresponding  points  equidistant  from  j-  in  the  projection.         a.  j.  m. 

Stober,  F.  Sur  un  precede  pour  tailler  des  On  account  of  the  objections  to  the 
grains  mineraux  en  lames  minces.  Bull.  opaque  cements  of  Thoulet  and  Mann, 
Soc.  Min.     22:61,1899.  ,  ,  ^11,  r 

the    author    uses    Canada   balsam    for 

cement  and  describes  a  quick,  convenient  way  of  mounting  and  grinding  the 

grains,  so  as  to  obtain  thin  sections.  l.  mci.  l. 

Wallerant,  F.    Perfectionnement  au  refracto-      Author  describes  apparatus   as   modi- 
metre    pour   les    cristaux    microscopiques. 
Bull.  Soc.  Min.  22:  67,  1899.  fied  by  Czapski. 

Mallet,  F.R.  On  Langbeinite  from  the  Punjab  Author  concludes  that  the  potassio- 
Salt  Range.    Min.  Mag.  12:  159,  1899.  magnesian  deposit,  at  the  Mayo  mines, 

consists    of   langbeinite   (2  Mg.  So^.  K2SO4),  intimately   mixed    with   kieserite,. 
picromerite  and  epsomite.  l.  mci.  l. 

Fletcher,  L.  On  a  Mass  of  Meteoric  Iron  from  This  is  the  first  meteorite  recorded 
the  neighbourhood  of  Caperr,  Rio  Senguerr,  found  in  Patagonia,  and  its  latitude 
Patagonia.     Min.  Mag.  12 :  167,1899.  .        ,         ^       ,  ,  ,     1     r 

IS    the    furthest    south    recorded    for 

meteoric  iron.  l.  mci.  l. 

Hillebrand,  M.  F.  Mineralogical  Notes.  Melon-  The  Melonite  (/)  gives  formula  Ni  Te.,, 
ite  (?),  Coloradoite,  Petzite,  Hessite.  Am.  y^^^^  ^^^  ^^^^  physical  characters  and 
Jour.  Sci.  iv.  8 :  295,  1 899.  .  ,,      ■,•,-,    ^        ,  , 

IS  supposed  to  be  identical  with  Genth  s 

melonite  (NijTe.j)  from  the  same  source  in  California.  L.  mci.  l. 


Medical  Notes. 

A  Point  in  the  Technique  of  Blood  Counting. — I  noticed  an  article  in  the 
December  number  of  the  Journal,  in  which  the  complaint  is  made  that  the 
cross  lines  in  the  Thoma-Zeiss  Blood  Count  Apparatus  are  indistinct  under  the 
microscope.  Will  you  allow  me  to  make  a  suggestion,  which  is  very  simple,  but 
which  I  have  found  to  obviate  this  difficulty  entirely  ?  It  is  to  lower  the  Abbe 
Condenser  far  below  the  usual  position  for  using  it,  until  the  lines  stand  out 
distinctly.  Possibly  every  one  has  discovered  this  for  himself,  but  as  I  have  not 
seen  it  mentioned,  and  it  took  me  some  time  to  formulate  it  into  a  rule  for 
myself,  I  hope  some  one  may  be  helped  by  the  hint. 
Cortland,  N.  Y.  •  Dr.  F.  W.  HiggiNS. 


and  Laboratory  Methods.  l^'-T 

Willebrand,    E.    H.     Stain    for    Simultaneous       Eosin,  0.5  per  cent,   alcoholic,      25  C.c. 
Staining  of   Blood  Smears  with  Eosin  and       nr  ^i     i        -r>i  i  .^k 

Methylen    Blue.       Deut.    Med.     Wochens.       Methylen  Blue,  COn.  aq.  SOl,         25  C.C. 

Jan.  24  and  31,  1901.  Acetic  acid,  1  per  cent.,      10-15  drops 

The  erythrocytes  are  stained  red,  nuclei  blue,  neuthrophile  granules  violet, 
the  eosinophile  granules  red,  and  those  of  the  mast  cells  an  intense  blue. 

c.  w.  J. 

Lewinson.     Method   of  Staining  Fat.   Vratch, 
21,  No.  39. 

1.  Fix  in  Miiller's  fluid  for  two  to  three  weeks. 

2.  Dehydrate  in  successive  changes  of   alcohol,   commencing  with   70   per 

cent. 

3.  Imbed  in  celloidin. 

4.  Stain  sections  of  10  to  15  yu  for  twelve  hours  in  following  solution  : 

Haematoxylin      .  -  .  .  2  grams 

Alcohol,  absolute     -     sufficient  to  dissolve  haematoxylin 
Acetic  acid,  2  percent,  solution  -  100  c.  c. 

5.  Wash  in  water. 

6.  Transfer  to  1  per  cent,  solution  permaganate  of  potash  and  leave  10  to 

15  minutes. 

7.  Wash  in  water. 

8.  Oxalic  acid,  2  per  cent,  solution  for  five  minutes. 

If  sections  remain  yellow  or  brownish  black,  carry  through  the  permanganate 
of  potash  and  oxalic  acid  solutions  again.  If  no  fat  is  present  the  sections  are 
colorless.  If  sections  contain  fat  they  are  slightly  ashy  or  gray-violet  in  color. 
Under  the  microscope  the  fat  appears  gray-violet,  while  all  other  structures  are 
unstained. 

The  following  counterstain  may  be  used  : 

1.  After  removal  from  oxalic  acid  solution,  wash  in  water  and  stain  for  24 

hours  in  an  ammonical  solution  of  borax-carmin. 

2.  Acid  alcohol  1  per  cent,  for  2  minutes. 

3.  Saturated  alcoholic  solution  of  picric  acid,  1  minute. 

4.  Clear  in  alcohol,  xylol,  or  oil  of  origanum. 

5.  Mount  in  Canada  balsam. 

Nuclei  are  stained  red,  protoplasm  yellow,  and  the  fat  dark,  almost  black, 

c.  w.  J. 

Kockel.     New   Stain  for  Fibrin.  Verhandl.  d. 
Deutsch.  Path.  Gesell.  11 :  320. 

1.  Stain  with  Weigert's  haematoxylin. 

2.  Counterstain  in  Weigert's  borax-potassium-ferricyanide   solution,   diluted 

with  three  times  its  volume  of  water. 
Fibrin  stains  dark  blue,  background  light  gray  or  bluish. 
It  is  recommended  that  tissues  be  hardened  in  alcohol,  sublimate  or  formalin 
before  being  stained  by  this  method,  c.  w.  j. 


1228 


Journal  of  Applied  Microscopy 


NEWS  AND  NOTES. 

At  the  January  session  of  the  New  Jersey  State  Microscopical  Society, 
Dr.  Byron  D.  Halsted  read  a  very  interesting  and  instructive  paper  on  "  The 
Movement  of  the  Sap  in  Plants."  The  paper  was  followed  by  a  fine  series  of 
lantern  slides.  During  the  past  year  it  occurred  to  Dr.  Halsted  to  prepare  a 
list  of  questions  concerning  sap,  any  one  of  which  might  naturally  occur  to 
the  "average  layman"  if  he  chanced  to  give  the  subject  a  little  consideration. 
"  What  causes  sap  to  rise  in  plants  ?  "  "  Is  there  more  than  one  kind  of  sap  ?  " 
"  Where  is  the  sap  in  winter  ?"  and  a  number  of  other  questions  of  a  like  nature. 
Answers  were  received  from  representatives  of  a  considerable  number  of  pro- 
fessions and  made  rather  interesting  reading.  Suffice  it  to  say  that  the  botanist 
was  led  to  believe  that  a  paper  on  the  subject  would  not  be  amiss. 

J.  A.  Kelsey,  Secretary. 


Recent  experiments  for  obtaining  pure  cultures  of  algae  have  shown  that 
Cyanophyceae  grow  rapidly  and  luxuriantly  in  a  decoction  of  Zamia,  with  the 
addition  of  peptone  and  sugar. 


The  accompanying  figure  rep- 
resents a  device  which  may  be 
used  as  a  suitable  lamp  for  field 
work,  and  also  as  a  substitute  for 
the  blast  lamp  in  laboratories 
where  gas  is  not  available.  A 
torch  designed  to  meet  the  same 
purpose  was  figured  and  de- 
scribed by  W.  J.  Morse  in  Vol. 
Ill,  p.  986,  of  the  Journal. 
The  "  Queen "  torch,  manu- 
factured by  the  Bridgport  Brass 
Co.,  has  two  burner  tips ;  one 
for  a  round  and  one  for  a  "  fish- 
tail "  flame.  There  is  an  attachment  for  regulating  the  flow  of  gas,  allowing 
the  flame  to  be  turned  down  while  not  in  use.  This  improved  torch  is  recom- 
mended by  Prof  Morse  as  one  which  satisfactorially  serves  all  purposes  for 
which  it  was  designed. 


THE       QUEEN  "  TORCH. 


In  Vol.  IV,  No.  1,  p.  1129,  the  name  of  Prof.  J.  G.  Adami  was  erroneously 
given  as  Prof.  Adann.  In  the  same  reference  the  first  sentence  of  the  third 
paragraph  should  state  that  the  pamphlet  "  How  to  Collect  Mosquitoes  "  was 
issued  by  the  British  Museum  and  sent  to  the  Montreal  Nat.  Hist.  Soc. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


Volume  IV. 


APRIL,  1901, 


Number  4 


The  Laboratory  Equipment  of  the  "  Bahama  Expedi- 
tion" from  the  University  of  Iowa. 

The  problem  which  confronted  the  originators  of  this  expedition  was  the  very 
common  one  of  getting  the  greatest  possible  educational  results  for  a  very  small 
expenditure  of  money.  The  plan  was  to  furnish  a  floating  laboratory  and  home 
for  a  class  of  university  students  which  should  lack  no  really  necessary  thing  for 
comfort  and  reap  the  best  results  from  a  scientific  standpoint.  It  was  deter- 
mined, moreover,  to  work  in  the  best  possible  field  and  to  extend  our  operations 


HAULING  UP  THE  DREDGE. 

down  to  a  sufficient  depth  to  reach  the  deep  water  fauna.  That  such  a  plan 
should  originate  in  a  university  that  is  almost  in  the  geographical  center  of  the 
United  States  is  not  so  strange  as  might  at  first  appear,  for  the  reason  that  we 
of  the  interior  feel  more  deeply,  perhaps,  than  our  brothers  of  the  coast,  the 
immense  advantage  of  study  of  marine  life  to  those  who  would  grasp  fundamen- 
tal biological  facts,  and  if  it  was  necessary  to  take  a  class  over  a  thousand  miles 
to  reach  salt  water  at  all,  we  argued  that  we  might  as  well  go  a  thousand  miles 

(1229) 


1230 


Journal  of  Applied  Microscopy 


THE       EMILY  JOHNSTON." 

CHARTERED    FOR    THE         BAHAMA    EXPEDITION. 


farther  while  we  were  about  it  and  reach  one  of  the  richest  marine  faunae  on 
earth,  that  found  in  the  West  Indian  region. 

In  the  choice  of  a  vessel  our  poverty  did  us  a  real  service  in  compelling  the 
selection  of  a  sailing  vessel  instead  of  a  steamer.     For  such  a  service  the  sailing 

craft  has  several  important  advantages 
over  the  more  modern  type,  and,  so  far  as 
our  experience  went,  very  few  disad- 
vantages. In  the  first  place  our  party 
would  have  required  a  much  larger  ves- 
sel if  it  had  been  necessary  to  provide 
room  for  engines  and  fuel  for  such  a 
cruise,  and  with  a  larger  vessel  we  could 
not  have  gone  to  some  of  the  most  de- 
lightful places  that  we  visited.  Again, 
the  smoke,  dirt  and  heat  of  a  steamer 
would  have  been  most  uncomfortable  in 
the  heat  of  the  tropics.  On  the  other 
hand,  it  is  remarkable  how  efficient  wind 
propulsion  is  when  the  skipper  knows  his  business,  and  ours  did.  A  biological 
party  is  never  at  a  loss  for  work  in  case  the  vessel  becomes  becalmed  in  the  West 
Indian  region.  In  such  a  case  there  is  always  enough  animal  life  at  hand  to  be 
secured  with  the  dip-net  or  the  tow- net  worked  from  a  row-boat  if  at  sea,  or  by  a 
landing  party  if  at  anchor.  As  for  dredging,  any  sort  of  a  wind  will  answer  for 
that,  and  our  work  was  fully  as  successful  as  it  could  have  been  with  steam. 

Our  vessel  was  an  ordinary  fruiting  schooner  from  the  Chesapeake,  of  the 
type  known  as  a  two-masted,  double-topsail,  centerboard  schooner,  with  a  net 
tonnage  of  116  tons.  She  was  95  feet  long,  with  26  foot  beam  and  a  depth  of 
hold  of  7  feet.  Although  not  notable  for  speed  she  was  not  slow,  and  was 
remarkably  staunch  and  "  dry  "  in  rough 
weather.  She  had  four  state-rooms  and  a 
toilet  room  opening  into  a  small  cabin  aft. 
The  rest  of  the  hold  was  ballasted  with 
pebbles  and  a  flooring  was  laid  over  the 
ballast.  Along  the  sides  of  the  hold  were 
arranged  the  bunks  for  the  men  in  two 
tiers  with  curtains  in  front  like  those  in 
a  sleeping  car. 

The  stores,  and  these  were  gradually 
replaced  by  the  collections,  were  stowed 
forward,  leaving  a  large  space  between 
the  stores  and  the  after  bulkhead  for  the 
laboratory,  library,  and  dining-room.  A  door  leading  from  the  cabin  to  the  hold 
divided  the  after  bulkhead  into  two  nearly  equal  parts.  On  one  side  of  this  door 
were  the  shelves  for  the  microscopes,  and  on  the  other  was  the  "  library  "  contain- 
ing the  "  Challenger  "  and  "  Blake  "  Reports,  a  number  of  text-books,  and  all  the 
works  concerning  the  sea  that  were  contained  in  the  university  library.     The 


VISITORS  ABOARD. 


and  Laboratory  Methods. 


1231 


SPOILS  FROM   LAND  AND  SEA. 


larger  volumes  were  covered  with  black  oil  cloth  and  lettered  with  white  paint,  a 
most  valuable  precaution. 

Two  deal  tables  20  by  4  feet,  with  ledges  around  them,  accommodated  the 
whole  party  either  as  dining  or  laboratory  tables,  the  light  from  the  sky-light 
being  ample  by  day  and  four  large  swing- 
ing lamps  serving  at  night,  although  but 
little  laboratory  work  was  done  after  dark. 
A  small  dark-room  for  photographic 
work  was  enclosed  on  the  starboard  side 
next  to  the  library  shelving,  and  this  room 
was  as  near  purgatory  in  the  heat  of  the 
tropics  as  one  would  be  willing  to  endure, 
even  in  the  cause  of  science. 

The  main,  and  altogether  the  most 
comfortable,  laboratory  was  on  deck.  The 
cabin  top  was  just  high  enough  to  serve 
as  an  excellent  table  to  work  at  in  a 
standing  position,  was  almost  flat,  and  large  enough  to  accommodate  the  entire 
party  at  once  when  necessary.  After  we  got  into  the  warm  region  of  the  West 
Indies,  this  cabin  top  served  as  a  bed  for  most  of  the  party,  and  the  bunks  below 
were  permanently  deserted  except  in  rainy  weather,  of  which  there  was  very  little. 
When  at  anchor  the  vessel  was  covered  with  an  awning  reaching  from  the 
foremast  to  the  stern,  and  a  cooler  or  more  convenient  laboratory  would  be  hard 
to  devise.  Of  course  it  was  not  provided  with  running  water,  but  the  very  purest 
of  sea  water  was  easy  to  secure  in  any  quantity  by  simply  dipping  it  up  in  buck- 
ets. An  abundance  of  tubs,  buckets,  tin  pans  and  glass  dishes  was  provided,  as 
well  as  suitable  instruments  for  dissection.  We  had  a  dozen  laboratory  micro- 
scopes of  convenient  type  for  dissection,  and  the  same  number  of  compound 

instruments,  although  it  seldom  happened 
that  all  of  these  latter  were  used  at  any 
one  time.  For  any  special  investigation 
demanding  a  better  instrument  we  had  a 
high-grade  microscope  with  a  1-1 2th  oil 
immersion  lens.  Histological  work,  be- 
yond the  examination  of  fresh  tissues,  was 
out  of  the  question,  under  the  circum- 
stances, and  would  have  been  less  pro- 
fitable than  the  study  of  living  organisms 
even  had  it  been  practicable. 

As  to  our   plan  of  work,  it  was,  as  is 
A  CATCH.  always    the    case    where   wisely    directed, 

determined  by  the  varying  and  unforseeable  conditions  that  daily  confronted  us. 
In  other  words,  we  studied  that  which  was  at  hand  in  greatest  abundance,  or  that 
which  seemed  most  instructive.  When  under  sail  we  depended  largely  upon 
the  dip-nets  for  material. 

One  day,  for  instance,  was  devoted  to  a  study  of  the  Sargosso  weed  and  its 


123-2 


Journal  of  Applied  Microscopy 


inhabitants.  A  better  object  lesson  in  the  matter  of  protective  coloration  it  would 
be  impossible  to  find  anywhere,  the  "weed  "  at  first  seeming  to  be  uninhabited, 
but  afterwards  disclosing  no  less  than  twenty-six  species  to  which  it  gave  shelter 
and  protection.  On  another  day  we  were  sailing  through  countless  millions  of 
the  little  medusa  Lincrges  mercurius,  carefully  described  and  figured  by  Dr. 
Fewkes.  Here  was  an  excellent  chance  to  become  acquainted  with  the  medusa 
structure.  Again,  we  made  a  careful  dissection  of  a  species  of  shark  which  was 
wonderfully  abundant  near  the  Dry  Tortugas ;  and  one  day  our  pilot  ran  the 
vessel  aground  on  a  sandy  bottom  thickly  strewn  with  an  immense  starfish,  Fen- 
taceros  reticulatus,  which  gave  us  the  best  possible  chance  to  study  the  anatomy 
of  the  Asteroidea.  Still  again,  we  had  the  unique  pleasure  of  studying  fully 
expanded  Millepoi-a. 

The  more  serious  work  of  the  expedition  was  in  the  line  of  dredging  in  com- 
paratively deep  water.  Those  who  were  informed  in  the  science  of  deep  water 
dredging  were,  for  the  most  part,  inclined  to  smile  quietly,  but  still  significantly, 


REELS  FOR  SOUNDING  LINE.     HAND       CRAB"  USED  IN   DREDGING. 


at  our  audacity  in  undertaking  such  work  without  steam  either  for  propelling  the 
vessel  or  hoisting  the  dredge.  Perhaps  it  was  a  case  where  "fools  rush  in,"  etc., 
but  we  were  nevertheless  entirely  successful,  and  probably  secured  as  many  inter- 
esting things  from  the  deep  water  fauna  as  any  expedition  has  obtained  in  the 
same  length  of  time. 

Our  equipment  for  dredging  was,  briefly,  as  follows : 

The  power  necessary  to  handle  the  dredges,  trawls,  tangles,  etc.,  was  fur- 
nished by  a  hoisting  machine  technically  known  as  a  "  crab,"  which  was  a  sort 
of  windlass  worked  by  hand,  constructed  after  plans  devised  by  Professor  Weld 
of  the  University  of  Iowa. 

It  consisted  essentially  of  a  horizontal  drum  fifteen  inches  in  diameter  and 
thirty  inches  long,  resting  on  a  heavy  iron  frame  bolted  to  the  deck.  This  drum 
was  provided  with  a  single  and  double  purchase  for  cranks,  by  which  a  sufficient 
power  could  be  applied  to  meet  every  demand  likely  to  be  made  upon  the  machine. 
The  lowering  of  the  dredge  was  regulated  by  a  powerful  friction  brake.     Upon 


and  Laboratory  Methods. 


1233 


LETTING  DOWN  THE  DREDGE. 


the  drum  was  reeled  something  over  three  hundred  fathoms  of  cast  steel  rope, 
not  a  foot  of  which  was  lost  during  the  entire  trip. 

Of  course  the  reeling  in  of  this  rope  with  a  heavy  dredge  at  its  end  under  the 
direct  heat  of  the  tropical  sun  was  no  child's  play,  and  taxed  the  endurance  of 
even  the  most  enthusiastic.  But  it  was 
done,  and  to  good  purpose.  Incidentally, 
it  may  be  remarked  that  in  my  opinion 
this  work  had  a  good  deal  to  do  with  the 
excellent  physical  condition  of  the  men 
during  the  cruise. 

Dredges  of  the  regular  "  Blake  "  pat- 
tern were  used.  These  and  the  trawls, 
also  practically  like  those  used  on  the 
Blake,  were  made  in  the  engineering  de- 
partment of  the  university  at  surprisingly 
small  expense.  Trawls,  however,  are  of 
little  use  where  the  bottom  is  rocky,  as 
was  the   case   almost  everywhere   in  that 

region  at  a  depth  of  from  fifty  to  two  hundred  and  fifty  fathoms.  By  far  the 
most  effective  instrument,  and  the  one  upon  which  we  eventually  depended  almost 
exclusively,  was  the  tangles,  made  after  a  pattern  suggested  by  Dr.  James  E. 
Benedict  of  the  National  Museum.  This  proved  such  a  decided  success  that  I 
may  be  excused  for  giving  its  construction  in  detail.  A  four-foot  length  of  1x2- 
inch  iron  bar  is  bent  in  the  middle  at  nearly  a  right  angle.  Five  iron  rings 
are  bolted  at  regular  intervals  to  the  inner  side  of  this  bar,  and  to  each  ring 
is  fastened  a  two-foot  length  of  fairly  heavy  chain.  Through  each  link  of 
these  chains  is  passed  a  six-foot  strand  of  2^ -inch  Italian  hemp  rope,  each 
strand  being  tied  in  the  middle  and  thoroughly  unraveled  throughout  its  length. 

The  dredging  cable  is  attached  to  a  hook 
bolted  to  the  outer  side  of  the  angle  of  the 
bar.  The  amount  of  material  secured  by 
this  device  was  astonishing,  and  included 
all  sorts  of  things  from  corals  to  fishes, 
quite  a  number  of  the  latter  being  secured 
in  this  way. 

Being  provided  with  several  of  these 
tangles,  we  were  able  to  economize  time 
by  detaching  one  from  the  cable  as  soon 
as  it  came  up,  and  sending  another  down 
to  be  at  work  while  we  were  picking  over 
the  first  one.  Many  hands  made  this 
usually  irksome  labor  light.  Each  student  had  a  particular  group  of  marine 
animals  to  look  after,  and  he  was  responsible  for  the  care  of  all  of  the  material 
in  his  group,  and  had  prepared  himself  to  do  just  that  work.  The  result  was  that 
the  collections  as  a  whole  came  through  in  very  good  shape. 

We  were  astonished  to  find  the  amount  of  knowledge  of  the  various  groups 


DREDGING  ON       POURTALES  PLATEAU. 


1284  Journal  of  Applied  Microscopy 

that  was  acquired  from  the  mere  handling  of  quantities  of  material,  and  sorting 
it  out.  Indeed,  this  seemed  to  me  to  be  the  most  thoroughly  educative  factor 
involved  in  the  expedition.  Again,  one  must  have  the  actual  experience  to  real- 
ize the  difference  in  the  instruction  derived  from  museum  specimens  and  those 
taken  fresh  from  their  proper  habitat.  For  instance,  we  were  all  astonished  at 
the  bright  colors  of  the  deep-sea  forms,  and  impressed  with  the  manifestly  adapt- 
ive nature  of  these  colors,  because  not  only  a  given  species  but  also  the  associa- 
ted forms  were  brought  up  together,  making  manifest  the  fact  that  these  colors 
were  very  often  protective. 

Such  facts  would  never  have  been  suggested  by  the  study  of  museum  speci- 
mens, no  matter  how  abundant  and  well  preserved  they  might  be. 

Of  course  we  had  to  depend  almost  exclusively  upon  alcohol  as  a  preserva- 
tive, formalin  not  having  yet  come  into  use.  An  excellent  device  for  saving 
alcohol  and  weight  was  carried  into  effect  at  the  suggestion  of  Dr.  Benedict. 
This  was  simply  to  take  specimens  after  they  had  been  for  a  few  days  in  alco- 
hol and  solder  them  up  in  large  tin  pans,  two  of  which  were  soldered  together 
by  their  broad  fiat  rims.  These  pans  were  both  square  and  round,  and  could  be 
conveniently  crated  when  sealed.  Specimens  preserved  in  this  way  often  came 
through  in  better  shape  than  when  left  in  alcohol. 

It  may  be  of  interest  to  some  of  your  readers  to  be  informed  that  the  entire  cost 
of  this  expedition  to  each  member  was  almost  exactly  two  hundred  dollars,  includ- 
ing fare  from  Iowa  City  to  Baltimore  and  return,  and  every  necessary  expense 
during  a  three  months  cruise.  There  was  no  accident  or  misfortune  of  any  kind, 
and  no  sickness  except  the  inevitable  sea-sickness.  C.  C.  Nutting. 

State  University  of  Iowa. 


A  Description  of  the  New  Wing  of  the  Laboratory  of  Hygiene 
at  the  University  of  Pennsylvania. 

As  an  immediate  result  of  the  increasing  interest  in  the  science  of  bacteri- 
ology, the  construction  of  suitable  laboratories  for  its  pursuit  has  come  to  be  a 
subject  of  no  little  importance.  At  present  there  is  of  necessity  more  or  less 
experimenting  in  this  line,  and  the  results  obtained  by  one  institution  are  of 
value  to  other  institutions  contemplating  the  erection  and  equipment  of  labora- 
tories and  lecture-rooms  for  the  work. 

Dr.  A.  C.  Abbott,  Professor  of  Hygiene  and  Bacteriology  at  the  ITniversity 
of  Pennsylvania,  has  recently  contributed  a  detailed  description  of  the  new 
addition  opened  a  year  ago  for  the  instruction  of  bacteriology  in  that  institution. 
In  planning  the  building,  the  primary  object  was  to  provide  a  lecture-room  with 
seating  capacity  for  not  less  than  three  hundred  students,  and  a  laboratory 
sufficiently  large  to  accommodate  not  less  than  seventy-five  students  working  at 
one  time.  Externally,  the  structure  as  completed  is  of  red  brick  trimmed  with 
brown  stone  and  terra  cotta,  and  two  stories  high ;  conforming  in  lines  and  finish 
to  the   original  building.     Internally  there  were  three  features  of  construction 


and  Laboratory  Methods. 


1235 


o5- 


^■ 


i" 


1236 


Journal  of  Applied  Microscopy 


which  were  insisted  on,  and  which  there  have  been  thus  far  no  reasons  to 
regret.  They  are :  hardwood  floors,  steel  ceiling  for  the  lower  room,  and  walls 
devoid  of  plaster.  Hardwood  floors  were  adopted  more  for  economy  than 
elegance,  maple  being  preferred  to  yellow  pine  as  it  is  less  apt  to  splinter  with 
hard  usage,  even  though  it  is  given  much  less  care.  The  floors  are  well  laid, 
stained,  oiled  and  varnished. 

Experience  has  shown  that  plaster  ceiling  is  liable  to  fall  at  any  time  should, 
through  accident,  which  is  not  rare,  the  floor  of  the  room  above  become  flooded 
with  water.  Plastered  walls,  unless  painted,  become  soiled  very  quickly,  and  are 
difficult  to  clean  ;  and  if  painted,  this  must  be  repeated  from  time  to  time,  thus 
incurring  constant  expense.  The  objections  that  bare  brick  walls  reflect  less 
light,  and  favor  the  condensation  of  moisture  upon  them,  are  readily  met  by  the 
use  of  light  colored,  smooth  brick,  and  laying  them  with  an  air  space  between 
external  and  internal  walls. 


Fig. 


-Lecture  room,     (s)  seats;  (t)  instructor's  table. 


The  first  floor  is  occupied  by  a  lecture-room  52  x  56  feet,  to  which  students 
gain  access  directly  from  without  through  a  vestibule  doorway,  while  the  instruc- 
tor may  enter  behind  the  lecture-table  directly  from  the  laboratory  hallway,  or, 
by  a  door  to  the  left  of  this,  from  the  adjoining  preparation  room.  (See  Figs. 
1  and  2  for  general  arrangement.) 

The  seats,  with  a  capacity  of  310,  extend  straight  across  the  room,  rising  on 
8-  to  9-inch  steps  from  the  lecture-table  to  within  9  feet  of  the  opposite  wall. 
They  are  comfortable  church  pews  of  oak,  with  antique  finish,  2'  8"  from  back 
to  back,  and  each  one  subdivided  by  cast-iron  arm  rests  19"  apart ;  the  object  of 
this  being  to  ensure  sufficient  room  for  comfort  to  each  individual,  and  also  to 
discourage  any  tendency  on  the  part  of  the  occupants  to  lounge.  There  is  a 
center  aisle  of  2'  8",  and  an  aisle  3'  6"  in  width  along  each  lateral  wall.  The 
ceiling,  17'  6"  high  at  the  point  occupied  by  the  instructor's  table,  is  of  paneled 
steel,  painted  with  zinc  paint  to  match  the  lighter  parts  of  the  walls,  which  are 


and  Laboratory  Methods. 


1237 


1238 


Journal  of  Applied  Microscopy 


of  light  buff  pressed  brick,  laid  in  mortar  of  corresponding  color,  and  smoothly 
finished.  For  a  distance  of  five  feet  above  the  floor  the  color  of  the  brick  is  a 
chocolate. 

Illumination  is  supplied  by  rows  of  windows  extending  up  to  the  ceiling  on 
the  east  and  west  walls  ;  in  addition  to  which  electricity  is  provided  for  use  on 
dark  days  and  at  night. 

Ventilation  is  through  a  large  stack  heated  by  steam  coils.  Heating  is  in 
part  by  direct  radiation  from  steam  radiators,  and  in  part  indirectly  from  large 
steam  coils  placed  beneath  the  perforated  stairways  entering  the  room.  Besides 
a  large  lecture-table  provided  with  gas,  water,  and  sinks,  the  instructor  has  at 
hand  movable  racks  for  the  exhibition  of  diagrams  used  in  the  lectures. 


Fig.  4. —  Laboratory,     (d)  desks  for  microscopical  work  ;  (t)  tables; 
(i.  t.)  instructor's  table ;  (h)  hood. 

A  laboratory  for  practical  work  in  bacteriology  occupies  the  second  floor, 
immediately  above  the  lecture-room.  The  walls  and  floor  are  similar  to  those  of 
the  room  below.  The  ceiling  increases  in  height  from  the  walls,  where  it  is  14 
feet,  to^the  center  of  the  sky -light,  where  it  is  about  24  feet.  With  a  large  sky- 
light, and  with^windows  in  the  three  walls,  the  illumination  of  this  room  is  all 
that  could  be  desired.  The  room  is  heated  by  steam,  is  well  ventilated,  and  has 
a  capacity  of  83  students  working  at  one  time.  The  arrangement  of  the  desks 
may  be  seen  in  Figs.  3,  4,  and  5.  Parallel  with  the  east  and  west  walls  are  two 
rows  of  desks  with  an  aisle  of  5  feet  between  them,  while  across  the  southern 
end  there  are  three  such  rows  separated  by  aisles  of  3'  6".  These  desks,  each 
one  3'  wide,  2'  3"  deep,  and  2'  8"  high,  are  made  of  poplar,  and  are  joined 
together  in  sets  of  from  four  to  six,  as  convenience  required.  On  the  low  parti- 
tion (about  2"  high)  dividing  one  desk  from  another,  are  gas  and  water,  the 
latter  being  syphoned  from  large  bottles,  held  in  suitable  iron  racks.  This  plan 
is  regarded  as  preferable  to  water-taps  from  the  regular  house  supply,  as  the 
latter,  even  though  filtered,  is  often  objectionable,  while  the  bottles  can  always  be 
kept  filled  with  distilled   water.     It  also  eliminates   the  frequent   annoyance  of 


and  Laboratory  Methods. 


1239 


1240  Journal  of  Applied  Microscopy 

obstructed  drain  pipes.  Each  desk  is  supplied  on  the  right  hand  side  with  a 
drawer  and  locker  one  foot  wide,  extending  through  the  entire  depth  of  the  desk ; 
while  beneath  the  top  of  the  desk  and  well  out  of  the  way  is  a  shelf,  inclined 
toward  the  back,  and  large  enough  to  accommodate  an  overcoat  and  a  hat,  thus 
obviating  the  necessity  of  special  coat  lockers. 

The  bodies  of  these  desks  are,  like  all  other  wood  fittings  of  the  room,  finished 
in  the  natural  color  of  the  wood,  oiled  and  varnished.  The  tops  of  all  desks  and 
of  the  tables  in  the  room  are  finished  in  black,  with  lamp-black  and  paraffin. 
This  finish  has  been  found  sirperior  for  laboratory  purposes  to  any  other,  and  is 
obtained  by  rubbing  into  the  freshly  dressed  desk  top  a  mixture  of  lamp-black 
and  turpentine  until  the  wood  is  thoroughly  soaked  with  it.  All  excess  of  the 
black  is  then  carefully  removed  by  thorough  rubbing  with  cotton  waste,  or  with 
old  rags.  After  this,  paraffin  of  a  high  melting  point  is  ironed  into  the  wood 
with  a  hot  iron.  The  excess  of  paraffin  also  is  finally  removed  by  thorough 
rubbing.  The  result  is  a  comparatively  dull  black  finish,  very  restful  to  the  eyes, 
an  excellent  background,  and  a  finish  that  is  not  injured  by  the  ordinary 
chemicals,  staining  solutions,  or  warm  objects  that  may  get  upon  it.  Under  no 
circumstances  should  a  laboratory  table  or  desk  be  varnished.  The  tops  of  the 
desks  are  not  screwed  or  nailed  to  the  bodies  in  the  ordinary  manner,  but  are 
held  in  place  by  screws  passing  through  slots  in  such  a  way  as  to  allow  the  wood 
to  shrink  without  cracking.  There  are  no  angles  to  tops  of  desks,  all  corners 
being  rounded  to  facilitate  cleaning. 

On  three  walls  of  the  room  are  glazed  lockers  for  microscopes,  each  one 
bearing  a  number  to  correspond  with  a  desk.  The  glazing  of  the  lockers  admits 
of  ready  inspection  of  contents  by  the  instructor,  without  his  being  obliged  to 
open  the  lockers.  Each  student  on  entering  the  laboratory  for  work  is  supplied 
with  a  desk,  a  locker  and  the  keys  for  the  same,  for  all  of  which  he  is  held 
responsible.  The  equipment  of  each  desk  consists  of  a  microscope  of  approved 
pattern,  including  an  oil  immersion  lens,  staining  reagents,  test-tubes,  dishes, 
funnels,  flasks,  a  gas  stove,  a  Bunsen  burner,  and  in  short  all  the  apparatus, 
except  slides,  coverslips,  towels,  notebooks,  etc.,  that  are  necessary  to  pursue 
the  course.     No  charge  is  made  for  any  apparatus  unless  injured   or  destroyed. 

In  addition  to  the  desks  there  are  four  large  tables  in  the  laboratory  that  are 
used  for  such  work  as  the  preparation  of  culture  media  and  the  demonstration 
of  autopsies,  dissections,  etc.  These  are  supplied  with  sinks,  hot  and  cold  water, 
and  gas.  Beneath  these  tables  are  lockers  for  the  use  of  students,  each  locker 
being  numbered  to  correspond  with  a  particular  desk. 

On  the  north  wall  of  the  room  are  the  necessary  shelves  and  closets  for 
apparatus  and  materials,  and  to  the  right  of  the  door  leading  into  the  room  is  a 
commodious  glass  hood,  the  framework  of  which  is  of  iron,  the  base  of  soapstone, 
and  the  back  of  brick,  This  hood  is  provided  with  gas,  water,  and  aspirating 
flues.  The  glass  inclosing  such  a  hood  should  never  be  cemented  firmly  in  the 
frames,  as  it  is  sure  to  crack  by  the  expansion  and  contraction  of  the  surround- 
ing metal.  It  should  be  either  loosely  set,  or  set  in  some  elastic  material,  that 
will  relieve  the  strain  upon  it.  The  total  cost  of  the  addition  to  the  original 
building  was  a  trifle  over  fifteen  thousand  dollars  ($15,001.2.'")).  c.  w.  j. 


and  Laboratory  Methods. 


1241 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 

THE  VALUE  OF  THE  TELEPHOTO  LENS. 

The  value  of  the  telephoto  lens  to  the  naturalist  has  been  shown  in  many 
ways.  Photography  has  reached  such  a  stage  that  it  is  impossible  for  a 
naturalist  to  do  without  it.  In  order  to  do  the  best  work  it  is  necessary  to  carry 
several  lenses,  or  combinations  which  will  produce  both  long  and  short  focus, 
depending  on  the  object  to  be  taken. 


PHOTOGRAPH     MADE    WITH    ORDINARY 
PHOTOGRAPHIC  LENS. 


PHOTOGRAPH   MADE  WITH  SAME  LENS 
AND  THE  TELEPHOTO. 


The  accompanying  pictures  show  how  a  telephoto  lens  was  necessary  to 
secure  a  picture  of  an  osprey's  nest  which  was  in  the  top  of  an  old  pine  tree, 
around  the  base  of  which  there  was  a  dense  jungle  of  underbrush.     The  view 


1242  Journal  of  Applied  Microscopy 

shown  across  the  river  is  the  only  view  to  be  had  of  the  nest  from  this  near  dis- 
tance without  having  the  foreground  so  full  of  limbs  as  to  obscure  the  nest. 

The  location  is  on  the  north  end  of  Flathead  lake,  in  western  Montana.  The 
nest  is  in  front  of  the  University  of  Montana  Biological  Laboratory,  which  is 
immediately  behind  the  illustration.  That  is,  it  is  at  the  photographer's  back. 
The  river  in  the  foreground  is  Swan  river,  or  Big  Fork.  It  is  swift  and  turbu- 
lent. There  is  no  place  on  the  opposite  shore  where  the  camera  may  be  placed 
so  as  to  take  in  the  nest.     The  nest  is  about  a  hundred  feet  from  the  ground. 

The  picture  with  the  nest  small  was  taken  on  a  Seed  orthochromatic  plate,  on 
a  cloudy  day  when  rain  was  falling.  Without  changing  the  camera  the  telephoto 
lenses  was  added,  and  the  camera  was  pointed  so  as  to  put  the  nest  in  the  mid- 
dle of  a  five  by  eight  plate,  and  the  magnification  raised  to  five.  Naturally  the 
exposure  was  porportionally  longer.  The  sky  was  overcast,  and  rain  was  falling. 
Indeed,  it  was  necessary  to  notice  that  water  did  not  get  on  the  lens.  Only  the 
small  opening  seen  between  the  trees  was  taken  by  the  telephoto,  and  the  back- 
ground was  clouds.  While  a  longer  exposure  would  have  produced  a  better  result 
for  definition,  the  two  pictures  show  how  inaccessible  objects  become  accessible 
by  the  use  of  this  lens. 

The  pictures  were  taken  in  August,  1900.     Many  of  these  nests  are  found  in 
this  vicinity,  and  it  is  said  this  nest  has  been  used  by  wild  geese  in  times  past. 
The  nest  has  been  used  by  ospreys  for  two  years  past,  the  birds  being  the  objects 
of  study  by  the  students  at  the  summer  laboratory. 
University  of  Montana.  MORTON   J.  ElrOD. 


MICRO-CHEMICAL  ANALYSIS. 
XII. 

THE  ANALYTICAL  REACTIONS  OF  GROUP  II. 

Ca,  Sr,  Ba,  —  Gl,  —  Mg,  Zn,  Cd,  Hg. 

CALCIUM. 

The  following  reagents  will  be  found  to  be  the  most  useful  of  those  which 
have  been  proposed  for  the  detection  of  this  element : 

I.  Sulphuric  acid. 

II.  Oxalic  acid. 

III.  Sodium  tartrate. 

IV.  Potassium  ferrocyanide. 
V.  Arsenic  acid. 

VI.     Primary  sodium  carbonate  (HNaCO^). 

/.  Dilute  Sulphuric  Acid  added  to  solutions  containing  salts  of  Calcium,  leads  to- 
the  separation  of  hydrated  Calcium  Sulphate. 

CaCl2  +  H2SO4  =  CaSO^  .   2H2O  +  2HC1. 
Method.— ^o  a  drop  of  the  solution  to  be  tested,  add  a  tiny  drop  of  sulphuric 
acid.     In  a  few  moments  monoclinic  crystals  of  calcium  sulphate  begin  to  form 


and  Laboratory  Methods. 


1243 


Fig-  43- 


\VVv.  "O.ol  w\>n 


near  the  circumference  of  the  test  drop  as  exceedingly  slender,  colorless,  trans- 
parent needles,  either  singly,  in  sheaves,  or  in  star-like  clusters  (Fig.  43).  When 
in  tiny  sheaves  near  the  edge  of  the  drop  the 
crystals  have  often  a  more  or  less  brownish  tint 
when  seen  by  transmitted  light.  Shortly  after 
the  appearance  of  the  bunches  of  needles  at  the 
periphery,  long,  thin,  slender  and  plate-like  prisms 
with  obliquely  truncated  ends  are  formed  through- 
out the  drop.  These  prisms  are  frequently 
twinned,  yielding  so-called  arrow-head  or  swallow- 
tailed  and  X-like  twins.  These  twin  crystals  are 
the  most  characteristic  of  the  forms  assumed  by 
calcium  sulphate  of  the  formula  CaSO^«  2H2O. 

Remarks.  —  The    sulphuric    acid     employed 
should  be   dilute  and   should   not   be  added  in 
excess.     Sulphate  of  sodium  or  of  ammonium  may  also  be  employed,  but  less 
advantageously. 

The  best  results  seem  to  be  obtained  when  the  reagent  is  added  to  a  dilute 
neutral  solution.  If  no  crystals  are  visible  after  waiting  a  short  time,  the  prepar- 
ation may  be  cautiously  concentrated.  This  procedure  (evaporation)  may,  how- 
ever, lead  to  the  separation  of  such  an  amount  of  other  salts  as  to  render  difficult 
the  detection  of  the  crystals  of  calcium  sulphate.  A  better  plan  is  to  hasten  the 
separation  of  the  calcium  salt  by  exposing  the  test  drop  to  the  vapor  of  alcohol. 
This  is  conveniently  performed  as  follows  :  place  a  small  bit  of  filter  paper  on  the 
slide,  a  few  millimeters  from  the  test  drop,  invert  a  25  mm.  watch  glass  over  the 

drop  in  such  a  way  that  part  of  the 

^,,,,^,;;^%„,x,&;;,,^,,,.^,,i,,,,,,,,.,,,,,,^^^^^^^  filter   paper  is  included    under  the 

glass  (see  diagram.  Fig.  44),  add 
^*^''''  sufficient  alcohol  to  the  part  of  the 

filter  paper  outside  the  glass  to  completely  saturate  it,  no  more.  If  the  test 
drop  is  situated  at  the  corner  of  the  slide,  as  is  usually  the  case,  place  another 
slip  alongside  to  support  the  watch  glass,  as  is  shown  in  the  diagram.  Allow 
the  preparation  to  stand  a  few  seconds,  remove  the  glass  and  paper,  and  examine. 

Strong  acids  should  be  absent.  In  the  event  of  their  being  present  add 
ammonium  acetate,  or,  better,  carefully  evaporate  the  solution  to  dryness,  if  possi- 
ble, and  take  up  the  residue  with  water. 

It  must  be  ever  borne  in  mind  that  in  the  presence  of  an  excess  of  salts  of 
Group  I,  the  solubility  of  calcium  sulphate  is  usually  so  greatly  increased  that 
the  detection  of  calcium  by  this  test  is  sometimes  difficult. 

A  more  serious  interference  is  that  of  the  chlorides  of  the  trivalent  metals. 
In  the  presence  of  these  salts  it  is  generally  advisable  to  proceed  as  follows : 
Add  to  the  somewhat  dilute  solution,  ammonium  acetate,  heat  to  boiling,  but 
avoid  long  or  violent  ebullition,  since  in  the  latter  case  the  precipitate  formed 
often  refuses  to  settle.  The  clear  liquid  is  then  separated  from  the  precipitate 
by  drawing  off  on  the  slide,  filtration,  or  by  means  of  the  centrifuge,  is  concen- 
trated if  necessary,  and  tested  for  calcium  with  sulphuric  acid. 


1244  Journal  of  Applied  Microscopy 

Sulphuric  acid  added  to  salts  of  strontium  may,  under  exceptional  conditions 
(if  the  preparation  be  examined  at  once),  yield  a  precipitate  which  closely  resem- 
bles that  given  by  calcium.  These  crystals  of  strontium  sulphate  rapidly  disin- 
tegrate, however,  and  there  results  a  fine  granular  deposit.  This  granular  or 
sandy  precipitate  is  the  form  assumed  by  strontium  sulphate  under  the  condi- 
tions which  ordinarily  obtain  in  this  test.  Barium  is  immediately  precipitated  in 
an  exceedingly  finely  divided  condition,  amorphous  in  appearance.  Any  lead 
which  may  be  present  will  also  be  precipitated  as  a  dense  white  amorphous 
powder. 

If  calcium  sulphate  be  heated  with  a  drop  or  two  of  sulphuric  acid  until  white 
fumes  (SO 3)  are  given  off,  and  the  preparation  allowed  to  cool,  the  calcium  will 
separate  either  as  the  salt  CaSO^  or  as  CaSO^*  H2SO4.  The  crystal  forms 
most  frequently  met  with  are  shown  in  Fig.  4.5.  This  modification  of  the  test  is 
not  satisfactory  for  calcium,  but  is  characteristic  for  barium  and  strontium  (q.  v). 

It  is  not  always  wise  to  conclude  that  calcium  is 
present  when  crystals  separate  on  the  addition  of  sul- 
phuric acid,  which  apparently  resemble  the  star  and 
sheaf-like  aggregates  of  calcium  sulphate,  even  if  the 
crystals  exhibit  oblique  extinction.  For  it  sometimes 
happens  that  other  compounds,  not  calcium  sulphate, 
separate  in  forms  not  to  be  distinguished,  at  first  sight, 
from  the  crystals  of  the  calcium  salt.  Such  instances 
are  fortunately  very  rare.  »pw-o.o.nv«v. 

It  has  been  proposed  to  check  this  test  as  follows  :  ^'s-  ^s- 

After  allowing  sufficient  time  for  the  separation  of  almost  all  the  calcium  as 
CaS04»  2H2O,  draw  off  the  supernatant  liquor;  add  to  the  residue  a  solution  of 
ammonium  carbonate ;  the  crystals  of  calcium  sulphate  are  dissolved  and  highly 
refractive  rhombs  and  grains  of  calcium  carbonate  appear,  which  are  easily 
found  by  examining  the  preparation  between  crossed  nicols.  A  high  power  is 
generally  required. 

In  the  presence  of  borates,  calcium  cannot  be  satisfactorily  detected  by  means 
of  sulphuric  acid.     In  such  an  event  Method  II  can  be  employed. 

Exercises  for  Practice. 

(See  methods  and  exercises  given  under  Strontium  and  Barium.) 

Try  reaction,  after  the  manner  given  above,  on  salts  of  calcium  in  neutral 
solution. 

Try  the  effect  of  precipitating  in  the  presence  of  free  hydrochloric  acid  ;  then 
in  the  presence  of  free  nitric  acid. 

Precipitate  with  dilute  sulphuric  acid,  then  heat,  adding  more  acid  if  neces- 
sary, until  white  fumes  are  given  off,  cool,  breathe  on  the  preparation  and 
examine. 

Try  testing  for  a  trace  of  calcium  in  the  presence  of  a  large  quantity  of  salts 
of  the  elements  of  Group  I. 

Try  effect  of  a  solution  of  ammonium  carbonate  on  crystals  of  calcium  sul- 
phate. 


and  Laboratory  Methods.  1245 

//.   Sa/ts  of  Calcium  give  with  Oxalic  Acid  a  difficultly  soluble  Calciiun  Oxalate. 

CaCl2  +  H2C20^  =  CaC20^«H20  +  2HC1. 

Method. — A  drop  of   a  solution  of  oxalic  acid  is  ^v^_ 

caused  to  flow  into  a  drop  of  the  solution  to  be  tested.  ^A  J~^ 

The  solution  of  the  substance  should  be  neutral  or  may  //  r-T,     ^  f~~i 


contain  a  trace  of  free  nitric  acid.     Calcium  oxalate  is  '^^^j^ 

almost  instantly  precipitated  in  the  form  of  tiny  highly  tb  ^V 

refractive   octahedra  or  rhombs.     Often   crosses   and         „  \^         '^%. 


more  or  less  irregular  bundles  of  minute  needles  are       ^^  ^v     C^        /S} 
obtained  (Fig.  46).  "^ 


Re^narks. — The  composition  of  the  salt,  with  respect  >pw.o.oi™„. 

to  the  amount  of  water  of  crystallization,  varies  accord-  ^'^'  '^^^ 

ing  to  conditions.  It  seems  to  be  quite  generally  accepted  that  when  precipitated 
from  neutral  or  alkaline  solutions  at  room  temperature  the  salt  formed  has  the 
formula  CslC^O^*  SHjO  and  is  to  be  referred  to  the  tetragonal  system;  while  if 
precipitated  from  hot  neytral  or  acid  solutions  or  from  acid  solutions  at  room 
temperature  there  is  obtained  an  oxalate  of  the  formula  CaC204»  H2O,  a  mono- 
clinic  salt.  This  latter  form  of  calcium  oxalate  seems  also  to  result  in  the  pres- 
ence of  an  excess  of  oxalic  acid.  It  follows,  therefore,  that  with  the  conditions 
which  usually  obtain,  there  may  be  precipitated  either  the  salt  with  three  mole- 
cules of  water  of  crystallization  or  the  salt  with  only  one  molecule. 

Free  nitric  acid  greatly  retards  the  reaction,  but  the  presence  of  a  very  little 
of  this  acid  gives  rise  to  the  formation  of  larger  crystals  (because  of  their  being 
more  slowly  formed),  which  are  therefore  more  easily  recognized. 

Calcium  oxalate  is  insoluble  in  acetic  acid  and  in  sodium,  potassium  and 
ammonium  hydroxides,  but  is  readily  dissolved  by  the  mineral  acids. 

Strontium  gives  with  oxalic  acid  an  identical  reaction,  save  that  the  crystals 
of  strontium  oxalate  are  generally  somewhat  larger. 

Barium  oxalate  takes  the  form  of  fibrous  bundles  of  needles  and  is  not  likely 
to  be  mistaken  for  either  calcium  or  strontium. 

Zinc  under  certain  conditions  may  yield  a  zinc  oxalate  difficult  to  distinguish 
from  the  oxalates  of  calcium  and  strontium. 

Magnesium  oxalate  will  separate  in  forms  not  to  be  distinguished  from 
calcium  oxalate  if  the  test  drop  contains  much  acetic  acid. 

Lead  oxalate  may  also  assume  forms  somewhat  resembling  those  of  calcium 
oxalate,  but  after  a  short  time  these  crystals  grow  into  large,  well  developed  prisms. 

Many  other  elements  are  also  precipitated  by  oxalic  acid.  If  such  elements 
are  present  in  large  amount  they  are  apt  to  interfere. 

Owing  to  the  minute  size  of  the  crystals,  testing  for  calcium  with  oxalic  acid 
is  not  always  satisfactory.  As  an  offset  to  this  disadvantage,  chlorides  of  the 
trivalent  metals  and  boric  acid  have  no  effect  other  than  a  retardation  of  the 
reaction. 

In  the  event  of  a  precipitate  of  doubtful  composition  being  obtained,  draw  off 
the  supernatant  liquid,  or  separate  by  means  of  the  centrifuge,  add  to  the  residue 
a  tiny  drop  of  dilute  sulphuric  acid.  Calcium  oxalate  is  dissolved  and  in  a  few 
seconds  the  characteristic  crystals  of  CaS04»  2H2O  make  their  appearance. 


1246  Journal  of  Applied  Microscopy 

Exercises  for  Practice. 

Try  reaction  after  the  manner  given  above,  on  a  salt  of  calcium  in  neutral 
solution.  Try  again  in  the  presence  of  free  HCl ;  then  in  the  presence  of  free 
HNO3. 

Precipitate  calcium  oxalate,  draw  off  the  supernatant  liquor,  and  treat  the  resi- 
due with  dilute  H2SO4.  After  examining  the  preparation,  add  more  acid,  and 
heat  until  white  fumes  appear,  cool  and  examine  again. 

(See  also  suggestions  under  Barium.) 

///.  Sodium  lartrate  added  to  neutral  or  acetic  acid  solutions  of  salts  of  Calcium 
catises  the  precipitation  of  Calcium  lartrafe. 

CaCl2  +  HNaC^H^Oe  =  CaC^H^O,.  4H2O  +  NaCl  +  HCl. 

Method. — To  a  drop  of  the  solution  to  be  tested  add  a  little  sodium  acetate 
and  a  little  acetic  acid,  then  add  a  fragment  of  sodium  tartrate.  In  a  few 
moments  crystals  of  calcium  tartrate  separate  near  the  spot  where  the  reagent 
was  added.  These  crystals  are  large,  colorless,  transparent,  and  well  developed 
prisms  belonging  to  the  orthorhombic  system  (Fig.  47). 

Remarks. — The  reaction  is  apt  to  fail  in  the  pres- 
ence of  free  mineral  acids  owing  to  the  solubility  of 
the  calcium  tartrate ;  hence  the  reason  for  the  addi- 
^^_  \\  <»  tion  of  the  sodium  acetate.     The  calcium  salt  is  also 

\     \    IS  t^^^  "^  soluble  in  sodium  and  potassium  hydroxides. 

A  little  free  acetic  acid  favors  the  formation  of 
well  developed  crystals. 

If  the  solution  is  too  dilute  no  crystals  will  appear 
Fig.  47.  for  some  little  time.     On  the  other  hand,  too  con- 

centrated solutions  give  rise  to  the  immediate  precipitation  of  crystallites  and 
imperfectly  developed  prisms. 

Strontium  gives  a  tartrate  isomorphous  with  that  of  calcium  and  hence  not  to 
be  distinguished  from  the  latter,  although  there  is  a  decided  tendency  on  the 
part  of  the  strontium  salt  to  form  shorter  and  therefore  proportionally  stouter 
prisms. 

Barium  is  precipitated  in  the  form  of  a  fine  powder. 

Lead  is  at  first  thrown  down  as  a  fine  sandy  precipitate  soon  crystallizing  in 
the  form  of  irregular  crystallites  not  to  be  confused  with  either  calcium  or 
strontium. 

In  the  presence  of  magnesium  the  formation  of  the  crystals  of  calcium  tar- 
trate is  greatly  retarded,  and  according  to  Behrens  the  crystals  then  formed  are 
more  slender  and  rod-like ;  in  the  experience  of  the  writer,  however,  the  forma- 
tion of  slender  mixed  crystals  is  seldom  observed. 

The  tartrates  of  potassium  and  ammonium  may  sometimes  be  precipitated  in 

forms  which  at  first  sight  are  difficult  to  distinguish  from  those  of  the  calcium  salt. 

The  testing  for  calcium  with  sodium  tartrate  is  of  little  value  when  dealing 

with  unknown  mixtures,  for  in  addition  to  the  fact  that  the  crystals  of  calcium 

tartrate  cannot  be  distinguished  from  those  of  strontium  tartrate,  salts  of  barium. 


and  Laboratory  Methods.  1247 

lead,  and  potassium  interfere.     The  salts  of  the  trivalent  metals  and  of  boric 
acid  prevent  the  formation  of  characteristic  crystals. 

With  simple  salts  of  calcium  the  reaction  is  a  beautiful  one,  leaving  little  to 
be  desired. 

Exercises  for  Practice. 

(See  under  Strontium.) 

IV.  Potassium  Ferrocyanide  added  to  solutions  of  Calcium  salts  in  the  presence 
of  amtnoniujn  chloride,  gives  rise  to  the  formation  of  a  Double  Ferrocyanide  of  Potas- 
sium and  Calcium. 

CaClg  +  K^Fe(CN)6  =  K2CaFe(CN)6.  SHgO  +  2KC1. 

Method. — To  the  drop  of  the  solution  of  the  substance  ,]   O     ^\ 
to  be  tested  add  a  trace  of  acetic  acid,  then  a  moderate      /^   ^'^   <^  j^ 
amount  of  ammonium  chloride,  stir  thoroughly  and  cause  a      ^^         n  /!>  /% 
drop  of  a  solution  of  potassium  ferrocyanide  to  flow  into  the    ^  I'     o  ^  ck:>  O 
test  drop.   Near  the  zone  of  union  tiny  rectangular  and  square  y>    O     °    ^.V:^ 

plates  are  immediately  precipitated  (Fig.  48).  ^  ^     V  ^ 

Remarks. — In  the  presence  of  free  mineral  acids,  first  add  '  vpvUo.otwnN 

ammonium  acetate  or  sodium  acetate  in  order  to  mitigate  Fig.  48. 

their  action. 

Concentrated  solutions,  with  respect  to  calcium,  lead  to  the  precipitation  of  an 
amorphous  product.  Too  much  ammonium  chloride  produces  a  like  result ;  but 
the  reagent  alone,  in  the  absence  of  the  ammonium  salt,  unless  added  in  consid- 
erable excess,  fails  to  yield  a  deposit  of  crystals.  Barium  gives  large  yellow 
rhombs  with  the  reagent  without  the  addition  of  NH^Cl,  while  strontium  fails  to 
yield  a  precipitate  in  either  case.  Potassium  ferrocyanide  is,  therefore,  some- 
times useful  in  dealing  with  mixtures  of  the  calcium  group,  but  as  a  character- 
istic test  for  calcium  in  simple  salts  it  is  of  but  little  value. 

All  elements  forming  insoluble  or  difficultly  soluble  ferrocyanides  interfere, 
and  in  most  cases  prevent  the  detection  of  calcium  by  the  above  method. 

Exercises  for  Practice. 

(See  suggestions  given  under  Barium.) 

V.  The  addition  of  Arsenic  Acid  to  ammoniacal  solutions  containing  Calcium, 
precipitates  Ammonium  Calcium  Arsenate. 

CaCl2  -i-  H3ASO4  +  SNH^OH  =  NH^CaAsO^.  6H2O  +  2NH4CI  +  3H2O. 

Method. — To  the  drop  of  the  solution  of  the  substance  to  be  tested  add 
ammonium  hydroxide  in  slight  excess,  and  cause  to  flow  into  the  test  drop  a  drop 
of  an  ammoniacal  solution  of  arsenic  acid.  There  is  immediately  produced  a 
heavy  precipitate  rapidly  growing  into  large  crystals  belonging  to  the  orthorhom- 
bic  system.  These  crystals  of  the  double  arsenate  of  calcium  and  ammonium 
generally  take  the  form  of  envelope-like  crystallites,  or  if  separating  from  dilute 
solutions  appear  in  hemimorphic  forms  like  those  of  ammonium  magnesium 
phosphate,  but  of  a  greater  size  (Fig.  49). 


1248 


Journal  of  Applied  Microscopy 


Remarks. — If  much  ammonium  chloride  is 
present,  the  crystals  at  first  formed  will  rapidly 
disappear,  or  there  may  be  no  separation  of  the 
calcium  salt  owing  to  its  marked  solubility  in 
solutions   of  ammonium  chloride. 

The  double  ammonium  arsenates  are  iso- 
morphous  with  the  double  ammonium  phosphates, 
a  fact  which  is  liable  to  give  rise  to  errors  in  the 
interpretation  of  results.  Moreover  it  happens 
that  the  usefulness  of  this  elegant  reaction  is 
unfortunately  restricted,  since  the  elements  of 
the  magnesium  group,  which  are  often  present 
in  mixtures  to  be  tested  for  calcium,  unite  to 
form  double  ammonium  arsenates  of  like  crystal- 
line appearance. 
Strontium  forms  minute  stars  and  tiny  crystalline  grains,  while  barium  yields 
a  dense  precipitate  amorphous  in  appearance. 


Exercises  for  Practice. 

Try  the  above  reaction  on  salts  of  calcium,  strontium  and  barium,  first  alone, 
then  in  mixtures. 

Try  on  salts  of  magnesium,  zinc  and  calcium. 

Try  a  salt  of  calcium  in  the  presence  of  much  ammonium  chloride. 

VI.  Priinary  Sodium  Carbonate  added  to  solutions  containing  Calcium  causes 
the  separation  of  crystalline  Calcium  Carbonate. 

CaCl.,  4-  2HNaC03  =  CaCOg  +  2NaCl    ;    CO,  +  H2O. 

Method. — Cause  a  concentrated  solution  of  the  reagent  to  flow  into  a  drop  of 
a  dilute  neutral,  or  ammoniacal,  solution  of  the  calcium  salt.  In  a  short  time 
very  small  disks  and  rhombs  of  the  compound  CaCOg  appear. 

Remarks. — The  addition  of  the  reagent  in  solid  form  gives  nearly  as  good 
results. 

Warming  the  preparation  increases  the  rapidity  of  tiie  reaction  and  leads  to 
the  formation  of  better  crystals. 

Unless  the  test  drop  is  quite  dilute  an  amorphous  precipitate  results. 

Ammonium  carbonate  can  be  substituted  for  the  sodium  salt,  the  crystals  then 
differ  but  little  if  any  from  those  obtained  as  above.  Normal  sodium  carbonate 
produces  an  amorphous  precipitate. 

Strontium  is  precipitated  in  the  form  of  dumb-bell  shaped  aggregates  and  in 
the  form  of  "  sphero-crystals."  Barium  gives  forms  somewhat  similar  in 
appearance. 

Elements  of  the  magnesium  group  interfere.  Lithium  likewise  interferes. 
But  the  chlorides  of  iron  and  aluminum  and  the  salts  of  boric  acid  have  n9  appre- 
ciable effect  on  the  reaction. 

When  in  doubt  as  to  the  nature  of  a  precipitate  formed  by  the  treatment  with 


and  Laboratory  Methods.  1249 

HNaCOg,  draw  off  the  supernatant  solution,  which  is  easily  done  since  the  crys- 
tals of  calcium  carbonate  adhere  closely  to  the  glass  slide,  wash  the  residue,  and 
then  add  dilute  sulphuric  acid.  If  the  precipitate  is  due  to  calcium,  character- 
istic crystals  of  CaSO^.  ^H.^O  appear. 

In  the  presence  of  a  great  excess  of  the  reagent  a  double  carbonate  of  cal- 
cium and  sodium  separates,  having  the  formula  CaCOg.  NaoCOg.  5H2O,  which 
crystallizes  in  stout  monoclinic  prisms  somewhat  resembling  the  short,  thin  prisms 
of  calcium  sulphate.  Strontium  and  barium  prevent  the  formation  of  the  double 
salt. 

Exercises  for  Practice. 

(See  suggestions  given  under  Zinc.)  E.  M.  Chamot. 

Chemical  Laboratory,  Cornell  University. 


Course  in  Biology  in  the  Horace  Mann  High  School.* 

Although  the" question  of  the  arrangement  of  courses  in  natural  sciences  in 
their  relation  to  other  courses  in  the  high  school  curriculum,  is  as  yet  open  to 
various  opinions  from  different  educators,  it  is,  however,  quite  generally  accepted 
that  the  courses  of  botany  and  zoology  should  come  early  in  any  plan.  In  the 
Horace  Mann  High  School,  the  courses  in  botany  and  zoology  are  given  in  the 
first  year,  and  are  followed  by  the  courses  in  physics  and  chemistry.  It  is 
believed  that  the  work  in  natural  history  appeals  more  strongly  to  pupils  in 
their  earlier  years  of  the  high  school,  than  to  those  who  pursue  the  same  work 
later.  An  objection  to  the  reverse  arrangement  lies  in  the  fact  that  the  applica- 
tion of  the  principles  of  physics  and  chemistry  to  botany  and  zoology  must  be 
repeated  in  the  biological  laboratory. 

The  course  in  zoology  occupies  the  first  half-year,  followed  by  the  botany  in  the 
second  half-year.  This  arrangement  seems  most  satisfactory,  both  because  of  the 
greater  interest  manifest  by  pupils  of  that  age  in  the  study  of  animals  than  plants, 
and  because  the  materials  for  botanical  work  are  more  available  in  spring  than 
in  fall.     Four  forty-five  minute  periods  each  week  are  devoted  to  the  work. 

The  courses  as  outlined  are  complementary  to  each  other;  for  instance,  the 
"cell"  is  studied  in  the  zoological  part  of  the  work  and  is  not  repeated  formally 
in  the  botany  course. 

Throughout  the  course  in  biology  it  is  the  aim  to  develop  the  scientific  method 
of  thought  and  at  the  same  time  impart  to  the  student  as  much  as  possible  of 
the  subject  matter  of  biology,  and  the  economic  importance  of  animals  and  plants. 
To  this  end,  attention  is  given  to  the  form  and  structure  of  living  organisms  and 
to  their  development,  relationships,  physiology  and  ecology. 

The  method  of  presentation  of  the  subjects  of  zoology  and  botany  is  a  depart- 
ure from  the  so-called  logical  method — -that  of  beginning  with  simple  forms  and 
proceeding  to  the  complex — for  the  reason  that  this  is  not  believed  to  be  the  best 
method  to  pursue  with  young  students. 

*  Lloyd,  F.  E.,  and  Bigelow,  M.  A.     Teachers  College  Record,  Vol.  2,  No.  i. 


1250  Journal  of  Applied  Microscopy 

Zop/oi^y  (Bigelow).  What  should  be  included  in  an  elementary  course  in 
zoology  for  secondary  schools,  is  a  problem  upon  which  no  two  persons  will 
exactly  agree.  Certainly  one  point  should  be  borne  constantly  in  mind,  viz., 
that  the  great  majority  of  pupils  will  be  unable  to  pursue  the  subject  further 
than  the  one  course,  for  which  reason  the  subject  matter  should  be  selected 
from  the  standpoint  of  a  liberal  education,  as  distinguished  from  special  and  tech- 
nical education. 

The  tendency  has  been  to  present  courses  embracing  the  detailed  compara- 
tive study  of  the  anatomy  of  animals  to  the  exclusion  of  other  phases  of  the 
subject,  as  the  natural  history,  physiology,  etc.  It  is  now  generally  recognised, 
that  this  imparts  an  extremely  narrow  view  of  the  animal  kingdom  in  its  varied 
aspects.  That  anatomy  should  form  a  part  of  any  course,  is  beyond  question, 
but  to  enter  into  anatomical  details  of  half  a  dozen  types  at  the  expense  of  all 
other  points  of  view  must  be  regarded  as  of  little  value  in  a  liberal  education, 
and  furthermore  as  using  time  which  should  be  devoted  to  undoubtedly  more 
important  phases  of  zodlogical  study.  The  physiological  side  of  animals  has  in  the 
past  received  but  little  attention  comparatively,  but  has  been  found,  in  the  ex- 
perience of  the  present  writer,  a  most  profitable  study  for  secondary  pupils. 
He  believes  that  no  other  phase  of  zoological  study  arouses  a  deeper  interest 
and  appreciation,  or  is  more  spontaneously  applied  by  the  pupils  in  connection 
with  study  of  their  own  life  activities. 

It  has  been,  therefore,  the  endeavor  of  those  who  outlined  the  course  in  zool- 
ogy for  the  Horace  Mann  High  School  to  combine  the  fundamentals  of  morphol- 
ogy, physiology  and  natural  history,  and  thus  give  the  pupils  the  most  valuable 
ideas  of  animals  and  the  widest  view  of  animal  life.  Structure  and  function  are 
studied  in  their  natural  relations.  The  principles  of  physiology  are  introduced 
as  the  different  animals  are  studied  morphologically,  each  principle  being 
exemplified  by  concrete  application.  Such  specific  and  comparative  studies  are 
made  to  lead  to  the  direct  application  of  the  principles  of  comparative  physiology 
to  the  activities  of  the  human  body. 

As  stated  above,  the  method  of  study  is  analytical,  that  is,  the  pupils  begin 
with  multicellular  animals  with  which  they  are  more  or  less  acquainted,  and 
proceed  down  the  scale  of  structural  and  functional  complexity  to  the  simplest 
forms.  By  this  method  pupils  are  introduced  gradually  to  the  compound  micro- 
scope and  are  therefore  able  to  use  it  with  a  degree  of  intelligence  when  they 
undertake  the  study  of  microscopic  organisms.  Furthermore,  the  pupil  is  better 
able  to  understand  the  principles  of  physiology  when  concretely  applied  to 
organs  of  an  animal  in  which  there  is  considerable  physiological  division  of  labor, 
than  were  he  to  begin  with  the  study  of  a  form  in  which  the  various  functions 
are  performed  by  the  single  cell.  From  the  standpoint  of  the  secondary  school, 
the  simple  animal  appears  to  be,  after  all,  the  most  complex  for  the  young 
beginner. 

The  course  therefore,  as  outlined  for  the  Horace  Mann  High  School,  begins  with 
the  complex  animal,  which  is  examined  from  the  several  view-points  of  zoology,  as 
anatomy,  histology,  embryology,  classification  in  connection  with  the  near  allies 
of  the  introductory  type,  distribution  and  ecology,  general   fundamental  princi- 


and  Laboratory  Methods.  1251 

pies  of  physiology,  habits  of  life  and  life  history.  To  be  sure,  none  of  these 
phases  go  far  into  details,  but  it  is  the  aim  to  lay  a  foundation  which  will  make 
later  study  of  animals,  from  whatever  standpoint,  more  interesting  and  more 
intelligible,  because  there  is  included  in  the  foundation  work  those  great  prin- 
ciples of  animal  structure  and  function  which  are  of  wide  interest  and 
application.  With  a  foundation  thus  gained  from  the  careful  study  of  a  suitable 
representative  type,  the  pupil  is  usually  eager  to  study  each  animal  as  it  is 
brought  before  him  as  thoroughly  as  the  introductory  type,  that  is,  from  the 
various  aspects  of  zoology. 

As  an  introductory  type,  the  crayfish  has  some  decided  advantages  over  other 
forms  frequently  used  for  beginners.  In  the  Horace  Mann  High  School  the  cray- 
fish is  viewed  from  the  view-points  indicated  above.  The  study  embraces  lectures, 
readings,  recitations  and  laboratory  work.  The  author  gives  a  complete  out- 
line of  the  subject  matter  as  presented  to  his  classes,  which  must,  unfortunately, 
in  this  review,  be  reduced  to  only  the  general  heads,  which  are  as  follows  : 
"  General  External  Structure  of  the  Crayfish,"  "  General  Internal  Structure," 
"  Introductory  Microscopic  Work  and  Elementary  Histology,"  "  Elementary 
Embryological  Study,"  "  General  Principles  of  Animal  Physiology  as  Illustrated 
by  the  Crayfish,"  "  Summary  of  the  Introduction." 

This  work  is  followed  by  a  more  limited  survey  of  forms,  both  invertebrate 
and  vertebrate,  which  are  studied  chiefly  from  the  standpoint  of  external  struc- 
ture, although  other  phases  are  considered  as  time  permits.  These  forms  are 
presented  in  the  following  order : 

1 .  Crustaceans. 

2.  Arachnids. 

3.  Insects. 

(a)  grasshopper ;    (/>)  butterfly ;  (c)  life   history    of    cricket,     beetle, 
bee,  ant,  fly,  may-fly,  cicada. 

4.  Worms. 

(a)  earth  worms  ;  (/-)  flat  worms  ;  (c)  round  worms. 

5.  Coelenterates. 

(a)  hydra ;  (<^)  hydroid   colony  (Pennaria,   Obelia,   Parypha  or  Cam- 
pan  ularia)  ;  (c)  corals, 
(j.      Sponges. 

7.  Protozoa. 

8.  Echinoderms. 

(a)  starfish;  (S)  sea-urchin. 

9.  MoUusks. 

{a)  gasteropods  ;  {F)  lamellibranchs  ;  {/)  cephalopods. 

10.  Vertebrates  {Jive  weeks ^ 

{a)  amphibians ;  {p)  fishes ;  {c)  reptiles ;  {(i)  birds ;  (<?)  mammals. 

The  above  outline  may  be  made  the  basis  for  a  full  year  course  with  much 
more  satisfactory  results,  perhaps,  than  for  a  half  year  course,  as  a  year's  time 
is  none  to  long  in  which  to  cover  the  field  indicated. 

Botany  (Lloyd). — In  general  the  methods  and  aims  pursued  in  the  course  in 
botany  are  similar  to  those  indicated  above  for  the  course  in  zoology.     It  is  the 


1252  Journal  of  Applied  Microscopy 

endeavor  to  view  plants,  in  all  their  phases,  giving  the  student  the  opportunity  to 
acquaint  himself  with  the  essentials  of  plant  structure,  physiology  and  ecology. 
The  work  is  begun  with  familiar  plants  and  is  carried  on  through  all  of  the 
groups  of  spermatophytes,  and  thallophytes.-  A  significant  feature  of  the  course 
lies  in  the  fact  that  those  subjects,  which  may  be  found  sufficiently  treated  in  the 
numerous  text-books  and  laboratory  guides  in  prevalent  use,  are  treated  of  only 
briefly ;  the  time  being  spent  on  those  subjects  which  are  not  so  satisfactorally 
presented  to  the  student  through  the  literature  within  his  reach. 

Another  point,  in  which  the  course  is  especially  advantageous  for  young 
pupils,  is  that  emphasis  is  placed  upon  the  study,  first  of  all,  of  the  fruit  rather  than 
the  seed,  thus  obviating  the  difficulties  which  present  themselves  in  the  study  of 
some  seeds.  The  fruit  is  studied  in  different  stages  in  order  to  impart  to  the 
student  the  idea  of  development  rather  than  a  statical  conception  of  the  matter 
in  hand. 

Attention  is  paid  to  foods  in  plants,  to  digestion  and  to  absorption,  the 
method  being  physiological  rather  than  microscopical. 

Especial  attention  is  devoted  to  the  problem  of  digestion,  in  which  the  essen- 
tial similarity  of  plants  and  animals  is  brought  into  prominence.  In  this  con- 
nection the  writer  recommends  the  cocoanut  and  the  date,  as  most  valuable 
material  for  demonstrating  the  morphological  facts  involved. 

The  subject  of  sexual  reproduction,  although  not  neglected,  is  deemed  less 
profitable  for  young  students  than  the  study  of  the  vegetative  body  and  the  more 
readily  observable  phenomena  of  adaptation.  However,  it  is  found  that  the  essen- 
tials of  the  subject  may  be  clearly  brought  out  in  a  study  of  such  forms  as  Spiro- 
gyra  and  Vaucheria.  In  the  study  of  seed  plants  in  this  connection,  somewhat 
more  attention  is  paid  to  details,  and  the  important  morphological  facts  involved 
are  demonstrated  by  means  of  charts  and  preparations.  At  this  time  also  is 
demonstrated  the  phanerogamic  embryo  in  earlier  and  later  stages  of  develop- 
ment, and  so  the  study  of  the  life  cycle,  which  was  commenced  in  the  study  of 
the  fruit,  is  rounded  out  to  completion. 

Like  the  outline  for  the  work  in  zoology,  the  course  in  botany  is  outlined  in 
detail.     Of  this  outline  only  the  headings  can  be  given  here : 

I.       The  Structure  and  Physiology  of  Plants. 

1.  The  Lima  Bean, 
(a)  fruit;  (/»)  seed. 

2.  T/ie  Indian   Com. 

{a)  fruit ;  {b)  embryo. 

3.  Tlie  Castor  Oil  Planf. 
(a)  fruit ;  (/')  seed. 

4.  T/ie  Pine. 

5.  Studies  in  Germination. 

(a)  absorption  of  water;  (/>)  rupture  of  seed  coats  ;  (r)  manner  in  which 
seedlings  break  through  the  ground  :  epicotyl  (pea),  hypocotyl  (castor 
oil),  cotyledon  (onion)  ;  (ci)  development  of  organs  in  embryo ;  (e) 
behavior  of  cotyledons  during  germination  ;  (y)  earlier  leaves  com- 


and  Laboratory  Methods.  1253 

pared  with  adult  form  ;  (^^)  production  of  otlier   shoots  (pea)  after 
destruction  of  plumule  ;    (//)  etiolation. 

6.  Respiratio)!. 

7.  A'ntritioii  {Foods). 

(a)  proteids  ;  (/>)  starch;  (c)  sugar;  (d)  cellulose;  (e)  oils;  (/")  min- 
eral substances. 

8.  T/ie  Digcstio)!  and  Absorption  of  Foods. 

9.  J  he  Stniiturc  and  Functions  of  Roots. 

(a)  root  system  of  ordinary  type  ;  (/>)  absorption  by  roots ;  (f)  mechan- 
ical fixation  of  the  plant  by  means  of  roots  ;  (d)  storage  of  food  in 
roots  ;  (e)  modification  of  roots  correlated  with  parasitic  habit  ;  (/) 
mycorhizal  roots,  root  tubercles;  (g)  air  roots;  (A)  modification 
of  roots  correlated  with  respiration  ;  (  /  )  contractile  roots. 
10.      7//e  Structure  and  Functions  of  the  Shoot. 

(a)  the  stem  ;  (d)  functions  of  the  stem  ;  (c)  the  leaf ;  (a)  functions 
of  a  typical  foliage  leaf  ;   (e)  the  bud. 

II.  A  Study  of  Types  of  the  Groups  of  Plants. 

1 .  Spermatophvta  :  Angiosper7ns. 

(a)  Willow  ;  {p)  Hazel  and  Alder,  Elm  and  Maple  ;  (<r)  Calla  Lily  ;  {d) 
Hyacinth;  {e)  Cypripedium  ;  (/)  Strelitzia  ;  (^)  Buttercup;  (//)  Ger- 
anium ;  (/)  Abutilon  or  Malvavistrum  ;  (  /)  some  leguminous  flower  ; 
(/')  Azalea  ;  (/)  a  cactus  flower  ;  (w)  Begonia  ;  (//)  Dandelion  or  field 
daisy. 

2.  Gymiiosperms. 

Fir,  spruce  or  pine. 

3.  Pteridophyta. 

(a)  Aspidium  ;  (/')  Equisetum  ;  (c)  Isoetes  ;  {d  )  Marsilia  ;  {e)  Pillularia  ; 
(/)  Salvinia  ;  {g)  Azolla  ;  (//)  Lycopodium  ;  (/)  Selaginella. 

4.  Bryophyta. 

{a)  Polytrichum  ;  {If)  Pogonatum  ;  {c)  Georgia  pellucida  ;  (//)  Funaria. 

5.  Hepaticie. 

(a)  Radula;  (/;)  Frullania ;  (c)   Scapania  ;  (d)  Marchantia  ;  (e)  Lunularia. 

6.  Fungi. 

(a)  Agaricus  ;  (/>)  Puccinia  or  Uromyces  ;  (c)  Morcella  ;  (d)  Sclerotinia  ; 
(e)  Lichens  ;  (/)  Claviceps  ;  (g)  Cordyceps  ;  (//)  Penicillium  ;  (/) 
Microsphaera  alni ;  (j)  Mucor ;  (/C')  Saprolegnia ;  (/)  Yeast;  (m) 
Algae ;  (n)  Schizophyta.  c.  w.  j. 


The  second  summer  session  of  the  Laboratory  of  Biology  of  Tufts  College 
will  open  at  South  Harpswell,  Maine,  on  June  fifteenth  and  continue  until  about 
September  first.  Courses  in  Invertebrate  Zoology,  Vertebrate  Zoology,  Botany, 
and  Embryology  will  be  given,  as  well  as  opportunity  for  special  research  work. 
The  laboratory  is  well  equipped  with  apparatus  for  ordinary  investigations  and 
has  a  library  of  several  hundred  volumes  and  pamphlets  selected  with  reference 
to  the  work  to  be  done.  These  advantages  are  placed  at  the  disposal  of 
students  for  the  consideration  of  a  very  reasonable  fee.  Communications  should 
be  addressed  to  the  director,  J-  S.  Kingsley,  Tufts  College,  Mass. 


1254  Journal  of  Applied  Microscopy 

Journal   of  Itis  very  evident  that  bacteriological 

A          1*     J     1\yi  •  methods  in  the  diagnosis  of  germ  dis- 

/\ppilCQ     IVllCr OSCOpy  eases  cannot  reach  their  highest  degree 

^"''  of  usefulness  in  the  prevention  of  epi- 

Laboratory  Methods.  ^^^^-      ^^  ^.^  ^^  i^^i^,,^  ^^^-^ 


Edited  by  L.  B.  ELLIOTT.  ^^^    general    public,    and    even    many 

medical  men,  are  better  informed  as  to 


Issued  Monthly  from  the  Publication  Department  (.1,^     a  Urwtn^  "     onH     "  «'Vnrc  "     /->f      fVi/=cp> 

of  the  Bausch  &  Lomb  Optical  Co.,  "^"^      HOWS      ana      \\  nys       ot    tnese 
Rochester,  N.  Y. methods,  and  are  thus  enabled  to  un- 


SUBSCRIPTIONS:  derstand  the  necessity  of   conforming 

One  Dollar  per ^Year.^^To^Fordgn^Countries,  $1.25       gtrictly      tO      their      requirements.         No 

more   convincing  proof   of  the  benefit 


The  majority  of  our  subscribers  dislike  to  have  their  i      •         i    i  •         r  i 

files  broken  in  case  they  fail  to  remit  at  the  expiration  denVCd    by    a  Community    irom  CUltUrC 
of  their  paid  subscription.     We  therefore  assume  that  no                 ,        ,               i  ■  n  i 

interruption   in   the   series   is   desired,  unless   notice   to  mCthodS    and  Strict    adherence    tO    baC- 
discontinue  is  sent. 


■  teriological  precautions  ought  to  be 
needed  than  is  shown  in  a  recently 
published  report*  of  some  instances,  where  the  evidence  of  the  bacteriologist  in 
the  diagnosis  of  diphtheria  was  taken,  in  spite  of  more  or  less  opposition  from 
practicing  physicians  not  thoroughly  acquainted  with  the  value  of  the  methods,  as 
a  basis  for  treatment  and  preventive  measures.  As  a  result  in  these  instances, 
the  positive  cases  were  promptly  identified  and  isolated,  and  proper  treatment 
applied  in  time  to  check  the  progress  of  the  disease;  while  negative  cases, 
however  suspicious  their  appearance  in  ordinary  clinical  diagnosis,  were  safely 
dismissed.  The  bacteriological  methods  recommended  for  the  control  of 
diphtheria  may  be  summarized  with  great  clearness  in  early  diagnosis,  early  use 
of  antitoxin,  strict  quarantine,  release  on  negative  cultures  only,  and  thorough 
disinfection.  This  procedure  is  based  on  the  natural  history  of  the  disease,  and 
is  the  most  logical,  well  defined  and  satisfactory  course  to  pursue  with  a  sus- 
pected case.  The  culture  methods  are  simple  but  most  reliable,  and  their  more 
rapid  introduction  and  universal  application  are  retarded  by  neglect  and  igno- 
rance on  the  part  of  physicians,  boards  of  health,  and  men  holding  positions 
pertaining  to  the  public  health,  and  prejudice,  due  to  ignorance,  on  the  part  of 
the  laity.  These  difificulties  must  be  overcome  by  the  thorough  instruction  of 
medical  men  in  the  use  of  the  methods  and  the  instruction  of  the  people  through 
the  public  schools.  The  latter  subject  is  just  now  receiving  much  attention 
from  men  in  charge  of  courses  of  study  for  pupils  in  high  schools. 

A  subject  of  so  much  importance  to  society  as  the  prevention  of  the  spread 
of  germ  diseases  should  receive  sufficient  attention  in  all  public  and  private 
schools,  to  inform  the  students  of  the  paramount  necessity  not  only  of  taking 
every  precaution  against  contracting  infectious  diseases,  but  when  once  infected 
of  submitting  to  the  methods  prescribed  by  the  physician  ;  the  result  of  which 
would  ultimately  produce  a  public  opinion  heartily  in  favor  of  better  sanitation, 
improved  methods,  and  strict  precautions  in  the  preservation  of  the  public  health. 


*  The  Control  of  Diphtheria  in  small  cities  and  country  districts  from  the  Bacteriological 
Standpoint.     Veranus  A.  Moore,  M.  D.,  Cornell  University. 


and  Laboratory  Methods.  1255 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Juel,  H.  0.    Vergleichende  Unterschungeniiber  A  preliminary  note  announcing  the  dis- 

typische  und  parthenogenetische  Fortpflan-  covery  of  parthenogenesis  in   Antenna- 

zung  bei  der    Gattung  Antennana.      Kongl.  ■'         '^                 "  _ 

Svenska    Ventenskaps-Akademiens     Hand-  >'ia  alpiiia   appeared  in   the    Botanisches 

lingar.  U'.  3-56,  pis.  1-6,1900.  Centralblatt  about   two  years  ago.      It 

was  also  noted  at  that  time  that  Antenuaria  dioica  presented  a  very  different 
developmental  history.  The  author's  subsequent  work  upon  these  two  species 
is  described  in  great  detail  in  the  present  paper. 

In  the  nucellus  of  Antcjina?'ia  dioica  the  sequence  presents  nothing  excep- 
tional, the  mother  cell  of  the  megaspore  producing  four  potential  megaspores, 
one  of  which  continues  to  develop  at  the  expense  of  the  other  three  and  becomes 
the  embryo-sac,  just  as  in  other  Compositse.  The  antipodal  cells  continue  to 
divide  and  form  a  tissue,  nineteen  cells  appearing  in  one  section  in  one  of  the 
author's  figures.  Fertilization  of  the  ^gg  takes  place  in  the  usual  manner,  but 
no  double  fertilization  could  be  detected.  At  the  first  division  of  the  nucleus  of 
the  megaspore  mother  cell,  a  reduction  in  the  number  of  chromosomes  takes 
place.  The  production  of  a  row  of  four  potential  megaspores  is  regarded  as  a 
true  tetrad  formation. 

In  Antennaria  alpina  the  mother  cell  of  the  megaspore  becomes  the  embryo- 
sac  directly,  just  as  in  Li/iion,  without  giving  rise  to  a  row  of  four  potential  mega- 
spores, but,  unlike  Liliiim  and  other  plants,  it  shows  no  reduction  in  the  number 
of  chromosomes.  Prof.  Juel's  previous  statement  that  the  embryos  develop  with- 
out fertilization  and  that  there  is  no  fusion  of  polar  nuclei,  is  repeated  with  more 
detailed  evidence. 

Only  one  plate  is  from  camera  lucida  drawings,  the  other  five  being  taken 
from  photographs  and  photo-micrographs.  The  latter  were  made  with  a  2  mm. 
oil  immersion  objective.  The  exposures  were  about  two  minutes  long  and  no 
ray  filters  were  used.  While  the  figures  show  the  stages  fairly  well,  they  also 
show  the  limitations  of  photo-micrography  in  its  present  stage  of  development. 

c.  J.  c. 

Smith,  R.  Wilson.     The  Achromatic  Spindle  in       The  object  of  this  work  was  tO   extend 

Z.^Zll'^tAt.t^r^^^ '"""'      ""'   '"'^^'-'^  knowledge  of  the  cytology 

of  the  vascular  cryptogams.  In  the 
spore  mother  cell  of  Ostnunda  rega/is,  Smith  finds  that  the  spindle  originates  out 
of  a  granular  zone  of  cytoplasmic  material  which  accumulates  about  the  nucleus. 
The  granules  of  this  material  arrange  themselves  into  short  rows  concentric  with 
the  nuclear  membrane.  These  rows  of  granules  become  massed  at  opposite  sides 
of  the  nucleus  and  eventually  become  the  cones  of  a  bipolar  spindle.     The  spin- 


1256  Journal  of  Applied  Microscopy 

die  appears  to  be  bipolar  from  the  beginning,  but  no  bodies  that  could  be  inter- 
preted as  centrospheres  were  found.  Although  tripolar  spindles  were  occasionally 
met  with,  Smith  is  certain  that  they  are  not  normal  stages  in  the  development  of 
the  spindle,  and  comes  to  the  interesting  conclusion  that  the  spindle  in  Osmunda 
does  not  pass  through  a  multipolar  stage.  To  the  reviewer  the  evidence  for  such 
a  conclusion  would  have  been  more  convincing  had  the  appearance  of  tripolar 
spindles  been  accounted  for  or  had  more  stages  in  the  formation  of  the  cones  of 
the  bipolar  spindle  been  figured. 

For  fixing  the  material  chrom-acetic  acid  and  Flemming's  weaker  solution 
were  employed.  Chloroform  was  used  to  precede  the  infiltration  of  paraffin. 
The  stains  that  gave  the  most  satisfactory  differentiation  were  iodine-green  and 
acid-fuchsin,  and  safranin  and  gentian-violet.  A.  A.  Lawson. 

Brown,  H.  T.,  and  Escombe,  F.     Static  Diffusion  The  authors  investigated  the  laws  gov- 

of  Gases  and  Liquids  in  Relation  to  the  As-  •           j-rc      •          ..u           i.                           n 

similation  of  Carbon  and  Translocation  in  ^''^^"g    diffusion    through    very     small 

Plants.     Phil.  Trans.  Roy.  Soc.  of  London,  apertures.       They  find  that  : 

193:  223-292,  1900.  ^^^  ^j^g  amount  varies  directly  as 

the  diameter  of  the  orifice.  This  holds  for  openings  5  or  6  mm.  or  less  in  diam- 
eter. It  follows  that  diffusion  through  holes  1  mm.  or  less  is  very  rapid  per  unit 
of  area. 

(2)  When  the  distance  between  the  holes  is  ten  times  the  diameter  of  the 
holes  themselves,  the  amount  of  diffusion  is  the  same  as  when  a  septum  is 
wanting. 

(3)  These  laws  hold  for  both  solutes  and  gases. 

By  analogy  with  the  lines  of  force  about  an  electrified  disc,  the  investigators 
have  reached  the  same  conclusions  mathematically. 

Applying  these  results  to  plant  structures  the  authors  conclude  that :  (a)  The 
open  stomata  of  a  normal  mesophyte  (^Helianthus  anmms)  are  sufficient  for  the 
diffusion  of  several  times  as  much  CO  2  as  the  plant  actually  uses.  There  is  no 
need  then  for  more  stomata.  (b)  The  limitation  of  the  amount  of  CO 2  absorbed 
is  to  be  looked  for  in  the  resistance  to  diffusion  offered  by  the  cell  wall,  (c) 
The  stomata  are  sufficient  to  account  for  transpiration,  (d)  The  translocation  of 
foods  is  probably  more  largely  a  phenomenon  of  diffusion  than  was  supposed, 
since  .7  per  cent,  of  opening  in  cell  walls  would  permit  30  per  cent,  of  free  dif- 
fusion. The  paper  confirms  Blackman's  results  (1895),  and  discusses  the  physi- 
cal laws  underlying  them.  T.  C.  Frye. 
Chicago. 


The  Martha's  Vineyard  Summer  Institute,  Hyde  Park,  Mass.,  announces  the 
summer  session  of  the  School  of  Nature  Study  to  be  held  during  July  and 
August,  1901.  Besides  the  School  of  Natural  Study  the  Institute  embraces 
Schools  of  Methods,  Oratory,  Languages,  Mathematics,  Science,  and  Art ;  in- 
formation concerning  which  may  be  obtained  from  the  President  of  the  Institute, 
Dr.  Wm.  A.  Mowry,  Hyde  Park,  Mass. 


and  Laboratory  Methods.  ^^^^ 


CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  Sage  College, 

Ithaca,  N.  Y. 

CURRENT  LITERATURE. 

Sjobring,  N.     Ueber  das  Formol  als  Fixinings-      -p^e  author  says  that  the  opinions  of 

fliissigkeit.      Allgemeines     ueber  den    Bau  ^ 

der  lebenden  Zellen.     Anat.  Anz.  17:  273-      formol  as  a  fixing  fluid  are  not,  in  gen- 

304,  3  Abb,  1900.    Abstract  in  Zeis.  f.  wiss.      gj-^i   favorable.     Many  writers  say  it  is 

Mikr.  u.  f.  Mikr.  Techn.  17:  337-340,  1900. 

decidedly  unfitted  for  the  finer  preserva- 
tion of  cell  tissue.  According  to  the  writer  formol  does  not  merit  this  condemna- 
tion, caused  by  the  fact  that  these  writers  have  failed  to  discover  the  small  point 
upon  which  the  successful  use  of  formol  depends.  A  distinction  is  made 
between  the  "  Formol  "  of  the  firm,  Meister,  Lucius,  u.  Briining,  Hochst  a 
Main,  and  the  "  Formalin  "  of  Actien  (Sobering)  of  Berlin.  Formalin  is  not 
so  suitable  for  histological  work  as  formol.  It  must  be  understood  that  formol 
is  only  a  fixing  agent,  not  a  hardener.  Material  fixed  in  formol  should  be 
hardened  in  95  per  cent,  alcohol  for  48  hours  or  longer,  at  least  mammalian 
tissue  should  be  so  treated.  For  tissue  containing  much  water,  different 
strengths  of  alcohol  are  desirable.  For  Anodonta,  50  per  cent,  alcohol  is  most 
favorable.  It  is  probably  this  point  that  causes  the  various  results  obtained  by 
authors  in  the  use  of  formol.  The  action  of  formol  on  tissue  is  probably  an 
oxidation  similar  to  that  of  osmic  acid.  The  first  requisite  for  a  successful  fixing 
fluid  is  that  it  should  be  approximately  isotonic  with  the  protoplasm.  Formol, 
in  comparison  with  the  tissues  of  mammals,  should  have  the  isotonism  of  8  to  10 
per  cent,  formaldehyde  (1  pt.  formol  to  4  of  water),  but  not  all  tissues  have  the 
same  tension.  For  mammals,  the  following  process  gives  the  best  results  :  Fixa- 
tion in  formol,  1  :4  water  for  48  hours  or  longer;  direct  into  95  per  cent,  alcohol 
for  at  least  two  days.  In  this  way  the  resting  nuclei,  red  blood  cells,  intracellular 
cement  between  epithelial  cells,  fibrin,  fibrinoid  degeneration  of  connective 
tissue,  gelatinous  and  other  albuminous  exudates,  are  especially  well 
preserved.  The  metakinetic  stages  of  mitosis  are  not  successfully  obtained. 
Formol  is  not  especially  good  for  nerve-tissue — preservation  is  good,  but  the 
staining  capacity  is  lessened — stronger  and  warmed  stains  are  necessary. 
Some  methods  of  staining  are  especially  applicable  after  formol  fixation.  Heiden- 
hain's  iron-alum-haematoxylin  is  especially  good  when  used  in  a  modified  way. 
Strong  solutions  were  found  most  effective.  Ha^matoxylin  of  a  concentrated 
aqueous  solution  and  iron-alum  for  the  mordant,  in  a  5  per  cent,  solution,  allowed 
to  act  for  three  hours.  For  differentiation,  the  same  strength  or  a  one-half  dilu- 
tion was  used.  The  stain  was  allowed  to  act  for  one  hour,  with  some  warming. 
Anilin  blue  was  used  for  a  preparatory  stain  ;  concentrated  alcoholic  solution  in 
50  per  cent,  alcohol   was  diluted   one-half  with  water.     Crystal-violet  in  a  1  per 


1258  Journal  of  Applied  Microscopy 

cent,  solution  in  50  per  cent,  alcohol ;  counterstaining  in  orange  or  eosin  is  very 
satisfactory.  This  brings  out  very  clearly  and  beautifully  almost  all  kinds  of 
granules.  Bordeau  red  is  not  so  good  as  with  sublimate  and  alcoholic  fixation. 
Another  method  which  gave  good  results  in  many  cases  where  iron-  haematoxylin 
failed,  is  anilin-water,  fuchsin-anilin  blue,  according  to  Lugol's  mixture.  Ehrlich's 
triacid  is  very  good  in  tissues  where  cell  infiltration  has  occurred,  but  is  uncer- 
tain in  action.  The  stain  must  be  made  very  concentrated  in  the  original  solu- 
tion by  warming  it  during  the  process.  Taking  up  the  stain  with  blotting  paper, 
and  decolorizing  with  95  per  cent,  alcohol  is  the  best  method.  The  results  after 
treatment  with  alcohol,  acid,  neutral  or  alkaline,  differentiate  connective  tissue 
cell  granules,  neutrophil  and  eosinophil  granules,  plasma-cell  granules,  and 
clasmatocyte  granules.  Formol  is  as  good  for  purposes  of  studying  the  cell 
body  as  is  Flemming's  solution  for  the  study  of  the  nucleus.  It  is  especially 
necessary  for  the  pathologist  to  make  himself  familiar  with  cell  morphology 
under  all  normal  and  post  mortem  conditions  before  the  method  is  applied  to 
pathology.  The  method  gives  very  fine  differentiations,  but  must  be  used  with 
great  care.  As  a  point  of  warning,  the  author  speaks  of  the  necessity  of  killing 
the  animals  used  for  cellular  physiology  by  other  means  than  chloroform,  since 
the  action  of  this  substance  is  to  decrease  the  staining  capacity  of  red  blood- 
cells  in  iron-hsematoxylin,  and  to  make  noticeable  changes  in  the  cell  granules, 
especially  those  of  the  liver  and  marrow  cells.  Guinea-pigs  are  killed  by  a  blow 
on  the  back  of  the  neck,  and  mice  by  cutting  off  the  head  with  shears,  etc. 
There  is  great  difference  in  the  staining  capacity  of  different  elements  of  the  cell ; 
liver,  kidney,  and  bone-marrow  are  the  easiest  to  stain,  while  those  of  the  intestine 
or  stomach  epithelium  are  difficult,  and  the  larger  granules  of  the  salivary  glands 
always  remain  unstained.  Those  elements  that  do  not  stain  in  iron-haematoxylin 
can  often  be  brought  out  by  anilin-fuchsin,  Ehrlich's  triacid,  etc.  If  large  and 
small  granules  are  present  in  the  cells,  they  often  stain  differentially. 

A.   M.  C. 

Jolly,  M.  J.     Recherches  sur  la   division   indi-  The    author    gives    first    an     extended 

recte  des  cellules    lymphatique  granuleuses  historical   review   of   the   subject.      His 

de  la  mole  des  os.     Arch,   d  Anat.  Microsc.  . 

3:  168-228,  2  plche.,  1900.    (Reviewed  in  studies  were  chiefly  on  adult  mammals. 

Zeit.  f^wiss.  Mikros.  u.  f.  Mikros.  Tech.  17:  i^  ^^g  laboratory,  the  Cobaya  rabbit, 

360-363,  1900).  ■'                     •' 

rat,   mouse,   dog,   and    cat ;    from  the 

market,  calf,  sheep,  hare.  Some  rarer  mammals,  the  bat  and  mole  ;  finally,  man  in 
several  stages,  were  studied.  Besides  these,  the  pigeon,  hen,  duck,  and  lizard  were 
also  examined.  Red  marrow  from  the  long  bones  was  always  used,  in  addition  to 
that  from  the  sternum  and  the  body  of  the  vertebrae.  In  the  lower  mammals 
there  is  a  distinct  separation  between  the  red  and  yellow  marrow,  which  is  less 
easily  found  in  man.  This  is  partly  due  to  the  uncertainty  as  to  whether  the 
marrow  was  perfectly  normal  in  the  material  in  use.  The  bone  marrow  of 
thirteen  infants  was  examined,  varying  in  age  from  eight  days  to  two  years,  which 
died  of  various  diseases.  The  structure  of  the  femur  was  always  the  same  ;  in 
the  middle  of  the  diaphysis  is  a  short  canal  filled  with  red  marrow  ;  farther 
towards  the  epiphysis  is  a  spongy  bone-tissue  filled  with  red  marrow.  This  was 
very  rich  in  cells,  excepting  in  the  case  of  two  individuals  who  died  of  hereditary 


and  Laboratory  Methods.  1-59 

syphilis.  In  these  the  marrow  was  comparatively  deficient  in  lymph  cells.  With 
the  adult  man  red  marrow  was  always  found  in  the  spongy  tissue  of  the  sternum 
and  the  bodies  of  the  vertebrae ;  and  being  more  easy  of  access,  this  was  usually 
the  source  of  material  for  this  investigation.  To  obtain  the  marrow  fresh,  the 
bones,  after  being  freed  from  all  other  tissue,  are  split  lengthwise  by  a  sharp 
stroke  on  a  strong  knife.  The  lyrnph-cells  of  the  marrow  were  examined  fresh  in 
blood  serum,  and  also  after  fixation  in  70  per  cent,  alcohol,  and  staining  in  picro- 
carmin,  and  mounting  in  glycerin.  For  nuclear  studies,  Malassez's  method  of 
1882  was  used.  A  slide  is  laid  gently  upon  the  fresh  marrow,  and  the  smear  is 
fixed  in  osmic  acid  fumes.  The  results  of  this  method  are  an  improvement  on 
the  old  smear  method,  since  it  avoids  tearing  and  distorting.  Such  a  spot 
shows  three  zones ;  a  central,  the  largest  of  considerable  thickness  not  available 
for  study,  a  peripheral,  very  thin,  of  a  single  layer  of  cells.  This  is  generally 
changed  by  drying  slightly.  Also,  there  is  a  middle  zone,  thin  enough  for 
observation  and  thick  enough  to  show  no  effects  of  drying.  Fixation  fluids  are 
poured  directly  upon  the  slide  ;  later  washing  loosens  the  thick  central  piece, 
but  the  rest  remains  in  place.  Besides  osmic  acid,  the  author  has  used  Flem- 
ming's  solution,  sublimate  with  platinum-chloride,  and  Zenker's  fluid.  Osmic 
acid,  1  per  cent,  solution,  for  30  to  60  seconds,  gave  good  preparations,  but 
Flemming  was  still  more  satisfactory  in  a  strong  solution.  For  staining,  the  fol- 
lowing combinations  were  used  :  haematein  and  eosin,  haematein  and  aurantia, 
hsematein  and  acid  fuchsin,  methylen  blue  and  eosin,  methylen  green  and  acid 
fuchsin.  The  Ehrlich-Biondi-Heidenhain  triacid  mixture  and  safranin,  with 
potassium  permanganate  (1  to  100),  as  a  mordant.  According  to  Henneguy's 
method,  gentian-violet,  thionin,  and  polychromic  methylen  blue  of  Unna,  were  all 
used.  The  general  method  of  preparation  was  as  follows :  a  small  spot  of 
marrow  is  fixed  in  Flemming's  fluid  (strong)  10  to  15  minutes,  washed  out  in 
running  water  for  15  minutes,  bleached  in  iodin  solution  (1  to  100.95  per  cent, 
alcohol)  for  one  second,  washed  in  95  per  cent,  alcohol  to  remove  the  iodin  ; 
wash  out  in  water,  stain  with  a  solution  of  eosin  containing  glycerin  (dry  eosin 
1  part,  95  per  cent,  alcohol  20  parts,  glycerin  50  parts,  and  water  50  parts)  for 
a  long  time,  so  as  to  over  stain.  Decolorize  in  alcohol,  and  stain  the  nucleus 
with  the  following  haematein  (haematein  1  part,  95  per  cent,  alcohol  25  parts,  5 
per  cent,  solution  of  ammonia  alum  200  parts) ;  wash  in  water,  alcohol,  clear  in 
clove  oil,  and  mount  in  Damar  balsam.  In  such  preparations  the  middle  of  the 
spot,  as  already  mentioned,  is  thick  and  badly  fixed.  This  is  removed  with  a 
needle,  if  it  has  not  already  fallen  out  in  the  various  washings.  The  cells  of  the 
peripheral  zone,  which  have  been  changed  already  through  drying,  show  a  weakly 
stained  and  diffuse  nucleus.  In  the  middle  zone  the  cells  are  well  fixed  and 
stained  ;  red  corpuscles  are  orange,  nuclei  of  lymph-cells  violet,  protoplasm  of 
these  is  grey,  eosinophil  granules  are  red.  Dry  preparations,  fixed  by  heat,  are 
useful  with  reference  to  histo-chemical  reaction,  but  are  useless  for  nuclear  study. 
By  drying  its  structure  is  altered  ;  it  stained  uniformly  and  slightly.  The  changes 
are  similar  to  those  in  peripheral  zone  of  spot.  The  appearance  of  these  altered 
nuclei  explains  the  definite  appearance  of  normal  blood.  The  diffuse  nuclei  of 
certain  leucocytes,  the  large  mono-nuclear  forms,  are  brought  out  by  drying.  The 
author  has  verified  his  results  by  sections.  The  marrow  was  imbedded  in  gum 
or  paraffin.  a.,  m.  c. 


1260  Journal  of  Applied  Microscopy 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Wilson,  H.  V.  Notes  on  a  Species  of  Pelo-  The  species  here  described,  P.  caroHn- 
myxa.     Amer.  Nat.  34:  535-550,  1900.  ^^^^y^^  ig  especially  favorable  for  labora- 

tory use  on  account  of  its  large  size  and  freedom  from  foreign  inclusions.  In 
sections  it  affords  fine  material  for  the  study  of  the  structure  of  protoplasm. 
Strong  acetic  carmin  (45  per  cent.)  was  used  in  killing  and  staining  for  whole 
mounts  in  glycerin.  The  external  form  and  the  internal  structure  were  better 
preserved  by  this  method  than  by  others.  The  author  regards  the  "  refractive 
bodies  "  as  globules  of  an  albuminous  nature.  The  culture  methods  employed 
in  rearing  this  rhizopod  are  of  especial  interest  since  they  are  applicable  to  other 
Rhizopoda,  such  as  the  various  species  of  Aviccba.  A  wooden  tub  is  filled  with 
ordinary  creek  sand  to  the  depth  of  four  inches  and  flushed  until  the  water  remains 
clear.  A  handful  of  Nitella,  two  or  three  opened  mussels,  and  fragments  of  a 
crayfish  are  partially  buried  in  the  sand  and  the  tub  is  placed  in  a  moderate 
north  light.  As  decomposition  progresses  a  stream  of  soft  water  is  turned  on  for 
a  short  time  every  few  days.  After  an  interval  of  tvi'o  to  eight  weeks  Avmba 
proteus  appears  in  numbers  on  the  surface  of  the  sand  and  sides  of  the  tub,  the 
smaller  forms,  A.  radiosa  and  A.  Umax,  appearing  earlier.  The  cycle  of  life  in 
such  a  culture  is  somewhat  constant.  Bacteria  appear  first  and  are  followed  by 
the  flagellate,  and  then  the  ciliate  infusoria,  especially  Stentor  ccerulens.  Later 
still  the  rotifers  and  Entomostraca  appear.  Cyclops  becomes  abundant  apparently 
at  the  expense  of  the  rhizopods.  Care  should  be  taken  not  to  introduce  the 
oligochaete  worm  Tubifex,  which  also  multiplies  rapidly  and  quickly  destroys  most 
of  the  bottom  forms.  The  brown  film  adhering  to  the  sides  and  bottom  of  the 
aquarium  harbors  the  rhizopods  and  Stentors  in  large  numbers.  c.  a.  k. 

Stole,  A.  Beobachtungen  und  Versuche  iiber  Pelojnyxa  was  collected  and  placed  in 
die  Verdanung  und  Bildung  der  Kohlenhy-  ^  i^rge  glass  dish  filled  with  SWamp 
drate  bei  einem  amobenartigen  Organismus,  °  * 

Pelomyxa  palustris  Greef.     Zeitschr.  f.  wiss.       water,  and  containing  the  other  swamp 
Zool.  68 :  625-668,  2  pis.  organisms  collected  at  the  same  time. 

The  evaporation  was  made  good  with  tap  water,  and  at  intervals  little  pieces  of 
gelatin  and  clean  filter-paper  or  cotton  placed  in  the  jar.  Under  these  condi- 
tions Pelomyxa  flourished,  the  individuals  being  usually  found  collected  about  the 
filter  paper  and  cotton.  The  oligochaete  Dero  and  the  sensitive  infusorian 
Spirostomum  also  did  well. 

To  isolate  the  animals  for  feeding  experiments  they  were  placed  in  small 
dishes  immersed  in  the  water  of  the  culture  jar  and  sometimes  covered  with  a 
cover-glass.  If  the  small  dishes  were  removed  from  the  culture  water  Pelomyxa 
developed  abnormally  and  soon  died.  The  food  was  always  solid ;  starch,  glu- 
cosides,   cellulose,   and    dried    and   powdered   proteids   being  injested   without 


and  Laboratory  Methods.  l'-^61 

difficulty.  With  glycogen  and  the  fats,  however,  special  methods  were  necessary. 
Glycogen  was  mechanically  united  to  albumin  by  dissolving  large  quantities  in 
egg-albumin  and  coagulating  with  heat.  The  mixture  was  then  dried,  powdered 
and  fed,  the  results  showing  that  glycogen  had  been  injested  with  the  albumin. 
In  the  case  of  the  fats  an  emulsion  of  fish  oil  in  albumin  was  treated  in  the  same 
way,  and,  after  feeding  the  dried  albumin,  oil  globules  could  be  seen  within  the 
cytoplasm. 

The  results  obtained  concern  almost  entirely  the  refractive  bodies  which  are 
present  in  great  abundance  within  the  cytoplasm,  and  are  easily  and  constantly 
affected  by  certain  kinds  of  foods.     The  principal  results  are : 

1.  The  refractive  bodies  are,  in  the  main,  composed  of  glycogen,  surrounded 
by  a  membrane  of  less  soluble  carbohydrate. 

2.  During  starvation  the  refractive  bodies  decrease  in  size,  the  glycogen  dis- 
appearing, until  finally  nothing  but  the  membrane  remains. 

3.  If  now  the  animal  be  fed  with  carbohydrate  food  (starch,  glucoside,  glyco" 
gen,  cellulose)  glycogen  is  stored  in  the  refractive  bodies  and  they  increase  in 
size. 

4.  Proteids,    gelatin,    and    fats  cause   no  change   in   the   refractive  bodies, 

although  the  injested  food  particles  are  gradually  dissolved. 

Frank  W.  Bancroft. 
University  of  California. 

„  „  ,„,,.„,  ,  „  ,  „  „.  Both  botanists  and  zoologists  will  be 
Senn,  0.     I'lagellata  in  Engler  and  Prantl  "  Die  ° 

Naturlichen    Pflanzenfamilien."    I    Theil,   i       interested    in    Dr.    Senn's    able    mono- 
Abth  Lief.  202, 203,  pp.  93-192, 1900.  Leip-  j.^  ^f  j^is  borderland  group  of  organ- 

zig.     W.  Englemann.  or-  &        r-  & 

isms.  The  following  orders  are  in- 
cluded :  Panto stomatijiece,  Protoviastiginecz,  DistomatinecB,  Chrysomonadinece,  Cryp- 
tomonadinecB^  Chloro7notiadine(e,  and  Eiigletihiece.  The  work  includes  a  compre- 
hensive biological  discussion  of  the  group  and  keys  to  the  genera,  with  very  full 
descriptions.  Abundant  illustrations  serve  to  characterize  many  of  the  species. 
Bibliographies  are  also  very  complete.  Investigations  since  the  publication  of 
the  monographs  of  Biitschli  and  of  Klebs  have  greatly  increased  the  number  of 
known  flagellates  so  that  this  revision  of  the  group  was  much  needed  and  will  be 
welcomed  by  all  who  have  to  deal  with  these  widely  distributed  and  biologically 
important  organisms.  c.  a.  k. 

Johnston,  J.  B.  A  Sealing  Stone  Jar  for  Zoo-  Stone  jars  eight  to  twenty-four  inches 
logical  Laboratories.  Amer.  Nat.  34:  969-  -^^  j^gj^^  ^^^  jg^  or  twelve  in  diameter 
971,  1900.  * 

are  made  by  the  Zanesville  Stoneware 

Co.,  Zanesville,  O.  The  rim  of  the  jar  bears  a  groove  to  be  filled  with  the  seal- 
ing fluid.  A  dependent  flange  on  the  lower  surface  of  the  lid  fits  into  the  groove, 
thus  sealing  the  jar.  The  edge  of  the  lid  projects  so  as  to  protect  the  rim  of  the 
jar  from  dust.  For  daily  class  use  water  may  be  used  as  a  sealing  fluid,  while 
for  permanent  storage  a  very  heavy  paraffin  oil  is  necessary.  Lighter  oils 
or  glycerin  do  not  make  good  sealing  fluids.  The  moderate  cost,  large  storage 
capacity,  slight  risk  of  breakage,  the  large  mouth,  and  above  all  the  ease  of 
opening  and  resealing  make  this  an  ideal  storage  jar  for  laboratories  and  museums. 

c.  a.  k. 


l'^6-  Journal  of  Applied  Microscopy 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Har\-ard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Vogel,    K.    Zur    Histologie    der    Pneumonia      The  author  studied  light  cases  in  which 

?79°r9oo!''°"''^'    ^''^^''''    ^''"''°''    ^*'      the  fibrinous  exudate  of  an  acute  lobar 

pneumonia  was  being  replaced  by  con- 
nective tissue.  The  origin  and  development  of  the  new  connective  tissue  was 
investigated. 

Several  staining  methods  were  employed.  Unna-Tanzer's  orcein  solution 
followed  by  Loeft"er"s  alkaline  methylen  blue  yielded  the  best  results.  Orcein 
colors  elastic  tissue  brown. 

(1)  Stain  (3  to  24  hours  in  the  following  fluid  : 

Orcein,           ------  0.5 

Absolute  alcohol,       -         .         -         .         .  40.0 

Distilled  water,     .         -         -         .         -  20.0 

Hydrochoric  acid,     -----  0.5 

(2)  Wash  in  water. 

(3)  Decolorize  about  30  minutes  in — 

Hydrochloric  acid,         -         -         -         .  5.0 

Alcohol  ----.-     100.0 

Distilled  water,       -----  20.0 

(4)  Wash  in  water. 

(5)  Stain  5  to  15  minutes  in  Loefier's  alkaline  methylen  blue  solution. 

(6)  Decolorize  for  a  few  minutes  in  70  per  cent,  alcohol. 

(7)  Absolute  alcohol. 

(8)  Oil  of  origanum. 

(9)  Canada  balsam. 

In  acute  pneumonia  fibrinous  strands  pass  from  the  masses  of  fibrin  in  the 
alveoli  to  the  alveolar  walls.  Some  strands  enter  Cohn's  pores  and  unite  with 
fibrin  plugs  in  other  alveoli,  others  are  attached  to  the  wall.  When  resolution  of 
the  exudate  fails  to  occur  the  plugs  of  fibrin  become  retracted  and  clear  spaces 
are  formed  in  the  peripher}'  of  the  alveoli.  In  early  cases  of  organizing  pneu- 
monia, spindle  shaped  connective  tissue  cells  are  found  on  the  surface  and  push- 
ing their  way  into  the  interior  of  the  fibrin  plugs,  and  spindle  cells  are  also  seen 
advancing  along  the  threads  of  fibrin  which  pass  through  Cohn's  pores.  The 
connective-tissue  fibrillse  form  at  first  a  loose  network  which  contains  in  its 
meshes  many  plasma  cells.  In  one  case  new-formed  elastic  fibers  were  demon- 
strable. 

Cohn  thought  that  the  connective-tissue  arose  from  the  inter-lobular  and  sub- 
pleural  tissues.  Ribbert  asserted  that  the  formation  began  in  the  smallest  bron- 
chi and  the  bronchioles.  Vogel  opposes  both  these  views.  In  the  first  stage  of 
organization  he  found  young  connective-tissue  outgrowths  springing  from  the 


and  Laboratory  Methods.  1263 

alveolar  wall  and  extending  along  the  fibrinous  strands.  He  concludes  that 
organization  proceeds  (1)  from  the  alveolar  wall  into  the  fibrin  plugs  :  (2)  from 
one  fibrin  plug  to  another  b}-  the  growth  of  connective-tissue  through  Cohn's 
pores.  J.  H.  p. 

c,  ex-  J  T^-      u    ■         r    u        i"  this  study  account  was  taken  of  the 

Flexner,  S.     xsature  and   Distribuuon    of   the  ^ 

New  Tissue  in  Cirrhosis  of  the  Liver.    ( Pre-       normal    and    pathological    distribution 

liminan-     Communication.)     Trans.     Asso.       ^f  ^^e  reticulum,  the    white  fibroUS   tis- 
Am.  Phys..  15:  523,  1900. 

sue  and  the  elastic  tissue. 

Two  methods  were  employed  for  the  demonstration  of  the  elastica.  The 
first  was  Unna's  orcein  stain,  the  other  that  of  Weigert,  which  emplo5-s  a  resorcin 
and  fuchsin  combination.  Unna's  method  was  unsatisfactorj^  because  the  stain- 
ing was  irregular  and  inconstant.     Weigert's  method  gave  uniformly  good  results. 

For  the  purpose  of  demonstrating  the  reticulum,  the  digestive  method,  as  first 
introduced  by  Wall,  as  well  as  the  modification  of  Spalteholz,  were  utilized.  By 
these  methods  both  fresh  and  preserved  tissues  in  sections  are  digested  in  alka- 
line solutions  by  means  of  pancreatin.  when  the  parenchvmatous  cells  and  elas- 
tic tissue  are  completely  removed.  There  remains  behind  a  framework  con- 
sisting of  white  fibrous  tissue  and  reticulum. 

In  the  study  of  the  white  fibrous  tissue  stained  sections,  both  before  and  after 
digestion,  were  employed.  Mallor}"s  specific  stain  was  found  of  especial  value 
in  demonstrating  the  fine  fibrils  of  white  fibrous  tissue  contained  within  the  liver 
lobules  ;  but  inasmuch  as  this  stain  also  colors  the  reticulum  its  use  is  somewhat 
more  limited  than  could  be  wished  ;  on  the  other  hand,  it  apparently  leaves  the 
elastic  fibers  unaffected. 

From  his  study  he  drew  the  following  conclusions : 

1.  In  all  forms  of  cirrhosis  the  white  fibrous  tissue  is  increased. 

2.  Along  with  the  increase  of  white  fibrous  tissue  there  is  a  new  formation 
of  elastic  tissue.  This  new  elastic  tissue  is  derived  from  pre-existing  tissue  in 
the  adventitia  of  blood  vessels  and  the  hepatic  capsules. 

3.  Both  white  fibrous  and  elastic  tissue,  in  all  forms  of  cirrhosis,  may  penetrate 
into  the  lobules.  This  penetration  takes  place  along  the  capillary  walls  or  follows 
the  architecture  of  the  reticulum.  The  chief  distinctions  between  the  histology 
of  atrophic  and  hypertrophic  cirrhosis  depend  upon  the  degree  of  extra  lobular 
growth  and  the  freedom  with  which  the  lobules  are  invaded.  In  hypertrophic 
cirrhosis  there  would  appear  to  be  less  interlobular  growth,  and  an  earlier  and 
finer  intralobular  growth. 

4.  The  alterations  in  the  reticulum,  per  se,  consist,  as  far  as  can  be  made 
out  at  present,  of  hypertrophy  rather  than  hyperplasia  of  the  fibers.  It  is  still 
uncertain  whether  any  of  the  differential  methods  now  in  use  suffice  to  distin- 
guish between  the  reticulum  and  certain  fibers  derived  from  the  white  fibrous 
tissue  of  the  periphery  of  the  lobules.  j.  h.  p. 


1264  Journal  of  Applied  Microscopy 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Holt,  E.  B.,and  Lee,  F.  S.  The  Theory  of  Pho-  In  the  literature  dealing  with  the  effect 
totactic  Response.  Amer.  Jour.  Physiol.  4:  ^f  jj  ^t  oil  the  movements  of  organisms 
460-481,  I90I.  °  ° 

two   modes   of  action  of  the  stimulus 

have  frequently  been  distinguished  ;  one  through  the  direction  of  the  rays,  and 
the  other  through  the  intensity  of  the  light.  It  is  the  purpose  of  Holt  and  Lee 
to  determine  to  what  extent  the  direction  of  ray  per  se  is  effective  in  producing 
the  orientation  of  an  organism  to  light.  The  two  leading  theories  of  light 
response,  those  of  Loeb  and  Verworn,  are  carefully  outlined,  and  that  of  Ver- 
worn  is  "  provisionally  adopted,"  because  it  seems  to  the  authors  to  be  more 
explicit  and  capable  of  being  applied  to  all  the  facts  than  the  other.  Four  typi- 
cal cases  of  light  reaction  are  examined.  The  first  and  simplest  reaction  con- 
sidered is  that  described  by  Strasburger  for  swarm-spores.  These  organisms 
move,  away  from  the  light  in  strong  illumination  and  towards  it  in  weak.  This  is 
explained  according  to  the  Verworn  theory  as  due,  on  the  one  hand  to  contrac- 
tion phenomena  induced  by  supra-optimal  stimulation  of  one  side  in  strong  light, 
and,  on  the  other  hand,  to  expansion  phenomena  induced  by  sub-optimal  stimu- 
lation of  one  side  in  weak  light.  This  results  in  movement  towards  the  optimum 
intensity  in  any  case.  The  second  point  considered  is  the  reaction  of  an  animal 
exposed  to  light  from  two  sources.  The  crustacean  Lyiiceus  was  experimented 
on  and  found  to  move  away  from  two  lights  of  equal  intensity  along  a  path  which 
equally  divided  the  angle  formed  by  the  light  rays  striking  the  animal  from  the 
two  sources.  This  again  is  evidently  what  would  be  expected  from  Verworn "s 
hypothesis,  since  the  path  taken  is  such  as  would  cause  the  two  sides  to  be  illum- 
inated by  light  of  equal  intensity.  The  third  case  of  phototactic  phenomena 
treated  is  the  response  of  an  animal  exposed  to  light  rays  coming  vertically 
through  a  prismatic  screen.  By  such  an  arrangement  one  end  of  the  vessel  is 
made  darker  than  the  other,  independently  of  the  direction  of  the  rays.  Lyiiceus 
and  Stentor  were  used  for  experimentation.  Both  went  to  the  dark  end  of  the 
vessel  along  a  more  or  less  diagonal  course.  The  explanation  is  that  the  ani- 
mal shows  contraction  phenomena  on  the  supra-optimally  stimulated  side  until 
the  body  is  in  such  a  position  that  both  sides  are  stimulated  with  equal  light 
intensities.  The  simple  diagonal  path  is  in  most  cases  moditied  by  the  fact  that 
the  animals  strike  the  back  wall  of  the  containing  vessel  and  are  veered  off  by 
it,  but  necessarily  in  the  general  direction  of  their  previous  course.  The  fourth 
type  of  reaction  considered  is  that  shown  by  animals  under  the  same  experimen- 
tal conditions  as  in  the  last  case  except  that  the  light  comes  obliquely  instead  of 
vertically  through  the  prism.  Under  these  conditions  a  negatively  phototactic 
animal  will  go  into  regions  of  brighter  illumination,  along  a  path  more  or  less 
parallel  to  the  direction  of  the  rays.     The  authors  show  that  it  is  possible  to 


and  Laboratory  Methods.  1265 

explain  this  reaction  as  a  result  of  the  attainment  of  a  position  of  equal  bilateral 
stimulation  by  the  same  sort  of  contraction  processes  on  the  supra-optimally 
stimulated  side  as  in  the  other  cases. 

The  principal  conclusion  is  that :  "  Light  acts  in  one  way,  that  is,  by  its  inten- 
sity. The  light  operates,  naturally,  on  the  part  of  the  animal  which  it  reaches. 
The  intensity  of  the  light  determines  the  sense  of  the  response,  whether  contrac- 
tile or  expansive  ;  and  the  place  of  the  response,  the  part  of  the  body  stimulated, 
determines  the  ultimate  orientation  of  the  animal."  Under  ordinary  circum- 
stances the  part  of  the  body  stimulated  is,  of  course,  a  direct  function  of  the 
direction  of  ray.  The  paper  shows  clearly  that  the  orienting  "  photopathic  " 
reaction  is  very  probably  the  same  thing  as  the  response  ordinarily  known  as 
"  phototactic."  R.  p- 

^  r,      ^     ,    ^,     .  ,         r  ,     „,  The  aim  of  this  work  is  to  determine 

Bardeea,  C.  R.     On  the  Physiology  of  the  Plan- 
aria  Maciilata  with  especial  reference  to  the       SOme    of     the     internal    conditions     of 
Phenomena  of  Regeneration.    Amer.  Jour,      regeneration  in  the  common  flatworm, 
Physiol.  5:   1-55,1901.  °  .,  r 

Planaria    maculata.     1  he    account    of 

the  regeneration  work  is  prefaced  by  sections  devoted  to  the  general  anatomy 
and  physiology  of  the  animal.  In  the  account  of  the  physiology  "  sensation  "  is 
discussed  in  a  very  general  way.  The  work  of  Loeb  on  the  light  reactions  of  the 
worm  is  mentioned  and  a  very  brief  description  is  given  of  the  reactions  to  con- 
tact stimuli.  In  this  section  the  author  makes  the  surprising  statement  that  he 
has  not  found  that  "  the  worm  is  sensitive  to  anything  but  light  and  contact." 
Under  "  Movement  "  two  sorts  of  progression,  "  swimming  "  and  "  crawling,"  are 
described.  The  ''  swimming,"  by  which  term  the  author  evidently  intends  to 
designate  the  motion  of  the  worm  ordinarily  spoken  of  as  "gliding,"  is  almost 
entirely  due  to  the  action  of  the  cilia  covering  the  ventral  surface  of  the  body. 
The  crawling  is  an  entirely  muscular  movement  brought  about  by  waves  of  con- 
traction passing  from  the  anterior  to  the  posterior  end.  Experiments  on  the  cen- 
tral nervous  system  showed  that,  at  any  level,  it  is  capable  of  governing  the  activ- 
ities of  all  parts  of  the  body  posterior  to  that  level.  Under  "  Internal  Activ- 
ities "  are  discussed  the  processes  of  deglutition,  food-dispersion,  defecation  and 
respiration.  Food  is  taken  in  by  peristaltic  contractions  of  the  pharynx  and  then 
distributed  evenly  through  the  branches  of  the  intestine  by  contractions  of  the 
body  wall.  Digestion  is  mainly  intracellular.  Defecation  is  brought  about  by  a 
series  of  sharp  contractions  of  the  whole  body  while  the  pharynx  is  held  open. 
There  are  a  few  brief  and  rather  loose  statements  in  regard  to  respiration  and 
excretion.  The  part  of  the  work  devoted  to  the  general  physiology  of  the  animal 
is,  on  the  whole,  weak  and  unsatisfactory.  The  remainder  and  larger  part  of  the 
paper  is  devoted  to  a  detailed  study  of  the  cellular  processes  taking  place  during 
regeneration.  Most  of  the  gross  forms  of  regenerated  animals  which  have  been 
obtained  by  other  workers  on  the  same  subject  are  carefully  described  with  refer- 
ence to  the  details  of  their  development.  The  processes  occurring  after  the 
removal  of  a  part  of  the  animal  are  briefly  as  follows  :  (1)  The  wound  becomes 
smaller  in  surface  area  on  account  of  the  contraction  of  the  surrounding  muscu- 
lature. (2)  The  cut  surface  remaining  is  protected  by  the  transformation  of  the 
cells  directly  exposed  to  the  water  into  mucoid  tissue.     Later  the  surface  epithe- 


1266  Journal  of  Applied  Microscopy 

Hum  grows  out  over  the  wound.  (3)  Along  the  cut  surface  and  in  the  region 
just  posterior  to  the  point  of  least  intestinal  pressure,  "  embryonic  tissue  "  is 
formed.  This  embryonic  tissue  comes  from  the  transformation  of  adult  paren- 
chyma cells.  (4)  The  next  stage  in  the  process  is  the  differentiation  of  the 
embryonic  tissue.  This  differentiation  depends  on  the  relation  of  the  tissue  to 
the  intestinal  apparatus  in  general,  and  to  the  axial  gut  in  particular.  Tissue  at 
the  anterior  end  of  the  axial  gut  forms  a  head,  at  the  posterior  end  a  pharynx, 
and  behind  the  pharyngeal  region  a  tail.  The  reason  for  this  relation  of  the 
differentiation  of  tissue  to  the  digestive  system  the  author  believes  is  to  be  found 
in  the  action  of  "  nutritional  currents  of  a  specific  direction,  intensity,  and  force." 
The  idea  is  a  suggestive  one  and  the  experimental  results  give  it  considerable 
probability.  r.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Jordan.     Some  observations  upon   the    Bac-      Dr.   Jordan    has   contributed  an   inter- 
terial    Self-purification    of    Streams.      Jour.       esting  and   timely   article   on  the  prob- 
xp.     e  .    .  _7  ,    9    .  j^^^  ^j  ^j^^  disappearance  of  bacteria  in 

flowing  streams  by  an  exhaustive  study  of  the  bacteria  in  the  Illinois  river, 
which  has,  in  the  last  year,  been  converted  into  a  drainage  system  for  Chicago, 
emptying  into  the  Mississippi  river,  after  flowing  318  miles.  The  pollution  of 
this  stream  with  the  sewage  of  Chicago  has  alarmed  the  people  along  its  banks, 
particularly  in  St.  Louis,  which  city  takes  its  supply  of  water  from  the  Missis- 
sippi river,  some  four  miles  below  the  outlet  of  this  system  of  sewage.  A  study 
of  the  bacteria  of  this  river  has  been  made  with  extreme  care,  and  the  bacterial 
contents  of  the  river,  at  different  distances  between  Chicago  and  the  Mississippi 
outlet,  have  been  determined.  The  result  shows  that  the  number  of  bacteria  in 
the  river  falls  rapidly,  and  at  its  outlet  apparently  all  of  the  bacteria  which 
came  from  the  sewage  of  Chicago  have  disappeared,  since  there  are  no  more 
bacteria  in  the  river  at  that  point  than  are  contained  in  the  ordinary  tributaries 
of  the  river.  The  river,  therefore,  purifies  itself  and  Chicago  sewage  does  not 
materially  pollute  the  Mississippi  river.  The  author  also  considers  the  causes  of 
this  disappearance  of  bacteria  without,  however,  reaching  very  positive  conclu- 
sions. He  is  inclined  to  believe  that  the  exhaustion  of  the  food  supply  is  one 
of  the  most  important  factors.  h.  w.  c. 

Ford.    The  Bacteriology  of  Healthy  Organs.      Bacteriologists,  in  the  past,  have  been 
Transactions  of  the  Association  of  American       of    the    opinion     that     the     organs     of 
ysicians.        -39-9    •  healthy  individuals  are  sterile,  and  that 

it  is  only  under  conditions  of  disease  that  bacteria  invade  the  tissues.  This 
opinion  has  been  questioned  occasionally,  but  no  very  definite  conclusion  has 
been  reached.  Ford,  desirous  of  settling  this  question,  performs  a  long  series 
of  very  careful  experiments.     His  method  of  work  appears  to  be  beyond  criti- 


and  Laboratory  Methods.  1267 

cism.  He  has  experimented  with  several  species  of  animals,  and  has  studied,  in 
all,  thirty-five  different  individuals.  His  conclusion  is  emphatic  and  decided. 
In  eighty  per  cent,  of  the  organs  studied,  positive  evidence  has  been  found  of 
the  presence  of  micro-organisms  in  the  normal  tissue  of  the  healthy  individual. 
Seventy-seven  per  cent,  were  demonstrated  by  growth  in  culture  media,  and  the 
other  cases  only  by  the  microscopic  presence  of  bacteria  in  the  organ.  He 
found  that  each  species  of  animal  showed  its  own  peculiar  bacteriology  ;  that 
each  animal  showed  a  distinct  bacteriology  ;  that  the  different  organs  showed 
the  same  bacteria  on  different  media,  although  different  culture  media  furnished 
a  variety  of  species.  The  bacteria  found  were  ordinary  species,  including 
staphylococci,  bacilli,  and  proteus  forms.  h.  w.  c. 

Wakker.  Wakker's  Hyacinth  Germ  Pseudo-  Dr.  Erwin  F.  Smith's  paper  on  Wak- 
monas  hyacinthi   (Wakker).      Bull.  Div.   of       ,       ,      tt        •    ,i     /-.  •  .  i 

Plant  Phys.  and  Path.  U.  S.  Dept.  of  Agri.      ^er  s  Hyacmth  Germ  IS  a  noteworthy 

26:  45.pl-  I-  contribution  towards  abetter  knowledge 

of  the  parasitic  bacterial  diseases  of  plants.  The  paper,  though  ready  for  publi- 
cation in  1897,  has  been  withheld  till  now  to  learn  why  such  meager  growth  was 
obtained  on  the  host  plant.  The  Pseudomonas  hyacinthi  (Wakker)  (E.  F.  Smith) 
is  a  yellow  rod-shaped  organism,  non-sporiferous,  color  distinctly  yellow  but 
somewhat  variable ;  old  cultures  on  some  media  darken  from  the  production  of  a 
soluble  pale-brown  pigment.  This  color  was  not  observed  in  acid  or  alkaline 
beef  broth,  on  cocoanut  flesh,  on  sugar  beets,  in  nutrient  starch  jelly,  in  agar,  or 
in  gelatin  with  or  without  sugar.  The  organism  is  pathogenic  to  hyacinths. 
The  host  plant  is  not  rapidly  destroyed,  the  cells  first  separate  by  solution  of  the 
middle  lamella.  The  cavities  contain  large  numbers  of  bacteria.  It  is  closely 
related  to  Ps.  campestris,  parasitic  or  cruciferous  plants,  B.  phaseoli  on  beans^ 
Ps.  stewartii  parasitic  on  corn,  especially  sweet  corn.  The  daughter  bulbs  con- 
tract the  disease  from  mother  bulbs.  The  bulbs  may  sometime  contract  the 
disease  from  germs  lodged  in  the  flowers.  A  more  extended  contribution  has 
been  promised.  The  paper  is  one  well  worthy  of  copying  as  a  model  for  this 
kind  of  work.  L.  H.  Pammel. 

Jones.  Soft  Rot  on  Carrot  and  Other  Vege-  This  paper  deals  with  a  soft  rot  of 
tables.  Bacillus  carotovorus  (Jones).  Rep.  ^^^rot  found  in  Vermont.  The  orga- 
Vt.  Agri.  Exp.  Sta.  13:  200-3-52,  fie.  11.  .         „      .,,  ,  /t  n 

x\\%\Xi  Bacillus  carotovorus  {)Qn^'&)c-3X\'S,&% 

a  rapid  soft  rot  of  carrots  which  resembles  Heinz's  white  rot  of  hyacinths  and 
Potter's  white  rot  of  turnip.  It  causes  the  rapid  disorganization  of  the  tissues 
apparently  due  to  an  enzyme  cytase,  excreted  by  the  bacteria,  which  softens  the 
middle  lamellae  of  the  cell  walls  and  causes  a  breaking  down  of  the  intercellular 
substance.  Wound  infections  led  to  decay  in  a  large  number  of  plants  such 
as  the  carrot,  parsnip,  salsify,  cabbage  head,  etc.  The  organism  producing 
this  disease  is  a  bacillus  having  vibratory  motion,  oscillating  or  darting  in  young 
liquid  cultures.  The  rod  is  provided  with  two  to  five  peripheral  flagella.  The 
author  suggests  that  we  have  a  considerable  number  of  groups  of  closely  related 
organisms  whose  differentiation  will  tax  the  skill  and  patience  of  the  bacterio- 
logist as  much  as  the  B.  coli  group.  The  organism  was  grown  in  a  large  number 
of  different  media.  Of  interest  is  the  fact  that  its  action  reduces  nitrates.  This 
paper  is  likewise  a  model  of  its  kind,  especially  in  regard  to  the  thoroughness 
of  testing  the  organism  in  different  media  while  studying  its  biological  and 
pathological  characters.  L.  H.  Pammel. 


1268  Journal  of  Applied  Microscopy 


Medical  Notes. 

Robin,  A.  Preservation  of  Sputum  for  Micro-  The  author  experimented  with  carbolic 
scopic  Examination.  Jour.  Bost.  Soc.  Med.  acid  5  per  cent,  solution,  trikresol  2 
Sci   5 :  7 

per  cent.,   formaldehyde   5    per   cent., 

and  hydrochloric  acid  10  per  cent,  to  determine  their  preservative  power  on 
sputum  containing  tubercle  bacilli.  The  sputum  treated  was  examined  at  the 
end  of  24  to  48  hours,  after  which  time,  weekly  and  then  monthly  examinations 
were  made  for  a  period  of  four  months.  Except  with  HCl  the  preservation 
was  good  and  the  bacilli  stained  deeply ;  HCI  seemed  to  disorganize  the  bacilli. 
The  author  recommends  the  addition  of  an  equal  volume  of  a  5  per  cent,  solution 
of  carbolic  acid  to  the  sputum,  which  should  be  vigorously  shaken  in  the  bottle, 
so  as  to  break  up  the  lumpy  coagulation.  c.  w.  j. 

Conn,  H.  W.  How  can  Bacteria  be  Satisfac-  I"  answer  to  this  question  the  author 
torily  Preserved  for  Museum  Specimens  ?  offers  the  following  method  :  A  two 
Jour.  Bost.  Soc.  Med.  Sci.  5:  7.  ^  ,^  ,.         .       ,         , 

per  cent,  agar  culture  medium  is  placed 

in  large  test-tubes  which  are  tilted  so  as  to  make  agar  slants.  The  tubes 
are  left  undisturbed  for  six  to  eight  weeks  to  allow  the  surplus  moisture  to 
evaporate.  They  are  then  inoculated  in  long  streaks  and  immediately  sealed 
wath  plaster  of  Paris  and  paraffin.  The  cultures  grow  for  a  few  days,  then  cease 
growing  and  remain  unaltered  indefinitely.  Only  one  unsatisfactory  feature 
presents  itself;  viz.,  moisture  within  the  tube  condenses  on  the  inside  of  the 
tube  with  changes  of  temperature,  thus  rendering  the  tube  cloudy  and  for  the 
time  injuring  the  value  of  the  display  specimen.  c.  w.  j. 

Eastes,  0.  L.    Note  on  the  Phenyl-Hydrazin      P^ace  60   c.   c.   of   filtered  urine  in   a 
Test  for  Sugar.    Brit.  Med.  Jour.,  Feb.  23,      beaker  of  100  c.  c.  capacity,  add  1  gm. 
^    ■  of  sodium  acetate,  and  a  little  less  of 

phenyl-hydrazin  hydrochlorate.  Stir  with  glass  rod,  which  is  left  in  the  mixture 
throughout  the  operation.  Place  beaker  on  water  bath  and  allow  the  mixture  to 
evaporate  gradually  down  to  10  or  15  c.  c,  occasionally  scraping  the  sediment 
from  the  sides  of  the  beaker  if  such  tends  to  collect.  When  reduced  to  the  bulk 
indicated,  remove  flame  and  allow  the  liquid  to  cool.  When  quite  cool  examine 
under  microscope.  Ozazone  crystals  will  have  formed  if  there  is  one  part  per 
thousand  or  more  of  sugar  in  the  urine.  If  no  crystals  are  formed  it  may  be 
safe  to  conclude  that  no  sugar  (glucose)  is  present.  c.  w.  j. 

Uhlenhuth.     Method  for  the  Differentiation  of       If,    at    intervals    of    six   to  eight  days, 

the  Blood  of  Various  Animals  with  especial      gn^^U  amounts  of  the  defibrinated  blood 

Reference  to  the  Demonstration  of  Human 

Blood.     Deutsche  Med.  Wochenschr.,  Feb.      of  any  animal  is  injected  into  the  rabbit, 

7'  '90I-  changes  are  produced   in  the   rabbit's 

blood  which  cause  it  to  give  a  reaction  with  the  blood  of  that  other  animal  alone 
and  with  no  other.  If  a  few  drops  of  the  serum  of  a  rabbit  that  has  been  treated 
with  ox  blood,  for  example,  are  dropped  into  each  of  a  row  of  test-tubes  contain- 
ing dilute  solutions  of  the  blood  of  various  animals,  absolutely  no  reaction  is 
produced  in  any  tube  except  that  containing  ox  blood,  which  at  once  shows  a 
slight  turbidity,  which  increases  on  standing  and  finally  develops  into  a  floccu- 
lent  precipitate. 

The  blood  of  a  rabbit  which  has  been  injected  with  human  blood,  furnishes 
an  infallible  reagent  for  detecting  human  blood  even  in  very  small  amounts,  and 
after  having  been  allowed  to  dry  for  four  weeks.  c.  w.  j. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


Volume  IV. 


MAY,  1901. 


Number  5 


The  University  of  Montana  Biological  Station. 

Most  of  our  Eastern  friends  who  have  not  been  through  this  Western  country 
and  have  not  seen  its  vastness  in  extent,  its  difficuhies  owing  to  the  absence  of 
roads  and  means  for  transportation,  can  scarcely  comprehend  the  work  necessary 
to  carry  on  any  amount  of  collecting  or  study  in  the  field.  The  idea  to  be  con- 
veyed through  this  paper  is  to  state  what  has  been  attempted  in  this  line,  the 
success  that  has  been  achieved,  and  the  suggestions  to  be  offered  from  the  expe- 
riences of  the  past  two  years. 


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~      >:  *~    Si^^^^C^^B 

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i 

1 

•^r... 


FIG.   1.     CAMP  AT  SIN-YALE-A-MIN   LAKE. 

In  the  spring  of  1899  plans  were  completed  for  the  establishment  of  out-door 
work  on  a  moderate  scale,  the  location  to  be  selected.  A  week  was  spent  in  the 
region  of  Flathead  lake,  Mont.,  and  all  available  sites  examined.  A  location  was 
secured  on  the  northern  end  of  the  lake,  on  the  bank  of  Swan  river,  close  to  the 
outlet.  The  location  was  chosen  as  possessing  the  following  advantages :  The 
mouth  of  Swan  river  offers  a  harbor  for  boats,  very  few  harbors  being  found  on  the 
lake.  Swan  river  affords  excellent  fishing,  and  the  region  round  about  is  a  dense 
forest,   practically   untouched.      This  is  one  of  the  most   convenient  places  to 

(1269) 


1270 


Journal  of  Applied  Microscopy 


reach  from  the  Great  Northern  railroad  on  the  north,  and  the  Northern  Pacific 
on  the  south,  is  on  the  regular  wagon  road,  is  easily  reached  by  steamboat,  and 
is  but  a  short  distance  from  the  mouth  of  the  Flathead  river,  which  has  abun- 
dance of  marshes  and  swamps.  This  is  one  of  the  few  places  on  the  lake  where 
suitable  accommodations  are  to  be  had  for  board  and  lodging. 

During  the  past  season  a  month  was  spent  in  the  Mission  mountains,  which 
extend  north  and  south  along  the  lake  and  Mission  valley  for  a  distance  of  nearly 
a  hundred  miles.  The  southern  end  of  the  range  has  a  number  of  high  peaks, 
the  highest  above  ten  thousand.  The  range  slopes  down  toward  the  northern 
end.  This  northern  end  has  been  ground  off  by  a  glacier,  which  has  left  undis- 
putable  proof  of  its  work  on  the  tops  of  the  high  hills.  The  range  ends  at  the 
Swan  river,  about  where  the  laboratory  is  situated. 

One  of  the  highest  peaks  at  the  southern  end  is  Sin-yale-a-min  mountain,  the 
Indian  word  meaning  "surrounded."     A  ten  days   camp  was  made  at  the  small 


FIG.  2.     CANVAS  BOAT       DAPHNIA"  WITH   COLLECTING  OUTFIT. 

lake  at  the  base  of  this  mountain,  and  called  also  Sin-yale-a-min  lake.  The  lake 
lies  in  the  heart  of  the  mountains,  with  high  peaks  on  all  sides  except  the  west, 
which  is  dammed  up  by  an  old  moraine,  though  it  is  of  recent  geological  origin. 

A  general  view  of  the  camp  at  Sin-yale-a-min  lake  is  given  in  Fig.  1.  The 
party  at  this  place,  all  told,  numbered  twenty-one,  and  with  one  or  two  exceptions 
all  were  engaged  in  some  work.  This  lake  is  about  fifteen  miles  from  the  near- 
est point  on  the  Northern  Pacific  railroad,  and  is  in  the  Flathead  Indian  reserva- 
tion.    It  is  therefore  primitively  wild  and  romantic. 

The  work  on  the  lake  was  accomplished  through  the  use  of  a  fourteen-foot 
canvas  boat,  which  was  taken  with  some  misgivings,  but  which  proved  all  that 
was  predicted  for  it  by  the  makers.  The  boat,  ready  for  use,  is  shown  in  Fig.  '2. 
the  photograph  being  taken  later  at  Swan  lake  when  fixed  ready  for  use.  The 
canvas  boat  carried  heavy  loads,  having  at  one  time  four  grown  people  and  one 
child,  guns,  ammunition,  nets,  and  other  material.     In  the  illustration  it  is  shown 


and  Laboratory  Methods. 


1271 


FIG.  3. 


FIELD  TABLE  AT  SIN-YALE-A-MIN   LAKE, 
FOR  MICROSCOPICAL  WORK. 


loaded  as  it  was  when  used  for  actual  work,  with  two  occupants  in  addition  to 

the  material.     At  the  front  is  seen  the  pump  after  plans  by  Ward,  for  taking  ento- 

mostraca  and  other  fresh  water  species.     Hanging  over  the  side  of  the  boat  is 

the  net  after  plans  by  Kofoid,  for  straining  the  pumpings.     To  the  right  of  the 

net  is  to  be  seen  the  apparatus  for 

measuring  depth,  which   is   an   in-     i  -»  -•  -^-^ 

strument    used    in    electric    light 

plants  and  other  establishments  for 

measuring  wire.     The  rubber  hose 

for  attachment  to  the  pump  is  also 

seen.     Of  this  hose   one   hundred 

and  forty  feet  were  carried. 

The  canvas  boat  was  used  con- 
tinuously, and  is  about  the  only 
available  means  for  work  in  these 
mountain  lakes,  so  remote  from 
civilization,  where  transportation 
is  a  grave  problem.  It  was  neces- 
sary to  use  common  garden  hose, 
owing  to  the  fact  that  no  other  kind 
was  kept  in  stock,  and  owing  to  the 
further  fact  that  large  hose  and  a 
large  pump  would  be  too  difficult  to  handle. 

In  Fig.  3  is  shown  the  laboratory  table  of  the  microscopist  in  his  study  of  the 
entomostraca  and  other  forms.  This  consists  of  two  sticks  nailed  to  a  fir  tree  at 
the  desired  height,  and  a  couple  of  rough  boards  nailed  to  the  top  of  these  sticks. 
The  location  is  selected  in  the  shade,  so  that  it  is  always  comfortable.  The  lake 
is  at  an  altitude  or  oS(l()  feet,  and  the  cold  water  makes  the  surrounding  air  cool, 
so  that  when  one  is  in  the  shade  one  is  always  comfortable.  Unfortunately  the 
microscopist  was  not  aware  the  picture  was  being  taken,  and  while  the  lower  part 
of  the  body  shows,  the  upper  part  is  lacking.  As  this  happens  to  be  the  only 
negative  worth  saving,  the  picture  is  shown  to  illustrate  the  ingenuity  in  making 
a  table.  The  eye  of  the  naturalist  will  readily  take  in  the  situation,  working  at  the 
fresh  material  from  a  lake  never  before  visited,  with  the  beautiful  sheet  of  water 
but  a  few  feet  away. 

In  this  work  a  small  microscope  was  carried,  with  a  battery  of  objectives,  and 
a  few  necessary  stains,  dishes,  slips,  covers,  and  the  like.  The  material  could 
be  taken  from  the  water  and  studied  immediately.  It  may  be  well  to  say  at  this 
time  of  the  year,  July,  rain  seldom  falls,  so  there  is  little  difficulty  from  that 
source.  When  there  was  danger,  or  when  the  sun  was  too  hot,  a  tarpaulin  was 
made  into  a  roof  with  ropes,  which  answered  as  protection.  In  case  of  emer- 
gency it  required  but  a  few  minutes  to  put  all  the  material  under  cover  of  the 
tents. 

A  similar  camp  was  made  at  McDonald  lake,  about  fifteen  miles  further  north, 
in  the  same  range,  and  in  the  same  reservation.  The  camp  at  this  lake  w^as  for 
the  purpose  of  collecting  additional  material  in  shells,  of  which  a  new  species  had 


1-27-2 


Journal  of  Applied  Microscopy 


been  found  the  year  previously,  and 
to  determine  the  microscopical  life 
of  the  waters  in  comparison  with 
those  of  Sin-yale-a-min  lake.  The 
view  shown  in  Fig.  4  will  give  to 
the  mountain  lovers  an  idea  of  the 
beautiful  peak  that  was  always  be- 
fore us.  This  peak,  McDonald, 
rises  to  a  height  of  over  ten  thousand 
feet.  The  view  here  given  was  taken 
from  the  mountain  side  near  camp. 
Photographers  may  be  interested  in 
knowing  that  the  picture  was  taken 
on  a  Seed  orthochromatic  plate,  the 
exposure  being  a  fiftieth  of  a  second. 
The  plate  was  somewhat  under  ex- 
posed, but  for  the  purpose  desired, 
which  was  to  bring  out  the  peaks 
with  the  clouds  above,  the  effect  was 
successful. 

McDonald  lake  is  much  similar 
to  Sin-yale-a-min  lake.  The  length 
is  about  a  mile  and  a  quarter,  the 
width  less  than  a  quarter,  the  depth 
68  feet.  Sin-yale-a-min  lake  was 
some  longer,  considerably  wider,  and  in  the  deepest  -")0  feet.  McDonald  lake 
is  surrounded  on  all  sides  by  high  and  rugged  mountains,  save  at  the  west,  where 
a  moraine  has  made  a  dam  as  in  the  case  of  the  lake  before  mentioned. 

Work  at  this  lake  was  conducted  much  as  in  the  first  case.     The  microsco- 


FIG.  4.  McDonald  PEAK  AND  LAKE. 


FIG.  5.  ORNITHOLOGISTS  AT  WORK  AT  McDONALD  LAKE. 


and  Laboratory  Methods. 


1-273 


pist  rigged  up  a  table  similar  to  the  one  described,  having  saved  the  lumber  and 
nails,  both  being  a  necessary  feature  in  the  unsettled  region.  The  ornithologists 
are  shown  at  work  in  Fig.  5.  This  table  and  roof  is  similar  to  that  made  use  of 
in  all  camps.  The  lake  lies  in  the  middle  foreground,  just  out  of  sight,  being 
lower  down.  Under  the  table  are  to  be  seen  various  sizes  of  zincs,  cylindrical 
in  form,  and  almost  closed.  These  are  used  for  placing  and  holding  the  made 
skins  while  they  dry.  It  is  often  necessary'  to  pack  the  skins  before  they  are  dry, 
and  even  afterward  the  jolting  the  mountain  roads  give  them  is  something  very 
difficult  to  understand  except  by  those  who  have  been  over  the  ground.  By 
placing  each  skin  in  a  zinc  cylinder,  the  cylinders  being  of  different  sizes  and 
lengths  to  accommodate  different  sized  birds,  it  is  then  an  easy  matter  to  pack 
the  skins,  and  at  any  time  get  them  out  to  dry  without  danger  of  injuring  the 
feathers  and  spoiling  the  shape.  It  is  true  the  zincs  are  heavy,  but  they  seem  to 
be  a  necessity  in  this  kind  of  work.  They  work  as  well  with  mammal  skins,  and 
are  also  employed  in  preserving  small  mimmal  skins. 


FIG     6.     VIEW  OF   UPPER    END  OF   FLATHEAD   LAKE,   SWAN    RIVER   OUTLET. 

For  the  ornithologists  long  excursions  were  unnecessary,  as  the  region  all 
about  is  dense  woods  up  to  the  mountain  sides,  and  it  was  necessar)'  but  to  take 
a  handful  of  shells  and  go  a  few  steps  from  camp  in  order  to  secure  enough  spec- 
imens for  a  half  day's  work.  The  picture  was  taken  on  a  Seed  orthochromatic 
plate  with  ray  filter. 

It  is  needless  to  relate  instances  of  camp  life,  or  to  describe  further  methods 
of  work.  It  is  in  order  to  say,  however,  that  to  change  camp  and  get  to  the  sta- 
tion, a  distance  of  only  about  fifty  miles,  one  must  descend  a  thousand  feet  over 
bad  road  with  all  the  paraphernalia  of  camp,  and,  with  all  material,  cross  the  res- 
ervation, a  distance  to  the  lake  of  twenty  or  twenty-five  miles,  taking  a  day.  dump 
the  material  off  at  the  lake  shore  and  again  pile  it  on  the  small  launch  or  the  large 
steamer,  whichever  is  taken,  cross  the  lake,  a  distance  of  about  thirty  miles,  again 
unload,  and  establish  camp  or  take  quarters  at  the  farm  house  near.      But  the 


1274 


Journal  of  Applied  Microscopy 


ride  is  one  never  to  be  forgotten,  especially  if  the  sun  is  shining  and  the  atmos- 
phere is  clear  so  as  to  bring  out  the  beauties  of  the  mountains  and  the  waters  of 
the  lake  bathing  the  base  of  the  range. 

Figure  6  gives  a  better  idea  of  the  country  adjacent  to  the  University  of 
Montana  Biological  Station  than  could  be  given  in  any  description.  The  view 
is  toward  Flathead  lake,  which  is  in  the  middle  of  the  illustration.  The  water  in 
the  foreground  to  the  left  is  Swan  river,  whose  outlet  into  the  lake  is  just  beyond 
the  bend.  The  location  of  the  station  is  on  the  bank  of  the  river  a  few  feet  to  the 
left  of  the  river  at  the  left  in  the  illustration.  The  narrow  point  of  land  behind 
the  trees  by  the  house  is  the  bar  made  by  the  sediment  from  Flathead  river, 
which  enters  the  lake  at  this  point,  and  which  is  some  two  and  a  half  miles  dis- 
tant.    The  mountains  in  the  background  are  the  Cabinet  range. 

The  field  laboratory  and  camping  ground  are  shown  in  Fig.  7,  seen  from  the 


FIG.  7.      EXTERIOR   UNIVERSITY  OF   MONTANA  BIOLOGICAL   LABORATORY 
AND  CAMPING  GROUND. 

rear,  the  only  place  from  which  a  picture  can  be  taken.  Immediately  in  front  of 
the  building  is  the  Swan  river,  which  has  a  bank  here  of  some  forty  or  fifty  feet. 
Directly  in  front  of  the  building,  and  at  the  water's  edge,  is  a  large  spring,  which 
furnishes  an  abundance  of  pure  water,  though  the  river  water  is  clear  and  pure. 
There  is  abundance  of  room  for  tents.  It  has  been  the  custom  to  live  in  tents  and 
take  meals  at  the  hotel  shown  in  Fig.  7,  though  since  the  picture  was  taken  a  large 
house  has  been  erected,  offering  excellent  accommodations  to  those  attending. 
The  field  laboratory  is  not  large.  It  was  planned  as  'a  convenient  outdoor 
laboratory  for  work.  It  will  be  understood  that  when  erected  the  building  was 
about  twenty  miles  from  Kalispell,  the  nearest  town.  Carpenters,  lumber,  and 
material  were  difiicult  to  secure,  and  the  attendance  upon  the  work  was  very 
problematical.  The  plan  was  to  make  a  building  suited  to  the  needs  of  a  few 
men  who  might  devote  a  month  or  more  annually  to  investigation  in  the 
immediate  region,  and  at  the  same  time  offer  the  privileges  to  any  who  might  wish 
to  take  advantage  of  the  offer. 


and  Laboratory  Methods. 


1275 


The  state  of  Montana  has  146,000  square  miles  of  territory.  There  is  a 
population  in  round  numbers  of  ■250,000  people.  Of  this  number  there  is  not  a 
large  number  who  wish  to  engage  in  such  study,  and  the  expense  of  getting 
around  is  no  small  item.  The  station  was  therefore  primarily  to  ofTer  a  haven 
for  a  few  enthusiasts  who  have  planned  to  do  something  toward  the  working  up  of 
the  material  of  the  state,  with  the  hope  that  the  enthusiasm  and  interest  would  be 
more  or  less  contagious,  and  that  in  time  there  would  be  work  of  considerable 
importance  and  by  considerable  numbers  at  the  laboratory. 

The  two  seasons  the  laboratory  has  been  opened  the  work  has  progressed 
well,  and  was  all  that  could  be  expected.  During  the  summer  of  1900  the  labora- 
tory was  taxed  to  its  utmost.  Figure  8  shows  a  portion  of  the  interior, 
with  the  students  at  work.  The  tables  are  rude,  and  the  chairs  have  been  con- 
structed from  raw  lumber  by  unskilled  hands,  but  the  material  with  which  they 


FIG.   8.      INTERIOR   OF   LABORATORY. 


work  is  from  the  university  laboratory,  and  is  the  best  the  country  alifords. 
Above  the  door  may  be  seen  rows  of  bird  skins.  To  the  left  is  the  working 
library  of  a  couple  of  hundred  volumes.  In  the  rear,  not  shown,  is  the  photographic 
dark  room  and  store  room.  With  this  small  building,  accommodating  no  more 
than  a  dozen  or  fifteen  at  a  time,  there  has  been  made  a  start  which  it  is  hoped 
will  later  develop  into  something  of  importance. 

Figure  9  is  an  illustration  that  will  interest,  if  not  please,  many  readers  of  the 
Journal.  Red-Horn,  an  Indian  who  had  been  on  a  visit  to  the  Blackfeet  in  the 
northern  part  of  the  state,  and  was  returning  to  his  home  on  the  Flathead  reserve, 
made  us  a  visit.  He  was  much  interested  in  our  work,  and  seemed  to  want  to 
know  what  was  being  done.  He  was  shown  various  things  through  the  micro- 
scope, which  pleased  him  greatly.  I  persuaded  him  to  let  me  take  his  picture, 
and  the  pleasure  he  is  having  is  shown  by  the  smile  on  his  countenance.  He 
was  then  taken  into  the  dark  room,  where  he  watched  the  picture  develop.    Later 


1276 


Journal  of  Applied  Microscopy 


he  received  a  print,  and  some  months  afterward  I  showed  him  his  picture  in  the 
daily  paper,  which  he  at  once  recognized  and  readily  understood. 

The  number  of  visitors  at  the  station  while  work  has  been  under  progress  has 


FIG.   9.     A  NEW  STUDENT  ARRIVES  AT  THE  LABORATORY. 

been  considerable,  including  the  governor  of  the  state,  many  school  men  of  prom- 
inence, a  number  of  government  men,  and  many  citizens  and  others  from  the 
region. 

The  equipment  of  the  station  as  regards  boats  is  shown  in  Fig.  10.  A  gaso- 
line launch  and  a  row-boat,  besides  the  canvas  boat,  offer  abundant  facilities  so 
far  for  all  who  have  attended,  and  for  those  having  charge  of  the  work.  In  addi- 
tion to  these,  the  launch  shown  in  the  illustration  to  the  rear  may  be  chartered  at 
any  time,  and  will  carry  several  tons,  being  o'J  feet  beam.     The  pump,  net,  hose, 

sounding  apparatus,  and  life  pre- 
servers have  been  put  out  to  dry. 
These  boats  are  in  the  harbor 
shown  in  Fig.  7,  being  just  below 
the  windmill   in  that  picture. 

By  means  of  these  boats  con- 
siderable work  has  been  done  on 
the  lake.     Soundings  have  been 
taken  in  many  places,  and  pump 
ings  made  from  various  depths. 
The    row-boat    is    also    taken    in 
wagon  to  the   smaller  ponds  ad- 
jacent to    the    station,  and   thus 
renders  the  work  there   effective. 
The  location  of  the  station  is  ideal  in  many  respects.     No  one  may  hope  for 
much  interest  to  be  taken  in  such  work  by  the  younger  element,  to  whom  we 
must  look  for  future  work,  without  making  ample  provision  for  recreation,  so  as 


FIG.  10.   STATION  BOATS  AND  EQUIPMENT. 


and  Laboratory  Methods. 


1277 


FIG.   11.     A  FLASH   LIGHT  AROUND  THE  CAMP  FIRE. 


to  combine  work  with  recreation.  This  is  especially  true  of  teachers  who  wish  a 
change,  and  are  seeking  a  place  where  they  may  have  a  chance  to  work,  and 
when  work  is  over  have  a  little  enjoyment.  During  the  first  summer  the  number 
of  plates  exposed  within  a  couple  of  miles  of  the  station  amounted  to  several 
hundred.  There  were  more  than 
a  half  dozen  cameras,  and  they 
were  in  almost  constant  use.  The 
dark  room  was  in  use  most  of  the 
time  both  day  and  night.  During 
the  second  season  the  number  of 
exposures  was  still  greater.  The 
rapids  above  the  station  are  a  de- 
light to  the  eye,  a  pleasant  place  to 
roam  when  there  is  nothing  to  do, 
and  a  great  resort  for  the  fisher 
men.  The  lake  beach  is  beauti- 
ful, and  many  romantic  bits  of  it  have  been  taken.  The  number  of  illustrations 
accompanying  this  paper  is  already  too  large,  or  more  would  be  shown.  It  is  suffi- 
cient to  say  that  the  writer  brought  home  from  the  summer's  trip,  including  the 
work  at  the  station,  more  than  a  hundred  and  twenty-five  good  negatives,  each 
illustrating  something  in  geology,  physical  geography,  or  biology. 

Bathing  in  the  lake  is  excellent.  The  bottom  is  smooth  and  sandy,  and  any 
depth  desired  may  be  obtained.  The  water  is  usually  comfortable,  the  cold 
water  from  the  rivers  not  reaching  this  portion  of  the  lake. 

Figure  11  is  given  as  an  illustration  of  an  attempt  to  take  a  flashlight  of  a 
group  around  the  camp  fire  at  night.  The  magnesium  was  placed  in  a  tin  pan 
with  a  paper  between  the  powder  and  the  pan,  the  paper  trailing  outside  so  as  to 
give  a  chance  for  lighting.  The  pan  was  placed  on  a  bench  with  a  tent  as  back- 
ground. Nearly  an  ounce 
of  magnesium  was  found 
necessary  to  produce  a  satis- 
factory picture.  The  camera 
was  placed,  and  the  person 
in  the  middle  of  the  group 
was  given  a  candle,  which 
was  used  to  determine  when 
a  sharp  focus  was  obtained. 
By  giving  the  candle  to  the 
party  at  one  end,  then  trans- 
ferring it  to  the  other  end,  a 
suitable  arrangement  was 
had.     The    shutter     was 

FIG.   12.     AN   EXPERIMENT  IN   REARING  DRAGON   FLIES.  ^        u        4-     4.U        4.-  iU 

opened  about  the  time  the 
trail  of  paper  was  lighted,  after  which  the  operator  walked  around  and  took  a 
place  in  the  group.  After  the  flash  he  returned  to  the  camera,  closed  the  shutter, 
and  later  made  development.  On  this  occasion  there  was  enough  smoke  to 
make  part  of  the  picture  a  trifle  hazy. 


127! 


Journal  of  Applied  Microscopy 


Figure  12  is  a  device  suggested  by  Calvert  for  rearing  dragon-flies,  suitable 
environment  having  been  obtained.  The  cylinders  of  wire  netting  are  placed  in 
the  water,  and  the  insects  placed  therein.  When  they  transform  it  is  possible  to 
identify  the  adults,  and  consequently  distinguish  between  young.  The  picture 
given  is  from  an  experiment  performed  at  the  laboratory. 

Figure  13  illustrates  to 
photographers  the  possi- 
bilities of  taking  bird  nests 
in-doors.  The  nest  is  that 
of  Wright's  flycatcher, 
E  mp  ido  n  a  x  iv  r  ig  h  t  i  /, 
Baird.  A  position  was 
taken  in  front  of  the  win- 
dow, though  out  of  the 
direct  sun.  A  black  felt 
cloth  was  used  as  a  back- 
ground, the  nest  being  set 
on  the  cloth  in  the  angle 
made  by  the  table  and  a 
pile  of  books.  A  mirror 
was  adjusted  so  as  to 
throw  light  into  the  nest, 
as  the  side  next  the  win- 
dow was  naturally  darker 
than  the  other.  The  nest 
was  several  inches  long,  but  was  incUned  so  as  to  be  parallel  to  the  lens,  hence 
the  observer  is  looking  into  the  nest.  The  plate  is  a  Seed  orthochromatic,  with 
ray  filter,  small  stop.  The  fluffy  appearance  of  the  surface  is  due  to  the  cottony 
material  with  which  the  nest  is  lined.  By  this  same  arrangement  a  series  of 
pictures  of  nests  was  taken,  without  moving  the  apparatus. 

The  biological  station  will  be  open  for  the  summer  of  1901  from  July  '22  to 
August  17.  The  six  weeks  preceding  will  be  spent  in  the  adjacent  region  col- 
lecting. Five  days  of  the  week  during  the  time  the  station  is  open  will  be  spent  in 
work,  the  sixth  will  be  taken  for  excursions.  As  is  usual,  there  are  no  fees  in 
connection  with  this  work,  all  of  the  material  being  provided  free,  and  the  teach- 
ing force  giving  their  time  gratuitously.  Those  attending  will  be  asked  to  pay 
for  what  is  broken  or  consumed,  and  to  pay  their  living  expenses. 

Accommodations  are  better  than  heretofore.  The  post-office,  Big  Fork,  has 
during  the  past  year  been  established  at  the  store  close  by  the  station.  The 
Kalispell  electric  light  plant  is  across  the  river,  and  several  houses  have  sprung 
up  during  the  summer.  Daily  mail,  electric  light,  a  railroad  just  built  a  short 
distance  away,  a  new  hotel,  and  other  conveniences,  make  living  less  wild  and 
more  natural,  and  will  give  greater  opportunity  to  those  who  wish  to  attend. 

As  the  result  of  the  two  years'  work  thus  far  accomplished,  several  bulletins 
are  ready  for  publication,  and  several  others  are  under  way.  There  is  a  fine 
opportunity  for  work,  and  plenty  of  material,  in  a  new  country,  with  practically  no 
opposition  ;  but  the  workers  are  too  few  and  life  too  short. 

Morton  J.  Elrod. 
University  of  Montana,  Missoula,  Mont. 


FIG.   13.     INTERIOR  OF  NEST  OF  WRIGHT'S  FLYCATCHER. 


and  Laboratory  Methods. 


1279 


The  Marine  Biological  Laboratory  at  Cold  Spring  Harbor,  L.  I. 

The  twelfth  annual  session  of  this  laboratory  will  be  held  during  the  months 
of  July  and  August,  of  the  present  year,  under  the  directorship  of  Professor  C.  B. 
Davenport.  The  regular  class-work  will  begin  Wednesday,  July  3,  and  will  con- 
tinue for  six  weeks  ;  the  laboratory  will  be  open  from  July  1  until  August  24, 
but  investigators  may  make  arrangements  for  using  it  from  the  middle  of  June 
until  the  middle  of  September. 

Cold  Spring  Harbor  is  about  thirty  miles  from  Brooklyn,  on  the  north  shore 
of  Long  Island.  It  is  a  deep,  funnel-shaped  inlet  of  Long  Island  Sound,  with 
steep,  wooded  shores,  about  five  miles  long,  and  one  and  a  quarter  miles  wide  at 
its  broad  end,  where  it  joins  the  sound.     It  is  divided  by  a  long  sand-spit  near  its 


COLD  SPRING  HARBOR,  WITH  A  VIEW  OF  THE  EAST  END  OF  THE  LABORATORY. 

inner  end  into  two  distinct  divisions,  an  inner  basin  about  half  a  mile  long,  upon 
which  the  laboratory  is  situated,  and  the  outer  harbor ;  near  the  middle  of  the 
western  shore  of  the  latter,  Oyster  Bay,  a  body  of  water  as  large  as  Cold  Spring 
Harbor,  opens  into  it. 

The  depth  of  the  water  in  the  harbor  varies  considerably.  The  mean  range 
of  the  tide  is  7.o  feet.  The  inner  basin  is  gradually  silting  up,  and  exposes 
about  half  of  its  bottom  at  every  low  tide  for  an  hour  or  so.  The  depth  of  the 
outer  harbor,  at  low  tide,  is  from  15  to  18  feet  above  the  entrance  of  Oyster  Ba}  , 
immediately  below  the  entrance  a  long  bar  extends  from  the  western  shore,  upon 
which  the  water  is  from  6  to  10  feet  deep  at  low  tide.  Beyond  the  eastern  end  of 
this  bar,  which  is  marked  by  a  small  light-house,  is  a  channel  72  feet  deep. 
Outside  the  bar  the  water  deepens  towards  the  sound. 


1280 


Journal  of  Applied  Microscopy 


The  country  surrounding  the  harbor  is  hilly  and  well  wooded.  The  soil  is 
moist  and  vegetation  in  the  woods  is  rank.  At  its  inner  end  a  small,  clear[stream. 
Cold  Spring  creek,  enters  the  harbor.  This  stream,  within  a  mile  of  its  mouth, 
runs  through  three  small,  deep  ponds,  all  of  which  are  surrounded  by  heavy 
woods  ;  a  portion  of  its  course,  also,  is  swampy. 


GROUND  PLAN  OF  THE  JOHN   D.  JONES  LABORATORY  BUILDING. 

The  situation  of  the  laboratory  is  an  especially  favorable  one,  inasmuch  as 
in  addition  to  the  marine  fauna  and  flora  at  hand,  a  rich  fresh-water  and  wood- 
land fauna  and  flora  are  also  easily  accessible,  l^he  harbor  and  the  adjoining  sound 
contains  a  variety  of  environments — marsh,  mud,  and  sand  flats,  hard  and  soft 
bottom,  each  with  its  peculiar  forms  of  life  ;  its  waters  are  very  rich  in  plankton. 


and  Laboratory  Methods.  1281 

The  same  is  also  true   of  the  fresh-water   ponds ;  deep  and  shallow  water  and 
marsh  are  present  with  abundant  life,  the  plankton  being  very  rich. 

The  most  characteristic  feature  of  the  marine  fauna  is  its  stability.  The 
animals  found  in  the  harbor  all  belong  there,  and  have  not  been  brought 
in  by  currents  or  tides  from  the  open  sea  ;  their  characteristics,  consequently, 
have  been  determined  by  their  relation  to  the  local  environment.  An  excellent 
opportunity  is  thus  given  of  studying  the  conditions  which  have  accompanied  the 
development  of  a  fauna. 

The  work  of  the  laboratory  is  divided  into  several  departments,  which,  with 
the  instructors  who  have  them  in  charge,  are  the  following  :  I.  Zoology.  In 
this  department,  the  following  courses  are  given :  high-school  zoology,  by  Profes- 
sors Davenport  and  S.  R.  Williams;  comparative  anatomy,  by  Professor  H.  S. 
Pratt ;  invertebrate  embryology,  by  Dr.  L.  E.  Griffin  ;  entomology,  by  Dr.  A.  G. 
Mayer  ;  variation  and  inheritance,  by  Professor  Davenport.  II.  Botany.  In 
this  department  the  following  courses  are  given  :  cryptogamic  botany,  by  Dr.  D. 
S.  Johnson  ;  ecology,  by  Mr.  H.  N.  Whitford ;  bacteriology,  by  Professor  N.  F. 
Davis.  III.  Microscopical  Methods,  by  Mrs  C.  B.  Davenport  and  Professor 
W.  L.  Tower.  IV.  Nature  Study,  by  Dr.  H.  A.  Kelly.  In  addition  to  these 
courses,  evening  lectures,  both  of  a  technical  and  of  a  popular  nature,  occur 
several  times  a  week. 

The  importance  of  excursions  and  collecting  trips  to  give  opportunities  of 
studying  the  fauna  and  flora  in  their  natural  environment  is  fully  appreciated. 
The  laboratory  has  a  launch  and  small  boats  with  dredging  and  other  collecting 
apparatus ;  a  large  oyster  boat  is  also  occasionally  used ;  and  trips  to  various 
parts  of  the  neighboring  waters  are  of  daily  occurrence.  A  trip  is  also  made  to 
Fire  Island  on  the  south  shore  of  Long  Island.  Every  facility  is  given  for  the 
collection  of  material  for  personal  use  and  for  the  use  of  the  institutions  with 
which  the  members  of  the  laboratory  are  connected. 

A  valuable  feature  of  the  laboratory  at  Cold  Spring  Harbor  is  the  quiet  and 
seclusion  of  the  place.  Situated  a  mile  from  the  village  of  the  same  name  and 
two  miles  from  the  railroad,  it  is  an  ideal  place  for  work  and  rest.  The  beauti- 
ful harbor,  the  fine  bathing  beach,  the  excellent  roads,  the  woods  and  fields,  the 
freshwater  ponds,  all  furnish  numerous  attractions  to  the  summer  visitor  outside 
the  work  he  accomplishes. 

The  laboratory  building  is  a  modern  structure,  72x36  feet,  lined  inside  with 
Georgia  pine,  and  with  excellent  ventilation,  due  to  the  height  of  the  roof ;  it  is 
provided  with  running  water,  both  fresh  and  salt,  and  a  complete  equipment.  A 
special  laboratory  for  investigators  is  also  now  being  completed.  The  lecture 
hall  is  a  large  building  lined  inside  with  Georgia  pine.  The  students  and  other 
members  of  the  laboratory  are  housed  in  three  dormitories,  one  for  men,  one  for 
women,  and  one  for  married  couples.  The  dining  hall  is  run  by  the  laboratory, 
and  board  is  furnished  at  cost. 

For  information,  application  should  be  made  to  Prof.  C.  B.  Davenport,  Uni- 
A^ersity  of  Chicago,  Chicago,  111.  H.  S.  Pratt. 

Haverford  College. 


1282 


Journal  of  Applied  Microscopy 


i^^ 


A  Method  for  Injecting  Small  Vessels. 

When  injecting  vessels  too  small  to  use  the  ordinary  removable  cannulas 
usually  provided  with  injection  syringes,  it  is  customary  to  employ  a  small  glass 
cannula  with  rubber-tube  connections.     This  has  the  great  objection  that  in  order 
to  avoid  forcing  air  into  the  vessel  it  is  necessary  to  fill  the  apparatus  before  it 
is  inserted  and  tied.     When  one  then  attempts  to  insert  the  cannula  it  is  very 
difficult  to  prevent  the  injecting  mass  from  coming  out  at  the  tip,  getting  into  the 
surrounding  tissues,  and  so  obscuring  things  that  it  is  next  to  impossible  to  see 
what  one  is  about ;  and  so  much  time  is  usually  consumed  in  the  process  that 
the  mass  is  apt  to  harden  and  clog  the  opening  of  the  cannula.     To  prevent  this  a 
clamp  is  usually  placed  upon  the  rubber  tube,  but  even  then  it  is  far  from  satisfactory. 
A  modification  of  this,  using  the  principle  of  the  removable  cannula,  has  been 
found  to  give  very  satisfactory  results.     A  piece  of  glass  tubing  of  a  little  larger 
diameter  than  would  ordinarily  be  used  is  taken  and  drawn  out  to  the 
—  c  desired  fineness,  depending  upon  the  size  of  the  vessel  for  which  it  is 
to  be  used.     It  is  then  cut  oiT  short  so  that  it  is  much  like  a  small 
funnel  (a).     The  tip  is  flared  slightly  in  the  ordinary  way  to  prevent 
the  ligature  from  slipping.     Another  piece  of  tubing   is  now  taken 
whose  outside  diameter  is  about  the  same  as  the  inside  diameter  of 
the  other — one  that  will  just  slip  within  the  other  nicely — ^and  is 
drawn  out  slightly  at  one  end  and  cut  off  so  as  to  leave  that  end 
somewhat  tapering.     A  short  piece  of  rubber  tubing  is  drawn  over 
this  tapering  end  (/?),  so  that  when  it  is  inserted  into  the  upper  end 
of  the  cannula  ( a)  it  makes  a  perfectly  tight  joint.     A  rubber  tube  from 
the  nozzle  of  the  syringe  leads  to  the  other  end  (r)  of  the  glass  tube. 
The  cannula  can  now  be  inserted  and  ligatured.     It  should  then 
be  filled  with  some  of  the  injection  mass,  either  with  a  pipette,  or  by 
allowing  it  to  drop  in  from  the  syringe.     By  using  a  small  wire  care- 
fully it  is  possible  to  get  practically  all  of  the  air  out  of  the  cannula 
and  to  get  it  well  filled  with  the  mass.     The  rubber-covered  end  of 
the  tube  can  then  be  placed  in  the  cannula  and  the  pressure  applied 
to  the  syringe,  care  being  taken  to  hold  the  joint  together  tightly. 
This  device  has  all  the  advantages  of  the  regular  injection  syringe 
over  the  glass  cannula  ordinarily  employed,  and  is  very  simply  and  easily  con- 
structed. Leon    |.  Cole. 
Zoiilogical  Laboratory,  University  of  Michigan. 


The  thirty-second  anniversary  meeting  of  the  New  Jersey  State  Microscopical 
Society  occurred  on  March  2.5th.  Mr.  F.  E.  Ives  of  Philadelphia  delivered  an 
illustrated  lecture  on  that  occasion,  his  subject  being  "  The  Kromskop  and 
Color  Photography."  J.  A.   Kelsey,  Secretary. 


and  Laboratory  Methods. 


1283 


The  Photo-Micrography  of  Tissues  with  Simple  Apparatus. 

The  growing  importance  of  photo-micrography  has  been  greatly  enhanced 
within  a  few  years  by  improvements  in  the  half-tone  processes  of  reproducing 
prints  ;  improvements  which  have  now  reached  such  a  degree  of  perfection  as  to 
make  the  reproductions  in  many  cases  excel  the  originals,  and  this  work  being 
done  at  a  very  trifling  cost,  places  in  the  hands  of  every  microscopist  ideal  facil- 
ities for  illustrating  the  result  of  his  labors  to  an  extended  audience,  provided  of 
course  he  is  familiar  with  photo-micrography,  and  can  make  micrographs  of  his 
subjects. 

The  appliances  for  doing  this  have  kept  pace  with  the  general  progress  in  all 


FIG.  1.     GIANT  CELL  SARCOMA.     XIOO. 

microscopical  manipulations  and  technique.  With  homogeneous  apochromatics, 
projection  oculars,  substage  condensers  of  high  numerical  apertures,  and  stands 
of  marvellously  perfect  workmanship  constructed  especially  for  the  purpose,  nnd 
having  every  conceivable  convenience,  it  would  seem  that  the  limit  of  optical 
possibilities  had  been  reached.  If  to  these  we  add  the  specially  designed  cam- 
eras, combining  in  one  the  suggestions  of  many  workers ;  orthochromatic  or 
"color  correct"  plates  and  the  many  new,  clearly  and  very  perfectly  developing 
reagents,  it  would  likewise  seem  that  the  photographic  branch  of  the  subject  has 
kept  pace  with  the  optical.  In  artificial  illuminants  we  are  equally  fortunate. 
The  electric  current  is  almost  universally  available  and  arc  lamps  of  great  sim- 
plicity and  steadiness  are  to  be  had  at  comparatively  moderate  cost.  The  new 
acetylene  light,  one  of  the  most  perfect  of  radiants  for  photo-micrography,  is  also 
available  everywhere  at  no  more  expense  than  the  old  coal  oil  flame.  In  short, 
to  the   man  with  a  desire  for  photo-micrography   and  a  full   purse,    the  world's 


1284 


Journal  of  Applied  Microscopy 


workshops  are  open  for  the  supply  of  an  unlimited  amount  of  perfect  apparatus 
for  his  purpose. 

But  to  most  students  and  the  great  mass  of  workers,  these  doors  are  closed. 
They  simply  have  not  the  money  to  spare  for  such  necessarily  costly  appliances 
and  doubtless  many  turn  aside  in  despair  at  the  impossibility  of  commanding 
their  use.  But  they  need  not.  It  is  quite  possible  to  do  most  of  the  work  they 
would  need  with  appliances  already  in  their  possession  or  quite  within  their 
means.  The  so-called  student's  microscope,  generally  in  use  in  our  colleges  and 
high  schools,  usually  has  an  inclinable  stand  with  two  eyepieces,  two  objectives 
of  about  1  or  2/3  inch  and  1/4  or  1/6  inch  focus  and  an  Abbe  condenser.  With 
such  an  instrument  it  is  quite  possible  to  do  most  excellent  photographing  with 
amplifications  ranging  from  about  50  to  600  diameters.  Very  little  tissue 
work  requires  over  .500,  while  most  of  it  may  be  acceptably  done  at  100  to  200. 


■^V 


FIG.  2.     MYXOMATOUS  TISSUE.     X  250. 


All  objectives  of  reputable  makers  are  now  so  well  corrected  chromatically  that 
there  is  little  need  to  give  the  old  bugaboo  of  focus  difference  consideration. 
Every  student  has  such  a  microscope  at  his  disposal,  so  we  find  the  optical 
part  of  the  question  needs  no  further  outlay. 

You  will  probably  ask,  "What  about  the  camera,  this  must  make  an  extra 
cost  ?"  Not  at  all.  While  very  convenient  and  highly  to  be  desired,  good  work 
can  be  done  without  a  camera  specially  designed  for  the  purpose.  In  fact,  any 
camera  provided  with  a  focusing  screen  and  from  which  the  lens  may  be  removed 
can  be  utilized  in  photo-micrography.  A  hand  camera  with  these  features  is  just 
as  good  for  the  purpose  as  any  other  form,  though  probably  not  so  convenient. 
It  merely  requires  to  be  firmly  fastened  to  some  support  at  such  a  height  as  to  per- 
mit the  tube  of  the  microscope  (when  inclined  to  the  horizontal  position)  to  enter 
the  lens  opening  and  project  the  image  of  the  object  on  the  stage  upon  the  foc- 
using screen.     The  latter  will  be  found  too  coarse  for  fine  focusing,  but  the  old 


and  Laboratory  Methods.  1285 

device  of  attaching  a  disk  of  cover  glass  to  the  ground  slide  with  Canada  balsam 
will  offer  a  perfect  surface  for  delicate  focusing.  A  sheet  of  plain  glass  may  be 
substituted  for  the  focusing  screen,  or  the  atrial  image  may  be  found  by  means 
of  a  hand  lens.  The  illumination  may  be  obtained  by  means  of  a  coal  oil  lamp 
standing  at  such  a  height  as  to  bring  the  center  of  its  flame  up  to  the  optical 
axis  of  the  microscope. 

For  many  years  I  have  been  profoundly  impressed  with  the  importance  of 
photo-micrography  as  an  educational  agent  which  the  successful  introduction  of 
the  half-tone  process  of  reproduction  greatly  intensified.  At  first  the  great 
cost  of  everything  necessary  for  the  work  was  no  doubt  a  bar  to  its  more  general 
introduction,  but  happily  this  no  longer  exists.  Quite  recently  one  of  a  type  of 
student's  microscopes  generally  adopted  by  our  best  institutions  of  learning  fell 
into  my  hands.  It  was  a  revelation  to  me  of  the  wonderful  progress  made  in 
the  mechanism  and  optics  of  the  microscope,  and  made  my  own  apparatus,  only 
some  two  decades  old,  seem  quite  ancient  in  comparison.  Yet  some  of  the 
objectives  of  my  outfit  represented  an  outlay  of  much  more  than  the  cost  of  this 
entire  apparatus. 

The  microscope  in  question  was  fitted  with  two  eyepieces,  2/.3  and  1/6-inch 
objectives.  Abbe  condenser  and  iris  diaphragm  ;  a  plain  working  stand,  as  will  be 
seen,  costing  very  little  money    but  of  admirable  workmanship  throughout. 

My  test  of  a  microscope  and  objectives  being  their  adaptability  to  photography, 
I  proceeded  to  apply  it  to  this  outfit,  but  came  a  shade  further  than  usual  in 
discarding  the  use  of  my  special  camera,  and  making  up,  instead,  an  impro- 
vised affair,  that  anyone  can  do  for  himself  in  a  very  few  moments.  A  small 
quarter-plate  camera  was  pressed  into  the  service,  secured  to  a  block  at  just  the 
proper  height  to  bring  its  axis  in  line  with  that  of  the  microscope.  An  old 
focusing  cloth  wound  around  the  tube  of  the  microscope  at  its  junction  with  the 
camera,  made  this  light  tight.  A  coal  oil  lamp  with  an  inch  flame  adjustable  to 
any  height  afforded  the  necessary  illumination  for  most  of  the  tests,  though  the 
far  more  actinic  light  of  an  acetylene  flame  was  also  used  at  times. 

With  this  very  simple  apparatus,  I  made  a  number  of  negatives,  mostly  of 
tissues  normal  and  diseased  and  varying  in  amplification  from  100  to  500 
diameters,  the  range  of  most  useful  enlargements  in  that  class  of  work.  While 
these  might  perhaps  be  exceeded  in  absolute  perfection  by  the  employment  of 
the  very  highest  attainable  excellence  in  optical  appliances,  my  conclusion  is  that 
they  are  good  enough  for  all  practical  purposes,  and  quite  within  the  means  and 
ability  of  every  student  to  make  for  himself  in  illustrating  his  own  microscopical 
work.     For  this  reason    it  is  urged  upon  everyone  to  make  the  attempt. 

Among  these  negatives  are  two  which  may  serve  to  illustrate  the  excellence  of 
the  optical  work  of  this  microscope.  Both  were  made  by  the  aid  of  the  usual 
Huyghenian  eyepieces  furnished  with  the  instrument,  a  form  that  we  are  told  in 
the  books  is  totally  unsuited  to  the  purpose.  One  was  made  with  the  Abbe 
condenser,  a  form  which  we  are  likewise  told  is  useless  in  photography.  But 
negatives  and  prints  tell  a  different  story.  It  is  obviously  impossible  to  give  a 
detailed  account  of  their  working,  within  the  limits  of  space  at  my  disposal  ;  but 
a  synopsis  may  prove  useful  to  many  seeking  information  on  the  subject. 


12S6  "  Journal  of  Applied  Microscopy 

No.  1. 
Giant  cell  sarcoma  x  100. 
Object,  thin  section,  carmin  stained. 
Objective,  2/3  inch,  achromatic. 
Ocular,  ordinary  Huyghenian  11/2  inch. 
Condenser,  none,  flat  side  of  flame  used. 
Light,  Acetylene  gas  flame,  1  foot  burner. 
Plate,  Seed's  Non-halation. 
Screen,  green  glass. 

Exposure,  six  minutes  (at  least  three  times  too  short). 
Developer,  metol — quinol. 

No.  2. 
Myxomatous  tissue  x  250. 
Object,  very  thick  section,  deeply  stained. 
Objective,  1/6-inch  achromatic. 
Ocular,  ordinary  Huyghenian  2-inch. 
Condenser,  Abbe,  iris  diaphragm. 
Light,  Acetylene  gas  flame,  1  foot  burner. 
Plate,  Wuestner's,  "Jersey  Beauty." 
Screen,  cobalt  blue  (Rainig's  Moderator). 
Exposure,  90  seconds. 
Developer,  eikonogen-hydroquinone. 
Prints  on  Velox  Glossy  Paper. 


W.  H.  Walmsley. 


COMBINED  UREOMETER  AND   SACCHAROMETER 

(IMPROVED.) 

FERMENTATION  TUBES  FOR  BACTERIOLOGIC  INVESTIGATIONS  OF 

FERMENTATION. 

Further  experiments  with  the  Ureometer  devised  by  the  writer  and  described 
in  the  Medical  Record,  59:  12,  477,  have  shown  that  the  evolution  of  gas  from  the 
decomposition  of  the  urine  by  the  hypobromite  can  be  greatly  facilitated  by  using 
the  following  modification  :  Instead  of  the  test-tube  for  the  hypobromite  solu- 
tion a  small  50  c.  c.  flask  is  used.  Twenty  c.  c.  of  the  hypobromite  solution  are 
put  into  the  flask,  the  urine  drawn  up  into  the  pipette,  which  is  inserted  into  the 
rubber  stopper  so  that  the  end  is  well  above  the  level  of  the  hypobromite  solution. 
One  c.  c.  of  urine  is  then  discharged  into  the  latter.  The  evolution  of  gas  takes 
place  at  once,  and  the  test  is  completed  in  a  few  minutes.  The  volume  of  air  in 
the  flask  displaced  by  the  1  c.  c.  of  urine  is  deducted  from  the  total  volume  gen- 
erated, and  in  order  to  avoid  calculations  1  c.  c.  of  air  space  should  be  allowed 
at  the  closed  end  of  the  graduated  limb  of  the  U  tube  and  the  graduations  begin 
at  zero.     The  accompanying  illustration  shows  the  improved  ureometer. 


and  Laboratory  Methods. 


1287 


Directions  for  Use. — Fill  the  ^tube  with  water  to  the  mark  A.  In  doing 
this,  put  the  index  finger  on  the  end  of  the  side-tube  rt'and  fill  the  limb  B.  By 
inclining  the  (7  tube,  the  water  is 
forced  into  limb  C,  and  any  air  bub- 
bles are  removed  in  a  similar  manner. 
As  soon  as  the  tube  is  filled  to  the 
mark  close  the  free  end  of  £  with  a 
cork  or  a  rubber  stopper.  This  pre- 
vents the  water  from  running  out 
through  the  side-tube.  Put  20  c.  c. 
of  the'  hypobromite  solution  into  the 
flask  £,  replace  the  double-perforated 
rubber  stopper  g,  insert  the  side-tube 
ti  into  one  perforation  and  the  pipette 
/  filled  with  urine  into  the  other. 
Remove  any  air  bubbles  from  the 
limb  C  by  inclining  the  apparatus, 
taking  care  that  none  of  the  hypo- 
bromite solution  comes  in  contact 
with  the  urine.  Remove  the  cork  from 
the  free  end  of  limb  B.  Open  the 
stop-cock  on  the  pipette,  allowing  1 
c.  c.  of  urine  to  flow  into  the  flask. 
The  iV accumulates  at  the  closed  grad- 
uated limb  C.  Gentle  shaking  of  the 
apparatus  will  greatly  hasten  the  reac- 
tion. 

The  hypobromite  solution  is  best  made  up  extemporaneously.  The  follow- 
ing method  will  be  found  most  serviceable :  Have  on  hand  a  saturated  solution 
of  sodium  hydrate.  Place  10  c.  c.  of  the  latter  into  the  flask  and  add  1  c.  c.  of 
bromin.  Shake  gently  until  reaction  is  complete  and  add  10  c.  c.  of  water.  The 
writer's  way  of  taking  up  the  bromin  will  no  doubt  be  appreciated  by  those  who 
have  had  their  Shneiderian  membrane  frequently  exposed  to  the  irritating  vapors 
of  this  dangerous  substance.  We  use  the  ordinary  1  c.  c.  pipette,  to  which  a  long 
piece  of  rubber  tubing  is  attached.  On  the  latter,  somewhere  near  the  end,  is 
placed  a  small  Hoffman  clamp.  The  bromin  is  sucked  up  to  the  mark  and  the 
clamp  at  once  closed  tightly  by  means  of  the  screw.  The  end  of  the  pipette  is 
then  carried  at  once  into  the  sodium  hydrate  solution,  and  the  bromin  discharged 
slowly  by  opening  the  clamp.  As  a  safe  precaution  we  keep  open  a  bottle  of 
ammonia  during  the  operation. 

To  use  this  apparatus  as  a  saccharometer  the  double-perforated  stopper  is 
replaced  by  one  with  a  single  perforation.  The  6^  tube  is  filled  with  water  as 
described  above,  10  c.  c.  of  diabetic  urine  put  in  the  flask,  1  grm.  of  Fleishman's 
yeast  added,  together  with  a  small  crystal  of  tartaric  acid,  and  the  apparatus  set 
aside  for  24  hours.     The  CO  2  generated  will  collect  at  the  closed  end  of  limb  C. 

Fermentation  Tubes. — On  the  same  principle  the  writer  devised  a  fermen- 


Fig.  I. 


Combined  Ureometer  and  Saccharometer 
set  up  as  Ureometer. 


1288 


Journal  of  Applied  Microscopy 


A    — 


Fig.  2.     (i.)     Fermentation  Tube. 


tation  tube  for  bacteriologic  purposes. 
As  seen  from  the  illustration  below  the 
6' tube  is  of  smaller  size,  the  stopper 
with  a  small  tube  drawn  out  to  a  capillary 
point,  and  a  short  tube  used  instead  of 
the  flask. 

The  side-tube  {■  is  plugged  with  non- 
absorbent  cotton  ;  the  6^  tube  is  filled 
with  mercury  to  the  mark  A,  the  cotton 
preventing  the  mercury  from  escaping. 
The  tube  B  containing  a  convenient 
quantity  of  sugar-bouillon  is  inoculated 
with  the  organism.  The  rubber  stopper 
is  inserted  into  S,  the  displaced  air  escap- 
ing through  c/.  This  done,  the  end  of  d 
is  sealed  in  the  flame  and  the  apparatus 
placed  in  the  incubator.  The  CO  2  collects  in  the  closed  end  of  the  ?7  tube 
under  mercury,  thus  assuring  the  complete  collection  of  the  gas,  which  in  the  ordi- 
nary fermentation  tubes  escapes  in  considerable  quantities  from  the  open  end. 
For  convenience  as  well  as  for  comparative  study  of  dififerent  fermenting  organ- 
isms a  trench  is  made  to  hold  6  tubes  (see  Fig.  2-2.)  Only  the  tubes  intended 
for  the  culture  need  be  sterilized.  The  rubber  stoppers  are  sterilized  (in  steam) 
in  a  wide-mouthed  bottle  and  kept  there  until  used.  The  rubber  stopper  devised 
by  the  writer  is  especially  useful  for  this  purpose  inasmuch  as  its  handling  does 
not  carry  with  it  contamination.  The  stopper  is  so  made  that  an  outer  jacket  is 
formed  which  fits  over  the  neck  of  the  container,  while  the  stopper  proper  is 
within.  The  illustration  in  Fig.  2  (3)  explains  itself.  The  writer  believes  that 
this  form  of  stopper  will  be  found  useful  wherever  an  ordinary  stopper  is  used,  as  it 
offers  the  additional  advantage  of  keeping  out  dust  and  preventing  the  escape  of 
gas.  Where  the  neck  of  the  bottle  is  unusually  thick  the  outer  jacket  is  reflected 
while  the  stopper  is  inserted.  A.  Robin,  M.  D. 

Delaware  State  Board  of  Health  Laboratory,  Newark,  Del. 


The  New  Jersey  State  Microscopical  Society. — At  the  February  session 
of  the  N.  J.  S.  M.  S.,  a  paper  on  "Pebbles"  was  presented  by  Dr.  A.  H.  Chester, 
professor  of  mineralogy  in  Rutgers  College. 

The  term  "pebble"  was  defined  as  a  more  or  less  rounded  piece  of  rock 
varying  in  size  from  that  of  a  particle  of  sand  to  a  boulder. 

The  three  chief  agents  in  the  formation  of  pebbles  are  the  small  streams  and 
rivers,  the  ocean  and  glaciers ;  the  last  named  being  by  far  the  most  important 
of  the  three. 

The  shape  of  a  pebble  depends  upon  the  shape  of  the  original  fragment,  and 
upon  which  of  the  three  above  named  agents  has  produced  it. 

A  number  of  lantern  slides  were  presented  illustrating  glaciers  chiefly,  and 
their  effects  upon  rocks.  A  large  and  exceedingly  interesting  collection  of  dif- 
ferent sorts  of  pebbles  was  also  placed  on  exhibition,  the  specimens  ranging 
from  the  most  common  forms  about  us  to  gold  nuggets  and  diamonds,  sapphires 
and  rubies  in  the  rough — in  their  pebble  state. 

J.  A,  Kelsey,  Secretary. 


and  Laboratory  Methods.  1289 

MICRO-CHEMICAL  ANALYSIS. 
XIII. 

STRONTIUM. 

We  can  employ,  for  the  detection  of  this  element :         , 
I.   Sulphuric  Acid. 
II.  Oxalic  Acid. 

III.  Sodium  Tartrate. 

IV.  Ammonium  Bichromate. 

V.  Primary  Sodium  Carbonate. 

None  of  these  reagents  can  be  considered  as  giving,  at  once,  a  characteristic 
and  reliable  test  for  strontium  in  the  presence  of  calcium  and  barium  or  mem- 
bers of  the  magnesium  group.  It  follows,  therefore,  that  the  detection  of  stron- 
tium is  often  a  matter  of  not  a  little  difficulty.  When  dealing  with  mixtures  of 
the  alkaline  earths  it  is  necessary  to  proceed  as  directed  wnA&x— Separation  of  the 
Calcium  Group — methods  which  will  be  found  immediately  following  the  reac- 
tions for  Barium. 

/.  Sulphuric  Acid  added  to  solutions  containing  salts  of  Strontium  leads  to  the 
separation  of  Strontium  Sulphate. 

SrCl2  -f  HaSO^  =  SrS04  +  2HC1 

Method. — To  the  drop  to  be  tested  add  a  drop  of  dilute  sulphuric  acid.  A 
granular  precipitate  results.  Add  another  large  drop  of  the  reagent,  heat,  and  if 
insufficient  liquid  remains  add  more  acid.  The  heating  is  continued  until  dense 
white  fumes  of  SO5  are  given  off  in  abundance.  Allow  the  preparation  to  cool 
and  examine  at  once.  At  first  globular  forms  and  rhombic  plates  appear,  later, 
these  develop  into  more  or  less  irregular  fusiform  crystals  which  generally  grow 
to  crosses  with  two  of  the  arms  very  short.     Fig.  50. 

Instead  of  recrystallizing  from  sulphuric  acid  we 
can  employ  hydrochloric  acid.  If  the  latter  method 
is  believed  to  be  preferable,  proceed  as  follows  :  after 
adding  the  reagent  in  sufficient  amount  to  insure  com- 
plete precipitation,  carefully  draw  off  the  supernatant 
solution  (or  filter  or  whirl  in  the  centrifuge).  Wash 
the  precipitate  with  hot  water  to  remove  any  free  acid 
and  soluble  salts,  then  add  several  drops  of  strong 
hydrochloric  acid.     Heat  the  preparation  to  boiling, 

draw  off,  allow  to  cool,  and  examine.     If  after  a  short 

1  11-1  I  I   I  I  I 

time  no  crystals  separate,  concentrate  the  solution  by  \-pvy..o.o\'nm. 

heating.     Strontium  sulphate  crystallizes  from  hydro-  '^'  ^°' 

chloric  acid  in  the  form  of  square  and  rectangular  plates,  long,  thin  prisms,  and 

sheaves  of  acicular  prisms.     Fig.  51. 

Remarks. — As  already  stated  under  Calcium,  the  addition  of  sulphuric  acid  to 


1290 


Journal  of  Applied  Microscopy 


Fig.  51. 


solutions  containing  strontium,  yields  bundles 
of  needles  rapidly  disintegrating  to  merely  a 
very  fine  granular  precipitate.  Unless  the 
preparation  is  examined  immediately  after  the 
addition  of  the  reagent  no  acicular  crystals  will 
be  seen. 

If  calcium  is  also  present  the  grains  of  stron- 
tium sulphate  are  generally  larger  and  often  ex- 
hibit a  tendency  toward  a  spindle  shape. 

In  all  cases  recourse  must  be  had  to  re- 
crystallization. 

It  is  probable  that  the  crystals  of  strontium 
sulphate  separating  from   hot  concentrated  sul- 
phuric  acid  have   a   composition   analagous  to 
calcium  sulphate  recrystallized  under  the  same  conditions. 

If  after  a  short  time  no  crystals  appear  in  the  drop  of  acid,  breathe  on  the 
preparation. 

It  is  imperative  that  the  drop  to  be  heated  be  placed  at  the  very  corner  of 
the  slide,  that  the  latter  be  inclined  so  as  to  keep  the  drop  at  the  corner,  and  that 
the  "  micro  "  flame  be  applied  a  little  to  one  side,  and  nearer  the  center  of  the 
slip.  This  procedure  is  necessary  in  order  to  avoid  (1)  the  breaking  of  the 
glass  slide,  and  (2)  the  spreading  of  the  sulphuric  acid.  This  tendency  of  the 
hot  liquid  to  flow  over  the  slip  when  it  is  placed  in  a  horizontal  position  is  so 
great  that  it  is  generally  advisable  to  transfer  a  part  of  the  acid  to  a  clean  slip. 
The  transfer  is  accomplished  by  gradually  raising  the  slip,  which  has  been  heated, 
until  it  assumes  an  almost  vertical  position  and  the  drop  has  flowed  to  the  extreme 
corner.  The  corner  is  then  brought  in  contact  with  a  clean  glass  slide  and  the 
drop  of  solution  caused  to  flow  onto  the  latter  by  means  of  a  glass  rod.  In  this 
way  a  clear,  well  rounded,  deep  drop  is  obtained  in  which  good  crystals  of  stron- 
tium sulphate  will  form. 

When  dealing  with  very  minute  quantities  of  material  it  is  better  to  heat  with 
sulphuric  acid  on  platinum  foil,  since  the  hot  acid  may  extract  sufficient  mate- 
rial from  the  glass  to  interfere  with  the  reaction. 

The  solubility  of  strontium  sulphate  in  strong  hydrochloric  acid  is  quite  low, 
hence  it  is  necessary  to  employ  a  considerable  quantity  of  the  solvent  in  order  to 
get  satisfactory  results.  The  resulting  crystals  are  quite  small  and  of  varied 
form.  The  results  are  less  satisfactory  than  with  sulphuric  acid,  but  there  is,  on 
the  other  hand,  the  advantage  that  barium  sulphate  is  insoluble  in  HCl.  It  is 
of  course  essential  in  recrystallizing  from  HCl  that  only  traces  of  free  H2SO4 
be  present.     Free  nitric  acid  should  also  be  absent. 

Before  any  attempt  is  made  to  recrystallize  the  precipitate  of  strontium  sul- 
phate, it  is  advisable,  and  usually  Accessary,  to  remove  any  calcium  which  may 
be  present.  This  is  accomplished  by  extracting  the  precipitate  with  hot  water. 
Unless  this  is  done,  peculiar  crystal  forms  are  obtained  which  are  difficult  to 
interpret. 

If  only  a  small  amount  of  barium  is  present,  characteristic  crystals  of  stron- 


and  Laboratory  Methods.  1291 

tium  sulphate  are  obtained  from  hot  H2SO4  ;  more  barium  is  apt  to  alter  the 
usual  crystal  form,  although  the  appearance  of  the  crystals  separating,  still  sug- 
gests the  strontium  sulphate  type.  An  excess  of  barium  seems  to  cause  the 
majority  of  the  crystals  to  assume  forms  somewhat  resembling  barium  sulphate. 
In  general,  crystals  of  both  strontium  and  barium  sulphate  can  be  distinguished 
in  mixtures  of  these  two  elements. 

Any  lead  which  may  be  present  will  be  precipitated  in  an  amorphous  condi- 
tion by  the  dilute  acid.  Recrystallized  from  hot  sulphuric  acid,  the  lead  sulphate 
will  separate  in  forms  which  at  first  closely  resemble  those  of  strontium  sulphate 
and  which,  later,    grow  to  forms  which  may  be  mistaken  for  barium  sulphate. 

Recrystallized  from  hydrochloric  acid  there  is  less  danger  of  confusion.  If 
in  doubt,  extract  the  precipitated  sulphates  with  a  solution  of  potassium  or  sodium 
hydroxide  in  which  lead  sulphate  is  soluble. 

As  in  the  case  of  calcium,  chlorides  of  the  trivalent  metals  and  salts  of  boric 
acid  may  sometimes  interfere  with  the  formation  of  typical  crystals  of  strontium 
sulphate. 

£xercises  for  Practice. 

To  a  drop  of  a  moderately  dilute  solution  of  SrCl2  add  dilute  H2SO4  and 
examine  at  once. 

Recrystallize  SrSO^  from  HgSO^;  and  from  HCl. 

Try  to  recrystallize  from  HCl  in  the  presence  of  H2SO4. 

Make  a  mixture  of  calcium  and  strontium  and  add  HgSO^.  Recrystallize 
the  product  from  H2SO4  without  having  removed  the  calcium.  In  another  por- 
tion remove  the  calcium  by  extracting  with  boiling  water  and  then  recrystallize 
the  residue. 

See  also  exercises  suggested  under  Barium. 

//.  Sfroiitinm  Oxalate  is  precipitated  from  solutions  of  salts  of  Strontium  by 
Oxalic  Acid. 

SrCl2  +  H2C0O4  =  SrC204  .   ;/H20  +  2HC1. 

Method. — Proceed  as  directed  under  Calcium,  Method  II.  Strontium  oxa- 
late is  precipitated  at  once.  The  crystals  of  this  salt  are  similar  to  those  obtained 
with  calcium,  but  are  somewhat  larger  and  crosses  are  more  pronounced  ;  yet 
when  dealing  with  mixtures  of  unknown  composi- 
tion, the  difference  is  scarcely  sufficient  to  permit 
of  strontium  being  distinguished  from  calcium. 

The  crystal  forms  of  strontium  oxalate  which 
are  most  frequently  met  with  are  shown  in  Fig.  52. 

Remarks.  —  Either    tetragonal    or    monoclinic 
crystals  are  obtained  as  in  the  case  of  calcium. 

The    remarks    under    Calcium    (q.    v.)    apply 
equally  well  to  strontium. 

It  is  always  advisable  to  draw  off  the  super- 
natant  solution  and  add  dilute  sulphuric   acid  to  Fig.  52. 


1292  Journal  of  Applied  Microscopy 

the  precipitate.  If  no  crystals  of  calcium  sulphate  appear,  add  more  acid,  heat 
until  white  fumes  appear,  cool  and  examine  the  preparation  for  crystals  of 
strontium  sulphate  (see  Method  I). 

Exercises  for  Practice. 

See  exercises  suggested  under  Barium. 

///.  With  Sodium  Tartrate  solutions  of  salts  of  Strontium  yield  difficultly  solu- 
ble Stroutium  Tartrate. 

SrCla  +  HNaC^H^Og  =  SrC^H^Og  .   4H2O  +  NaCl  +  HCl. 

Method. — Proceed  as  directed  under  Calcium,  Method  III.  Strontium  tar- 
trate is  isomorphous  with  calcium  tartrate  and  is  not  to  be  distinguished  from  the 
latter  (see  Fig.  47).  There  is,  perhaps,  a  tendency  on  the  part  of  the  strontium 
compound  to  form  shorter  and  stouter  prisms  and  thin  plate-like  crystals  in 
greater  abundance  than  is  the  case  with  the  calcium  salt. 

Remarks. — See  remarks  under  Calcium.  It  is  not  possible  to  distinguish 
between  calcium  and  strontium  by  this  test. 

Behrens  suggests  the  addition  of  magnesium  acetate  and  acetic  acid  to  the 
mixture  thought  to  contain  both  elements,  before  introducing  the  reagent.  This, 
he  states,  retards  the  reaction  and  prevents  the  normal  development  of  the  cal- 
cium salt  while  the  strontium  tartrate  grows  to  its  usual  size.  Such  a  modifica- 
tion of  the  test  requires  considerable  experience  in  order  that  just  the  proper 
conditions  shall  be  obtained  ;  for  this  reason  the  modification  is  seldom  success- 
ful in  the  hands  of  a  beginner. 

The  test  is  useless  in  the  presence  of  barium  and  many  other  elements ;  the 
most  important  of  these  being  lead,  iron  and  aluminum  as  chlorides,  and  boron 
as  borates. 

IV.  Ammonium  Dichromate  in  alkaline  solution  precipitates  Strontium  Chromate. 
2SrCU  +  (NH j2Cr207  -|-  2NH4OH  =  2SrCr04  +  4NH4CI  +  H2O. 

Method. — To  a  dilute  neutral  or  very  slightly  acid 

xV  S^  solution  of  the  substance  to  be  tested  add  a  fragment 

^fi^/y  of  ammonium  dichromate  (or  potassium   dichromate). 

jg,         A  No  precipitate  should  result  if  only  strontium  is  present. 

i^<^^  Should  a  precipitate  result,  draw  off  the  clear  liquid 

^  ^'tf       after  all  the  reagent  has  dissolved;  then  add  to  it  a 

^  *J?^      ^  small  drop  of  ammonium  hydroxide.     Strontium  chro- 

•^^        *<>^^^^^^  mate  immediately  separates  in  tiny  yellow  globulites  or 

11,1  dumb-bell-like  forms.     Near  the  circumference  of  the 

"    *"  drop  short    rods    appear   later    (Fig.    53).      Warming 

gently,  hastens  the  separation. 

Remarks. — Unless  care  is  taken  to  employ  a  sufficiently  dilute  solution,  the 

precipitate  obtained  will  consist  of  such  minute  granular  masses  as  to  appear  to 

be  amorphous. 

The  addition  of  sodium  acetate  in  excess  will  also  cause  the  precipitation  of 
strontium  chromate. 


and  Laboratory  Methods.  1293 

Normal  potassium  chromate  (KoCr04)  on  the  other  hand,  will  precipitate 
strontium  at  once  from  neutral  or  slightly  acid  solutions,  as  SrCr04,  in  the  form  of 
slender  rod-like  prisms  of  the  orthorhombic  system.  The  crystals  obtained  with 
KoCrO^  are  usually  better  than  those  produced  by  KgCrgO^  or  (NH4)2Cr20.  ; 
unfortunately  barium  is  precipitated  by  both  these  reagents  in  either  acid  or 
alkaline  solution.  It  thus  becomes  a  decided  advantage  to  use  a  dichromate 
in  a  solution  acidified  with  acetic  acid ;  under  these  conditions  only  barium  will 
be  precipitated,  the  supernatant  liquid  can  then  be  drawn  off,  and  to  it  ammo- 
nium hydroxide  added,  when  strontium  will  be  precipitated. 

Salts  of  calcium  yield  no  precipitate  with  ammonium  dichromate,  whether  the 
solution  be  acid  or  alkaline. 

Testing  for  strontium  with  dichromate  is  impossible  in  the  presence  of  zinc, 
cadmium  or  the  rare  earths. 

Lead  and  other  elements  forming  insoluble  chromates  will  be  precipitated 
before  the  ammonium  hydroxide  is  added,  but  may  escape  complete  precipitation 
and  interfere  with  the  subsequent  test  for  strontium. 

Exercises  for  Practice. 

See  exercises  and  suggestions  given  under  Barium. 
V.  Primary  Sodium  Carbonate. 

This  reagent  precipitates,  from  very  dilute  solutions,  strontium  carbonate  in 
the  form  of  spherulites,  often  of  considerable  size. 

When  simple  salts  of  the  elements  Ca,  Sr,  Ba  are  employed  it  is  not  at  all 
difificult  to  distinguish  between  them  by  testing  with  primary  sodium  carbonate 
(or  ammonium  carbonate).  A  drop  of  the  almost  saturated  solution  of  the 
reagent  being  caused  to  flow  into  the  dilute  neutral  test  drop,  calcium  will  give 
well  defined,  highly  refractive  grains  and  rhombohedra,  strontium  sperulites  exhib- 
iting the  usual  black  cross  between  crossed  nicols,  and  barium,  spindle  shaped 
crystallites  and  fibrous  masses.  But  if  two  or  more  of  these  elements  are  pres- 
ent the  reaction  fails,  characteristic  crystals  being  the  exception. 

Elements  of  the  magnesium  group  must  be  absent. 

Primary  sodium  carbonate  is  of  more  value  as  a  group  reagent  than  as  an 
identification  test. 

BARIUM. 

The  most  important  reagents  available  for  the  microchemical  detection  of 
barium  are  as  follows  : 

I.   Sulphuric  Acid. 
II.  Oxalic  Acid. 

III.  Potassium  Ferrocyanide. 

IV.  Ammonium  Fluosilicate. 
V.  Ammonium  Dichromate. 

VI.  Potassium  Antimonyl  Tartrate. 
VII.  Primary  Sodium  Carbonate  or  Ammonium  Carbonate. 


1294 


Journal  of  Applied  Microscopy 


\P\V.=  O.Olr 


Fig.  54. 


/.  Sulphuric  Acid  added  to  solutions  containing  Bariu7n  p)-ecipitaTcs  Barium 
Sulphate. 

BaCU  +  H2SO4  ==  BaS04  +  -HCl. 

Method.— A.dd  to  the  test  drop  dilute  sulphuric  acid  as  long  as  any  precipi- 
tate is  formed ;  draw  off,  treat  the  residue  with  a  large  drop  of  the  reagent,  and 
heat  until  copious  white  fumes  are  given  off.     Cool,  breathe  on  the  preparation,  and 

examine.  Barium  sulphate  separates,  first  as 
tiny  rectangular  plates  and  X-shaped  skeletons ; 
then,  in  a  short  time,  much  larger  crystallites 
appear  with  more  or  less  feathery  arms  which 
still  retain  the  X-form.  See  Fig.  54.  These 
crystals  apparently  belong  to  the  orthorhombic 
/J7  "^    1^^  system. 

\J  "i^T^  ^3-   ^  Remarks. — Owing    to    the    low    solubility   of 

barium  sulphate,  a  considerable  amount  of  sul- 
phuric acid  is  necessary  and  the  preparation  must 
be  strongly  heated  in  order  to  obtain  a  solution  of 
the  precipitate.  In  this  operation,  the  precau- 
tions mentioned  under  Strontium  must  be  ob- 
served. 

In  the  event  of  a  heavy  precipitate  being  obtained  with  the  reagent,  it  is  wise 
to  remove  a  small  portion  to  another  slide  for  recrystallization,  rather  than 
attempt  to  dissolve  the  whole  mass. 

Recrystallization  in  the  presence  of  much  calcium  is  to  be  avoided.  First 
extract  the  calcium  sulphate  with  hot  water. 

In  the  presence  of  moderate  amounts  of  strontium  the  crystallites  of  barium 
sulphate  are  generally  not  well  formed.  If  strontium  is  in  excess,  the  crystals 
separating  from  the  hot  sulphuric  acid  have  the  general  type  of  strontium  sul- 
phate, but  are  not  well  developed  and  exhibit  an  inclination  to  approach  X-forms 
of  barium  sulphate.  For  this  reason  it  is  advisable  to  remove  any  strontium 
which  may  be  present  by  repeatedly  heating  with  hydrochloric  acid,  in  which 
strontium  sulphate  is  soluble  while  the  barium  compound  remains  undissolved 
and  can  then  be  recrystallized  by  heating  with  sulphuric  acid. 

Any  lead  sulphate  which  may  be  present  will  appear,  first,  in  crystals  very 
suggestive  of  strontium  sulphate,  then,  in  a  short  time,  in  larger  crystallites  which 
may  at  times  be  mistaken  for  barium  sulphate.  Treatment  with  hydrochloric 
acid  or,  better,  with  sodium  hydroxide  will  remove  the  lead,  leaving  the  barium 
salt  unacted  upon. 

It  is  sometimes  desirable  to  apply  other  tests  to  the  precipitated  sulphate  in 
order  to  confirm  the  presence  of  barium.  In  such  an  event,  transfer  the  washed 
precipitate  to  platinum  foil  or  to  a  platinum  cup  and  fuse  with  potassium  carbon- 
ate. The  fused  mass  is  then  extracted  with  water  and  the  residue  of  barium  car- 
bonate dissolved  in  hydrochloric  acid.  This  solution  can  then  be  tested  for 
barium  by    any  of  the  tests  given  below. 

Since  chlorides  of  the  trivalent  .metals  sometimes  interfere  with  the  formation 


and  Laboratory  Methods. 


1295 


of  characteristic  crystals  of  barium  sulphate,  it  is  advisable  to  draw  off  the  super- 
natant liquor  after  the  addition  of  the  reagent  and  before  heating  with  an  excess 
of  the  acid.  When  dealing  with  mixtures  it  is  always  best  to  proceed  in  this 
manner. 

Exercises  for  Practice. 

Try  above  method  on  a  simple  salt  of  Ba. 

Make  a  mixture  of  Ca  and  Ba,  recrystallize  at  once  without  removing  the  Ca. 
From  another  portion  remove  the  Ca  with  hot  water  and  recrystallize  the  residue. 

Try  a  mixture  of  Sr  and  Ba.  Remove  the  Sr  by  treating  with  HCl  and  recrys- 
tallize the  residue. 

Try  a  mixture  of  Ca,  Sr,  and  Ba ;  first  recrystallizing  at  once,  then  removing 
in  turn  the  Ca  with  hot  water  and  the  Sr  with  HCl. 

After  having  tried  the  other  reactions  for  barium,  described  below,  fuse  some 
BaSO^  with  K2CO3  and  proceed  as  directed  above. 

//.    Oxalic  Acid  precipitates  Barium  Oxalate  from  solutions  of  salts  of  Barium. 
BaCl2  +  H2C2O4  =  BaC^O^  .   //H2O  +  2HC1. 

Method. — To  a  drop  of  a  very  dilute  solution  of  the  barium  salt  add  sodium 
acetate  and  then  oxalic  acid  in  the  same  manner  as  in  testing  for  calcium  and 
strontium.  In  a  few  seconds  large  branching  aggre- 
gates in  the  form  of  radiating  bundles  and  sheaves 
of  fibrous  needles  are  seen.  These  radiating  masses 
occasionally  assume  forms  resembling  snow  crystals. 
Rarely  well  developed  monoclinic  prisms  are  ob- 
tained. 

The  usual  forms  of  barium  oxalate  are  shown  in 
Fig.  55. 

Remarks.— The.  solution  to  be  tested  should  be 
neutral.'  A  slight  trace  of  acid  is  apt  to  prevent 
the  separation  of  the  characteristic  crystals. 

If  no  crystals  appear  after  a  short  time,  add  a 
fragment  of  sodium  or  ammonium  acetate. 

When  calcium  or  strontium  are  present  the 
characteristic  crystal  forms  of  barium  oxalate  will 
not  be   obtained.     Recourse    may   then  be  had  to 

testing  in  dilute  nitric  acid.  From  nitric  acid  solutions  the  barium  salt  will  not 
separate,  while  the  oxalates  of  calcium  and  strontium  will  slowly  crystallize  in 
their  usual  form.  After  allowing  sufficient  time  for  the  complete  separation  of 
calcium  and  strontium,  draw  off,  concentrate  the  solution,  and  add  sodium 
acetate.  Barium  oxalate  now  appears,  usually  in  the  form  of  rosettes  of  thin 
prisms. 

Barium  oxalate,  like  the  oxalates  of  calcium  and  strontium,  assumes  different 
crystal  forms  according  as  the  test  drop  is  hot  or  cold.  Hot  solutions  give  rise 
to  the  production  of  strongly  polarizing  orthorhombic  plates. 

Since,  in  order  to  facilitate  the  separation  of  barium  oxalate,  sodium  acetate 


1296 


Journal  of  Applied  Microscopy 


has  been  added,  it  is  well  to  bear  in  mind  that  there  is  danger  of  interference 
from  members  of  the  magnesium  group. 

Boric  acid  present  in  the  test  drop  may  prevent  the  formation  of  characteristic 
crystals  of  barium  oxalate. 

Although  chlorides  of  iron  and  aluminum  have,  as  has  been  stated,  no  dele- 
terious iniiuence  on  the  precipitation  of  the  oxalates  of  calcium  and  strontium, 
we  meet  in  the  case  of  barium  with  a  most  interesting  and  remarkable  reaction. 
Owing  to  the  formation  of  a  double  oxalate,  instead  of  the  forms  shown  in  Fig. 
55,  there  are  now  obtained  tufts  and  bunches  of  very  long,  fine,  curving,  hair-like 
crystals  of  exceedingly  characteristic  appearance.  The  chemical  composition 
and  formula  of  this  compound  is  not  yet  clear.  In  order  to  obtain  this  interest- 
ing compound,  proceed  as  follows :  To  the  test  drop  containing  barium,  add 
ferric  chloride  in  sufficient  amount  to  impart  a  faint  but  distinct  yellow  color ; 
then  add  a  fragment  or  two  of  sodium  or  ammonium  acetate  ;  stir.  The  yellow 
should  now  have  changed  to  a  reddish  tint.  Into  the  drop  thus  prepared  cause 
a  drop  of  oxalic  acid  to  flow.     Tufts  and  sheaves  of  very  fine  needles  soon 

appear.  The  needles  rapidly  grow 
longer  and  longer  and  soon  begin  to 
curve  in  a  most  peculiar  manner. 
See  Fig.  56.  The  presence  of  cal- 
cium or  strontium,  or  both,  in  even 
large  amount  does  not  appear  to  have 
any  serious  influence  on  the  forma- 
tion of  this  double  oxalate  of  barium 
and  iron,  save  that  its  separation  is 
often  somewhat  retarded.  In  such 
mixtures  the  oxalates  of  calcium  and 
strontium  first  appear  in  their  usual 
form,  then  after  a  time  the  hair-like 
tufts  of  the  double  oxalate  appear. 
If  the  quantity  of  barium  is  quite 
small,  little  rosettes  of  radiating 
needles  are  obtained,  separating  near 
the  edges  of  the  drop. 

Aluminum  gives  rise  to  the  for- 
mation of  a  similar  product,  but  the  crystal  masses  are  colorless,  while  those  of 
the  iron  salt  are  light  brown. 

Chlorplatinic  acid  interferes  with  the  formation  of  barium  oxalate  in  a  manner 
similar  to  iron  and  aluminum.  Hence  it  is  inadvisable  to  test  a  preparation 
with  oxalic  acid  for  borium,  which  has  already  been  tested  for  potassium. 

For  a  list  of  the  elements  with  which  oxalic  acid  may  give  a  crystalline  pre- 
cipitate, see  the  list  of  reagents*. 

Exercises  for  Practice. 

Try  the  reaction  of  oxalic  acid  on  salts  of  Ca,  Sr,  Ba,  in  neutal  solution,  first 
*Jour.  App.  Micros.  Ill,  8 1 8. 


Fig.  56. 


and  Laboratory  Methods. 


1297 


cold  then  hot.     Draw  off  the  mother  Uquor  and  test  the  precipitate  with  Ho  SO  4. 

Try  the  three  elements  in  test  drops  acidulated  with  nitric  acid.  To  the  drop 
from  which  barium  oxalate  does  not  separate  add  sodium  acetate. 

Try  oxalic  acid  on  a  salt  of  magnesium,  then  add  an  excess  of  acetic  acid  to 
the  test  drop  and  examine  again. 

Test  salts  of  Zn,  Cd,  and  Pb. 

Make  a  mixture  of  Ca,  Sr,  Ba.  Add  H2C2O4.  Repeat  the  experiment  in 
HNO,  solution;  after  a  few  moments,  draw  off  the  clear  solution,  concentrate 
slightly  and  add  sodium  acetate. 

Try  the  effect  of  the  presence  of  ferric  chloride  on  the  precipitation  of  the 
oxalates  of  Ca,  Sr,  Ba;  first  each  element  separately,  then  in  mixtures  of  Ca  and 
Ba  ;    Sr  and  Ba  ;   Ca,  Sr,  Ba. 

If  barium  borate  is  at  hand,  try  testing  it  for  Ba.  E.  M.  Chamot. 

Cornell  University. 


Simple  Washing   Device. 

A  copper  tube  twenty-four  inches  long  and  two  inches  in  diameter,  placed 
horizontally,  is  connected  with  the  faucet  through  a  half-inch  tube  let  in  midway 
above.  The  ends  are  closed.  Below  are  let  in  twenty  quarter-inch  pipes  one 
inch  long.  Over  these  are  slipped  rubber  tubes,  each  carrying  a  nozzle  of  glass 
brought  nearly  to  a  point.  The  nozzle  is  pushed  through  the  cheese-cloth 
fastened  by  rubber  bands  over  the  mouth  of  the  bottle  containing  material  to  be 
washed.  The  bottle  is  made  to  stand  in  a  hole  in  the  plank  forming  the  base  of 
the  support  for  the  main  pipe.     The  water  then  turned  on  descends  through  the 


twenty  feed-pipes,  washing  through  any  bottles  which  may  be  set  into  the 
apparatus.  The  whole  arrangement  stands  in  the  sink.  No  pinch-cocks  are 
needed  for  feed-pipes  not  in  use. 

The  apparatus  was  designed  by  Mr.  Ames,  is  not  expensive,  and  proves 
very  handy. 

The  bottles  used  and  to  be  highly  recommended  are  "  sample-tubes "  of 
rather  thick  glass,  straight  all  the  way  up,  two  and  three-fourths  by  one  and  one- 
fourth  inches.  Material  is  carried  in  the  same  bottle  without  removal,  from 
collection  up  to  the  paraffin  bath.  Robert  G.  Leavitt. 

Ames  Laboratory,  North  Easton,  Mass. 


1298  Journal  of  Applied  Microscopy 

Journal   of  Seaside,   lakeside,   and   field  labo- 

ratories  will   soon   open,  and,  judging 

Applied    Microscopy         from  the  preparations  being  made,  the 

3"'^  attendance  this  year  will  be  larger  and 

Laboratory   Methods.  more  representative   than   ever.     It  is 

interesting  to   note   in   this  connection 
— ::     the  progress  which  has  recently  been 


Edited  by  L.  B.  ELLIOTT. 


Issued  Monthly  from  the  Publication  Department       made  in  the  establishment  and  expan- 

of  the  Bausch  &  Lomb  Optical  Co.,  ^ 

Rochester,  N.  Y.  sion  of  summer  laboratories.     It  is  but 


SUBSCRIPTIONS:  ^  f^^  years  since  Agassiz  and  his  pupils 

One  Dollar  per  Year.     To  Foreign  Countries,  $1.25       began    their    investigations     in    the    CX- 
per  Year,  in  Advance.  °  ° 


==  tremely    unpretentious     laboratory    at 

The  majority  of  our  subscribers  dislike  to  have  their  "PoniL-oco  Tlici      oi->mi     o-        a-ycr\  "11 

files  broken  in  case  they  fail  to  remit  at  the  expiration  .reniKese.  1  ne      COlIling      SeaSOn     Will 

of  their  paid  subscription.     We  therefore  assume  that  no  r:„j    „,„ii    „„.,;„„„j     i„u <- :„„     „    „"i 

interruption   in   the   series  is  desired,  unless  notice  to  "nd    WCll    equipped     laboratories,  CaSlly 
discontinue  is  sent. 


.     accessible  from  all  parts  of  the  country, 

with  hundreds  of  teachers  and  students, 

many  of  them  entering  for  the  first   time   into  the  real   spirit  of  research  work 

and  gaining  a  clearer  view  of  the   possibilities  for  development  in  their  own 

laboratories,  where  of  necessity  the  most  time  is  to  be  spent. 

The  opportunity  to  come  in  personal  contact  with  the  various  forms  of  life  in 
their  native  places,  to  study  them  under  these  most  favorable  conditions  with 
the  assistance  of  experienced  and  enthusiastic  instructors,  and  to  meet  as  co- 
laborers  a  similarly  interested  company,  is  one  which  ought  to  be  taken  advantage 
of  by  every  teacher  of  biological  science,  especially  since  the  cost  is  made  so 
very  moderate.  Specially  prepared  short  courses  are  now  offered  at  most  of  the 
laboratories  which  are  suitable  for  those  beginning  this  work,  and  the  informa- 
tion gained  is  of  such  a  nature  as  to  be  of  practical  assistance  for  class  use. 

The  life  at  a  summer  laboratory  is  conducive  to  physical  recuperation,  and 
the  new  ideas  and  impulses  gained  will  be  an  antidote  for  the  fossilizing 
tendency  of  sticking  too  closely  to  the  native  heath. 

Some  have  helped  defray  expenses  by  collecting  at  the  seaside  laboratory 
sufficient  material  for  class  use  during  the  ensuing  year — star  fish,  sea  urchins, 
crustaceas,  worms,  sea  anemones — which  can  be  easily  preserved  and  sent 
inland  by  freight. 

There  are  in  every  state  many  science  teachers  and  others  preparing  for 
teaching  who  could  spend  two  or  three  months  at  a  summer  laboratory  at 
scarcely  greater  expense  than  any  ordinary  vacation  costs,  and  reap  benefits 
which  could  be  had  in  no  other  way.  "No  doubt  many  who  would  spend  the 
summer  vacation  at  some  laboratory,  do  not  do  so  from  a  lack  of  confidence  in 
the  practical  value  /o  //lem  of  the  work  and  an  exaggerated  idea  of  the  expense 
involved.  We  would  suggest  in  such  instances  correspondence  with  the  directors 
of  the  various  laboratories. 


and  Laboratory  Methods.  1299 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Grout,  A.  J.    Mosses  with  a  Hand-lens.    8vo,      This   convenient   little  book   certainly 

pp.  xi  +  74. 1900.    Published  by  the  author,      supplies  a  long  felt  need.     It  is  a  non- 

360  Lenox  Road,  Flatbush,  New  V  ork  City.  . 

technical  handbook  of  the  more  com- 
mon and  more  easily  recognized  mosses  of  the  Northeastern  United  States.  Two 
general  keys  are  given,  one  based  mainly  upon  structural  characters  and  the 
other  based  mainly  upon  habitat.  With  the  aid  of  these  keys,  the  descriptions 
and  Miss  Thayer's  numerous  excellent  illustrations,  the  student  is  enabled  to 
recognize  about  one  hundred  mosses.  An  illustrated  glossary  of  bryological  terms 
is  an  important  feature.  It  is  a  matter  of  common  observation  that  experienced 
bryologists  make  a  liberal  use  of  the  hand-lens,  while  beginners  are  much  more 
dependent  upon  the  compound  microscope.  All  who  would  become  familiar 
with  the  mosses  are  indebted  to  the  author  for  the  clear  presentation  of  those 
characters  which  will  enable  one  to  recognize  so  many  forms  in  the  field  without 
the  necessity  of  bringing  them  to  the  laboratory  and  making  mounts  for  the  com- 
pound microscope.  However,  it  is  very  probable  that  a  student  who  uses  this 
little  book  will  soon  find  his  interest  increasing  and  will  be  led  to  use  the  more 
extended  and  technical  works  which  would  never  have  attracted  him  at  the 
beginning.  c.  j.  c. 

Campbell,  D.  H.  The  Embryo-sac  of  Peperomia.      In  this,  his  third  paper  on  Peperomia, 

Annals  of  Botany.    15:  lovuS,  pi.  6,  iqoi.       ,,  .,  ,  ,     ,  .,  ^ 

^  J  r         :r  |.jjg  writer  acknowledges  that  some  of 

his  previous  interpretations  must  be  abandoned  and  that  Johnson's  results  are 
substantially  correct.  It  will  be  remembered,  however,  that  Johnson  confirmed 
the  most  important  point  in  Dr.  Campbell's  preliminary  paper,  namely,  that  there 
are  sixteen  nuclei  in  the  embryo-sac  instead  of  eight,  the  usual  number  in  angio- 
sperms.  The  principal  results  of  the  present  work  are  as  follows  :  All  species 
of  Peperomia  seem  to  agree  in  having  sixteen  nuclei  in  the  embryo-sac,  and  there 
is  no  polarity  as  in  other  angiosperms.  The  Q.gg  cell  is  somewhat  differentiated 
by  an  accumulation  of  cytoplasm  about  it,  but  there  are  no  well  marked  syner- 
gids.  Several  (usually  eight)  nuclei  fuse  to  form  the  endosperm  nucleus. 
These  are  regarded  as  homologues  of  the  polar  nuclei  of  typical  angiosperms. 
One  of  the  male  nuclei  from  the  pollen  tube  fuses  with  the  egg  nucleus,  but  the 
fate  of  the  other  male  nucleus  could  not  be  determined.  The  embryo  is  small 
and  shows  no  differentiation  into  organs  when  the  seed  is  ripe.  The  divisions 
of  the  endosperm  nuclei  are  always  accompanied  by  the  formation  of  cell 
walls. 

The  writer  still  believes  that  the  embryo-sac  of  Peperomia  represents  a  primi- 
tive condition  and  that  the  presence  of  sixteen  nuclei  is  not  a  derived  feature. 


1300  Journal  of  Applied  Microscopy 

He  agrees  with  Johnson  and  Strasburger  in  not  regarding  the  fusion  of  polar 
nuclei  as  a  sexual  process,  but  merely  a  physiological  phenomenon.  The  whole 
endosperm  as  well  as  the  antipodal  cells  are  regarded  as  gametophytic  struc- 
tures. Gnefii?fi,  as  described  by  Lotsy,  furnishes  the  nearest  approach  to  the 
embryo-sac  structures  of  Peperomia. 

The  author  had  several  species  of  Peperomia  germinated  at  Kew  and  they 
proved  to  be  genuine  Dicotyledons.  Attention  is  called  to  significant  resem- 
blances to  the  lower  Monocotyledons,  especially  the  Araceae.  The  conclusion  is 
reached  that  Peperomia  is  the  most  primitive  type  of  the  Dicotyledons  and  that 
the  resemblances  between  the  Piperacese  and  lower  Monocotyledons  suggests 
that  the  divergence  of  the  two  groups  may  have  occurred  very  early. 

c.  J.  c. 

Timberlake,  H.  G.     Swarm  Spore  Formation  in       ^"^   this    short    preliminary    note    Prof. 
Hydrodictyon  utriculatum  Roth.    Bot.  Gaz.      Timberlake   announces   some  interest- 
^'   ^    ■  ing  results  of  his  work  on  Hydrodictyon. 

Material  was  fixed  in  a  fluid  recommended  by  Eisen. 

(1)  Iridium   chloride  (0.5  per  cent,  aqueous  solution)    -    100  c.  c. 
Glacial  acetic  acid.     -----------         1  c.  c. 

(2)  Iridium  chloride  (1  per  cent,  aqueous  solution)     -     -     100  c.  c. 

Glacial  acetic  acid,  -       ----------         3  c.  c. 

The  second  solution  gave  better  results.  There  are  no  dififerentiated  chrom- 
atophores,  but  the  chlorophyll  is  distributed  throughout  the  cytoplasm.  The 
nuclei  have  the  structure  of  those  of  higher  plants.  When  the  segments  of 
older  nets  are  to  give  rise  to  swarm  spores,  cleavage  furrows  are  run  in,  at  first 
cutting  out  large  multinucleated  portions  of  cytoplasm,  which  are  then  divided 
and  subdivided  until  each  mass  contains  only  a  single  nucleus.  Each  mass  then 
gives  rise  to  a  single  uninucleated,  biciliated  spore.  c.  j.  c. 

Palisa,  J.      Die   Entvvickelungsgeschichte   der  Among  many  ferns  the  power  of  regen- 
Regenerationsknospen,      welche      an      den  >^-         i         i  i  i  tt    • 

Grundstiicken  isolirter  Wedel  von  Cystop-  oration   has   long  been   known.      Hem- 

teris-Arten  entstehen.    Ber.  d.  deutsch.  bot.  reicher,  in  studying   the   resistance  of 

Gesell,  18 :  398-410,  pi.  14,  1900.  ,        ....        ,      ,      r  /-^     -  . ^     ■   i    jl-j- 

'        jv    t    >  r     t.    y  adventitious  buds  of  Cystoptcris  hulbifera 

to  draught,  found  that  after  the  central  apical  part  of  the  bud  had  decayed,  small 

plantlets  often  arose  from  the  outer  parts,  and  he  ascertained  by  experiments 

that  they  arose  from  the  bud-scales.     He  also  found  that  similar  buds  arose  from 

the  basal  part  of  the  fronds  of  other  ferns.     The  developmental  history  of  the 

adventitious  buds  of    Cystoptcris  and  other  ferns  has  been  determined  by  Hein- 

reicher,  and  in  the  present  article  Palisa  gives  an  account  of  the  development  of 

these  regeneration  buds.     He  worked  mainly  on  two  forms,  Cystoptcris  hulbifera 

and  C.  Montana.     On  the  former,  the  buds  arise  from  the  outer  scales  of  the 

adventitious  buds,  and  on  the  latter  from  the  basal  portion  of  the  fronds.     The 

scales  of  the  former  were  removed  and  placed  in  moist  sand  under  glass  tubes, 

while  in  the  latter  case  the  formation  of  buds  was  invoked  by  cutting  off  from  the 

underground  rhizome  the  still  unrolled  frond  blade. 

Palisa  endeavored  to  answer  two  questions  ;  first,  are  there  any  predetermined 


and  Laboratory  Methods.  1-^01 

cells  from  which  the  buds  arise  ?  Of  four  hundred  scale  leaves  tried,  over  half 
regenerated.  The  location  of  the  regeneration  buds  was  mostly  on  the  flanks 
at  the  leaf  base.  On  older  scales  there  is  a  greater  tendency  for  the  buds  to 
appear  on  the  median  line  of  the  scale.  If  the  scale  be  divided  in  two  by  a  cross 
section,  regeneration  only  occurs  from  the  basal  half.  The  power  of  regenera- 
tion diminishes  with  the  distance  from  the  leaf  base.  Many  hnd  prhnoi'di a  may 
start  together  and  only  one  survive.  If  a  pfimordia/  outgrowth  be  removed 
by  cutting,  then  numerous  primordia  arise  about  the  margin  of  the  cut  surface. 
No  anatomical  diiTerence  could  be  noticed  between  the  epidermal  cells  from 
which  the  buds  arise,  and  the  adjoining  ones,  and  Palisa  concludes  that  they 
may  arise  from  any  of  the  epidermal  cells. 

The  second  question  concerns  the  development  of  the  buds.  They  always 
arise  from  a  group  of  epidermal  cells.  Sections  through  the  scales  show  the 
hypodermal  cells  to  take  no  part  whatever  in  the  development.  The  first  appear- 
ance is  a  dome-shaped  elevation  on  the  surface,  which  soon  becomes  prominent 
above  the  surrounding  tissue.  The  outline  of  the  original  epidermal  cells  remains 
quite  distinct  after  many  divisions  have  occurred.  When  the  outgrowth  reaches 
a  considerable  size,  an  apical  cell  is  organized  and  further  growth  proceeds  from 
it.  From  its  segment  the  frond  is  formed  and  from  the  lower  part  of  the  frond  the 
roots  spring.  In  the  case  of  outgrowths  which  arise  later  and  are  more  scattered 
and  thus  have  more  space,  the  growth  from  each  epidermal  cell  may  organize  an 
apical  cell  and  originate  a  bud.  Between  these  methods  there  is  every  stage  of 
gradation.  A  number  of  apical  cells  may  start,  close  to  one  another,  but  one 
usually  develops  more  rapidly  than  the  rest,  draws  the  nourishment  from  them, 
and  they  cease  to  function. 

Palisa  compares  the   developmental   history  of  these  buds  with  that  of  the 
normal  adventitious  buds  which  always  arise  from  a  single  epidermal  cell. 
Chicago.  W.  B.  MacCalLUM. 


CYTOLOGY,   EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  shoiild  be  sent  for  review  to 

Agnes  M.  Claypole,  Sage  College, 

Ithaca,  N.  Y. 

CURRENT  LITERATURE. 

Nussbaum,  J.,  u.  Prymak.  T.    Zur  Entwickel-  This  work  demonstrates  that  the  leuco- 

ungsgeschichte   der   eymphoiden    Elemente  cytes    of     the   thymus    in     bony    fishes 
der  Thymus  bei  den  Knochenfischen.  Anat.  .        ,  ,       -r         ,  ■     ^      r  .1 

Anz.  19:  6-iQ,  iqoi.  arise  largely   if   not   entirely  from  the 

epithelium ;  this  is  a  point  of  very 
general  significance  in  regard  to  the  germ  layer  origin  of  the  lymphoid  elements. 
The  special  point  of  interest  and  importance  is  the  entire  harmony  of  this  work  with 
that  of  J.  Beard  reviewed  in  March,  1901,  of  the  Journal  of  Applied  Micros- 
copy AND  Laboratory  Methods.  a.  m,  c. 


1302  Journal  of  Applied  Microscopy 

Eismond,  J.    Ueber  die  Natur  der  Sogenann-  The  question  of  the  significance  of  the 

ten  Kinetischen  Centren  der  Zellen.     Anat.  centrosome    is   one   of   the  most  promi- 
Anz.    Centralblt.    fiir   die  Gesammte  Wiss.  ,  ,  .    i      •      i  ,  i 

Anat.  Erganz.  18:  125-141,1900.  "^nt   among  the  cytological  problems 

of  the  present  day  ;  recently  it  is  espe- 
cially considered  in  reference  to  cell  mechanism  as  it  shows  itself  to  be  of 
importance  in  this  connection.  The  view  generally  held  is  that  the  centrosomes 
are  a  distinct,  granular,  primary  element  of  the  cell,  a  permanent  part  of  the 
cell,  and  like  the  nucleus  multiplying  by  self-division.  Opposed  to  this  is  the 
view  of  possible  spontaneous  origin  of  centrosomes.  At  the  same  time  it  is 
possible  the  centrosome  may  be  a  cell-organ,  having  to  do  primarily  with  the 
processes  of  division.  Many  authors  say  that  the  centrosome  collects  around 
itself  a  specially  active  kind  of  protoplasm — kinoplasm- — and  permeates  this  in 
the  form  of  the  different  achromatic  threads  of  the  nuclear  division  figures. 
Further  work  has  developed  the  resemblances  between  this  "  cell  organ  "  and 
basal  bodies  in  ciliated  cells  and  the  blepharoplasts  in  the  plant  antherozoid. 
The  question  is  still  whether  the  centrosome,  the  middle-piece  of  the  spermato- 
zoan,  the  blepharoplasts  and  the  basal  bodies  are  truly  kinetic  centers  of  the 
cell ;  whether  they  originate  the  power  for  such  kinetic  process. 

Previous  work  by  the  author  has  developed  the  view  that  the  centrosome  is 
not  a  pre-formed  organ  in  the  cell,  multiplying  by  division,  but,  at  least  in  embry- 
onic cells,  more  likely  arises  de  novo  and  persists  until  changes  enter  into  the  cell- 
mechanism  to  destroy  the  mechanical  reasons  for  its  existence.  Comparing  the 
"  kinoplasmic  fibers "  of  the  mitotic  figures,  in  so  far  as  they  actually  show 
distinctly  differentiated  parts  of  the  plasmatic  network,  with  the  radiating  fibers 
of  pigment  cell  and  the  "  muscle-threads  "  of  the  protozoan,  compels  the  opinion 
that  these  structures  are  somewhat  similar.  The  special  character  of  the 
muscle-threads  as  a  peculiar  elastic  structure  acts  to  make  them  not  cause  con- 
traction, but  keep  the  body-form  of  the  animal  by  their  elastic  property.  Proof 
of  elasticity,  not  contractility,  lies  in  the  effect  of  reagents  on  these  fibers.  The 
axial  fibers  coil  spirally.  The  comparable  nature  of  the  elastic  supporting 
apparatus  of  Heliozoa  and  tissue  cells  can  be  stated  as  follows :  In  embryonal 
cells  there  is,  as  in  some  tissue  cells  (pigment)  and  protozoa  (Heliozoa),  a 
permanent  structure  which  forms  a  support  for  the  cell-mechanism  and  can  be 
considered  an  elastic  cyto-skeleton.  Considering  the  centrosome  of  embryonic 
cells  in  this  connection,  the  cause  for  division  is  apparent.  The  centrosome  is 
the  inert  central  knot  of  this  elastic  skeleton  and  must  be  divided  by  the 
division  of  the  cell  body.  It  is  a  passive  division.  The  evidence  from  Schaud- 
inn's  work  on  Ancanthocysiis  acukata  shows  clearly  the  origin  of  the  centro- 
some in  the  new  cells,  no  division  of  the  original  centrosome  takes  place.  The 
great  variations  in  the  form  of  the  centrosome,  from  small  central  granules  to 
large,  irregular  axes  as  long  as  the  cell,  or  as  vacuolated  vesicles,  support  the 
de  twvo  view,  and  show  that  the  kinoplasmic  apparatus  near  the  centrosome  is  in 
general  a  supportive  structure,  whose  center  has  different  space  relations  accord- 
ing to  the  mechanical  conditions  involved. 

The  basal  bodies  of  ciliated  cells  are  next  considered.  Cilia  in  most  cases 
have  no  power  of  independent  motion,  but  are  passive,  often  stiff  cell  appendages. 


and  Laboratory  Methods.  1303 

Hence  the  motive  force  lies  outside  these  structures.  The  structures  are  com- 
pared with  the  supporting  bones  of  a  fish's  fin,  and  a  comparison  is  made  to 
bring  out  the  resemblances  caused,  of  course,  by  the  similar  mechanical  condi- 
tions. Finally,  the  author  states  his  belief,  that  the  ciliated  cell  apparatus,  the 
supportive  structures  of  the  mouth  cirri  of  amphioxus  and  the  blepharoplasts 
of  antherozoids  are  all  similar  structures.  a.  m.  c. 

Regaud,  CI.    Quelques  details  sur  la  division  I"  1^99  several  articles  were  published 

amitotique    des   Noyaux  de  Sertoli  chez  le  by  the  same  author  to  show  that  the  cells 

rat.        Sort    du    nucleole        Deux    varietes  r  r-     ^   i-    t           ^     i          •        i              ^  •^- 

d'amitose:      Equivalence    ou    non  -  equiva-  of  Sertoh  do  not  play  Simply  a  nutritive 

lence  des  noyaux  fils.  role  for  sperm  cells,  but  are  cells  cap- 

Anat.  Anz.    Centralblatt  f.  d.  Gesammte  Wis-  ,  ,         ,  .^   ^.        i-    •   •  11 

sen.    Anatomie.     Erganzungsheft    zum  xvii       able   of    amitotic    division,     and    hence 
Bd.,  1900,  p.  110-124,  15  fig.  im  text.  of     producing     spermatogonia.       The 

evidence  for  this  is  based  not  only  on  the  nuclear  figures  clearly  amitotic,  but 
also  on  observations  on  the  stages  of  development  of  the  spermatogonia  and  on 
transition  forms  of  nuclei  between  those  of  the  Sertoli  cell  and  the  spermatogonia  ; 
finally  on  the  impossibility  of  explaining  the  renewal  of  spermatogonia  by  the 
karyokinesis  of  the  other  cells  present.  The  method  used  for  the  study  of 
chromatic  parts  of  the  seminal  epithelium,  is  a  double  stain  of  hsematoxylin  and 
safranin.  This  process  gives  very  good  results  after  fixation  in  Baum's  picro 
aceto-formol  mixture,  and  Lenshossek's  of  sublimate  alcohol  and  acetic  acid. 
The  most  exact  results  follow  the  use  of  Tellyesniczky's  bichromate  of  potash 
and  acetic  acid.  The  sections  are  stained  rather  deeply  with  alum  haematoxylin, 
then  washed  in  water.  If  the  sections  appear  too  deeply  colored  under  an 
immersion  lens  in  water  without  a  cover  glass,  decolorization  follows  with  an 
aqueous  solution  of  formic  acid  (1-100).  Washing  in  ordinary  water  restores 
the  blue  color.  After  this  the  sections  are  stained  for  twenty-four  hours  or 
more  in  Zwaardemaker's  solution  of  safranin.  A  rapid  washing  in  water  is 
followed  by  decolorizing  in  ninety  per  cent,  weakly  acidulated  alcohol  (1  HCl- 
1000  Ale).  The  safranin  is  removed,  but  the  haematoxylin  is  unaffected  : 
neutral  ninety  per  cent,  alcohol  is  followed  by  absolute  and  then  xylol  and 
Canada  balsam.  If  the  two  stains  have  acted  with  just  equal  intensity,  a  con- 
dition easily  obtained  by  practice,  the  cytoplasm  is  stained  a  pale  rose-violet 
and  the  chromatic  parts  are  very  intensely  colored,  sometimes  a  purple-violet, 
sometimes  a  red-purple,  sometimes  intermediate  between  these  colors.  The 
chromatic  granules  in  the  accessory  nucleolus  of  the  nucleus  of  the  Sertoli  cells, 
the  surface  chromatin  of  the  spermatogonia  and  young  spermatocytes,  the  chro- 
matin of  the  nuclear  mass  of  the  spermatocyte  during  the  first  part  of  their 
development,  the  nucleus  of  the  spermatids  during  first  period  of  their  transfor- 
mation into  spermatozoa,  are  all  colored  a  violet-purple.  The  extra  nuclear 
chromatic  bodies  of  the  spermatocyte  and  spermatid,  the  nucleolus  of  the  nucleus 
of  Sertoli  cells,  certain  parts  of  the  nuclear  chromatin  of  the  spermatocyte 
(body  of  Lenshosse'k  at  certain  stages),  the  nucleolus  of  the  spermatocyte,  the 
nucleus  of  the  spermatid  during  the  last  period  of  transformation,  etc.,  are 
colored  a  red-purple.  Intermediate  between  these  are  certain  chromatic  bodies 
of  the  young  spermatogonia  and  the  chromatin  of  the  nuclear  filament  of  the 
spermatocyte  in  certain  stages.     During  the  karyokinesis  of  spermatogonia  their 


1-304  Journal  of  Applied  Microscopy 

chromatin  is  a  violet-purple ;  during  the  karyokinesis  of  the  spermatocytes  their 
chromatin  is   always  a  red-purple.     The  author  sums  up  his  results  as  follows  : 

1.  The  amitotic  division  of  the  nucleus,  which  in  most  cases  indicates  a 
degeneration  of  the  cells,  does  not  always  show  the  approach  of  final  degener- 
ation. The  nuclei  of  the  Sertoli  cells  divide  a  considerable  number  of  times, 
perhaps  indefinitely,  by  amitosis.  The  spermatogonia  resulting  from  amitosis 
are  the  founders  of  a  line  of  cells  which  show  ultimately  more  karyokinesis  and 
finally  develop  into  spermatozoa. 

'1.  Amitosis  in  the  case  noted  is  the  same  as  that  in  many  others  :  a  phenom- 
enon of  the  nucleus  only,  without  an  immediate  division  of  the  protoplasm. 
Much  later  the  protoplasm  divides. 

3.  The  nucleolus  of  the  Sertoli  cell  appears  to  be  a  cellular  organ  of 
primary  importance,  it  is  possibly  the  carrier  of  a  reserve  of  hereditary  sub- 
stance. 

4.  It  is  remarkable  that  the  nuclei  of  the  Sertoli  cells,  the  stem  nuclei 
which  carry  the  determinants  (Weissmanh)  of  the  species,  are  really  the  poorest 
in  chromatin  of  all  the  nuclei  of  the  germinal  epithelium.  The  quantity  of 
chromatin  which  passes  into  the  nucleus  of  the  spermatogonia  is  infinitesimal  in 
comparison  with  that  contained  by  the  spermatocyte  at  the  moment  of  karj'O- 
kinesis.  The  chromatin  of  the  spermatocyte  is  then  acquired,  at  least  in  its 
mass,  and  is  not  hereditary.  a.  m.  c. 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Ewing,  J.    Malarial  Parasitology.    Journal  of      Recent  advances  in  our  knowledge  of 
Experimental  Medicine,  S:  420-401,  iqoi.  ,  ,     ,  ^     ,  ,      •    , 

the  morphology  of  the  malarial  para- 
sites have  been  largely  due  to  improved  staining  methods.  Ewing  restricts  the 
use  of  fresh  blood  to  the  study  of  various  vital  phenomena  in  the  parasite,  such 
as  amoeboid  movement,  vibratory  motion  of  pigment,  and  ex-flagellation.  The 
discovery  of  parasites  is  so  much  more  certain  and  rapid  in  stained  dry  speci- 
mens that  a  negative  result  with  fresh  blood  invariably  requires  verification  by 
search  through  a  dry  specimen,  stained  preferably  by  Nocht's  method. 

For  all  ordinary  purposes  staining  by  eosin  and  methylen  blue  is  recom- 
mended. The  solutions  used  are  :  (a)  a  saturated  alcoholic  solution  of  alcoholic 
eosin  diluted  with  an  equal  quantity  of  95  per  cent,  alcohol,  and  (b)  a  saturated 
watery  solution  of  Ehrlich's  rectified  methylen  blue  at  least  one  week  old. 

Methylen  blue  does  not  stain  the  young  ring  forms  well.  For  this  purpose 
Nocht's  method  is  especially  useful.  The  method  of  Benario  and  Marchoux  as 
modified  by  Futcher  and  Lazear  is  also  of  value,  as  the  rings  are  densely  stained 
and  the  preparations  are  permanent.  It  is  employed  as  follows  :  Fix  the  speci- 
mens five   minutes  in  95  per  cent,  alcohol,  100  c.  c,  to  which  has  been  added 


and  Laboratory  Methods.  1305 

1  c.  c.  of  formalin.  Stain  one  to  three  minutes  in  the  following  mixture  :  satu- 
rated alcoholic  solution  thionin,  '20  c.  c,  "JO  per  cent,  carbolic  acid,  100  c.  c. 
The  fixing  solution  must  be  fresh,  and  the  staining  fluid  at  least  one  week  old. 

Nocht's  modification  of  Romanowsky's  method  consists  in  the  addition  of  a 
few  drops  of  neutralized  Unna's  polychrome  methylen  blue  (Griibler)  to  the  1  per 
cent,  solution  of  ordinary  methylen  blue.  The  author  obtained  uniformly  good 
results  by  the  following  procedure :  (1)  To  1  oz.  of  polychrome  methylen  blue 
(Griibler)  add  5  drops  of  3  per  cent,  solution  of  acetic  acid  (U.  S.  P.  33  per  cent). 
(2)  Make  a  saturated  (1  per  cent.)  watery  solution  of  methylen  blue,  preferably 
Ehrlich's  rect.  (Griibler),  or  Koch's,  dissolving  the  dye  by  gentle  heat.  This 
solution  improves  by  age,  and  should  be  at  least  one  week  old.  (3)  Make  a 
1  per  cent,  solution  in  water  of  Griibler's  aqueous  eosin.  The  mixture  is  prepared 
as  follows  :  To  10  c.  c.  of  water  add  4  drops  of  the  eosin  solution,  6  drops  of  the 
neutralized  polychrome  methylen  blue,  and  '2  drops  of  1  per  cent,  methylen  blue, 
mixing  well.  The  specimens,  fixed  in  alcohol  or  by  heat,  are  immersed  for  two 
hours,  specimen  side  down,  and  will  not  overstain  in  24  hours.  The  red 
corpuscles  are  stained  light  pink,  the  body  of  the  parasite  blue,  while  the  chro- 
matin particles  of  the  nucleus  appear  deep  red. 

Nocht's  procedure  is  also  of  value  in  studying  the  nuclear  structures  in  other 
micro-organisms. 

Goldhorn  (N.  Y.  Path.  Soc,  Feb.  13,  1901)  has  succeeded  in  increasing  the 
amount  of  the  red  staining  principle  by  digesting  polychrome  methylen  blue  with 
lithium  carbonate.  He  stains  the  specimen  for  a  few  seconds  in  n.l  per  cent, 
watery  solution  of  eosin,  then  in  digested  polychrome  blue  30  to  t50  seconds. 

Ewing  found  no  evidence  for  the  view  that  there  is  more  than  one  species  of 
the  aestivo-autumnal  parasite.  The  nucleus  of  the  malarial  parasite  belongs  to 
the  "  distributed  type  "  of  protozoan  nuclei,  consisting  of  granules  of  chromatin. 
While  not  a  true  nucleus  in  the  metazoan  sense,  it  possesses  all  the  nuclear 
structures  required  in  some  protozoa.  He  believes  that  conjugation  of  malarial 
parasites  occurs.  His  observations  seem  to  him  to  admit  of  no  other  explana- 
tion ;  but  he  does  not  regard  conjugation  as  an  essential  feature  of  the  growth  of 
the  parasite.  He  regards  the  existence  of  several  species  of  malarial  parasites 
as  not  yet  proven,  and  adheres  to  the  theory  that  there  is  a  single  polymorphous 
species.  j.  h.  p. 

Rosenberger,  R.  C.    A  New  Blood  Stain.  Phil-      Rosenberger   discovered    that    phloxin 
adelphia  Medical  Journal,  7:  448,  1 90 1.  stains    the    granules    of   the    leucocytes 

remarkably  well.     He  recommends  the  following  solution  as  a  differential  stain 
for  the  cells  in  the  blood  : 

Saturated  aqueous  solution  of  methylen  blue,     -         -         5  parts 
Saturated  aqueous  solution  of  phloxin,  -         -  -     2  parts 

Alcohol  (95  per  cent.),  -----         3  parts 

Distilled  water,     --------     6  parts 

These  are  mixed  together.  A  precipitate  generally  forms.  The  stain  should 
be  well  shaken  before  using.  It  works  well  after  fixation  by  heat,  alcohol  and 
ether,  or  absolute  alcohol.  Stain  one  to  four  minutes,  wash  freely,  dry,  mount 
in  balsam.  j.  h.  p. 


1306  Journal  of  Applied  Microscopy 

Baum.  E.  Ueber  die  punktformigen  Kalkkor-  ^^^  yellowish-white  points  sometimes 
perchen   (sogen    verkalkte  Glomerule)    der       seen  on  the  surface  of  the  kidney  have 

Nierenrinde.  Virchow's  Archiv,  162  :  85-93,  ^een  regarded  as  calcified  glomeruli. 
1900.  '^  ° 

In  the  great  majority  of  cases  they  are 

not  glomeruli,  but  cysts  containing  lime-salts.  These  cysts  are  of  two  kinds. 
They  are  present  in  kidneys  in  which  there  is  no  evidence  of  chronic  interstitial 
changes.  The  larger,  irregularly  shaped  cysts  arise  from  the  uriniferous  tubules. 
Their  walls  are  lined  in  places  with  high  epithelium.  The  cysts  of  the  other 
variety  are  round,  about  the  size  of  a  glomerulus  and  confined  to  the  cortex. 
They  represent  capsular  spores  of  malpighian  corpuscles,  in  which  glomeruli  have 
not  developed.  The  lining  epithelium  when  present  is  of  a  low  type.  The 
lime  is  deposited  in  the  colloid  material  which  fills  the  cysts.  The  lime  occurs 
as  small  granules  and  as  concentric  masses.  Only  rarely  does  a  sclerosed  glom- 
erulus become  calcified.  j.  h.  p. 

whii^^.,  w  c  A  r^  1  J  c-  1  A/r  .u  J  The  writer  states  that  the  various  meth- 
Wnitney,  W.  t.     A  Quick  and  Simple  Method 

for  Fixing  the  Blood  Corpuscles  for  Differ-  ods  for  the  rapid  fixation  of  blood 
ential  Staining.  Jour.  Boston  Soc.  Med.  Sci.  g^^^ars  that  have  been  devised  are  all 
5:  341,  1901. 

uncertam.  The  method  has  given  uni- 
formly good  results.  It  consists  simply  in  the  use  of  a  modified  Zenker's  fluid. 
This  solution  consists  of  a  mixture  of  potassium  bichromate  two  parts,  sodium 
sulphate  one  part,  water  100  parts,  saturated  with  corrosive  sublimate,  plus  5 
per  cent,  of  glacial  acetic  acid.  In  Whitney's  modification  5  per  cent,  of  strong 
nitric  acid  is  substituted  for  the  5  per  cent,  of  acetic  acid. 

The  blood  is  drawn  and  spread  in  the  usual  way  and  dried  thoroughly  in  the 
air  or,  if  preferred,  by  a  gentle  heat.  The  cover-glass  is  taken  with  the  forceps, 
the  prepared  surface  covered  with  a  few  drops  of  the  fluid  and  held  while  twenty 
are  counted  slowly.  It  is  washed  off  with  running  water  and  blotted.  The 
action  depends  upon  the  combination  of  corrosive  sublimate  with  potassium 
nitrate  and  chromic  acid  which  are  formed  in  the  solution. 

Ehrlich's  triacid  stain,  Unna's  polychrome  methylen  blue  and  Chenzinski's 
eosin  work  well  after  the  fixation.  j.  h.  p. 

Goldhorn,  L.  B.  A  Rapid  Method  of  Staining  1-  Fix  fresh  preparation  by  immersion 
the  Chromatin  of  the  Malaria  Parasite.  in  pure  methyl  alcohol  for  15  seconds. 
Med.  Rec.  59:   ii.  ^    ,.r     ,    •  • 

z.  Wash  m  runnmg  water. 

3.  Stain  for  7  to  30  seconds  in  0.1  per  cent.  aq.  sol.  of  eosin. 

4.  Wash  in  running  water. 

5.  Stain  for  80  to  60  seconds  in  polychrome  solution. 

6.  Wash  again  and  dry  in  air ;  no  filter  paper  or  heat  being  used. 

If,  on  exposure  to  air,  the  dye  becomes  too  alkaline,  a  few  drops  of  a  4-5  per 
cent,  solution  of  acetic  acid  may  be  added.  If  this  amount  of  acid  proves  too 
much,  add  a  few  drops  of  a  saturated  aqueous  solution  of  lithium  carbonate. 
The  stain  improves  on  keeping.  c.  w.  j. 


and  Laboratory  Methods.  130" 


GENERAL   PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Schenck,  F.     Physiologische  Charakeristik  der      This  work  aims   to  determine   in   how 
Zelle.     Wiirzburg,    A.   Stuber's   Verlag  (C.       ,       ,,  ,,  ,  .  ,         ,       ,,     , 

Kabitzsch).  8vo,  pp.  viii  and  123,  1899.  ^^r  the  cell  may  be  considered  a  "  phys- 
iological unit  "  or  "  elemental  organ- 
ism," and,  having  settled  this  point,  to  examine  the  real  physiological  signifi- 
cance of  the  cell  and  its  parts.  The  method  taken  is  a  critical  examination  and 
analysis  of  a  considerable  part  of  the  important  literature  bearing  on  the  subject. 
The  author  is  strongly  opposed  to  the  view  that  the  cell  presents  the  simplest 
condition  of  life  phenomena,  and  that  a  physiological  study  of  the  cell  ought  to 
precede,  and  form  a  basis  for  the  "  organ  physiology "  of  higher  animals. 
Nearly  a  third  of  the  book  is  occupied  with  a  criticism  of  this  view,  the  criticism 
being  mainly  directed  towards  Verworn.  The  principal  points  made  in  this  por- 
tion of  the  work  are  :  1.  That  all  cells  are  not  capable  of  independent  existence 
and  hence  are  not  physiological  individuals.  '2.  That  since  some  multicellular 
forms  are  physiological  individuals,  this  sort  of  individuality  must  be  independ- 
ent of  the  formation  of  the  organism  out  of  cells.  3.  That  in  a  multicellular  or- 
ganism the  cells  are  in  organic  connection  with  one  another  by  means  of  proto- 
plasmic strands  and  that,  therefore,  the  whole  must  be  considered  as  the  individ- 
ual. 4.  That  the  study  of  the  contraction  phenomena  in  unicellular  animals 
does  not  lead  to  any  better  understanding  of  the  contractility  of  muscle,  and 
furthermore  that,  in  all  probability,  the  phenomena  are  simpler  and  lend  them- 
selves more  readily  to  analysis  in  the  latter  than  in  the  former  case. 

The  more  distinctly  constructive  contributions  have  as  their  purpose  to  find 
whether  the  whole  cell  or  only  parts  of  it  are  necessary  in  the  carrying  on  of  the 
fundamental  life  processes.  These  processes  are  discussed  under  four  heads. 
The  first  to  be  considered  is  the  relation  of  the  cell  to  "  physiological  combus- 
tion "  or  oxidation,  and  to  the  phenomena  which  primarily  depend  upon  the  oxi- 
dation of  the  living  substance.  The  author  makes  citations  from  the  literature 
which  show,  according  to  his  belief,  that  movement,  processes  of  dissimilation, 
electrical  phenomena,  irritability,  etc.,  may  take  place  in  enucleated  cell  frag- 
ments. He  concludes  that  "  physiological  combustion  "  does  not  depend  upon 
the  combined  action  of  all  the  parts  of  the  cell,  and,  therefore,  that  so  far  as  this 
process  is  concerned  the  cellular  structure  of  the  organism  is  without  significance. 
The  next  processes  to  be  considered  are  those  of  assimilation,  growth  and  mor- 
phogenesis. The  point  brought  out  here  is  that,  while  enucleated  protoplasm  is 
capable  to  some  slight  extent  of  carrying  on  the  processes  of  assimilation,  growth 
and  regeneration,  yet  these  processes  can  only  go  on  continuously  when  both  the 
characteristic  cell  parts,  nucleus  and  cytoplasm,  act  together.  It  is  maintained, 
however,  that  the  cell  is  dependent  in  its  formative  activity  on  the  whole  organ- 
ism of  which  it  is  a  part.     The  division  of  labor  between  nucleus  and  cytoplasm 


1308  Journal  of  Applied  Microscopy 

is  the  next  topic  discussed.  It  is  believed  that  the  cytoplasm  has  to  do  prima- 
rily with  the  relation  of  the  organism  to  the  external  environment,  i.  e.,  with  the 
reactions  to  stimuli ;  while,  on  the  other  hand,  the  nucleus,  as  a  result  of  its  as- 
similatory  activity  which  conditions  growth  and  reparation,  enables  the  organism 
to  continue  living.  In  this  section  the  author  offers  some  very  interesting  and 
suggestive  theories  as  to  the  physiological  significance  of  the  division  of  organ- 
ism into  cells,  and  the  phylogenetic  origin  of  the  nucleus.  In  a  short  section  on 
cell  and  nuclear  division  it  is  maintained  that  the  centrosome  is  the  part  of  the 
cell  essentially  concerned  with  these  processes. 

In  concluding,  Schenck  condemns  the  view  that  "  cell  physiology  "  is  synony- 
mous with  "  general  physiology,"  and  even  considers  that  the  attention  which 
has  been  paid  to  the  former  has  tended  to  hinder  the  progress  of  the  latter.  The 
work  is  throughout  one  of  interest,  and  in  many  respects,  of  value.  The  style  is 
uncommonly  clear  and  straightforward.  The  thing  which  most  mars  the  work 
is  the  polemic  character  which  pervades  the  whole  and  at  times  descends  to  ab- 
surd personalities.  r.  p. 

Cole,  L.  J.    Notes  on  the  Habits  of  Pycnogo-      This   paper,   although  brief,  furnishes 
nids.     Biol.  Bull. 2:  iqi;-207.  iqoi.  .  •,     ,• 

an      important     contribution    to    our 

knowledge  of  the  general  physiology  of  the  somewhat  neglected  group,  the 
Pycnogonida.  The  principal  points  treated  are  the  movements  and  the  reactions 
to  light,  both  of  which  are  described  in  detail.  Two  curious  facts  brought  to 
light  in  connection  with  the  movements  are  :  (a)  the  action  of  the  legs  is  pre- 
cisely the  same  in  both  the  swimming  and  crawling  movements  ;  and  (b)  the 
stroke  of  the  anterior  legs  is  found  to  be  stronger  than  that  of  the  posterior,  and, 
since  the  action  of  both  is  qualitatively  the  same,  it  results  from  the  structural 
relations  of  the  body  that  the  posterior  legs  act  as  hindrance  to  forward  move- 
ment when  the  animal  is  crawling.  The  pycnogonids  studied  are  shown  to  be 
positively  phototactic,  but  the  precise  form  of  the  orientation  differs  according 
as  the  animal  swims  or  crawls.  When  crawling  towards  the  light  the  anterior 
end  of  the  body  precedes,  while  when  sivimming  towards  the  light  the  posterior 
end  is  in  advance.  The  reaction  is  the  same  in  the  two  cases,  but  the  result  is 
conditioned  by  the  mechanical  relations  of  the  animal  to  a  solid  object,  i.  e.,  the 
bottom.  This  point  is  of  considerable  theoretical  interest  as  indicating  that  the 
essential  thing  in  an  orientation  is  not  the  getting  of  one  end  of  the  organism  to- 
wards or  away  from  the  source  of  the  stimulus,  but  is.  on  the  contrary,  the 
placing  of  the  axes  of  the  body  in  definite  relations  to  the  lines  of  action  of  the 
directive  stimulus.  The  transfer  of  the  eggs  from  the  female  to  the  male  was  ob- 
served and  found  to  be  a  comparatively  simple  process,  apparently  involving  no 
psychic  activity  on  the  part  of  the  animals.  R.  p. 

Ritter,  W.  E.,  and  Congdon,  Edna  M,  On  the  In-      The  question  as  to  the  effect  of  an  arti- 
hibition  by  Artificial  Section  of  the  Normal       r    •   \         ^-  c  -i-^l^^ 

Fission  Plane  in  Stenostoma.  Proc. California  ^Cial  section  of  an  animal  JUSt  about  tO 
Acad.  Sci.  (Third  Series).  Zoo).  2  :  divide  by  fission  was  suggested  to  the 
3  5-37  '  P  •  xv">    9    •  senior   author    of    this   paper,    in    the 

course  of  his  work  on  monogenesis  in  ascidians.  A  partial  answer  is  gained  from  this 
study  of  the  common  rhabdocoele  turbellarian  Stenostoma  leucops  O.  Schm.    The 


and  Laboratory  Methods.  1309 

method  taken  was  to  cut  across  with  a  small  scalpel  an  individual  in  which  the 
normal  fission  plane  had  become  well  formed,  at  some  other  point  of  the  body 
than  that  at  which  the  fission  plane  was  appearing.  It  was  found  that  when  the 
cut  was  anterior  to  the  normal  fission  plane,  the  formation  of  this  was  inhibited 
and  the  ganglionic  masses  which  mark  its  position  moved  forward  till  they  came 
to  the  cut  anterior  end,  where  a  head  was  formed.  In  other  words,  the  tissue 
which  was  to  form  a  head  migrated  as  a  result  of  the  operation  to  a  position 
where  head  tissue  would  normally  never  occur.  When  the  cut  was  posterior  to 
the  normal  fission  plane  the  latter  was  not  inhibited,  but  the  operation  had  a  dis- 
tinct retarding  influence  on  its  formation.  The  rule  in  this  case  appears  to  be 
that  the  normal  fission  plane  is  not  completely  formed  until  the  posterior  piece 
which  has  been  cut  away  is  wholly  regenerated. 

In  the  theoretical  discussion  a  comparison  is  made  between  the  regulation 
shown  in  this  case  and  that  of  the  blastomeres  of  the  segmenting  egg.  It  is 
thought  that  the  fundamental  cause  of  the  migration  of  the  ganglionic  cell  mass 
is  to  be  found  in  the  "-specific  form  of  the  animal,''  or,  to  quote  the  exact  words-of 
the  authors :  "  We  may  conceive  all  the  tissues  of  the  individual  animal  to  be  in 
a  state,  not  of  ^^?<!/-librium,  but  of  Stenostoma-\\\iX\\mi ;  and  that  when  this  is  dis- 
turbed in  any  way  the  whole  system  together  tends  to  re-establish  it ;  and  this 
may  be  done  through  physico-chemical  means.' 

The  paper  is  one  of  great  interest  and  suggests  many  possibilities.       R.  p. 

„..,,,,      ^.  ^         ,      T  ,         •  In  these  papers  are  given  the  results 

Qauie,  J.     Ueberden  Einfluss    der   Jahreszeit  i     i                   o 

auf  das  Gewicht  der  Muskeln  bei  Froschen.  of    a    Series    of    weighings    of    certam 

Arch.  f.  d.  ges.  Physiol.    Bd.  83,  p.  81,  82.  muscles  of  frogs  of  both  sexes,  at  dif- 

Taf.  V,   1900.  ^                                  •    r          1 

Ueber  die  geschlechtliche  Differenz  der  Mus-  ferent  seasons  of  the  year.      It  IS  found 

keln  bei  Froschen.      Ibid.  p.  83-88.    Taf.  ^^at    during    the     summer    while    the 

V,   1900.  ^     ,. 

frogs  are  feedmg  the  muscles  take  on 

weight,  while  in  the  winter  months,  when  there  is  no  feeding  and  the  sexual  pro- 
ducts are  being  formed,  the  muscles  lose  weight.  The  muscle  weight  of  the 
males  is  at  all  times  greater,  per  unit  of  body  weight,  than  that  of  the  females. 
Physiological  measurements  were  also  made  of  the  relative  amounts  of  work 
done  by  the  isolated  gastrocnemius  muscles  from  the  two  sexes,  when  they  were 
put  under  the  same  experimental  conditions  and  stimulated  in  the  same  way. 
The  results  here  show  that  the  muscle  of  the  male  shortens  more  in  contracting 
than  that  of  the  female,  but,  on  the  other  hand,  the  muscle  from  the  female 
raises  a  slightly  greater  weight  than  that  of  the  male.  The  product  gained  by 
multiplying  the  height  through  which  the  weight  is  raised  by  the  amount  of  the 
weight,  gives  a  measure  of  the  work  done,  and  from  determinations  made  in  this 
way  it  appears  that  the  muscle  of  the  male  frog  is  capable  of  doing  more  work 
than  that  of  the  female.  The  author  believes  that  the  material  for  the  formation 
of  the  sexual  products  is  taken  directly  from  the  muscles  where  it  has  been 
stored  during  the  summer  feeding  season.  These  differences  in  the  condition 
of  the  muscles  between  the  females  and  the  males  are  thus  thought  to  be  due  to 
the  greater  amount  of  energy  required  in  the  forming  of  the  sexual  products  in 
one  case,  over  that  required  in  the  other.  R.  p. 


1-^10  Journal  of  Applied  Microscopy 

CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

1.  Moore,  V.  A.     Directions   for   Beginners    in  -phe  very  rapid  development  of  courses 
isactenology.     Ginn  &  Co.,  Boston,  Mass.  j       r                    r 

2.  Frost,  W.  D.    A  Laboratory  Guide  in  Ele-  ^^  bacteriology  has  led  to  the  appear- 
mentary  Bacteriology.     Madison,  Wis.  ance  of   quite  a  number  of   outlines  for 

3.  Curtis,  fi.  J      Essentials   of   Practical    Bac-       practical  bacteriological  courses.      The 
teriology.      Longmans,   Green  &  Co.,  New       ^  ° 

York.    '  three    books   here   referred  to  are   all 

designed  as  laboratory  guides.  The  first,  by  Dr.  Moore,  gives  a  series  of 
sixty-four  lessons  in  bacteriological  technique,  covering  the  general  topics  which 
students  study  in  practical  bacteriology.  The  Laboratory  Guide,  by  Frost,  is 
more  specially  designed  as  a  laboratory  note-book  in  the  study  of  bacteriology. 
About  half  of  it  is  taken  up  with  blanks  to  be  used  by  students  in  describing  species 
of  bacteria,  while  the  other  half  is  devoted  to  various  exercises  in  general  bacteri- 
ology. The  Practical  Bacteriology,  by  Curtis,  is  a  somewhat  more  extended  work, 
and  contains  a  great  deal  more  information  than  the  other  two  books.  It  is  full  of 
illustrations  of  apparatus  and  laboratory  devices,  as  well  as  of  the  chief 
species  of  bacteria.  In  addition  to  laboratory  technique  it  gives  a  large  amount 
of  information  in  regard  to  various  bacteria,  and  their  relations  to  disease.  This 
work  is  especially  useful  as  giving  the  student  not  only  a  knowledge  of  laboratory 
technique,  but  also  a  great  amount  of  information  in  regard  to  the  significance 
of  the  organisms  which  he  is  studying.  The  work  is  especially  full  in  its  descrip- 
tions of  some  of  the  more  unusual  forms  of  bacteria.  Ringworms  and  cancer 
are  considered  in  very  considerable  detail,  with  the  purpose  of  indicating  lines 
of  research  for  advanced  students.  A  more  detailed  study  of  the  Actinomyces 
and  its  allies  is  given  than  can  be  found  in  most  works  of  bacteriology.  For 
these  reasons  the  student  will  obtain  from  the  work  much  more  than  would  be 
indicated  by  a  common  course  in  laboratory  bacteriology.  h.  w.  c. 

Dinwiddie.      The    Relative    Susceptibility    of       The    author   has   conducted   some  very 
Domestic    Animals    to    the    Contagion    of  .  .  , 

Human  and  Bovine  Tuberculosis.     Bui.  No.       suggestive  experiments   to  test  the  con- 
63  Ark.  Agri.  Exp.  Sta.  clusion  as  to  whether  the  bovine  and 

human  tubercle  bacillus  are,  as  has  been  claimed  by  Smith,  different  in 
their  pathological  properties.  For  this  purpose  he  experiments  not  only  with 
cattle,  but  with  other  animals.  The  result  of  the  experiment  is  not  only  to 
verify  Smith's  conclusions,  but  to  extend  them.  He  finds  that  the  bovine 
bacillus  is  always  more  virulent  than  the  human  bacillus,  and  that  this  is  true 
whether  the  tests  are  made  on  cattle,  sheep  or  pigs.  So  far  as  his  experiments 
go,  they  appear  to  indicate  that  the  bovine  bacillus  is  more  virulent  for  all  sus- 
ceptible animals.  It  certainly  appeared  to  be  for  all  animals  experimented  with, 
and  the  author  infers  that  it  is  also  more  virulent  for  men.  This  inference, 
which  is  of  extreme  importance,  the  author  recognizes,  however,  as  not  yet 
proved.  h.  w.  c. 


and  Laboratory  Methods.  1311 

Obermuller.     Ueber  neuere'  Untersuchung  des      Having  been  for  many  years  engaged 
Vorkommen    echter  Tuberkuloseereger   in       .        ^,  .     ,         r    i.   v.        i        u      -ir     • 

der  Milch  und  den  Molkereiproduktin  be-       1"     the     Study    of    tubercle     bacilh    in 
treffend.    Hyg.  Rund.,  10:  845,  1900.  dairy  products,  the  author  in  this  arti- 

cle summarizes  his  general  conclusions  in  regard  to  the  proper  relation  which 
should  be  taken  towards  this  highly  important  problem.  These  conclusions  are 
hardly  capable  of  brief  summary.  The  most  important  are  as  follows :  Milch 
cows  should  be  subjected  to  obligatory  inoculation  by  tuberculin  under  state  law. 
Bovine  tuberculosis  can  be  reduced  and,  perhaps,  largely  gotten  rid  of  by  the 
gradual  destruction  of  tuberculous  animals  which  show  signs  of  the  disease, 
especially  those  with  udder  tuberculosis.  For  infants  and  invalids  especial  care 
should  be  taken  to  use  milk  from  sound  cows  only.  Cream  freed  from  tubercle 
bacilli  should  alone  be  used  for  butter  making.  General  mixed  milk  from  the 
market  is  a  source  of  danger,  unless  such  milk  is  pasteurized.  The  author 
advocates  the  establishment  of  governmental  bacteriological  stations,  whose  duty 
it  shall  be  to  test  market  milk  for  the  tubercle  bacillus  and  other  pathological 
bacteria.  h.  w.  c. 

Tobler,  Maria.    Beitrag  zur  Frage  des  Vorkom-      The  author  takes  up  the  investigation 
mens    von   Tuberkel  bacillen   und  anderen  ,   ^,       ^    ,         1      u      -u  j    -^        11  • 

Saurefesten    Bacillen    in    der    Marktbutter.       O^   the   tubercle   bacillus   and   its    allies 
Zeit.  f.  Hyg.  u.  Inf ec,  36 :  120,1901.  in  the  market  butter  of  Zurich.     The 

conclusions  reached  are,  in  general,  in  accordance  with  those  obtained  by  others, 
inasmuch  as  true  tubercle  bacilli  are  found  in  a  certain  number  of  the  samples 
of  market  butter.  The  special  point  of  interest  in  the  investigation  is  the  dis- 
covery of  five  new  species  of  bacilli  in  the  butter,  which  microscopically 
resemble  the  tubercle  bacillus  and  have  the  same  power  of  holding  stains 
against  the  action  of  acids.  These  five  "  sauerfest  "  bacteria  are  pathogenic  for 
various  animals,  but  they  are  wholly  difTerent  from  the  tubercle  bacillus  and 
different,  also,  from  the  similar  organisms  described  by  Rabinowitsch  and  others. 

H.  w.  c. 

Rabinowitsch,  Lydia.     Ueber   die  Gefahr  der      The  author  has  continued  the  investi- 
Uebertragung  der  Tuberkulose  durch  Milch  ^.  ^    l        1      u      -n-  •       j    • 

und  Milchprodukte.     Cent.  f.  Bak.  u.  Par.  i,      g^tions   upon    tubercle   bacilh  m   dairy 

29,  p.  309,  1 90 1.  products,  in   which   she  has  for  some 

Befund  von  Sauerfest  tuberkelbacillenannhch-  ,  j        tt  1 

en  Bakierium  bei  Lungen  gangran.    Deutsch       Y^ars  been  engaged.      Her  general  COn- 
Med.  Woch,  1900.  elusions    are     expressed    as    follows : 

Three  dairy  supply  companies  which  regularly  test  their  cows  with  tuberculin, 
and  whose  milk  she  has  carefully  studied,  furnish  a  product  entirely  free  from 
tubercle  bacilli.  Other  dairies,  that  depend  entirely  upon  clinical  examinations 
by  veterinarians,  furnish  milk  which  frequently  contains  living,  virulent  bacilli. 
The  conclusion  is,  of  course,  that  a  clinical  examination  of  cows  is  insufficient 
to  guarantee  the  freedom  of  the  milk  from  tubercle  bacilli.  The  author  recom- 
mends the  sale  of  milk  from  herds  tested  with  tuberculin  at  a  price  higher  than 
that  of  ordinary  milk. 

In  the  second  article  the  author  discovers  in  the  sputum  of  persons  suffer- 
ing from  gangrene  of  the  lungs,  a  bacillus  which  is  microscopically  identical 
with  the  tubercle  bacillus.  The  organism  in  question,  upon  careful  study, 
proves  not  to  be  the  tubercle  bacillus,  but  one  of  the  "  sauerfest  "  bacilli  which 
are  coming  now  to  be  recognized  as  so  abundant  in  dairy  products. 

The  author  makes  no  attempt  to  draw  any  casual  connections  between  the 
disease  and  the  bacillus.  h.  w.  c. 


131'i  Journal  of  Applied  Microscopy 

NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Clarke,  p.  W.  The  Constitution  of  Tourma-  Recent  investigations  of  Penfield  and 
line.    Am.  Jour.  Sci.  iv,  8 :  in,  189Q.  Yoote  have  led  the  author  to  modify  in 

some  particulars  his  formula;  proposed  in  1895. 

Clarke  regards  all  tourmalines  as  derived  from  the  alumino-boro-silicate  acid 
Hj^Al^BgSigOg,  with  all  the  H  atoms  replaceable  by  bases.     Using  the  formula : 

AlgCSiOJe  (B02)2.  BO3H2.H,., 
which  is  applicable  to  the  discussion  of  the  analyses,  the  author  shows  that  the 
results  of  analyses  can  be  expressed  by  the  combination  of  diiiferent  molecules, 
which  are  all  derived  from  the  general  formula  by  the  substitution  of  different 
bases  for  H.  For  example,  the  composition  of  the  brown  Gouverneur,  N.  Y. 
tourmaline  corresponds  to  the  following  molecular  mixture  : 

5.  A\,  (SiOJe  (60^)2  •    B03Ca  .    Mg,H„ 
3.  Al2(Si04)6(B02)2  .    BO.Mg.   Mg^H^, 
2.  Al2(Si04)6(B02)2  .    BOgNaH  .    Al2Na2H4. 
An  elaborate  chemical  discussion  is  given  and  many  examples  cited. 
Future  investigations  may  prove  tourmaline  to  be  derived  from  a  complex 
boro-silicic  acid ;  hence  the  constitution  must  still  be  regarded  as  unsettled. 

L.  McI.  L. 

„    ,     „,  .,     -,  TVT      i,T        •    T  The  occurrences  are  of  the  usual  type, 

Foote,  W.  M.     Note  on  a  New  Meteoric  Iron,  -'. 

found  near  the  Tombigbee  River,  in  Choc-       the    disintegration    of    the    iron    being 
tow  and  Sumter  Counties,  Alabama.    Am.      rather  marked.   Schreibersite  was  found 

Jour.  Sci.  IV,  0:    153,  1099. 

to  be  present.  The  plessite  in  one 
specimen  exhibited,  under  etching  test,  a  beautiful  phenomenon  suggestive  of  a 
metallic  sun-stone. 

The  course  of  the  meteorite  must  have  been  from  N  to  S  ;  as  after  breaking 
up  the  fragments  were  found  in  this  direction,  the  smaller  having  fallen  first. 

L.  McI.  L. 

Foote,  W.  M.  Note  on  a  New  Meteoric  Iron  Resembles  most  siderites,  but  Wid- 
found  near  Iredell,  Bosque  Co.,  Te.xas.  Am.  mannstatten  figures  did  not  appear  on 
Jour,  iv,  8:  415,  1899.  .         ,  ,     ,,  ,•     •         r  ,, 

etching,  hence  probably  a  distinct  tall 

from  the  other  meteorites  of  that  region.  L-  McI.  l. 

Termier,  P.     Sur  la  composition  chimique  at  les      The  general  description   is  given   in  a 
proprieties  optique  de  la  leverrierite.     Bull.       previous  article. 
Soc.  Min.  22,  27,  1899.  .        .  ' 

Analysis  gives : 

SiOj    AI2O3   FegOg   MgO  CaO  K2O      loss  by  calcination  (nearly  all  H2O) 

49.90    37.02       3.65     0.30    tr.        1.13        8.65  =  100.65, 

yielding  formula  (H2,  K2)0  .    AI2O,  •    2Si02,    somewhat    similar  to    that    of 

Muscovite.     G.  =  2.6.     Axial  angle  varies  from  0°-50°,but  is  generally  small. 


and  Laboratory  Methods.  1313 

Optical  character  negative,  y  (pract.  equal  /i)  =  1.582,  a  =  1.554,  y — a  = 
0.28  (by  Wallerant  on  Quarter-Gaillard  material.  Author  finds  double  refraction 
variable  from  0.02  to  .0.03. 

The  mineral  occurs  in  an  argillaceous  band  in  a  bed  of  coal  at  the  Fontannes 
pits.  L.  McI.  L. 

Farrington,  0.  C.     Publications   Field   Colum-      Liesite.   Crystals  of  the  rare  mineral,  ine- 

bian   Museum.     Geol.   Series,!:    221-241.      site,  from  San  Cayetano  mine  near  Villa 
Feb.  1900.  ^ 

'    Corona,  Durango,  Mexico,  exhibit  the 

following  forms  (100),  (010),  (001),  (201),  (Ott),  (11.0.12),  and  (946),  of  which  the 

two  latter  are  new.     An  analysis  recalculated  to  100  per  cent,  gave  the  following  : 


Analysis  Calc.  to  100. 

Theory. 

SiO^ 

44.76 

42.91 

MnO 

38.86 

.     40.51 

CaO 

8.21 

8.00 

H2O 

8.17 

•      8.58 

100.  100. 

from  which  the  formula  H^CMn  .    Ca)gSigOit,  +  SHgO  is  deduced. 
^       Caledonite.     Crystallographic  examination  of  the  rare  mineral,  caledonite,  from 
the  Stevenson-Bennett  mine,  Organ  Mts.,  near  Las  Cruces,  New  Mexico.     Eight 
forms  were  identified  and  from  measurements  in  the  zone  of  the  basal  and  macro- 
pinacoids,  the  author  concludes  that  the  crystals  are  orthorhombic. 

Gay-liissite.  Examination  of  microscopic  crystals  of  gay-lussite  from  Sweet- 
water Valley  near  Independence  Rock,  Wyoming. 

Epso7nite.  Crystals  of  epsomite  from  Wilcox  Station,  Wyoming,  described. 

Golden  Calcite.  Calcite  crystals  from  concretions  of  the  Fort  Pierre  shale  of 
the  Bad  Lands,  South  Dakota,  described.  Distorted  crystals  with  the  rhombohe- 
dron,  -2  as  the  dominant  form. 

Dolomite  used  as  I)idian  Money.  Perforated  cylinders  of  dolomite  from  near 
Lakeport,  Lake  county,  California.  A  partial  analysis  gave  CaO  28.27 ;  MgO 
22.46,  Fe  1.18,  which  percentages  are  near  those  of  normal  dolomite. 

Crystal  Forms  of  Calcite,  frofn  Joplin,  Mo.  A  crystallographic  study  of  the 
calcites  of  this  interesting  mineral  locality.  The  author  distinguishes  five  types 
of  crystals  with  the  following  forms.  R,  4,  -i,  -f,  -|,  -2,  -4,  -11,  -20,  If,  1^ 
■|  2,  i  ^  of  which  il,  -20,  is  new  for  calcite.  Twin  crystals  are  described  with  O, 
and  -\  as  twinning  planes,  those  with  the  latter  twinning  planes  resembling  some 
from  Guanajuato,  Mexico,  described  by  Pirrson.  a.  f.  r. 

Palache,  Charles.  The  Crystallization  of  Cal-  An  exhaustive  Study  of  the  calcite  crys- 
cite,  from  the  Copper  Mines  of  Lake  ^^^^  ^f  l^j^^  Superior,  which  perhaps  in 
Superior.  Geol.  Surv.  Mich.  0 :  161-184,  1900.  ^  ^  r  r 

symmetry  and  beauty  rival  those  of 
any  other  locality.  Eighty-seven  forms  with  eight  doubtful  ones  are  described 
in  detail.  Of  these  thirty-two  are  described  as  new,  but  //'  <j  ll.l.To.O  \-  was 
previously  given  by  Schnorr.  (Abstr.  Zeit.  f.  Kryst.  30.660)  and  (J  -\  4.20.27.17  \- 
minus  \^  to  f  power  by   Sansoni  (Giorn.  Min.  1.136). 

Crystals  twinned  according  to  the  tw^o  laws  O,  and  —\  are  described. 
A  gnomonic  projection   of   all  the  forms  adds  to  the  value  of  the  paper. 
Details  of  the  measurements,  which  were  for  the  most  part  made  on  the  two-circle 
goniometer,  are  to  be  given  in  a  forthcoming  number  of  the  Zeitschrift  fiir  Krys- 
tallographie.  a.  f.  r. 


1314  Journal  of  Applied  Microscopy 


MEDICAL   NOTES. 

Ehrlich's  Triacid  Mixture  for  Staininc;  Blood. 

Orange   G,  sat.    aq.  sol.,    ....  30. 

Saurefuchsin,     "     "  ....     20. 

Methyl  Green,  "     "  ....  33. 

Alcohol,  absolute,  .....     25. 

Glycerin,   .......  50. 

Water,  dist., 75. 

Unless  the  solutions  are  absolutely  saturated  before  mixing,  results  will  be 
unsatisfactory.  In  making  up  the  mixture,  the  orange  G  and  acid  fuchsin  are 
first  thoroughly  mixed;  then,  drop  by  drop,  the  methyl  green  is  added,  the 
solution  being  well  shaken  after  each  addition.  The  other  three  elements  are 
then  added,  and  the  whole  shaken  thoroughly. 

When  the  stock  solution  is  once  prepared  it  should  never  be  shaken,  but  the 
desired  amount  drawn  from  the  top  by  means  of  a  pipette.  The  specimen  to  be 
stained  is  prepared  in  the  usual  manner  by  heat,  and  over  the  cover-glass  spread 
is  placed  a  drop  of  the  stain,  which  is  allowed  to  remain  for  two  or  three  minutes 
or  longer,  if  desired,  after  which  the  specimen  is  washed  in  water.  Care  must 
be  taken  in  the  application  of  heat  during  the  staining  process,  for  if  too  much 
heat  is  applied  the  specimen  becomes  pale  yellow  and  is  indistinct  under  the 
microscope ;  while  if  not  enough  heat  is  applied  the  specimen  is  too  dark. 

After  being  thoroughly  washed  in  water,  the  specimens  are  dried  with  filter 
paper  and  mounted  in  Canada  balsam.  If  the  mixture  is  properly  made  it  will 
keep  for  years.  c.  w.  j. 


Method  for  Cultivating  and  Staining  the  Diphtheria  Bacillus  (  Weiner 
Medical  Wocheiischi-ift,  No.  10,  1900). — Twist  a  small  piece  of  absorbent  cotton, 
impregnated  with  glucose  glycerinated  agar-agar,  around  the  end  of  a  sterilized 
glass  rod.  A  supply  of  these  rods  thus  prepared  may  be  kept  on  hand,  each  in 
a  sterilized  test  tube.  To  make  a  culture  the  cotton  is  swabbed  over  the  affected 
material,  and  the  rod  returned  immediately  to  the  test  tube.  After  being  kept  in 
the  incubator  at  a  temperature  of  36°  to  37°  for  four  or  five  hours,  enough  bac- 
cilli  will  have  developed  to  make  a  smear.  This  is  stained  with  the  following 
methylen  blue  solution  : 

Methylen  blue,       .....  1  gm. 

Alcohol,  20  c.c. 

Water,  dist., 420  gms. 

Acetic  acid,       .....  50  gms. 

This  solution  must  not  remain  on  the  specimen  more  than  two  or  three  sec- 
onds, after  which  time  the  slide  should  be  thoroughly  washed  with  water.  For 
a  counterstain  the  following  is  used  : 

Vesuvin,       ......  2  gms. 

Water, 1000  gms. 

which  is  heated,  and  filtered  while  still  warm. 

The  vesuvin  should  act  on  the  specimen  for  fifteen  to  twenty  seconds,  being 
then  washed  off  with  water. 

In  absence  of  true  bacilli,  the  smear  appears  brown.  If  both  true  and  pseudo 
bacilli  are  present,  a  blue  and  brown  color  is  visible.  The  true  bacilli  stain 
brown  with  their  polar  ends  blue,  while  the  pseudo  bacilli  stain  wholly  brown. 

c.  w.  J. 


and  Laboratory  Methods.  1315 


NEWS  AND  NOTES. 

The  Academy  of  Science  of  St.  Louis. — At  the  meeting  of  the  Academy 
of  Science  of  St.  Louis,  on  April  1,  1901,  thirty-three  persons  present,  a  memo- 
rial notice  of  the  late  Judge  Nathaniel  Holmes,  a  charter  member  of  the  Acad- 
emy, was  presented  by  a  committee  composed  of  Professor  Nipher,  Dr.  Sander 
and  Dr.  Baumgarten. 

Dr.  John  S.  Thurman  delivered  an  address  on  the  many  industrial  uses  now 
made  of  compressed  air,  illustrating  his  remarks  by  apparatus  in  operation, 
including  electric  motor  air  compressor,  compressed  air  auger,  drill,  disinfecting 
atomizer,  sculptors'  and  stone-cutters'  tools,  carpet  renovators,  etc.,  and  a  set  of 
lantern  slides  showing  the  practical  uses  made  of  these  and  other  implements 
and  machines  operated  by  means  of  compressed  air. 

Dr.  Theodore  Kodis  exhibited,  under  the  microscope,  slides  illustrating  a 
new  method  of  staining  brain  tissue,  whereby,  in  four  or  five  days,  it  has  proved 
possible  to  prepare  single  or  double  stained  preparations  containing  nerve  cells 
with  the  dendrites  of  the  latter  brought  out  by  a  direct  stain,  instead  of  being 
differentiated  merely  as  amorphous  silhouettes,  as  is  the  case  with  the  much 
slower  Golgi  process  commonly  employed.  It  was  stated  that  the  material  is 
treated  before  sectioning,  for  about  twenty-four  hours,  with  cyanide  of  mercury, 
followed  for  approximately  the  same  length  of  time  by  a  formaldehyde  solution, 
after  which  sections  are  cut,  stained  with  phosphomolybdate  haematoxylin  and,  if 
desired,  a  contrasting  stain,  such  as  one  of  the  anilin  greens,  and  mounted  in 
the  usual  way.  William   Trelease, 

Recording  Secretary. 


We  have  received  an  announcement  of  the  Summer  School  for  Apprentices 
and  Artisans,  which  will  be  held  at  the  University  of  Wisconsin,  from  July  1st  to 
August  9th  of  this  year.  The  school  has  been  established  for  the  benefit  of 
machinists,  carpenters,  or  sheet  metal  workers ;  stationary,  marine,  or  locomo. 
tive  engineers  ;  shop  foremen  and  superintendents  ;  superintendents  of  water- 
works, electric  light  plants,  power  stations,  factories,  large  offices  and  store  build- 
ings in  cities ;  and  for  young  men  who  wish  to  qualify  themselves  for  such  posi- 
tions. The  purpose  is  to  give  to  apprentices  a  certain  amount  of  theoretical  and 
practical  instruction  in  the  line  of  their  trade,  which  they  would  not  get  in  the 
shops. 

No  detailed  educational  requirements  are  specified  for  entrance,  the  fitness  of 
the  applicant  being  determined  by  a  series  of  questions  to  ascertain  whether  or 
not  he  seems  likely  to  be  benefited  by  the  work,  and  not  be  a  hindrance  to  others. 

It  is  believed  that  employers  can  well  afford  to  give  intelligent,  ambitious 
young  men  leave  of  absence  from  actual  employment  in  order  that  they  may 
increase  their  efficiency  by  availing  themselves  of  the  advantages  offered  in  such 
sessions  as  the  one  outlined  for  this  summer  at  the  University  of  Wisconsin. 
Persons  desiring  to  attend  this  school  during  the  coming  summer  are  asked  to 
make  application  on  or  before  June  1,  1901,  to  J.  B.  Johnson,  Dean  College  of 
Engineering,  University  of  Wisconsin. 


1316  Journal  of  Applied  Microscopy 

The  Cold  Spring  Harbor  Biological  Laboratory. — -The  twelfth  session 
of  the  biological  laboratory  of  the  Brooklyn  Institute  of  Arts  and  Sciences  will 
be  held  at  Cold  Spring  Harbor,  L.  I.,  from  Wednesday,  July  3d,  until  August 
13th.  The  following  courses  are  offered  :  Professor  C.  B.  Davenport,  University 
of  Chicago,  high  school  zoology ;  variation  and  inheritance.  Professor  H.  S. 
Pratt,  Haverford  College,  comparative  anatomy.  Dr.  L.  E.  Griffin,  Western 
Reserve  University,  invertebrate  embryology.  Dr.  A.  G.  Mayer,  Brooklyn 
Institute  Museum,  entomology.  Professor  E.  B.  Copeland,  West  Virginia  Uni- 
versity, cryptogamic  and  physiologic  botany.  Mr.  H.  H.  Whitford,  University 
of  Chicago,  plant  ecology.  Professor  N.  F.  Davis,  Bucknell  University,  bacteri- 
ology. Mrs.  C.  B.  Davenport,  microscopic  methods.  Dr.  H.  A.  Kelly,  Ethical 
Culture  Schools,  nature  study.  Professor  S.  R.  Williams,  Miami  University, 
will  give  instructions  in  animal  bionomics;  Professor  W.  L.  Tower,  Antioch 
College,  assists  in  entomology,  and  Mr.  A.  F.  Blakeslee,  Harvard  University,  in 
botany.  Louise  B.  Dunn,  Barnard  College,  assists  in  ecology.  A  new  labora- 
tory, exclusively  for  investigators,  is  announced.  The  dining  and  rooming 
accommodations  are  under  the  immediate  control  of  the  laboratory.  There  is  a 
uniform  fee  of  twenty- five  dollars  for  study  at  the  laboratory;  private  rooms  are 
fifty  dollars  for  the  entire  season.  Board  and  room  costs  six  dollars  per  week. 
Correspondence  should  be  addressed  to  the  director,  Professor  C.  B.  Davenport, 
University  of  Chicago,  Chicago,  111. 


QUESTION    BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

What  is  meant  by  the  '' growing  t\^''  in  Allium — when  used  for  mitosis  as 
'figured  in  Wilson's  book  on  the  Cell  ? — v.  a.  l. 

Can  synthol  be  used  in  place  of  carbolic  acid  ? — v.  a.  l. 

What  are  the  three  formulae  of  Kaiserling's  solution,  so  often  quoted  ? — v.  a.  l. 

Can  a  Welsbach  light  be  used  in  photo-micrography  with  ^  H.  I.  objective 
and  an  achromatic  ocular,  and  give  light  enough  to  focus  on  ground  glass  screen  ? 

V.  A.  L, 

Can  H.  I.  lenses  yL,  1^2-  to  >  ^^c.,  be  used  in  photo-micrography  with  a  Welsbach 
gas  light  only,  using  Abbe  condenser  and  bulls-eye  with  an  ocular  in  place  ?  The 
light  on  the  focusing  screen  is  so  dim  as  to  render  the  object  indiscernible.  Is 
an  aplanatic  ocular  prone  to  give  more  light  for  this  purpose  ? — c.  l.  p. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


VOLUME   IV. 


JUNE,  1901.  NUMBER  6 


Improved  Automatic  Microtomes. 

The  two  microtomes,  to  which  I  wish  to  call  attention,  are  modifications  of 
the  two  forms  of  automatic  microtomes  described  by  me  in  1897.^  In  neither 
instrument  have  the  essential  features  of  the  construction  been  changed,  but  the 
alterations  made  were  introduced  partly  to  increase  the  accuracy  of  the  cutting, 
partly  to  facilitate  the  manipulation  of  the  apparatus.  Messrs.  Bausch  &  Lomb 
undertook  the  series  of  improvements  at  my  request,  and  I  am  indebted  espec- 
ially to  Mr.  Edward  Bausch  for  the  time  and  thought  he  has  given  both  to  plan- 
ning and  executing  the  work  involved.  Every  detail  has  been  the  subject  of 
extended  consultation,  but  I  wish  the  pleasure  of  acknowledging  that  several 
valuable  innovations  were  first  suggested  by  Mr.  Bausch.  The  new  feed  for  the 
precision  microtome  was  devised  and  worked  out  by   Messrs.  Bausch  &  Lomb. 

I.  THE  AUTOMATIC  WHEEL-MICROTOME. 
Perhaps  the  most  important  improvement  in  this  instrument  is  its  increased 
accuracy,  which  has  been  secured  by  the  use  of  the  finest  machine  tools  for 
planing  the  sliding  surfaces,  cutting  the  micrometer  screw  and  cutting  the 
teeth  of  the  feed  wheel.  The  accuracy  is  now  so  great  that  one  can  cut  easily  a  un- 
iform series  of  sections  of  two  microns  in  thickness,  and  presumably  of  one  micron, 
but  the  one  micron  sections  I  have  not  sufficiently  tested.  In  the  former  instru- 
ments, both  American  and  German,  the  sections  would  skip  occasionally,  and 
then  the  following  section  would  be  of  nearly  double  thickness  and  the  uniform- 
ity of  a  series  ruined.  With  the  present  instruments,  three  of  which  I  have 
tested,  this  vexatious  irregularity  does  not  occur,  at  least  with  ordinary  objects. 
I  have  not  yet  tried  the  microtome  with  objects  specially  difficult  to  cut. 

Other  improvements  have  rendered  the  microtome  more  convenient  to  use. 
The  following  five  changes  are  most  important  :  First— i:h&  toothed  wheel 
which  supplies  the  automatic  feed  has  been  enlarged  and  cut  to  have  five- 
hundred  teeth,  so  that,  as  the  micrometer  screw  has  a  half-millimeter  pitch,  each 
tooth  equals  a  feed  of  one  micron.  6'^^^//^/— The  automatic  feed  has  been  so  con- 
trived that  it  will  give  any  desired  thickness,  from  one  to  twenty-five  microns, 
and  can  be  changed  in  a  moment.     This  is  accomplished  by  having  the   pawl- 

1  Science,  Vol.  V,  No.  127,  pages  857-866,  June  4,  1897. 

(1317) 


1318 


Journal  of  Applied  Microscopy 


bearing  lever  strike  against  an  eccentric  cam,  which  has  twenty-five  stops  in  it. 
To  diminish  the  wear  and  friction,  the  lever  where  it  strikes  the  cam  is  furnished 
with  a  steel  wheel.  To  prevent  the  dislocation  of  the  pawl  it  is  made  in  the 
form  of  a  fork,  the  prongs  of  which  fit  over  the  edges  of  the  toothed  wheel,  so 
that  the  pawl  cannot  slip  to  either  side.  Third— To  prevent  the  overthrow  of  the 
wheel,  which,  in  rapid  section  cutting,  is  apt  to  cause  a  greater  feed  and  there- 
fore sections  of  greater  thickness  than  intended,  a  simple  and  effective  brake  has 
been  added,  the  tension  of  which  is  easily  regulated.  The  brake  consists  of 
two  steel  springs,  each  with  a  leather  pad,  which  press  against  the  rim  of  the 
wheel  by  a  set-screw;  these  pads  may  be  pressed  together  more  or  less,  thus 
regulating  their  pressure  upon  the  wheel  between  them.  Fourth — A  split  nut  has 
been  provided  for  the  micrometer  screw,  the  two  pieces  of  the  nut  are 
attached  to  levers,  which  work  like  pincers,  so  that  by  pressing  the  levers  the 
nut  is   opened  and   object  carrier  may  be  run  forward  or  back  rapidly  without 


disturbing  the  screw  ;  by  releasing  the  levers  the  nut  closes,  and  as  it  closes 
snaps  into  place  automatically.  The  device  is  such  that  when  the  carriage  is 
run  way  back  to  the  beginning  of  its  excursion,  the  nut  snaps  into  place  of  itself, 
and  the  machine  is  ready  to  work.  Fifth — The  main  wheel  by  which  the  machine 
is  worked,  has  been  so  carefully  balanced  that  the  microtome  may  be  stopped  at 
any  point,  and  will  remain  in  the  same  position  without  change. 

There  are  other  minor  alterations,  which  do  not  call  for  special  description. 
The  resulting  apparatus  produced  by  Messrs.  Bausch  &  Lomb  is  a  fine  instru- 
ment of   precision,   very  convenient   and  satisfactory  in  use,  and   attractive   in 

appearance. 

II.     THE  PRECISION  MICROTOME. 

The  original  form  of  this  instrument  was  found  by  long   continued  use  to" 
have  certain  minor  defects,  three  of  which  caused  inconvenience.     Perhaps  the 


and  Laboratory  Methods. 


1319 


most  serious  of  them  was  the  HabiUty  of  the  automatic  feed  to  get  out  of  order 
unless  great  and  constant  care  was  taken  in  the  use  of  the  machine.  The  sec- 
ond defect  was  the  wear  on  the  ways,  which  took  place  chiefly  in  the  middle,  and 
very  little  or  not  at  all  at  the  ends,  so  that  the  carriage  was  liable  to  bind  at 
the  end  of  a  longer  excursion  than  usual.  A  third  defect  was  that  the  object 
holder  could  be  lowered  only  by  the  slow  process  of  turning  the  micrometer 
screw  backwards.  Messrs.  Bausch  &  Lomb  have  sought  to  remedy  the  first 
defect  by  a  new  feeding  device,  which  cannot  be  clearly  described  without  spec- 
ial illustrations.  The  general  principle  is  to  have  a  lever  bearing  a  pawl,  which 
moves  the  toothed  wheel ;  the  backward  motion  is  so  arranged  that  the  pawl  is 
lifted  free  from  the  wheel  altogether,  but  at  the  end  of  the  backward  motion  the 


pawl  is  brought  into  place  against  the  wheel  again,  by  an  action  of  the  lever. 
For  this  purpose  the  lever  is  hinged  in  its  middle,  so  that  its  outer  arm  can 
bend  independently  in  one  direction  without  displacing  the  whole  lever.  The 
motion  of  the  outer  arm  is  utilized  to  bring  the  pawl  into  place  against  the 
toothed  wheel.  This  wheel  is  provided  with  five  hundred  teeth,  each  tooth 
equaling  a  feed  of  one  micron.  It  is  also,  to  prevent  overthrowing,  supplied 
with  a  brake  similar  to  that  on  the  wheel  microtome.  Of  the  new  automatic 
feed  it  is  proposed  to  publish  a  separate  account,  with  figures,  on  another  occa- 
sion. The  second  defect,  that  of  the  ways,  has  been  obviated  very  simply  by 
shortening  the  ways  themselves   so  that  the  whole  of  the  ways  will  be  worn 


1320  Journal  of  Applied  Microscopy 

during  each  ordinary  excursion  of  the  carriage.  The  third  defect  has  been  met 
by  the  addition  of  the  split-nut ;  it  is  only  necessary  to  press  upon  the  levers 
which  open  the  nut,  in  order  to  allow  the  object  holder  to  sink  gently  to  a 
lower  level.  Other  minor  improvements  have  been  made,  of  which  I  will  men- 
tion only  the  spring-buffers,  which  prevent  the  carriage,  if  it  be  moved  a  little  too 
far  or  fast,  from  hitting  too  violently  against  the  frame  of  the  microtome. 
The  other  improvements  have  been  intended  chiefly  to  increase  the  rigidity  of 
the  apparatus. 

The  two  instruments,  above  described,  seem  to  me  better  suited  to  meet 
the  severer  requirements  of  microtomic  work,  than  any  others  which  I  have 
hitherto  tested.  The  "  Wheel"  microtome  will  probably  be  more  used  than  the 
"  Precision,''  partly  because  it  works  more  rapidly.  It  is,  however,  adapted  only 
to  paraffin  cutting.  When,  on  the  other  hand,  the  finest  grade  of  section  work 
and  a  larger  variety  of  imbedding  substances  are  demanded,  the  precision  micro- 
tome is  preferable,  since  it  can  furnish  not  only  the  finest  but  also  the  thickest 
sections,  and  will  give  perfect  sections  of  objects  which  cannot  be  cut  satis- 
factorily with  any  other  microtome,  and,  finally,  it  can  be  used  for  either  dry  par- 
affin or  wet  celloidin  sectioning.  We  consider  the  precision  microtome  so 
much  more  accurate  than  any  other,  that  we  use  it  almost  exclusively  for  cut- 
ting the  series  for  the  permanent  collection  of  the  Harvard  Embryological 
Laboratory.  Charles  S.  Minot,  LL.  D. 


New  Freezing  Microtome  for  Use  with  Carbon-Dioxide  Tanks. 

A  freezing  microtome  offers  two  great  advantages  to  the  student  of  micro- 
scopic anatomy.  By  its  use  thin  sections  of  animal  tissues  can  be  prepared  more 
quickly  and  in  many  respects  in  a  less  altered  condition  than  is  possible  by  other 
methods.  Freezing  was  one  of  the  earliest  methods  discovered  of  rendering 
animal  tissues  hard  enough  to  be  cut  readily  into  thin  slices.  Thus,  Stilling,  in 
1843,  was  enabled  to  prepare  thin  sections  of  the  central  nervous  system.  Since 
that  date  freezing,  as  a  method  of  hardening,  has  always,  to  a  greater  or  less  extent, 
been  utilized  by  histologists. 

In  the  earlier  days,  freezing  mixtures  were  made  use  of.  The  stand  on  which 
the  object  to  be  cut  was  placed  was  surrounded  by  ice,  salt,  and  water  until  the 
tissue  became  frozen.  Freehand  sections  were  made  with  a  razor.  This  method 
of  freezing  tissues  for  microscopic  work  was  superceded  by  methods  which  involve 
the  use  of  volatile  fluids  like  ether.  Instruments  for  the  utilization  of  these  fluids 
were  devised  as  attachments  to  the  precision  microtomes  which  were  invented 
after  the  use  of  celloidin  and  paraffin  as  embedding  agents  was  discovered. 
These  instruments  are  still  in  use  among  histologists.  In  the  hands  of  careful 
workers  they  give  satisfaction.  They  are,  however,  slow  in  action,  expensive  to 
use,  and  easily  put  out  of  order.  For  these  reasons,  although  almost  all  biolo- 
gists have  freezing  attachments  to  their  sliding  microtomes,  few  make  much  use 
of  them.  Of  late,  carbon-dioxide  has  been  much  utilized,  especially  by  patholo- 
gists, as  a  means  of  freezing  tissues  for  sectioning.     The  convenience  with  which 


and  Laboratory  Methods. 


1321 


fluid  carbon-dioxide  may  be  obtained  in  tanks,  and  its  power  of  rapid  freezing, 
have  caused  it  to  be  preferred  to  ether  and  similar  fluids.  In  every  active  patho- 
logical laboratory  the  freezing  microtome  is  in  daily  use.  Perhaps  its  greatest  value 
lies  in  the  fact  that  thin  sections  may  be  made  within  a  few  minutes  after  the 
removal  of  tissue  from  the  body,  and  in  a  few  minutes  more  these  sections  may 
be  hardened,  stained,  cleared,  and  mounted.  The  surgeon  may  thus  be  given  a 
positive  diagnosis  of  the  microscopic  condition  of  diseased  tissues  while  he 
proceeds  with  an  operation. 

The  carbon-dioxide  microtomes  commonly  used  have,  however,  several  draw- 
backs which  have  served  to  render  them  far  less  useful  than  they  should  be. 
From  the  practical  standpoint  their  most  serious  drawbacks  are  a  tendency  to 


Fig.  I. 

A.  Cover  of  freezing  stage  ;  B.  Glass  track  for  carrying  knife  ;   E.  Spiral  spring  ; 

F.  Tubal  base  of  knife-stage  ;    I.  Wheel  :  J.  Nut  for  attaching  axial 

tube   to  tank;  M.   Handle   of   tank-valve;  N.  Pointer. 

become  clogged  and  a  great  wastefulness  of  gas.  From  the  scientific  standpoint, 
lack  of  control  over  the  temperature  of  the  freezing  stage  serves  to  give  rise  to 
an  "  over-freezing,"  which  produces  marked  alterations  in  the  tissues.  In  order 
to  remedy  these  defects  the  machine  described  below  was  devised.  In  designing 
a  practical  machine  I  had  the  able  assistance  of  Mr.  E.  F.  Northrup.  The 
Bausch  &  Lomb  Company,  who  have  undertaken  its  manufacture,  have  also 
offered  suggestions  that  have  proved  of  much  value. 

Figure  1  shows  the  machine  as  it  stands  ready  for  use.  It  is  supported 
directly  by  the  nozzle  of  the  carbon-dioxide  tank.  This  offers  a  firm  and  con- 
venient means  of  attachment,  but,  if  desired,  a  heavy  tubing  may  be  utilized  to 


1322 


Journal  of  Applied  Microscopy 


connect  tank  and  machine.  When  the  microtome  is  screwed  directly  upon  the 
carbon-dioxide  tank  it  is  necessary  that  the  latter  lie  in  a  horizontal  position.  On 
the  other  hand,  if  an  L-shaped  piece  of  tubing  be  utilized  to  connect  freezing 
microtome  and  tank,  the  tank  may  be  placed  at  any  desired  angle. 

The  valve  of  the  tank  is  used  to  control  the  escape  of  gas  into  the  machine. 

The  axis  and  main  support  of  the  instrument  consists  of  a  stout  tube  with  a 

narrow  lumen  (K-D,  Fig.  2).     This  axial  tube  is  united  by  a  nut  (J,  Figs.  1  and  2), 

either  directly  to  the  nozzle  of  the  tank,  or,  in  case  a  connecting  tube  is  used,  to 

the  latter. 

On  the  top  of  the  axial  tube  the  freezing  stage  (A,  Fig.  1,  A-C,  Fig.  2),  is 
screwed.  This  stage  piece  consists  of  two  parts,  a  base  and  a  cover.  The  base 
is  the  part  screwed  into  the  upper  end  of  the  axial  tube  (C,  Fig.  2).  To  this  base 
the  cover  piece  is  screwed  (A,  Fig.  2).  Between  the  base  of  the  stage  and  the 
axial  tube  is  placed  a  thin  brass  plate  (D,  Fig.  2),  with  a  very  narrow  aperture  at 
its  center.  Through  this  narrow  aperture  the  carbon-dioxide  escapes  into  the 
lumen  of  the  stage  piece  (C,  Fig.  2).  The  difference  in  pressure  on  the  two  sides 
of  the  brass  plate  causes  a  very  rapid  expansion  of  gas  between  the  cover  and 
base  of  the  freezing  stage.  The  passage  open  for  the  escape  of  gas  from  the 
lumen  of  the  base  (C,  Fig.  2)  to  the  external  world  is  in  the  form  of  a  spiral  pas- 
sage which  finally  opens  out  through  the  side  of  the  cover,  as  shown  in  Fig.  1,  A. 
Between  the  cover  and  base  of  the  freezing  stage  an  asbestos  washer  is  placed. 

The  expanding  gas,  therefore,  can  absorb 
little  heat  from  the  base  of  the  stage.  Almost 
all  heat  absorption  must  take  place  from  the 
cover.  This  heat  absorption  is  greatly  facili- 
tated by  the  metallic  spiral,  which  projects 
down  from  the  cover  so  as  to  give  rise  to  the 
spiral  passage  through  which  the  gas  escapes. 
Through  the  mechanism  here  described 
far  the  greater  part  of  the  heat  absorbing 
power  of  the  expanding  gas  is  utilized  to 
lower  the  temperature  of  the  surface  of 
the  cover  of  the  freezing  stage.  The  tem- 
perature of  the  rest  of  the  machine  is  but 
little  altered.  Good  control  of  the  tempera- 
ture of  the  freezing  stage  can  be  thus 
maintained.  This  control  is  further  rendered 
possible  by  the  valve  of  the  tank.  If  this 
valve  be  turned  on  full  the  temperature  of 
the  cover  of  the  freezing  stage  will  be  quickly 
reduced  to  a  very  low  point.  Tissue  placed 
on  it  is  quickly  frozen.  On  the  other  hand, 
if  the  gas  is  not  permitted  to  escape  from  the 
tank  with  full  force,  the  difference  in  pressure 
on  the  two  sides  of  the  brass  plate  is  less  and 
heat  absorption  from  the  cover  is  less  marked. 


A 


CiJ 


H 


Fig.  2. 


A.  Cover  of  freezing  stage  ;  B.  Glass  track  for 
carrying  knife  ;  C.  Aperature  in  base  of  freez- 
ing stage  ;  D.  Aperture  in  tliin  brass  plate  ; 
E.  Spiral  spring  ;  F.  Tubal  base  of  knife  stage  ; 
G.  Check  for  limiting  movements  of  knife-stage  ; 
H.  Groove  for  G  ;  I.  Wheel  ;  J.  Nut  for  at- 
taching axial  tube  to  tank  ;  K.  Opening  into 
lumen  of  axial  tube. 


and  Laboratory  Methods.  1323 

In  this  way  tissues  placed  on  the  cover  may  be  slowly  frozen  without  subjecting 
them  to  severe  cold.  Thus,  too,  a  constant  low  temperature  may  be  maintained 
by  opening  the  tank  valve  to  the  required  point. 

The  mechanism  for  controlling  the  thickness  of  the  sections  is  equally 
simple.  On  the  lower  end  of  the  axial  tube  a  movable  wheel  (I,  Fig.  1  and 
Fig.  2)  is  placed.  This  wheel  moves  up  and  down  the  axial  tube  on  a  screw 
thread,  cut  twenty-five  threads  to  the  inch.  A  complete  revolution  of  the  wheel, 
therefore,  raises  or  lowers  it  a  millimeter.  The  margin  of  the  wheel  is  divided 
into  fifty  spaces,  each  of  which  therefore  represents  twenty  microns.  A  pointer 
(N,  Fig.  1)  serves  to  indicate  the  number  of  spaces  passed  in  a  partial  revolu- 
tion of  the  wheel,  and  thus  to  show  the  thickness  of  the  sections  cut. 

The  knife-stage  (F-B,  Fig.  1  and  Fig.  2),  consists  of  a  tubal  base  (F),  which 
surrounds  an  axial  tube  and  rests  on  the  movable  wheel ;  and  of  two  flanges  (B), 
which  extend  above  the  freezing  stage  on  each  side  for  the  support  of  the  cutting 
blade.  The  base  of  the  knife  stage  is  moved  up  the  axial  tube  by  screwing  the 
wheel  upwards.  It  is  forced  down  the  axial  tube  by  the  spring  (E,  Fig.  1  and 
Fig.  2)  whenever  the  wheel  is  turned  so  as  to  be  carried  downwards.  The  flanges 
of  the  knife-stage  support  parallel  glass  tracks  upon  which  the  cutting  blade  is 
carried  to  and  fro. 

For  cutting  sections  a  razor  or  a  plane,  or  almost  any  good  steel  blade  with  a 
straight  edge,  may  be  used. 

The  advantages  of  the  machine  are  as  follows  : 

1.  But  little  carbon-dioxide  is  wasted. 

2.  The  temperature  of  the  freezing  stage  can  be  controlled. 

3.  The  machine,  including  the  tank,  may  be  readily  carried  about. 
This  should  render  it  of  especial  value  to  surgeons. 

4.  Above  all,  it  is  simple  in  design,  strong,  and  exceedingly  unlikely 
to  get  out  of  order.* 

Charles  Russell  Bardeen. 
Anatomical  Laboratory,  Johns  Hopkins  University,  Baltimore. 


MICRO-CHEMICAL    ANALYSIS. 
XIV. 

BARIUM— Continued, 
///.  Barium  unites  with  Potassiu7n    Ferrocyanide  to  fonn   a   Ferrocyanide  of 
Barium  and  Potassium. 

BaClj  -h  K^FeCCN)^  =  BaK2Fe(CN)g  .  5H2O  +  2KC1. 

Method. — To  the  test  drop,  which  should  contain  no  free  mineral  acid,  add  a 
little  acetic  acid,  then  a  little  potassium  ferrocyanide,  and  warm  the  preparation 
very  gently.     In  a  few  seconds  rhombs  of  the  double  ferrocyanide  will  appear 


*The  description  of  the  microtome  here  given  is  essentially  similar  to  one  that  will  appear  in 
the  May- June  number  of  the  Johns  Hopkins  Bulletin,  1901. 


1324  Journal  of  Applied  Microscopy 

near  the  edge  of  the  test  drop  (Fig.  57).  These  crystals  are  clear  and  trans- 
parent. By  transmitted  light  they  appear  to  be  colorless,  but  if  examined  by 
reflected  light  they  will  be  seen  to  have  a  very  faint,  almost  imperceptible  yellow 
tint. 

Remarks. — The  reagent  crystallizes  in   prisms  belonging  to  the  monoclinic 
system,  while  the  barium  salt  is  to  be  ascribed  to  the  orthorhombic  system.     The 
danger  of  confusing  the  two  salts  is,  therefore,  slight. 
The  crystals  of  the  ferrocyanide  of  barium  and  potas- 
sium extinguish  parallel  to  the  diagonal  bisecting  the 
acute  angles  of  the  rhombs.     Many  of  the    crystals  of 
w  \  -^X       ^   ^  >r^\^^^  barium  salt  appear  to  be  rectangular  plates  or  even 
■-■    \-^  a  -  \«r^ cubes,   according  to    the    position  in  which  they  are 

i — *^  Q  I '  /  ^/^  seen.  An  examination  with  crossed  nicols  will  dispel 
^— ^y  p^  ^^  the  illusion.     When  the  test  drop  is  concentrated  with 

respect  to  barium,  the  crystals  of  the  double  ferro- 
cyanide separate  at  the  point  where  the  reagent  was 
introduced. 

Potassium  ferrocyanide,  though  giving  a  neat  re- 
action with  pure  salts  of  barium,  is  of  little  value 
when  dealing  with  mixtures.  It  is  then  often  difficult  to  avoid  the  precipita- 
tion of  calcium  with  the  barium,  particularly  if  much  ammonium  chloride  is 
present,  or  if  much  sodium  acetate  has  been  added  to  mitigate  the  action  of 
mineral  acids. 

From  mixtures,  strontium  may  sometimes  be  precipitated  if  the  solution  is 
quite  concentrated,  and  may  thus  interfere  with  the  test.  Pure  salts  of  strontium 
give,  in  very  concentrated  solutions,  only  a  granular  deposit  consisting  of  globular 
masses,  exhibiting  no  distinguishable  crystal  form. 

Magnesium  is  precipitated  from  ammoniacal  solutions,  but  neither  from  acid 
nor  from  neutral  solutions ;  hence  the  presence  of  this  element  will  not  mask  the 
test  for  barium. 

In  addition  to  calcium  and  strontium,  there  are  a  number  of  other  elements, 
which,  if  present,  will  either  be  precipitated  in  insoluble  form  or  will  interfere 
with  the  formation  of  the  barium  crystals.  In  this  list  the  most  frequently  met 
with  will  be  lead,  iron,  zinc,  rare  earths,  and  less  often  copper,  mercury,  uranium, 
titanium. 

Exercises  for  Practice. 

Crystallize  a  little  of  the  reagent  alone,  and  determine  its  optical  properties. 

Try  reagent  on  pure  salts  of  Ca ;  Sr ;  Ba  ;  using  both  dilute  and  concen- 
trated solutions.     Try  again,  this  time  proceeding  as  directed  under  Calcium. 

Try  the  reagent  on  mixtures,  say  of  Ca  and  Sr;  Ca  and  Ba;  Sr  and  Ba. 

Try  effect  of  the  reagent  on  salts  of  Pb,  Zn  and  Fe.  Then  make  mixtures  of 
Ba  and  these  elements,  and  test. 

Make  a  preparation  of  BaK2Fe(CN)g  •  5  HgO,  measure  the  angles  of  the 
crystals,  and  determine  the  optical  properties  of  the  compound. 


and  Laboratory  Methods. 


1325 


Fig.  58. 


IV.  Amnwuiuni  Fluosilicate  precipitates  Barium  Fluosilicate. 

BaCU  +  (NH^)2SiF6  =  BaSiFg  +  2NH4CI. 

Method. — Place,  on  a  celluloid  slip,  a  drop  of  the  moderately  dilute  solution 
to  be  tested.  Acidify  with  acetic  acid  ;  spread 
out  the  drop  a  trifle  ;  add  a  fragment  of  am- 
monium fluosilicate  and  warm  gently.  There 
will  immediately  form,  throughout  the  test 
drop,  fusiform  crystals,  either  singly,  in  crosses, 
or  in  more  or  less  irregular  masses  (Fig.  58). 

If  the  solution  is  quite  dilute,  instead  of 
the  usual  fusiform  crystals,  well-defined 
rhombohedra  and  prismatic  crystals  are  ob- 
tained. 

Remarks. — See  Sodium,  Method  III.  It 
is  important  to  avoid  testing  concentrated 
solutions,  since  fluosilicates  of  calcium  or 
strontium  may  possibly  separate,  although 
neither  of  these  elements  will  be  precipitated  under  the  conditions  which  usually 
obtain  in  testing.  This  caution  as  to  concentration  is  necessary,  because  when 
crystals  of  calcium  fluosilicate  CaSiFg  •  'iH^O  do  appear,  the  forms  obtained 
may  resemble  the  barium  salt  quite  closely.  Calcium  fluosilicate  is  to  be 
referred,  however,  to  the  monoclinic  system.  The  corresponding  strontium  salt, 
SrSiFg  .  2H2O,  is  isomorphous  with  the  calcium  compound,  and  is  slightly  less 
soluble  than  the  latter. 

The  form  of  barium  fluosilicate  varies  quite  a  little  according  to  the  concen- 
tration of  the  test  drop,  and  to  its  state  of  acidity. 

Much  free  mineral  acid  is  apt  to  interfere  slightly  with  the  precipitation. 

When  employing  celluloid  slips,  it  is  of  course  essential  to  use  great  care  in 
warming  the  preparation,  owing  to  the  inflammability  of  the  material.  Under 
proper  precautions  there  is  very  little  danger  of  losing  the  test.  The  warming 
should  be  slight. 

If  barium  alone  is  to  be  searched  for,  a  glass  slip  may  be  employed,  as  the 
formation  of  any  sodium  fluosilicate  will  not  materially  affect  the  test  for  barium. 

In  the  absence  of  ammonium  fluosilicate,  ammonium  fluoride  and  a  little 
silica  may  be  added  to  the  test  drop,  or  the  silica  may  be  suppressed  and  the 
test  performed  on  a  glass  slip. 

Exercises  for  Practice. 

Test  pure  salts  of  Ba ;  Sr ;  Ca ;  first  in  dilute,  then  in  concentrated,  in 
neutral,  and  in  acid  solutions. 

Make  a  mixture  of  Ca,  Sr,  Ba,  and  test  as  above. 

Try  reaction  on  mixtures  of  Na  and  Ba ;  then  on  Na,  Ca,  Ba ;  Na,  Sr,  Ba ; 
varying  the  concentration  of  the  test  drops. 

Test  a  mixture  of  Ba  and  Mg ;  then  one  of  Ba  and  Fe. 

Try  the  reagent  on  a  salt  of  Pb. 


1326  Journal  of  Applied  Microscopy 

V.  Ammonhim  Dichromate  added  to  solutions  containing  Barium  precipitates 
Barium  Chromate. 

2BaCl2  +  (NH4)2Cr207  +  H2O  =  2BaCr04  +  2NH4CI  +  2HC1. 

Method. — Employ  a  very  dilute  solution  of  the  barium  salt.  Add  a  little 
acetic  acid,  and  then  a  small  fragment  of  the  reagent.  Do  not  stir.  In  adding 
the  reagent  avoid  scratching  the  glass  slide  with  the  glass  rod  or  platinum  wire. 

Barium  chromate    separates   in  the   form   of  very 
^    ^-^,  minute,  light  yellow,  globular  masses,  and  tiny    rods 

c^         &        ^  with  rounded  ends.     These  rods    are  often  arranged 

^^<0  (j^  ^  -^  crosses  and  Xs  (Fig.  59). 

(m     ^^  ^  c\  /p  O  Occasionally  rectangular  plates  are  obtained. 

^^      ^     o^         "^  ^^?^  J^^^^'^^^^- — The   complete   precipitation   of   all   the 

^  ^^  barium  present  is  slow. 

^e>>  <SP      III 

\p>v  Jo.ov.is»,.  Strontium  will  not  separate  in  acid  solutions,  and 

Fig.  59.  calcium   not    even    in    the    presence    of     ammonium 

hydroxide. 

When  strontium  is  to  be  tested  for  as  well  as  barium,  the  test  drop,  after  the 
addition  of  the  reagent  and  examination  for  barium,  is  gently  heated,  then 
allowed  to  stand  for  some  time.  The  supernatant  solution  is  drawn  off,  and  to 
it  a  tiny  fragment  of  the  reagent  is  again  added,  and  the  preparation  warmed  ; 
if  no  precipitate  results,  showing  that  all  the  barium  has  been  removed,  strontium 
can  be  tested  for  by  adding  ammonium  hydroxide. 

In  warming  the  preparation  to  accelerate  the  separation  of  the  barium  salt, 
great  care  must  be  exercised  in  order  to  avoid  concentrating  the  drop  to  a  point 
where  strontium  might  be  precipitated. 

It  is  often  better  to  allow  a  drop  of  ammonium  hydroxide  to  flow  slowly  into 
the  side  of  the  test  drop,  rather  than  add  it  at  once  to  the  center  of  the  prepara- 
tion. 

Normal  potassium  chromate  produces,  with  barium  salts,  a  precipitate  similar 
to  that  obtained  with  dichromate,  but  is  not  to  be  recommended  as  a  reagent 
because  of  its  property  of  also  precipitating  strontium  compounds  in  acid 
solution. 

Ordinarily  the  precipitate  of  barium  chromate  is  largely  amorphous  in 
appearance.  Here  and  there,  however,  will  be  found  spots  where  there  are 
recognizable  crystals.  A  high  power  is  always  required  for  the  recognition  of 
the  form  of  the  crystals,  hence  the  drop  to  be  studied  must  be  spread  out  quite 
thin. 

Free  mineral  acids  interfere  with  the  test. 

Exercises  for  Practice. 

Try  reaction  on  salts  of  Ba ;  Sr ;  Ca ;  in  acid,  neutral  and  ammoniacal  solu- 
tions, both  in  concentrated  and  in  dilute  solutions. 

Try  mixtures  of  Ca  and  Ba ;  Sr  and  Ba  ;  use  solutions  acidified  with  acetic 
acid,  draw  off  the  clear  solution,  and  to  it  add  ammonium  hydroxide. 


ipW.  :0.olntm. 

Fig.  60. 


and  Laboratory  Methods.  1^'^^ 

VI.  Potassium  Antijnonyl  Tartrate  precipitates,  from  solutions  containing 
Barium^  a  Double  Tartrate  of  Barium  and  Antimony!. 

BaCla  +  2[K(SbO)C4H40e  •  l/'iHjO]  =  [BaC^H^Og  •  {^"oO) ,_^Q, ^YL ^O ^• 

2H20J+2KC1. 

Method. — To  the  neutral  and  moderately  concentrated  drop  to  be  tested  add 
a  small  drop  of  acetic  acid.  Place  close  to  the  test  drop  a  drop  of  distilled 
water  containing  the  reagent.  Warm  the  reagent  drop,  and  stir  until  all  the 
tartar  emetic  has  dissolved.  Warm  the  test  drop, 
and  while  both  drops  are  warm  cause  the  reagent 
to  flow  into  the  solution  being  examined.  On 
cooling,  crystals  of  the  double  tartrate  will  separate 
in  masses  near  the  edges  of  the  drop.  As  soon  as 
crystals  appear  draw  a  platinum  wire  through  one 
of  the  crystal  masses,  and  thence  across  the  drop. 
This  will  induce  crystallization  along  the  path  of 
the  wire  and  will  lead  to  the  formation  of  well 
formed  single  crystals.  When  this  procedure  is 
followed,  beautiful,  clear  cut,  thin,  transparent,  color- 
less crystals  of  the  orthorhombic  system  are  obtain- 
ed. The  usual  forms  are  rhombs  and  hexagons  (Fig.  60)  ;  the  latter  result 
from  the  cutting  ofiE  of  the  acute  angles  of  the  rhombs.  Multiple  twins  are 
frequent.     There  is  a  great  tendency  toward  the  formation  of  aggregates. 

Remarks. — Free  mineral  acids  must  be  absent. 

It  is  unfortunate  that  this  neat  reaction  is  of  quite  limited  application. 

Strontium  may  be  precipitated  in  like  forms  if  the  conditions  are  favor- 
able. 

Calcium  interferes,  as  do  also  members  of  Group  I,  and  of  the  magnesium 
group. 

When  dealing  with  small  amounts  of  barium  in  a  mixture,  it  is  necessary, 
owing  to  the  solubility  of  the  barium  double  salt,  to  concentrate  the  solution  to 
such  a  point  as  to  render  it  practically  impossible  to  obtain  satisfactory  results, 
because  of  the  crystallization  of  other  compounds. 

Lead  forms  a  double  antimonyl  tartrate  isomorphous  with  that  of  barium  and 
strontium.  * 

Of  the  other  heavy  metals,  silver  is  the  only  one  which  will  yield  a  crystal- 
line precipitate  with  the  reagent,  but  under  the  conditions  of  the  test  as  described, 
the  precipitate  with  silver  is  usually  amorphous. 

Exercises  for  Practice. 

Dissolve  a  little  of  the  reagent,  and  allow  it  to  crystallize.     Study  the  crystal 
forms,  and  determine  their  optical  properties. 
Try  the  reaction  on  BaClg  ;  CaClo  ;   SrC^- 
Test  mixtures  of  Na  and  Ba  ;  Mg  and  Ba;  Ca  and  Ba. 
Try  action  of  the  reagent  on  salts  of  Pb. 


*  Traube,  Zeit.  Kryst.  26:  if 


1328  Journal  of  Applied  Microscopy 

VII.    With  Primary  Sodium  Carbonate  or  Ammonium  Carbonate. 

The  latter  reagent  gives  much  better  results,  but  even  at  its  best  the  reaction 
yields  unsatisfactory  crystals. 

Neutral,  very  dilute  solutions  are  necessary  in  order  that  recognizable  crystals 
shall  be  obtained.  The  sodium  salt  tends  to  produce  minute,  spider-like 
aggregates  and  spherulites. 

Ammonium  carbonate  yields  tiny  spindle-shaped  crystallites,  dumb-bells,  and 
irregular  masses. 

The  test  is  not  applicable  in  the  presence  of  Ca,  Sr,  Mg,  Li,  etc. 

SEPARATION    OF  THE   CALCIUM   GROUP. 

Brief  outlines  of  the  methods  for  the  separation  and  identification  of  calcium, 
strontium,  and  barium  have  already  been  given  in  the  discussion  of  the  various 
tests  for  these  elements.  There  remains,  therefore,  only  the  necessity  of  sum- 
marizing the  various  processes. 

To  separate  this  group  from  other  elements,  three  reagents  can  be  employed  : 
/,  Ammonium  Carbonate ;  II,  Oxalic  Acid ;  III,  Sulphuric  Acid.  For  conven- 
ience each  of  these  reagents  will  be  discussed  separately  and  in  turn. 

/.  Ammonium  Carbonate  in  Ammoniacal  Solution. 

In  addition  to  Ca,  Sr,  and  Ba ;  there  can  also  be  precipitated  a  number  of 
other  elements  and  compounds.  Chief  among  these  should  be  mentioned,  rare 
earths,  Mn,  Cr,  Al,  Fc,  Pb,  Magnesium  group,  phosphates,  borates,  arsenates, 
molybdates,  oxalates,  tartrates,  etc. 

Inasmuch  as  the  tests  for  the  elements  other  than  those  of  Group  I  and  the 
Calcium  group  have  not  yet  been  described,  it  is  not  deemed  expedient  at  this 
point  to  enter  into  a  discussion  of  the  methods  for  dealing  with  complicated 
mixtures. 

The  clear  liquid  is  drawn  off,  or  otherwise  separated  from  the  precipitate 
produced  by  the  reagent.  The  precipitate  is  washed,  and  dissolved  in  hydro- 
chloric acid. 

Test  one  portion  of  the  hydrochloric  acid  solution  with  sulphuric  acid  for  Ca, 
if  an  amorphous  or  granular  precipitate  results,  Sr  or  Ba  (or  Pb^  is  present,  or 
the  substance  may  contain  both. 

Test  a  second  portion  with  ammonium  fluosilicate  for  Ba. 

If  no  Ba  is  found,  test  for  Sr  with  ammonium  dichromate  and  ammonium 
hydroxide. 

If  Ba  is  present,  precipitate  this  element  with  dichromate  in  acid  solution, 
Draw  off  and  test  for  Sr  with  ammonium  hydroxide. 

//.    Oxalic  Acid. 

Three  modifications  can  be  satisfactorily  employed,  the  choice  being  gov- 
erned by  the  nature  of  the  material. 

a.  Precipitation  with  oxalic  acid  in    nitric  acid  solution. 

b.  Precipitation  with  oxalic  acid  in  the  presence  of  ferric  chloride. 

c.  Precipitation  with  oxalic  acid  in  the  presence  of  stannic  chloride. 


and  Laboratory  Methods. 


1329 


0    0 


0 


O   0°  '^ 


^o 


o^ 


^ 


§ 


o 


0 


a.  To  the  test  drop  add  a  little  nitric  acid,  then  the  reagent.  Ba  is  not 
precipitated.  Ca  and  6"^  separate  slowly  in  the  usual  form  of  their  oxalates. 
After  allowing  sufficient  time  for  the  complete  separation  of  Ca  and  Sr,  separate 
the  clear  solution  and  to  it  add  sodium  or  ammonium  acetate.  Ba  is  now  pre- 
cipitated, and  can  be  identified  from  the  crystal  form  of  its  oxalate.  The  precipi- 
tated oxalates  of  Ca  and  Sr  can  be  tested  at  once  by  adding  sulphuric  acid,  or 
they  can  be  dried,  heated,  and  thus  converted  into  carbonates.  The  carbonates 
can  be  dissolved  in  acid,  and  the  solution  thus  obtained  tested. 

d.  To  the  test  drop  add  ferric  chloride  sodium  acetate,  and  then  the  reagent. 
Ca  and  Sr  appear  in  their  normal  form,  and  hence  cannot  be  distinguished  one 
from  the  other  ;  but  Ba  separates  as  a  double  oxalate  in  the  form  of  long  fili- 
form crystals  of  characteristic  appearance. 

c.  Oxalates  of  Ca,  Sr,  Ba  undergo  a  marvelous  change  when  precipitated  in 
the  presence  of  stannic  chloride.  This  beautiful  method  of  distinguishing 
between  these  elements  is  due  to  Behrens. 

To  a  drop  of  the  moderately  concentrated  solution  to  be  tested,  which  should 
be  neutral,  or   at  the   most   only  very  faintly  acid,   add   a 
little  stannic  chloride  ;  stir,  then  add  a  fragment  of  oxalic 
acid. 

Instead  of  the  usual  crystal  forms,  the  oxalates  separat- 
ing in  the  presence  of  stannic  chloride  undergo  a  remark- 
able change.  Ca  yields  rounded  and  oval  grains  and  thin 
disks,  with  here  and  there  crystals  showing  unmistakable 
evidence  of  trying  to  develop  into  octahedra.  The  crystals 
are  never  of  large  size,  though  larger  than  those  of  the  nor- 
mally formed  oxalate,  and  apparently  never  grow  into  clear 

cut  octahedra  (Fig.  61).     Sr  under  the  same  conditions  yields  large  octahedra 
(Tetragonal),  clear  cut  and   beautifully    developed  (Fig.    62).     These    crystals 

soon  become  corroded,  and  may  eventually  disap- 
pear ;  hence  it  is  necessary  to  examine  the  prepara- 
tion immediately  after  the  addition  of  the  oxalic  acid. 
Too  much  free  mineral  acid,  or  an  excessive  amount  of 
the  stannic  salt,  interferes  with  the  reaction. 

Ba  is  precipitated  by  oxalic  acid  under  the  above 
conditions  as  neat,  well  developed  prisms,  singly,  in 
crosses,  and  in  radiating  masses  (Fig.  63).  If  much 
Ba  is  present,  long,  very  pointed,  fusiform  crystals  re- 
sult, and  bundles  of  slender,  pointed  needles. 

Mixtures  of  the  alkaline  earths  do  not  yield,  as 
a  rule,  the  characteristic  forms  above  figured.  The 
form  of  the  oxalates  separating  from  such  solutions 
is  then  dependent  upon  the  dominating  element. 
Since  it  is  diflicult  to  properly  describe  the  peculiar  changes  to  be  observed,  the 
student  is  advised  to  try  the  reaction  on  mixtures  containing  the  elements  of 
the  calcium  group,  taking  care  to  have  first  one,  then  another  of  these  elements 
in  slight  excess. 


„    00®   #• 

I  I  I  I  I  I 

^J3\v.=  o.o»vT>»«i. 

Fig.  6i. 


\'p\M.-z.O-Oinif<^. 

Fig.  62. 


1330 


Journal  of  Applied  Microscopy 


<:::^ 


When  dealing  with  mixtures  of  Ca,  Sr,  and  Ba, 
Behrens  suggests  the  addition  of  a  little  acetic  acid 
prior  to  that  of  the  oxalic  acid,  then  by  cautiously 
neutralizing  with  magnesium  carbonate,  one  element 
after  another  can  be  caused  to  separate.  This  method 
of  procedure  requires  great  care  and  considerable 
experience.  For  this  reason  it  generally  fails  in  the 
hands  of  the  beginner. 

///.   Sulphuric  Acid. 

The  method  of  procedure  has  already  been 
thoroughly  discussed.  Attention  has  been  called  to 
the  fact  that  from  mixtures  of  the  sulphates,  Ca  can 
be  extracted  with  hot  water;  Sr  (and  Pb)  with  hot  hydrochloric  acid;  Ba 
remaining  unacted  upon. 

Concentration  of  the  water  extract  will  give  crystals  of  calcium  sulphate. 
Evaporation  of  the  hydrochloric  acid  solution  yields  crystals  of  strontium 
sulphate. 

The  residual  barium  sulphate  can  be  recrystallized  from  sulphuric  acid,  or  can 
be  converted  into  barium  carbonate,  dissolved,  and  tested. 


l*ig-63. 


With  simple  mixtures  it  is  often  unnecessary  to  proceed  according  to  the 
above  methods.  Combinations  of  the  different  tests  can  be  resorted  to.  For 
example,  Ba  can  be  precipitated  in  acetic  acid  solution  by  means  of  ammonium 
dichromate.  The  clear  liquid  is  decanted,  ammonium  chloride  and  potassium 
ferrocyanide  added,  and  the  Ca  precipitated  and  identified.  The  clear  liquid  is 
again  separated  and  tested  for  Sr  with  sulphuric  acid  or  potassium  sulphate. 
The  precipitated  strontium  sulphate  can  then  be  washed  and  recrystallized. 


In  addition  to  the  above  methods,  it  is  possible  to  effect  a  fair  separation  by 
converting  into  nitrates,  evaporating,  and  drying  carefully.  The  perfectly  dry 
nitrates  can  then  be  extracted  with  a  mixture  of  absolute  alcohol  and  ether.  Ca 
nitrate  is  quite  soluble,  Sr  nitrate  much  less  so,  while  Ba  nitrate  is  practically 
insoluble. 

The  alcohol-ether  extract  is  evaporated  to  dryness,  the  residue  dissolved  in 
water,  and  tested  for  Ca  with  sulphuric  acid,  arsenic  acid,  or  potassium  ferro- 
cyanide. The  residual  nitrates,  insoluble  in  the  alcohol,  can  then  be  tested  for  Sr 
and  Ba  by  the  dichromate  method,  or  with  stannic  chloride  and  oxalic  acid,  or 
ferric  chloride,  sodium  acetate,  and  oxalic  acid ;  or  oxalic  acid  in  nitric  acid 
solution. 


In  all  cases  the  choice  of  method  must  be  governed  by  the  nature  of  the  sub- 
stance being  examined.  The  ability  to  select,  at  once,  the  proper  method  of  pro- 
cedure which  will  yield  the  requisite  information  in  the  shortest  possible  time, 
and  without  error,  is  to  be  acquired  only  by  experience  and  much  practice. 

E.  M.  Chamot. 
Cornell  University. 


and  Laboratory  Methods.  1331 

Journal   of  Although  the  list  of  scientific  jour- 

nals  and  periodicals  is  already  so  long 

Applied    Microscopy  that   it   is   quite  impossible  for  one  to 

and  gain  even  a  casual  review  of  the  sub- 

LabOratOry   iVletnOaS.  jects  they  contain,  the  growing  impor- 

tance    of    investigation    in   which   the 
principal   feature   is    the   collection  of 


Edited  by  L.  B.  ELLIOTT. 


Issued  Monthly  from  the  Publication  Department         large    serics    of    Statistical    information 
of  the  Bausch  &  Lomb  Optical  Co.,  ° 

Rochester,  N.  Y.  has  Opened  a  new  field,  and  it  is  pro- 


SUBSCRIPTIONS:  posed    to   establish    a     journal    to   be 

One  Dollar  per  Year.     To  Foreign  Countries,  $1.25       devOted  tO  the  publication  of  biological 
per  Year,  in  Advance.  ^  " 


=^==     data    and    known    as    the  Journal    of 

The  majority  of  our  subscribers  dislike  to  have  their  rj-^/^^v    /     Cv^y.V.'.V^         C/^U     ^     ^..Kllr^^ 

files  broken  in  case  they  fail  to  remit  at  the  expiration  JilOlOglCal    statistics.       5UCn    a    pUDllCa- 

of  their  paid  subscription.     We  therefore  assume  that  no  f:„„   o/->i,l/^   /^ovfoinKr  Ko  morlo  t  TnlnoKlo 

interruption   in   the   series  is  desired,  unless  notice  to  LlOn  COUia  Ceriainiy   ue  maue  a  vaiuauie 

discontinue  is  sent.  ^.^  j^  ^^^  distribution  of  the  results  of 

research  work. 

There  are  at  present  no  journals  of  biological  science  that  wish  to  fill  their 
pages,  beyond  a  very  limited  degree,  with  long  series  of  tabulated  observations, 
which  often  form  the  basis  of  most  important  theories  and  conclusions.  It  is 
rather  the  rule  to  accept  only  the  conclusions  drawn  by  the  investigator,  and 
rely  upon  his  judgment  to  interpret  correctly  the  significance  of  the  mass  of  facts 
he  has  collected.  To  be  sure,  this  is  necessary  in  most  publications,  as  the  great 
majority  of  readers  cannot  devote  the  time  necessary  to  review  carefully  the 
ground  covered.  However,  those  studying  the  same  or  similar  questions  desire 
the  most  detailed  reports  of  other  workers  in  the  same  field. 

It  often  occurs  that  men  most  capable  of  handling  large  series  of  statistics 
are  not  in  position  to  collect  them  ;  and,  on  the  other  hand,  men  who  collect  data 
often  fail  to  see  the  full  significance  of  the  facts  before  them.  There  are  few  men 
who,  like  Darwin,  can  collect  facts  and  at  the  same  time  are  able  to  give  them 
the  most  accurate  interpretation.  It  is  therefore  most  desirable  that  data  upon 
which  theories  and  conclusions  of  general  interest  are  based  should  have  a 
medium  through  which  they  may  have  unlimited  circulation. 

* 

*  * 

The  setting  aside  of  the  week  in  which  January  1st  falls,  as  a  time  for  the 
session  of  scientific  societies,  will  certainly  receive  general  approval.  It  will 
undoubtedly  be  the  means  of  increasing  the  attendance  at  the  meetings.  It 
will  also  certainly  add  to  the  good  derived  by  those  who  go,  for  the  heat  of 
summer  and  the  relapse  that  usually  comes  after  the  year's  work  naturally  tend 
to  lessen  the  enthusiasm  of  the  members. 

It  is  to  be  hoped  that  the  universities  and  colleges  throughout  the  country 
will  cooperate  in  establishing  the  convocation  week,  thus  making  it  possible  for 
scientific  men  to  assemble  at  a  time  favorable  to  the  most  profitable  sessions. 

* 

*  * 

Owing  to  the  necessary  insertion  of  other  matter,  the  department  of 
Laboratory  Photography  has  been  omitted  from  this  issue. 


1332  Journal  of  Applied  Microscopy 

CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

„         c     ^       .,     •        „.     ,     ,    ,     r-  This  paper  contains  a  detailed  descrip- 

Ikeno,  a.     Contribution  a  letude  de  la  fecon-  ^    ^  ^ 

dation  chez  le  Ginkgo  biloba.  Ann.  Sci.  tion  of  fertilization  and  related  phe- 
Nat.Bot.  Ser.  VIII,  13:  305-318,  pi.  2-3,  nomena  in  Ginkgo,  from  the  formation 
1901.  "^ 

of    the    ventral  canal    cell    up   to  the 

first  division  of  the  oospore  nucleus.  The  nucleus  of  the  ventral  canal  cell 
rapidly  disorganizes,  while  its  sister  nucleus  increases  in  size  and  moves  toward 
the  center  of  the  oosphere.  In  preparations  stained  with  methyl  blue  and  acid 
fuchsin,  the  metaplasmic  ground  substance  of  the  nucleus  stains  red,  and  the 
chromatin,  which  forms  a  small,  irregular,  granular  mass,  also  takes  the  red, 
while  the  nucleoli  stain  blue. 

The  nucleus  now  undergoes  a  great  change  in  its  structure,  so  that  the 
metaplasm  and  chromatin  can  no  longer  be  distinguished  from  each  other.  The 
further  development  of  the  nucleus  of  the  oosphere  agrees  with  the  description 
of  the  corresponding  phenomena  in  Pifuis  laricio  as  described  by  the  reviewer 
some  time  ago.  In  one  instance  Professor  Ikeno  noted  an  abnormal  develop- 
ment of  the  nucleus  of  the  ventral  canal  cell,  resembling  the  cases  described  for 
Pinus  laricio. 

The  tube  nucleus  and  the  nucleus  of  the  stalk  cell  disorganize  within  the 
pollen  tube  and  do  not  enter  the  oosphere,  and  it  is  very  probable  that  only  one 
of  the  antherozoids  is  discharged,  the  other  disorganizing  without  being  able  to 
enter.  The  nucleus  of  the  antherozoid  slips  out  from  the  cytoplasmic  mantle 
before  conjugating  with  the  nucleus  of  the  oosphere.  The  mode  of  fusion  is 
like  that  already  described  for  Cycas  revoliita,  i.  e.  the  male  nucleus  gradually 
penetrates  into  the  nucleus  of  the  oosphere  and  lies  within  this  nucleus  before 
losing  its  own  membranes.  At  the  time  of  fusion  the  sex  nuclei  are  very 
unequal  in  size,  the  female  being  about  ten  times  as  large  as  the  male.  The 
behavior  of  the  chromatin  during  the  fusion  is  not  described. 

The  spindle  in  the  first  division  of  the  fusion  nucleus  is  very  broad  and 
multipolar  and  is  never  parallel  with  the  longitudinal  axis  of  the  oosphere.  In 
the  case  figured  the  spindle  is  tranverse.  Fertilization  takes  place  before  the 
ovules  fall  from  the  tree.  c.  j.  c. 

The  zygospore  of  Sporodinia  is  sur- 
Gruber,   Eduard.      Ueber   das   Verhalten   der  1 

Zellkerne  in  den  Zygosporen  von  Sporodinia  rounded  by  three  COatS,  the  outer  of 
grandis  Link.    Ber.  d.  deutsch.  bot.  Gesell.      ^hich  is  dark  brown,   warty,  and   cut- 

19:     51-55,   pi-    2,     I9OI.  .      .  ,  ,     .  r  1       r  1 

mized,  and  is  formed  from  the  mem- 
brane of  the  conjugating  gametes,  while  the  two  inner  coats  belong  to  the 
zygospore  itself.  The  middle  coat  is  somewhat  thickened  and  has  a  lamellate 
appearance,  while  the  innermost  is  a  mere  Hautschicht. 


and  Laboratory  Methods.  1333 

Leger,  who  worked  on  Sporodinia  six  years  ago,  found  that  both  gametes  con- 
tain hundreds  of  small  nuclei  which  become  scattered  in  the  mingling  cytoplasm 
after  the  membrane  separating  the  gametes  has  broken  down.  Double  staining 
showed  two  kinds  of  nuclei,  smaller  ones  near  the  periphery  and  much  larger 
ones  nearer  the  center.  At  a  later  stage,  all  the  nuclei  disappear  and  at  each 
pole  of  the  zygospore  a  spherical  mass,  the  "embryonic  sphere,"  is  seen,  each 
sphere  containing  a  large  number  of  granular  bodies.  These  spherical  masses 
increase  in  size  and  fuse  with  each  other,  and  soon  afterward  numerous  nuclei 
again  appear,  which  pass  into  the  germ  tube  as  the  zygospore  germinates. 

The  present  writer  also  finds  a  large  number  of  nuclei  in  the  zygospore,  and 
finds  that  the  nuclei  are  more  numerous  at  the  periphery,  but  there  are  also 
many  nuclei  in  the  center  and  all  of  the  nuclei  are  approximately  alike  in  size. 
This  condition  persists  for  a  long  time,  and  subsequent  stages  were  hard  to 
follow.  No  fusion  division  or  disorganization  of  nuclei  could  be  established 
with  any  certainty.  On  germination  the  nuclei  appear  in  greater  numbers  in 
the  germ  tube.  The  presence  of  "  embryonic  spheres  "  is  regarded  as  doubtful. 
Although  the  writer  was  not  able  to  observe  any  fusion  of  nuclei,  he  believes 
that  a  fusion  of  the  nuclei  at  the  center  of  the  zygospore  is  very  probable. 

c.  J.  c. 

Davis,  Bradley  Moore.  Nuclear  Studies  on  This  work  was  undertaken  with  the 
Pellia.  Annals  of  Botany,  IS:  147-180,  object  of  extending  our  knowledge  of 
pis.  lo-ii,  1901.  ^j^g  cytology  of  the  Hepaticae,  and  with 

the  hope  of  throwing  some  light  on  the  morphological  relationships  between  the 
various  manifestations  of  kinoplasm.  Three  phases  in  the  life  history  of  the 
plant  were  examined,  namely,  sporogenesis,  the  germination  of  the  spore,  and 
the  vegetative  activities  in  the  seta.  In  the  spore-mother-cell  the  spindles  are 
developed  in  the  same  fashion  as  that  which  prevails  in  the  spore-mother-cell  of 
the  Pteridophytes  and  pollen-mother-cells  of  Spermatophytes.  In  the  stages  of 
spore  germination,  asters  with  centrospheres  were  observed  in  the  prophase. 
These,  however,  appear  to  be  transitory  structures  as  they  disappear  before  the 
daughter  nuclei  are  formed.  In  the  vegetative  cells  the  type  of  spindle  forma- 
tion is  essentially  similar  to  that  described  by  Hof  and  Nemec  for  the  vegetative 
cells  of  the  flowering  plants.  Davis  also  states  that  "  it  is  probable,  of  course, 
that  there  is  likewise  a  blepharoplast  at  the  time  of  spermatogenesis."  He  con- 
cludes, however,  that  the  kinoplasmic  fibrillae,  the  centrospheres  and  kinoplasmic 
caps  are  all  secondary  developments  from  the  primal  granular  protoplasm,  which 
is  the  only  form  of  kinoplasm  in  any  sense  permanent  in  the  cell. 
Chicago.  •  A.  A.  LawsON. 

.,..„,.,       J-     TT    ,   u  ,        •       In  this  paper  Noll   takes  up  again  the 

Noll,  F.  Ueber   die   Umkehrungsversuche  mit  ^       <=> 

Bryopsis,  nebst  Bemerkungen  iiber  ihren  much  discussed  subject  of  polarity 
zelligen  Aufbau  (Energiden).  Bar.  d.  among  marine  algae.  Beginning  with 
deutschbot  Gesell.  18:  444-451,  1900.  *  °.  . 

the  statement  that  m  Bryopsis  miiscosa, 

on  which  he  worked,  the  polarity  was  as  pronounced  as  in  Piniis,  he  gives  us 
some  interesting  results  of  his  experiments  ;  namely,  that  very  few  indeed  of  his 
plants  reversed  their  root  and   shoot  pole   when   inverted.     Measurements  and 


1334  Journal  of  Applied  Microscopy 

dates  show  that  the  young  and  actively  growing  plants  were  so  strongly  polarized 
as  to  resume  the  original  manner  of  growth ;  that  only  older,  more  slowly  grow- 
ing forms  succumbed  to  the  external  conditions,  and  turned  root  into  shoot  and 
shoot  into  root.     These  results  agree  with  those  of  Winkler  of  an  earlier  date. 

Noll  takes  exception  to  the  definition  of  "  Energid  "  as  given  by  Sachs,  and 
calls  the  Siphoneae  "  single  but  multinucleate  energids,"  laying  stress  upon  the 
"  Hautschicht  "  rather  than  upon  the  nucleus  and  its  dominated  mass  of  proto- 
plasm. He  therefore  defines  the  energid  as  a  "  one  or  many  nucleate  plasmatic 
body  enclosed  in  a  definite  wall."  Philip  G.  Wrightson. 

Chicago. 

Life,  A.  C.    The  tuber-like  rootlets  of  Cycas      The  coral-like   outgrowths   on   the  up- 
revoluta.     Bot.  Gaz.  31  :  265-271,  lo  figures      ward    rising    roots    of    Cjras    revolida 
'         '  have  long  been  known,  and  the  endo- 

phytic alga  and  fungus  have  also  been  described.  Life  has  made  a  careful  study  of 
the  subject  from  thin  microtome  sections  and  has  been  able  to  give  a  more  precise 
account.  In  regard  to  the  reputed  dichotomy,  he  finds  that  not  all  of  the  meri- 
stem  passes  over  into  the  two  branches,  but  that  a  small  portion  is  left  as  a 
bridge  between  them.  This  small  portion,  however,  does  not  continue  the  main 
axis,  and  very  soon  disappears  so  that  sections  of  roots  in  which  the  branching 
can  be  seen  at  the  surface  show  no  trace  of  meristem  between  the  two  branches. 
The  development  of  the  algal  zone  is  clearly  figured  and  described.  Three 
forms  of  fungi  were  observed.  They  make  their  appearance  in  advance  of  the 
algal  zone  and  seem  to  prepare  the  way  for  the  algae,  which  are  referred  to  the 
genus  Nostoc.  The  presence  of  the  fungi  affects  the  intercellular  spaces  so  that 
they  become  the  rather  large  chambers  occupied  by  the  algae.  The  Nostoc 
probably  enters  through  the  numerous  lenticular  areas.  It  is  suggested  that 
the  tubercles  serve  for  aerating  and  also  assist  in  nitrogen  assimilation. 

c.  J.  c. 


CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  Sage  College, 

Ithaca,  N,  Y. 

CURRENT  LITERATURE. 

^    _,      „     „     ,.         ,       ,.     .    ^    ,        ,         The  work  gives  the  results  of   studies 

Overton,  h.     Studien  ueber  die  Aufnahme  der  ° 

Anilinfarben  durchdielebendeZelle.  Prings-  upon  the  action  of  anilin  Stains  on 
heim's  Jahrb.f.wiss.  Bot.  Bd.  34:  669-701,  animal  and  plant  cells.  Basic  anilin 
1899. 

stains    are   readily  taken   up  by  both 

kinds  of  cells.  Four  classes  of  these  stains  were  studied  in  detail.  (1) 
Triphenylenethane  stains  :  rosanilin  (chlorhydrate,  nitrate,  sulfate),  gentian  violet, 
methyl  violet,  dahlia,  anilin  blue  soluble  in  alcohol,  toluidin  blue,  victoria  blue, 
malachite  green,  methyl  green,  iodine  green,  auramin,  rhodamin  ;  (2)  CJmionhnid 


and  Laboratory  Methods.  1335 

stains:  thionin,  methyl  blue,  methylen  green,  safranin,  toluylen  red  (neutral  red), 
nigrosin  soluble  in  alcohol,  indulin  ;  (3)  Azo  stains :  chrysoidin,  vesuvin,  bis- 
marck  brown,  the  last  two  being  probably  the  same  ;  and  (4)  Acridin  stains  : 
chrysanilin.  All  these  stains  penetrate  the  living  cell  most  rapidly,  only  rhodamin 
is  somewhat  slower.  Quite  different  is  the  effect  of  the  sulphuric  acid  stains. 
(1)  Acid  fuchsin,  acid  green,  acid  violet,  anilin  blue  soluble  in  water  ;  (2)  Ni- 
grosin soluble  in  water,  and  indulin  ;  (3)  Congo  red,  ponceau  red,  bordeaux 
red,  biebricher  scarlet;  and  (4)  Indigo  carmin,  all  penetrate  neither  animal  nor 
plant  cells.  Only  the  acid  stains  belonging  to  group  three  {Azo  stains),  methyl 
orange  and  tropaolin  00  and  000  are  an  exception,  since  these  act  in  some  cases 
after  long  immersion.  Eosin  and  acid  carmin  are  in  general  not  taken  up, 
curcuma  acts  quickly,  carthamin  more  slowly.  Since  all  the  studies  agreed  that 
all  the  substances  easily  soluble  in  fatty  oils  and  similar  substances  were  taken 
up  quickly  by  living  cells,  while  those  insoluble  or  soluble  with  difficulty  did 
not  penetrate  living  cells,  the  conclusion  was  obvious  that  the  osmotic  con- 
dition of  living  cells  rests  on  the  phenomena  of  selective  solution.  Especially 
was  the  author  drawn  to  this  conclusion  since  the  plasma-skin  of  cells  is  im- 
pregnated with  cholesterin  or  a  mixture  of  cholesterin  and  lecithin.  It  especi- 
ally concerns  these  anilin  colors,  since  all  the  commercial  salts  are  basic  anilin 
stains  mixed  with  cholesterin,  or  else  are  easily  soluble  in  a  strong  solution  of 
cholesterin  or  lecithin  in  organic  fluids.  Also  these  liquids  in  a  pure  condition 
have  no  solubility  for  these  dyes.  Tannic  acid-methylen  blue,  which  does  not 
penetrate  the  living  cell,  is  also  wholly  insoluble  in  cholesterin  and  lecithin  solu- 
tions. With  a  few  exceptions  all  the  sulphuric  acid  stains  and  acid  carmines  are 
completely  insoluble  in  these  liquids.  Methyl  orange  and  tropaolin  are  excep- 
tions and  penetrate  very  slowly  and  slightly  into  the  living  cell.  a.  m.  c. 

_  ^,         ^     ^       ,         .       ,    ,       „  ,  For  this  work  sublimate  solution,  Zen- 

Retterer,  t.     Transformation  de  la  cellule  car- 

tilagineuse   en    tissue    conjunctiv    reticule.       ker's  and   Flemming's    fluids   and    also 
Comp.   Rend.  See.  de  Biol.  51 :  904-907,      an  aqueous  solution  of  platinic  chloride, 

1  pt.  to  1000,  were  used.  Without 
previous  decalcification,  objects  such  as  the  ribs  of  rabbits  and  guinea  pigs, 
are  embedded  in  paraffin.  The  following  combination  of  stains  gave  the  best 
results :  after  leaving  the  sections  for  a  few  hours  in  a  solution  of  safranin  in 
anilin  water,  they  are  washed  out  in  water,  stained  for  a  few  minutes  in  Boehmer's 
haematoxylin,  and  decolorized  in  alcohol  to  which  a  very  little  picric  acid  is 
added.  a.  m.  c. 

Petroff,  N.    Neue   Farbungmethod  ziir  rothe      Up    to    now    the     contrast-staining    of 
Blutkorperchen  in  Schnittpreparaten.     Bol-       blood  corpuscles  has  been  done  by  the 

nicznaia  Gazeta  Botkina,  iSgg.     (Russian.)  r     ^    •        r  .1  1      u-^ 

■*  >     vv     V  '      ygg  Qf  stams  from  the  malachite-green 

group,  which  differentiate  by  virtue  of  the  special  characters  of  red  blood  cells. 

This  process  is  as  follows  :  material  previously  fixed  in  Miiller's  fluid  or  formalin, 

or  Orth's  mixture,  is  embedded  in  paraffin,  not  collodion,   cut  into  the  thinnest 

sections  possible  of  regular  thickness,  and  fastened  to  the  slide.     The  paraffin  is 

then  taken  out    with  xylol   and  the  sections  washed  in  alcohol  and  water.     (2) 

Nuclear  staining  is  done  by  putting  the  sections  for  10-15  minutes  in  a  concen- 


1336  Journal  of  Applied  Microscopy 

trated  solution  of  bismarck  brown  in  one  per  cent,  acetic  acid,  or  in  lithium  or 
borax  carmin  for  20-30  minutes  ;  in  case  of  using  borax  carmin  it  should  be 
washed  out  in  hydrochloric  acid  alcohol.  Washing  with  water  follows.  (3) 
Stain  next  for  10-15  minutes  with  20  per  cent,  aqueous  solution  of  malachite 
green,  also  brilliant  green  or  victoria  green.  The  solution  is  made  by  diluting 
the  alcoholic  solution  five  times.  Wash  out  in  water.  (4)  Stain  for  i-1  minute 
long  according  to  Van  Gieson's  tincture  method  or  with  concentrated  aqueous 
picric  acid  solution,  which  is  diluted  4-5  times  with  water.  Wash  in  water.  (5) 
The  quickest  possible  dehydration  and  decolorization  in  absolute  alcohol,  mount- 
ing in  xylol  and  balsam.  Turpentine  or  bergamot  oil  may  be  used  in  the  place  of 
xylol.  All  the  decolorizing  necessary  is  easily  managed,  and  can  be  allowed  to  con- 
tinue for  a  long  time.  In  preparations  made  in  this  way  the  beryl-green  corpus- 
cles are  distinguished  from  all  other  structures,  which  are  a  gold-brown  from 
bismarck  brown  or  red-gold  from  carmin.  a.  m.  c. 

Godlewski,  E.  O.  rozmnazanin  jader  w  nies-  I"  O^'^er  to  learn  to  recognize  the 
niach  prazkowanych  zwierzat  kregowych  multiplication  of  nuclei  in  Striated 
(Ueber    Kernvermehrung    in   den  querges-  ,       ^  ,  ,      .  , 

treiften    Muskelfasern    der    Wirbelthiere.),      muscle  of  vertebrates  durmg  embryonic 
Bull,  de  I'Acad.  des  Sci.  de  Cracovie,  Avril,      and    postembryonic   development,   the 

author  studied  the  striated  muscle  of 
newly  born  guinea  pigs  and  mice,  also  those  of  salamander  larvae.  The  extrem- 
ities of  embryos  taken  from  the  mother  or  of  narcotized  newly  born  young  were 
put  in  toto  into  the  fixation  fluid.  Perenyi's  fluid  or  concentrated  sublimate 
solution  with  the  addition  of  two  per  cent,  acetic  acid  was  used,  followed  by 
increasing  strengths  of  alcohol.  After  hardening,  small  pieces  of  muscle  were 
separated  from  the  bone.  During  these  fixing  and  hardening  processes  a  great 
deal  of  contraction  takes  place  in  the  muscle  fibers.  Muscles  were  cut  in 
paraffin  in  longitudinal,  transverse,  and  oblique  sections  five  yu  in  thickness. 
These  were  stained  in  thionin,  also  in  Heidenhain's  haematoxylin,  double 
stained  with  bordeaux  red  or  eosin.  In  preparations  so  made  the  nucleoli  are 
sharply  tinted  with  red,  so  that  a  clear  contrast  is  obtained  between  these  and 
the  blue  chromatin  bodies.  a.  m.  c. 

His,  W.  Ueber  Sogenannte  Amitosis.  Anat.  Since  the  discovery  and  demonstration 
Anz.  Centralblt.  f.  d.  Gesam.  Wiss.  Anat.  of  bipolar  mitosis  it  has  been  known 
18:  52-60,  iQoo.  ,  1         ^  •         ,  •  ,     , 

that  nuclear  figures  exist  which  do  not 

agree  with  the  newly  discovered  principles.  Flemming  gives  a  second  type  of 
division,  direct  or  amitotic.  The  characters  of  this  kind  are  negative,  absence 
of  the  spindle  and  splitting  of  the  chromosomes.  This  form  was  considered 
degenerate  or  pathological,  but  recent  work  shows  that  by  changing  the  con- 
ditions of  growth  the  cells  of  Spirogyra  may  be  made  to  pass  from  mitotic 
division  to  amitotic  and  back  again,  without  disturbing  the  normal  conditions  of 
growth.  This  places  the  difference  in  the  two  types  of  division  in  the  province 
of  physiology  and  the  problem  is  to  determine  in  how  far  the  two  processes  follow 
a  common  law,  and  in  what  way  they  are  related  to  each  other.  It  has  already 
been  suggested  (His)  that  amitosis  may  be  plenipolar  mitosis  and  be  related 
to  the  growth  of  multinuclear  giant  cells  and  syncytial  formation.  For  these 
processes  His  suggests  the  name  "  syncaryosis." 


and  Laboratory  Methods.  1337 

In  the  division  of  the  periplast  cells  of  Selachians  two  types  are  recognizable, 
one  in  which  the  nuclei  have  central,  regularly  arranged  chromatin,  and  another 
in  which  the  chromatin  is  rod-shaped  in  separate  pieces.  Those  of  the  first  type 
are  found  earlier  in  development.  The  nuclei  contain  several  small  granules  to 
which  small  furrows  run  radially.  These  granules  grow  to  large  masses  without 
loosing  their  relations  to  the  radial  grooves.  Later  a  giant  cell  contains  six, 
eight  or  more  large  nucleoli.  The  nuclei  of  the  second  type  with  separate 
chromatin  rods  are  distinguished  by  their  transparency  and  even  staining. 
Transition  forms  are  found  forming  two  lines,  one  in  the  direction  of  dissocia- 
tion and  one  towards  synthesis.  The  former  process  is  a  simple  breaking  up 
of  the  chromatin  rods  into  fine  granules.  Many  stages  of  these  are  found. 
Reconstructive  processes  follow  definite  steps  :  (1)  Breaking  up  of  the  plasma 
bodies,  carrying  the  chromatin  into  several  small  balls  ;  (2)  separation  of  these 
balls,  still  remaining  connected  by  a  thread ;  (3)  re-appearance  of  chromatic 
rods  ;  (4)  radial  structure  appears  in  connection  with  chromatin  ;  (5)  formation 
of  enclosed  nuclei,  thickening  of  nuclear  wall.  The  process  continued  still 
farther  and  showed  itself  that  of  a  syncytial  formation  ;  it  is  a  kind  of  nuclear 
division.  Comparing  this  process  with  regular  bipolar  mitosis,  we  find  in  com- 
mon the  phases  of  dissociation  of  chromatin — prophases ;  the  formation  of  the 
chromatin  rods  and  their  radial  arrangement  are  the  anaphases.  The  meta- 
phase  would  correspond  to  the  dissociated  mass  of  granules.  As  long  as  the 
plasma  of  the  nucleus  retains  a  connection  with  the  dissociated  chromatin  a 
"  spirem  "  is  present.  The  equivalent  of  spindle  fibers  are  the  plasma  threads 
stretched  between  the  nuclear  balls.  The  origin  and  relations  of  polycentric 
giant  cells  are  understandable  on  general  cellular  laws.  It  is  known  that  a 
central  force  acts  in  such  phenomena  as  division.  Its  nature  is  unknown,  but 
simple  exhibitions  of  "  pull  "  and  "  push  "  are  to  be  seen. 

The  process  of  mitosis  can  be  divided  into  five  steps  :  (1)  The  division  of 
the  pre-existing  centrosome  ;  (2)  the  separation  of  these  parts  ;  (3)  the  changed 
influence  of  these  centrosomes,  due  to  their  changed  position,  shown  in  the 
appearance  of  double  radiations ;  (4)  the  grouping  of  the  chromatin  bodies 
and  arrangement  in  the  daughter  nucleus  ;  (5)  the  formation  of  cell  walls. 
Each  of  these  processes  requires  a  separate  time  ;  but  any  change  in  the  time 
requisite  for  these  steps  may  change  the  appearances  entirely.  If  the  division 
of  centrosomes  is  relatively  too  rapid,  new  ones  arise  without  the  correlated 
changes,  and  the  subsequent  steps  are  those  of  cells  with  many  centrosomes. 
The  distribution  of  the  chromatin  is,  hence,  difficult  to  follow.  The  relation  of 
the  nucleolus  to  these  centrosomes  and  the  plasma  cells  remains  yet  to  be 
studied.  a.  m.  c. 

Moll,    A.      Zur     Histochemie    der     Korpels.      From  the  results  of  the  author.  Tan  zer's 

Centralbl.  f.  Physiol.  13 :  2215-226,1800.  .         ...        ,         •     ak  ^i^^i^^i 

■'  3        '     yy  orcein  solution  (orcein  0.5  gram,  alcohol 

absolute  40.0  c.  c,  dist.  water  20.0  c.  c,  hydrochloric  acid  10  drops)  is  an  instructive 

double  stain   for  embryonic  cartilage.     The  preparations  (embryos  or  parts  of 

them)  must   be  hardened  in  alcohol  (not  in  chromic   acid),  and  then  in  thin 

celloidin  sections  be  put  into  the  above  staining  solution  for  6-24  hours,  then 

washed  in  80   or  90   per   cent,   alcohol   until   the  celloidin   is  nearly   colorless. 


1338  Journal  of  Applied  Microscopy 

dehydrated  in  ninety-eight  per  cent,  alcohol,  cleared  in  origanum  oil,  and  mounted 
in  balsam.  All  preformed  hyaline  cartilage  shows  itself  distinct  microscopically, 
through  its  blue  violet  color,  from  the  rest  of  the  brownish  red  tissue.  Micro- 
scopic investigation  shows  that  the  blue  color  has  its  place  in  the  ground  sub- 
stance of  the  cartilage.  This  blue  cartilage  network  makes  a  strong  contrast  with 
the  red  nuclei  of  the  merely  light  blue  or  non-colored  cartilage  cells.  In  the  embry- 
onic fibro-cartilage  of  the  intervertebral  discs,  as  yet  undifferentiated,  the  central 
cartilage  cells  stain  blue,  the  nuclei  red.  The  cells  always  become  paler  toward 
the  margin.  With  orcein,  embryonic  elastic  cartilage  gives  no  double  stain. 
The  change  in  the  color  from  that  of  adult  cartilage  is  worth  mention. 
Here  the  ground  substance  is  reddish,  the  cartilage  cell  with  its  surrounding 
area  intense  blue,  so  that  the  red  nuclei  can  only  be  seen  in  the  thinnest  sec- 
tions. Similarly  changed  is  the  adult  fibro-cartilage  ;  only  a  few  fibro-bundles 
are  blue.  The  elastic  cartilage  also  shows  no  double  stain  in  the  adult  condi- 
tion. Developed  bone,  both  decalcified  and  non-decalcified,  likewise  shows  no 
double  stain.  e.  j.  c. 

Linser,  P.  Ueberden  Bau  und  die  Entwicklung      The  Weigert  method  was  used  to  dem- 
des     Elastischen    Gewebes    in     der    Lung.  ^  .         i      ,•     ,•  •      ,i       i 

Anat.  Hefte.  H.  42,  43:  307-336,  m.  3  Tfln ,      onstrate  the  elastic  tissue  in  the  lung. 

1900-  The  stain  acted  usually  in  2-3  hours, 

yet  a  longer  time  for  staining,  even  to  24  hours  did  not  do  much  harm.  If  a 
shorter  time  was  used,  15-20  minutes,  no  usable  results  could  be  obtained.  By 
the  longer  staining  one  had  this  advantage,  with  others,  to  stain  the  adjoining 
kinds  of  connective  tissue  in  contrast.  Usually  a  simple  washing  out  in  strong 
alcohol  sufficed  to  make  the  bundles  appear  separate.  After  a  longer  continu- 
ance of  the  staining  process  it  is  necessary  to  differentiate  in  hydrochloric  acid 
alcohol  if  one  wishes  to  have  the  elastic  fibers  stained.  For  a  nuclear  stain, 
alcoholic  borax  carmin  and  lithium  carmin  were  used  ;  control-stains  were 
carried  on  with  haemalum-eosin  after  Van  Gieson's  method.  For  investigation 
12  embryos,  3.3  cm.  long  (Kopf  Steiss),  to  the  oldest  fcetal  stages,  were  used. 
Further,  fourteen  children  up  to  five  years  of  age  and  eight  older  human  lungs 
of  different  ages.  Further,  eight  different  stages  of  the  rat,  both  before  and  after 
birth,  lungs  of  young  and  old  cattle,  of  new  born  and  older  guinea  pigs,  of  hare, 
dog,  horse,  pig,  roe,  stag.  Tissues  were  preserved  in  formalin  or  alcohol  and 
imbedded  in  celloidin.  e.  j.  c. 

Foote,  K,and  Strobell,  E.  C.    Egg  of  Allolo-      A  series  of  micro-photographs  of  this 
bophora    foetida.      Journ.    of     Morph.    16:       egg  is   published    to    illustrate   the    fol- 
07-  I  ,  3ps.,  1900.  lowing  points:  (1)  The   effect  on   the 

cytoplasm  of  the  different  fixation  fluids  now  in  common  use.  (2)  The  charac- 
ter of  the  fertilization  cone.  (3)  The  position  of  the  middle  piece  of  the  male 
aster.  (4)  The  origin  of  the  sperm  granules.  (5)  The  early  stages  in  the 
development  of  the  pronuclei.  (6)  The  presence  of  osmophile  granules  in  the 
nucleoli  of  the  germinal  muscles.  The  photographs  have  been  taken  at  two 
magnifications,  G60  and  050,  and  it  is  believed  that  proof  has  been  offered  of 
some  of  Foote's  earlier  conclusions,  in  regard  to  the  cytoplasmic  origin  of  centro- 
some  of  the  male  aster.  .  a.  m.  c. 


and  Laboratory  Methods.  1339 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Fulleborn,  Dr.    Ueber  Formalinconservierung.      After  several  years'  experience  in   col- 
Zool.Anz.  24:  42-46,  1901.  lecting  in  temperate  regions   and  the 

tropics,  Dr.  Fulleborn  gives  his  unqualified  approval  of  formalin  as  a  preserva- 
tive of  zoological  material.  Its  portability,  cheapness,  ease  of  application,  and 
its  qualities  as  a  preservative  for  histological  purposes,  combine  to  commend  it 
for  use  in  preference  to  alcohol  on  collecting  expeditions.  Large  objects  for 
anatomical  work  should  be  hardened  in  5-10  per  cent,  formalin  for  8-14  days. 
For  transportation,  objects  thus  hardened  may  be  packed  in  excelsior  moistened 
in  formalin,  in  zinc  cases  which  are  soldered  up  when  filled.  These  zinc  cases 
are  made  up  in  sizes  which  "nest"  readily  for  transportation  into  the  field. 
Large  fish  should  be  opened  along  the  ventral  side  and  along  the  vertebral 
column,  or  the  skin  should  be  freed  from  the  musculature  in  places  and  a  wad- 
ding saturated  in  formalin  thrust  beneath  it.  Small  fish  may  be  thrown  into  the 
formalin  solution  or  injected  in  the  digestive  tract.  Formalin  is  especially  recom- 
mended for  fish  whose  scales  are  easily  rubbed  off.  Large  fish  kept  for  six 
years  in  formalin,  in  relatively  weak  solutions,  are  still  in  a  state  of  excellent 
preservation. 

As  a  preservative  of  natural  colors,  formalin  has  not  fulfilled  the  high  hopes 
which  it  first  called  forth.  Dr.  Fulleborn  reports  that  it  preserved  color  well  in 
some  tropical  Amphibia,  and  in  many  other  instances  specimens  reached  European 
museums  from  the  tropics  in  unfaded  condition.  On  the  other  hand,  the  iridescent 
colors  of  fishes  fade  as  quickly  in  formalin  as  they  do  in  alcohol.  The  brilliant 
pearly  sheen  found  on  certain  beetles  was  preserved  in  specimens  in  formalin, 
though  it  faded  at  once  in  dried  and  in  alcoholic  material.  The  egg-masses  of 
Necturus  with  their  gelatinous  coverings  have  kept  well  in  formalin,  the  form,  the 
eggs,  and  transparency  of  the  membranes  remaining  unchanged.  Tropical 
plankton  was  preserved  in  2-5  per  cent,  formalin,  the  algae  retaining  the  green 
color  of  the  chlorophyll  and  the  smaller  Entomostraca  keeping  their  natural  form 
as  a  rule.     Some  species,  however,  are  distorted  by  the  formalin. 

Small  birds  were  mummified  by  injecting  a  solution  of  5-10  per  cent,  forma- 
lin saturated  with  sodium  arsenate  with  a  hypodermic  syringe  into  the  thoracic 
and  abdominal  regions,  the  musculature  of  the  breast  and  shoulders,  the  eyes, 
and  the  brain  (through  the  orbit).  Injections  should  not  be  made  between  the 
musculature  and  skin.  The  small  openings  made  by  the  syringe  do  not  permit 
the  fluid  to  escape  and  soil  the  feathers,  if  care  is  taken  in  handling  the  birds. 
This  fluid  is  to  be  preferred  to  15  per  cent,  carbolic  acid,  sometimes  used  in 
mummifying  birds,  since  it  does  not  destroy  the  color  of  the  feathers  wet  by  it. 
Large  birds  may  be  treated  (in  addition  to  the  injection)  by  removing  the 
viscera  and  packing  the  body  cavity  with  wadding  saturated  with  the  injecting 


1340  Journal  of  Applied  Microscopy 

fluid.  After  injection  the  feathers  should  be  properly  arranged  and  the  birds 
hung  by  the  bills,  when  they  will  dry  rapidly.  Specimens  thus  treated  may  be 
softened  subsequently  and  mounted  in  the  usual  manner.  This  method  is  not 
only  a  rapid  one,  facilitating  field  work,  but  it  also  preserves  the  skeletons. 

c.  A.  K. 

Saint-Remy,  G.     Contributions  a  I'etude  du      The   small   size  and  the  very  resistant 
developpement  des  Cestodes.    Arcli.  de  la      membranes   of  the  eggs  of  tapeworms 

Parasit.  3:   292-315,0!.  7,  1900.  ,       ^u     ^      u    •  r^-u-       ..ju 

^    J  J'  r         y  render  the  technique  of  their  study  by 

modern  methods  a  matter  of  considerable  difficulty.  These  difficulties  have 
been  surmounted  to  some  extent  by  Professor  Saint-Remy,  who  has  studied  the 
development  of  two  species  of  Anoplocephala,  parasites  of  the  horse.  After 
removal  from  the  host,  the  worms  were  kept  in  normal  salt  solution.  Examina- 
tion of  the  living  eggs  reveals  but  little,  and  the  study  of  sections  of  the 
proglottids  for  the  development  of  the  eggs  contained  therein  is  even  less  satis- 
factory. The  eggs  are  freed  from  the  worm  by  compression  or  laceration,  and 
are  collected  upon  slides  in  sequence  from  the  last  proglottid,  forward  as  far 
as  they  can  be  found,  thus  securing  successive  stages  in  development.  The 
eggs  are  killed  upon  the  slide,  and  the  coagulated  fluid  in  which  they  lie  serves 
to  fix  them  to  the  glass.  A  large  number  of  reagents  were  tried,  and  good 
results  were  obtained  with  the  aqueous  solution  of  corrosive  sublimate,  and  also 
with  Carnoy's  fluid  (absolute  alcohol  6  vol.,  chloroform  3  vol.,  glacial  acetic  acid 
1  vol.).  The  eggs  were  not  sectioned,  but  were  mounted  i?i  toto  in  balsam.  For 
this  purpose  it  was  found  that  alum-carmin  and  also  bleu  de  toluidin  eosin  gave 
the  finest  results. 

The  development  of  Anoplocephala  resembles  that  of  Taenia.  A  small  egg- 
cell  and  a  large  yolk  mass  are  enclosed  in  the  egg  shell.  The  former  gives 
rise  to  two  minute  polar  cells.  Two  of  the  cleavage  cells  invade  the  yolk,  grow 
at  its  expense  into  two  giant  cells  which  form  the  outer  covering  surrounding 
the  embryo,  which  is  ultimately  cast  off.  Three  or  four  other  cells  form  a 
second  envelope,  a  pyriform  cap  provided  with  branching  filaments  in  the  form 
of  a  grapnel,  and  the  balance  of  the  embryonic  mass  forms  the  onchosphere  or 
embryo  proper,  within  which  the  characteristic  hooks  are  formed  before  the  em- 
bryonic membranes  are  shed.  No  decisive  evidence  is  contributed  to  the 
solution  of  the  problem  of  the  germ  layers  in  Cestodes.  c.  a.  k. 

^    _     ^,     ^        ,.,.         T  •      J         J      Students    of    our    native    reptiles    will 
Cope,    E.    D.     The  Crocodiiians,  Lizards,  and 

Snakes  of  North  America.  Report  U.  S.  welcome  this  posthumous  work  of 
Nat  Mus..  1898,  153-1270,  36  pis.  with  Professor  Cope,  for  it  is  a  very  com- 
347  figures  in  the  text,  1900.  ... 

prehensive  manual,  includmg  all  of  the 

nearctic  species  of  the  orders  of  Loricata  and  Squamata.  It  is  based  upon  the 
extensive  collections  of  the  author,  of  the  Philadelphia  Academy  of  Natural 
Sciences,  and  the  U.  S.  Natural  Museum.  While  it  deals  mainly  with  the 
taxonomy  of  the  group,  it  also  gives  many  facts  pertaining  to  the  external  and 
internal  anatomy,  especially  the  osteology  of  the  species  described.  Incidentally 
reference  is  made  to  many  interesting  points  in  the  biology  and  natural  history 
of  the  animals  discussed.     Ample  synoptic  keys  are  provided  for  the  purposes 


and  Laboratory  Methods.  1341 

of  identification  of  species,  while  abundant  illustrations  facilitate  the  recognition 
of  diagnostic  characters.  The  fact  that  this  monograph  is  issued  in  the  Report 
of  the  U.  S.  National  Museum  makes  possible  a  much  wider  distribution  to  the 
public  than  was  given  Professor  Cope's  earlier  bulletins  upon  the  Batrachia.  It 
is  to  be  hoped  that  the  monograph  of  the  turtles,  in  preparation  by  the  late 
Professor  Baur,  will  soon  be  issued  to  complete  the  manual  of  the  North 
American  Reptilia.  c.  a.  k. 

Sayce,  0.  A,     A  Method  of  Preserving  Crusta-       Suppleness  in  dried  specimens  of  such 

cea.     Victorian  Nat.  17:  71-78,  igoo.  .        ,  ^1        /^        ^ 

-^  ^  animals   as   the   Crustacea    is  a  great 

desideratum,  especially  in  laboratory  demonstrations.     Mr.  Sayce  secures  this  and 

also  preserves  to  a  considerable  degree  the   natural   appearance   of  the   animal, 

and  at  the  same  time  obviates  preservation  in  fluids,  by  the  following  treatment: 

The  specimens,  either  fresh  or  from  TO  per  cent,  alcohol,  are  immersed  for  some 

days,  ten  will  suffice  for  crayfish,   in   a  fluid  which,   in  metric  equivalents,  has 

approximately  this  formula : 

Glycerin  -- .-...     375  c.  c. 

Methylated  spirit  ----------     250  c.  c. 

Water      ------------     -250  c.  c. 

Corrosive  sublimate    ---------      0.5  gm. 

Slight  punctures  in  inconspicuous  parts  of  the  carapace  will  facilitate  pene- 
tration. After  thorough  soaking  in  this  fluid,  the  specimens  are  removed  and 
drained  and  allowed  to  dry.  They  can  be  stored  in  boxes  or  wrapped  in  water- 
proof paper.  To  avoid  too  much  drying  and  also  to  prevent  the  accumulation 
of  moisture  due  to  hygroscopic  action  of  the  glycerin,  specimens  should  be  given 
a  thin  coat  of  gelatin  and  then  immersed  in  10  per  cent,  formalin  for  a  few 
minutes.  This  hardens  the  gelatin,  renders  it  impervious  to  water,  but  does 
not  interfere  with  its  transparency.  c.  a.  k. 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

~  ..       .     ,.  ,       ,.     T,    •  ,  ,      ,1,  The  circumscribed  areas  of  acute  par- 

rujiDami.     Leber  die  Beziehungen   der  Myo-  '^ 

carditis  zu  den  Erkrankungen  der  Arterien-       enchymatOUS    myocarditis    are    always 

wandungen.    Virchow's  Archiv.,  159 :  447"      associated   with  the  narrowing  or  oc- 
490,  1900.  _  _       ° 

elusion  of  the  small   arterial  branches 

which  supply  them.  This  is  the  only  form  of  myocarditis  which  bears  a 
close  and  constant  relation  to  sclerosis  of  the  coronary  arteries.  In  fibrous 
myocarditis  sclerotic  changes  are  found  in  the  course  of  the  coronary  arteries, 
but  not  usually  in  immediate  connection  with  the  fibrous  areas. 

Arterio-sclerosis,  without  complete  occlusion  of  the  vessels,  leads  to 
disturbances  of  nutrition  in  quite  large  portions  of  the  muscle-wall.  Degenera- 
tion of  the  muscle-fibers  results,  followed  by  a  reparative  growth  of  connective 
tissue. 


1342  Journal  of  Applied  Microscopy 

A  destruction  of  the  muscle  does  not  always  take  place.  The  author  de- 
scribes a  primary  interstitial  non-purulent  form  of  myocarditis  in  which  collec- 
tions of  cells  pushed  aside  the  intact  muscle-fibers.  He  regards  this  variety 
as  toxic  in  origin.     The  foci  of  cells  are  at  length  replaced  by  fibrous  nodules. 

The  seat  of  the  disease  in  the  blood-vessels  can  be  in  the  main  branches  of  the 
coronary  arteries,  or  outside  the  heart  in  the  root  of  the  aorta,  or  in  the  orifices 
of  the  coronary  arteries. 

Fujinami  concludes  that  fibrous  myocarditis  originates  in  a  variety  of  ways. 
It  is  simply  the  final  outcome  of  a  number  of  different  pathological  processes. 
The  thickenings  of  the  vessel-walls,  demonstrable  microscopically,  are  not  al- 
ways to  be  regarded  as  the  cause  of  the  formation  of  the  fibrous  areas.  The 
vascular  changes  can  occur  as  a  result  of  the  fibrous  myocarditis  just  as  the 
vessels  in  the  scar  tissue  of  healing  ulcers  become  sclerosed. 

Fragmentatio  myocardii  is  frequently  associated  with  sclerosis  of  the  coron- 
ary arteries  and  fibrous  myocarditis.  j.  h.  p. 

„,    .. .         .  .  ,       ,   T,    ,     ■       ,         According  to  Schmorl,  accessory  ad- 

Wartnin.      Accessory    Adrenal    Body   in    the  °  ■' 

Broad  Ligament  (Adrenal  of  Marchand).  renals  are  found  in  the  neighborhood 
American  Journal  of  Obstetrics,  42 :  797-  ^f  ^j^g  adrenals,  in  the  adrenal  and 
805,  1900. 

solar  plexuses,  and  along  the  adrenal 

and  spermatic  veins,  in  92  per  cent,  of  all  autopsy  cases.  Accessory  adrenal 
tissue  has  been  found  in  the  kidney  capsule  and  cortex.  Along  the  spermatic 
vein,  in  the  pampiniform  plexus,  between  the  testis  and  epididymis,  in  the  cor- 
pus Highmori,  pancreas,  liver,  and  broad  ligament. 

The  author  was  able  to  collect  from  the  literature  only  23  cases  of  accessory 
adrenals  in  the  broad  ligament.  Marchand  in  1883  was  the  first  to  report  this 
anomaly.  The  diagnosis  is  made  by  the  characteristic  epithelial-like  cells,  and 
the  relation  of  these  cells  to  the  connective  tissue  and  capillaries.  Usually  the 
structure  of  the  body  is  uniform  throughout,  in  some  cases  resembling  the  cortex 
and  in  others  the  medullary  portion.  As  a  rule  the  accessory  adrenals  of  the 
broad  ligament  consist  of  cortical  tissue  only.  The  accessory  adrenal  found  by 
Warthin  was  a  pale  yellow,  fat-like  body  of  the  size  of  a  pea.  It  was  situated 
behind  the  ovary,  near  the  plexus  of  veins.  j.  h.  p. 

Abbott,  M.  E.    Pigmentation  Cirrhosis  of  the      An  advanced  cirrhosis  of  the  liver  and 
Liver    in    a    Case"  of     Hasmochromatosis.  ,  ,  ^      1  •      • 

Journal  of  Pathology,?:  55-69,  1900.  ^  moderate  degree  of  chronic  mtersti- 

tial  pancreatitis  were  associated  with 
an  extensive  deposit  of  iron-containing  pigment  in  the  tissues.  There  was  a 
bluish  gray  slaty  tinge  of  the  skin,  and  a  rusty  brown  discoloration  of  the  inter- 
nal organs.  Sections  of  the  liver  and  pancreas  were  loaded  with  golden-brown 
pigment,  responding  with  a  deep  blue  color  to  Perl's  test  for  iron,  which  was 
present  also,  though  in  a  lesser  degree,  in  the  spleen,  suprarenals,  and  heart 
muscle.  There  was  more  or  less  fibrosis  of  all  the  organs  except  the  kidney. 
In  both  liver  and  pancreas  the  heavy  pigmentation  of  the  connective  tissue  had 
its  source,  in  part  at  least,  in  the  broken-down  pigmented  cells  of  the  paren- 
chyma. A  fairly  advanced  chronic  interstitial  pancreatitis  existed  without  the 
clinical  picture  of  diabetes  so  common  in  cases  of  advanced  ha^mochromatosis. 


and  Laboratory  Methods.  1343 

Sections  of  the  organs  were  tested  for  iron  with  ammonium  sulphide  and 
with  potassium  ferrocyanide,  with  affirmative  results.  In  the  closer  study  of  the 
case  Perl's  test  only  was  used.  The  routine  method  at  first  employed  was  as 
follows :  Potassium  ferrocyanide,  2  per  cent,  solution,  three  minutes  ;  hydro- 
chloric acid  1  per  cent,  watery  solution,  two  to  five  minutes  ;  wash  with  distilled 
water.  The  bulk  of  the  material  was  hardened  in  Miiller's  fluid  to  which  2  per 
cent,  formalin  had  been  added,  and  was  preserved  in  methylated  spirits.  With 
the  fresh  tissue  the  reaction  was  prompt,  but  after  two  months  no  typical  reac- 
tion occurred,  the  granules  turning  green  or  a  greenish  yellow ;  many  did  not 
react  at  all.  That  the  iron  was  not  liberated,  but  that  the  reaction  was  only 
delayed,  was  proved  by  the  fact  that  sections  left  in  the  hydrochloric  acid 
solution,  two  to  twenty-four  hours,  gave  a  typical  Prussian  blue  color,  while 
when  the  test  was  performed  with  hot  hydrochloric  acid  the  reaction  was  almost 
instantaneous.  Sections  kept  in  4  per  cent,  formalin  gave  a  typical  reaction  in 
two  minutes  with  cold  hydrochloric  acid.  Bits  of  tissue  hardened  in  alcohol 
reacted  readily.  Miiller's  fluid  seems  thus  to  have  been  the  cause  of  the 
delayed  iron  reaction. 

Four  other  cases  of  haemosiderosis  were  studied  by  the  author.  She  con- 
cludes that  in  general  haemachromatosis  some  primitive  agency,  as  yet  unknown, 
is  at  work  leading  to  (a)  an  increased  destruction  of  haemoglobin  taking  place 
either  in  localized  haemorrhages,  or  within  the  blood  stream,  or  perhaps  some- 
times within  the  parenchymatous  cells  themselves ;  (b)  a  degeneration  of  the 
cells  of  certain  organs  by  which  they  become  unable  to  throw  ofif  the  granular 
pigment  deposited  in  them,  and,  becoming  loaded,  finally  disintegrate.  The 
cirrhosis  would  seem  to  be  the  nature  of  a  chronic  interstitial  inflammation, 
secondary  upon  the  presence  in  the  tissues  of  pigment  set  free  after  the  destruc- 
tion of  the  parenchymatous  cell.  j.  h.  p. 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Muhlmann,  M.  Uber  die  Ursache  des  Alters.  This  discussiop  of  the  general  physiology 
Grundzuge  der  Physiologie  des  Wachsthums  ^f  growth  begins  with  a  comparison  of 
mit  besonderer  Beriicksichtigung  des  Men-  o  o 

schen.    Wiesbaden  (J.  F.    Bergmann),  pp.      thebiological  relations  of  unicellular  and 
xii  und  195,  mit  15  Abbildungen,  1900.  multicellular  organisms.     From  the  two 

premises  that,  on  the  one  hand,  the  differences  between  unicellular  and  multicel- 
lular organisms  are  the  results  of  the  close  proximity  of  the  cells  to  one  another 
in  the  latter  as  compared  with  the  former,  and,  on  the  other  hand,  that  one  most 
important  difference  between  the  two  is  that  the  multicellular  organism  dies, 
the  author  arrives  at  the  conclusion  that  growth  causes  death.  This  preliminary 
statement  of  the  general  standpoint  opens  the  way  for  an  analysis  of  the  laws  of 
growth.     The   subject  is  developed  in  the  following  way  :  Growth   is  primarily 


1344  Journal  of  Applied  Microscopy 

related  to  nutrition.  In  the  case  of  the  Protozoa  each  cell  is  able  to  take  nutri- 
ment and  carry  on  respiration  over  its  whole  surface.  When,  however,  the  cells 
resulting  from  the  division  of  a  single  one  remain  permanently  in  contact  with 
one  another,  it  necessarily  follows  that  a  smaller  portion  of  each  cell  can  come 
into  contact  with  and  absorb  nutriment.  Such  developmental  forms  as  the 
blastula  and  gastrula,  and  in  fact  all  folds,  furrows,  invaginations  and  evagina- 
tions  appearing  during  development,  are  explained  as  the  result  of  the  insuffi- 
cient nourishment  which  is  unable  to  keep  pace  with  the  growth.  Since  only 
the  cells  immediately  in  contact  with  nutritive  substances  are  sufficiently  nour- 
ished to  support  growth,  while  the  cells  within  and  away  from  nutriment  are 
correspondingly  insufficiently  nourished,  these  latter  very  soon  begin  to 
degenerate  and  show  necrobiotic  phenomena. 

The  author  considers  that  practically  all  processes  of  cell  differentiation  are, 
from  the  standpoint  of  the  cell,  regressive  in  nature.  The  primitive  cells  or 
"  Blastzellen,  "  from  which  are  developed  all  other  kinds  of  cells,  are  seen  in 
the  embryo  before  any  beginning  of  differentiation,  and  are  characterized  by  their 
large  size,  their  richness  in  cytoplasm,  and  their  large  nuclei.  Such  cells  are 
also  found  in  the  adult  body  in  the  Malphigian  layer  of  the  epidermis,  the 
mucous  lining  of  the  alimentary  canal,  the  endothelium  of  the  blood  vessels,  the 
germinal  epithelium,  the  osteoblasts,  etc.  All  changes  which  occur  in  these 
"Blastzellen"  are  regressive  in  nature.  The  only  progression  is  found  in  their 
viiiJtiplication,  which  is  possible  up  to  a  very  old  age. 

The  theory  is  next  applied  in  detail  to  the  processes  of  ontogeny  and  histo- 
genesis. It  is  believed  that  the  development  of  the  individual  begins  with  the 
formation  of  the  ovum  within  the  ovary.  The  maturation  of  the  ovum  marks 
the  beginning  of  the  degenerative  changes  which  ultimately  lead  to  the  death  of 
the  individual.  The  egg  is  left  poorer  in  protoplasm  and  nuclear  material  after 
maturation.  The  development  of  the  body  form  in  all  its  details  is  explained 
as  a  result  of  the  better  nutritive  conditions  of  cells  on  the  periphery  over  those 
in  the  center  of  an  embryo  or  an  organ.  The  same  principle  is  applied  to  the 
differentiation  of  the  various  tissues.  Muscle  cells  or  ganglion  cells  are  degen- 
erated because  they  have  lost  the  cuboidal  or  polygonal  form  of  the  "Blastzellen" 
and  are  less  rich  in  the  sort  of  protoplasm  that  makes  up  the  body  of  Amoeba. 
The  chemical  as  well  as  the  morphological  aspect  of  histogenesis  is  developed. 

The  relation  of  function  to  structure  and  the  origin  of  the  functional  differen- 
tiations and  adaptations  are  next  discussed.  The  author  is  strongly  opposed  to 
a  teleological  consideration  of  life  phenomena  and  in  order  to  escape  some  of 
the  difficulties  along  this  line  which  his  theory  involves,  he  advances  some 
astonishing  physiological  principles.  An  example  will  indicate  the  nature 
of  these.  It  is  stated  that  the  saliva  is  a  product  of  the  regressive  met- 
amorphosis of  the  poorly  nourished  cells  of  the  salivary  gland,  and  is  useless  to 
the  organism.  The  ptyalin  is  useless  because  the  sugar  into  which  it  converts 
the  starch  of  the  food  has  to  be  reconverted  into  "  animal  starch.  " 

The  remainder  of  the  book  is  devoted  to  a  collection  of  data  relating  to  the 
growth  in  size  and  weight  of  the  human  body  and  its  organs,  from  birth  to  old 
age.     These  data  support   the  author's  view  that  there  is  during  the   course  of 


and  Laboratory  Methods.  1^^^ 

life  a  steady  diminution  in  the  relative  amount  of  the  additions  to  the  size  and 
weight  of  the  body  in  successive  years  after  birth.  The  same  is  true  of  the 
organs  and  tissues  except  in  the  case  of  those  which  are  largely  made  of 
"  Blastzellen."  Such  tissues,  as  for  instance  the  epithelial  lining  of  the  alimentary 
canal,  continue  to  grow  by  the  multiplication  of  cells  until  nearly  the  time  of 
death.  The  reason  for  the  greater  absolute  weight  of  an  adult  man  over  that 
of  an  infant  is  found  in  the  fact  that  the  muscular  and  skeletal  systems  take  on 
weight  by  processes  of  cell  metamorphosis  essentially  regressive  in  character, 
there  being  in  these  cases  no  growth  by  cell  multiplication  after  early  life. 

The  work  is  one  of  value  on  account  of  the  mass  of  data  on  the  growth  of 
the  human  body  which  it  presents.  While  it  is  probable  that  few  would  agree 
with  all  the  theories  proposed,  the  discussion  nevertheless  brings  out  strongly  the 
possible  importance  of  cell  nutrition  as  a  factor  in  developmental  processes. 

R.  p. 

Bataillon,  E.    La  pression  osmotique  et  les      The  general  standpoint  of  the   author 
grands  problemes  de  la   Biologie.     Arch.  f.  " 

Entwick.-mech,  II:  149-184,  pi.  5,  igoi.  is  that  osmotic  pressure  is  a  general 

and  fundamental  factor  in  biological  phenomena,  and  should  furnish  the  basis 
for  the  investigation  of  such  important  problems  as  the  resistance  of  organisms 
to  dehydration  (latent  life,)  teratogeny,  the  production  of  multiple  embryos,  and 
artificial  parthenogenesis.  On  all  these  points  experimental  results  are  offered. 
The  first  experiments  discussed  are  on  the  extraordinary  ability  of  Ascaris  eggs  to 
resist  the  action  of  fixing  agents  and  other  poisonous  fluids.  The  reasons  for 
this  resistance  capacity  are  found  in  the  facts  that  the  egg  is  surrounded  by 
a  membranous  chorion  which  is  semi-permeable,  and  that  the  fluid  of  the  interior 
of  the  egg  is  of  such  a  concentration  as  to  furnish  a  very  high  osmotic  pressure. 
On  account  of  this  high  osmotic  pressure  ordinarily  harmful  substances  cannot 
enter  the  egg.  There  is  no  plasmolysis  of  the  egg  in  the  fluids  of  less  osmotic 
pressure  than  that  of  a  15  per  cent,  solution  of  NaCl.  The  eggs  are  unable  to  with- 
stand the  dehydration  produced  by  a  30  per  cent,  solution  of  NaCl.  The 
author  thinks  that  cases  of  "  latent  life,"  of  which  desiccated  rotifers  form  a  good 
example,  are  to  be  explained  as  a  result  of  the  great  osmotic  pressure  of  their 
body  substance  which  resists  the  extraction  of  water  beyond  a  certain  point. 

Loss  of  water  is  found  to  have  a  retarding  influence  on  development  and 
may  completely  stop  it.  The  eggs  of  Petromyzon  Planeri  show  no  segmentation 
in  a  1  per  cent,  solution  of  NaCl,  while  in  a  .2  per  cent,  solution  their  development 
proceeds  normally.  In  solutions  of  intermediate  concentrations  there  are  varying 
degrees  of  retardation.  Solutions  of  CaCl2  and  sugar  isotonic  with  the  NaCl  were 
tried  and  the  same  results  were  obtained,  indicating  that  the  osmotic  pressure  is 
the  important  factor  rather  than  the  chemical  composition.  Twin  larvse  of 
Petromyzon  were  obtained  by  placing  the  fertilized  eggs  for  a  certain  time  (about 
eighteen  hours)  in  solutions  isotonic  with  1  per  cent.  NaCl  and  then  removing 
them  to  water,  in  which  the  development  took  place.  Fertilized  eggs  of  the 
teleost  Leucisciis  rutilus  treated  in  the  same  way  (except  that  they  were  kept  in 
the  solution  only  one  hour)  developed  into  multiple  monstrosities. 

By  the  use  of  the  same  method,  with  variations  in  the  time  of  action  of  the 
solution,  the  author  obtained  segmentation  of  unfertilized  eggs  of  some  fish  and 


1346  Journal  of  Applied  Microscopy 

amphibians.  Leiiciscus  riifilus  and  Raiux  escnlenta  gave  the  best  results.  The 
explanation  offered  for  all  of  these  phenomena  has  its  basis  in  the  osmotic  effect 
of  the  different  solutions.  r.    p. 

^.j.  „..  .       ,     ,      „r  ■.        TT  .         u  In     a    series    of  papers    which    have 

Kijanitzin,    J.    J.      Weitere    Untersuchungen  ^    ^ 

iiber  den  Einfluss  sterilisirter  Luft  auf  appeared  at  intervals  during  several 
Thiere.    Virchow's    Archiv,    162:515-533,  j.g         ^     Kijanitzin    has    given    an 

Taf.   14,   1900.  •'  .  . 

account    of    his    experiments    on   the 

physiological  effects  of  sterilized  air  on  animals.  The  paper  here  considered 
sums  up  the  results  which  have  thus  far  been  gained  and  announces  a  conclu- 
sion which,  if  proven  by  future  investigation  to  be  true,  will  be  of  great  signifi- 
cance with  reference  to  the  general  subject  of  animal  metabolism.  The  author 
maintains  that  in  addition  to  the  oxygen  of  the  air  there  are  necessary  for  the 
normal  metabolism,  and  consequently  the  life  of  the  animal,  certain  micro-organ- 
isms. These  micro-organisms,  entering  into  the  blood  in  the  process  of  respira- 
tion, are  taken  up  by  the  leucocytes  and  digested.  In  the  course  of  their  diges- 
tion an  oxydising  enzyme  is  given  off.  By  the  action  of  this  enzyme  under  nor- 
mal conditions  oxidation  processes  in  the  tissues  are  brought  about.  The  exper- 
iments seem  to  show  that  in  the  absence  of  this  enzyme  the  normal  process  of 
oxidation  in  the  animal  quickly  declines,  and  soon  ceases  altogether.  The  ani- 
mal then  dies  on  account  of  the  formation  of  large  quantities  of  incomplete, 
intermediate  products  of  metabolism  (leucomaines).  r.  p. 

Weinland,  E.     Ueber  den  Glykogengehalt  ein-       I"  ^n  analysis  of  specimens   of  a   tape- 
iger  parasitischer  Wiirmer.     Zeitschr.  f.  Biol.       \^OXVa.  {Tceuia  expafisd)  ZXi^  oi   the    par- 

'  asitic    nematode     Ascaris,     Weinland 

found  a  very  high  glycogen  content  in  both  cases.  In  Tcenia  the  glycogen 
amounted  to  1.5-4.7  per  cent,  of  the  fresh  animal,  while  in  Ascaris  the  amount 
was  4.2-7.1  per  cent,  of  the  fresh  animal.  The  amount  of  glycogen  in  the 
dry  substance  was  found  to  be  in  the  case  of  Tcenia  1,5-47  per  cent,  while  in 
Ascaris  it  was  from  20-34  per  cent.  The  highest  previously  known  glycogen 
content  was  in  the  mussel  Cardinm,  14  per  cent,  of  the  dry  substance  of  which 
is  glycogen.  In  mammals  the  glycogen  content  is  rarely  more  than  ,3  per  cent, 
of  the  dry  weight.  The  author  discusses  the  chemical  nature  of  the  glycogen 
obtained  from  these  worms.  R.  p. 

„  ,,  ,  J  >,,  .  u  «,  »  ^  ,  In  this  text-book  the  authors  have 
Macy,   M.    L.,  and    Norris,   H.  W.     A  General 

Physiology  for  High  Schools,  Based  upon  endeavored  tO  bring  all  the  conven- 
the  Nervous  System,  pp.408.  (No  date.)  ^ioj^^j  subject  matter  of  the  "high 
American  Book  Company,  New  York.  _  _      ° 

school  physiology  "  under  one  point  of 

view,  and  so  treat  its  individual  phases  in  their  relation  to  a  common  basis.  The 
idea  is  commendable,  but  the  choice  of  a  basis,  or  view  point,  is  not  altogether 
fortunate.  The  attempt  is  made  to  discuss  all  the  structures  and  activities  of 
man's  body  as  things  primarily  related  to  the  nervous  system.  It  will  readily  be 
seen  that  such  a  method  is  a  purely  artificial  one,  and,  from  a  physiological  stand- 
point, unsatisfactory.  The  detailed  treatment  of  most  of  the  topics  is  very  good. 
The  most  excellent  features  of  the  work  are  the  sections  devoted  to  laboratory 
and  demonstration  methods  for  the  use  of  the  teacher.     Some  of  the  methods  of 


and  Laboratory  Methods.  1347 

demonstration  with  simple  apparatus  are  ingenious  and  valuable.  Especially 
worthy  of  mention  in  this  connection  are  the  methods  given  for  illustrating  the 
processes  of  circulation  and  respiration.  The  text  figures  are  numerous  and  for 
the  most  part  copied  from  standard  works.  As  a  whole  the  book  makes  a 
very  good  impression  and,  in  the  hands  of  a  competent  teacher,  ought  to  prove 
an  excellent  high  school  text.  R.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Migula,  W.    System  der  Bakterien.    Vol.  II.      A  second   volume  of  Migula's  System 

Gustav  Fisciier.     Jena,  looo.  ,         -r,   ^  .      •         i  j       v 

^  der    Bakterien   has    made   its   appear- 

ance. The  author's  original  purpose  was  to  collect  all  species  of  bacteria  which 
had  been  described,  and,  by  testing  them  in  culture  media  in  his  own  laboratory, 
to  make  comparative  studies  and  descriptions.  This  task  proved  to  be  wholly 
impracticable.  Many  of  the  species  he  could  not  obtain,  and  many  of  those 
sent  him  were  not  in  condition  for  study.  The  book  is  therefore  simply  a 
compilation  of  descriptions  of  species  as  given  by  the  original  authors.  It  is  a 
large  work  of  1068  pages,  with  35  figures,  and  indicates  an  immense  amount  of 
labor  in  compilation  on  the  part  of  the  author.  h.  w.  c. 

Beijerinck.    Anhaufungsversuche  mit  Ureum-      The  very  great  importance  of  the  fer- 
bakterien.    Cent.  f.  Bak.  u.  Par.  II,  VII,      mentation  of  urea  makes   it   somewhat 
p.  33,  1901.  .  .... 

strange  that  the  bacteria  producing  this 

phenomenon  have  not  been  more  carefully  studied.  Until  the  appearance  of  this 
work  of  Beijerinck  very  little  has  been  known  in  regard  to  the  micro-organisms 
concerned  in  urea  fermentation,  a  few  observations  made  some  time  ago  com- 
prising our  sole  information.  The  author,  however,  has  investigated  the  sub- 
ject, and  has  described,  with  excellent  figures,  five  new  species  of  micro-organisms 
associated  with  this  universal  and  significant  fermentation.  These  specimens 
include  bacilli,  some  of  which  have  flagella  and  others  not,  and  it  also  includes 
a  Sarcina  species  which  is  motile  and  abundantly  provided  with  flagella ;  a 
somewhat  unusual  relation.  The  author  also  studied  the  subject  from  a  phys- 
iological standpoint  and  concluded  that  the  decomposition  of  urea  is  produced 
by  an  enzyme,  urase.  This  enzyme  is  completely  insoluble,  and  is  so  intimately 
bound  to  the  body  of  the  bacterium  that  it  cannot  be  separated  from  it. 

H.  w.  c. 

Stutzer.     Die   Organismen   der    Nitrifikation.       For  some  years  there   has  been   a   dis- 

Cent.  f.  Bac.  u.  Par.  II,  VII,  p.  i68,  igoi.  ^     v    i.  c^.   i.  j    ^u     is 

'        '  ^        '    ^  pute  between  Stutzer  and  the  Russian 

bacteriologist,  Winogradsky,  in    regard   to   the    nature    and    the    physiological 

properties  of  the  extremely  important  soil  organisms  known  as  nitrifying  bacteria. 

Winogradsky,  who  originally  discovered  them,  has  described   them  as   bacteria 

having  a  most  extraordinary  sensitiveness  to  the  presence  of  organic  substances, 


1348  Journal  of  Applied  Microscopy 

being  prevented  from  growth  by  the  presence  of  the  smallest  amount  of  organic 
material  or  ammonia.  In  previous  work  Stutzer  has  taken  grounds  quite  in 
opposition  to  many  of  the  points  which  were  held  by  Winogradsky.  The  present 
article  is  practically  a  withdrawal  on  the  part  of  Stutzer  of  all  of  his  previous 
claims.  In  an  introduction  he  explains  how,  in  the  press  of  engagements,  he 
was  led  into  error  by  leaving  the  work  to  an  assistant,  and  he  now  reports  the 
results  of  more  careful  work.  In  practically  all  respects  Stutzer  now  agrees 
with  Winogradsky,  so  that  the  relation  of  the  nitrifying  bacteria  to  the  various 
conditions  of  nature  which  had  previously  been  so  carefully  described  by 
Winogradsky,  must  be  taken  as  confirmed  by  this  latter  work  of  his  opponent 
Stutzer.  Stutzer  studies  both  the  nitrate  and  the  nitrite  forming  bacteria,  and 
in  most  respects  comes  to  identical  conclusions  with  those  of  the  Russian 
bacteriologist.  h.  w,  c. 

Hiltaer.      Ueber    die    Ursachen,   welche    die       As    is    well   known,  all   species   of    leg- 
Grosse,    Zahl,   Stellung   und    Wirkune    der  .        .  ,.  .,  vi     i 

Wurzelknollchen  der  Leguminosen  bedin-  "^es  growmg  m  ordmary  soil  are  likely 
gen.  Cent.  f.  Bak.  u.  Par.  II,  VII,  p.  to  develop  tubercles  on  their  roots 
"°^'  ^^    ■  through  the  agency  of  bacteria.     Out- 

side of  the  family  of  legumes  only  three  or  four  families  of  plants  are  known  to 
produce  similar  tubercles,  and  these  only  in  exceptional  cases.  The  author 
raises  the  question  as  to  the  reason  why  the  soil  bacteria  have  this  power  of 
growing  in  the  roots  of  legumes.  Thinking  that  it  was  possible  that  the  organ- 
isms produce  some  secretion  which  affects  the  roots  of  the  legume,  he  instituted 
experiments,  the  result  of  which  was  to  show  him  that :  (1)  the  changes  in  the 
root  hairs  of  the  legumes  accompanying  the  production  of  a  tubercle  are 
produced  by  some  soluble  substance  secreted  by  bacteria ;  (2)  this  substance  is 
present  in  great  quantity  in  the  tubercles  ;  (3)  the  older  root  hairs  are  immune 
against  the  action  of  this  substance. 

The  author  finds  that  different  cultures  of  the  tubercle  organisms  have 
considerable  difference  in  their  power  of  producing  tubercles.  When  a  plant 
which  already  possesses  tubercles  is  inoculated  with  culture  of  a  bacteria  of  a 
higher  virulence,  it  is  noticed  that  there  is  a  very  considerable  increase  in  the 
number  and  size  of  the  tubercles.  If,  however,  a  plant  possessing  tubercles  is 
inoculated  by  a  culture  of  the  same  virulence  there  is  no  increase  of  number  of 
tubercles.  In  other  words,  according  to  the  author's  conclusions,  the  presence 
of  the  tubercles  renders  the  legume  immune  against  the  further  action  of  cul- 
tures of  the  same  grade  of  virulence,  although  they  are  not  immune  against  a 
culture  of  a  higher  virulence.  h.  w.  c. 

„.    .       . ,       .    .  ,.        .  ,.    ,    ..        The    authors    investigate  a  somewhat 

Piorkowski  and  Jess,     r.actenum  coli  als  Ur-  ° 

sache  eines  seuchenartigen  Fferdesterbens  unusual  epidemic  among  horses  occur- 
in  Westpreussen.  Cent.  f.  Bac.  u.  Par.  I,  j-ing  in  West  Prussia,  and  causing  the 
XXIX,  p.  285,  1901.  ^  '  ° 

death  of  quite  a  number.     The  disease 

was  accompanied  by  fever  and  intestinal  troubles,  and  lasted  from  two  hours  to 
eight  weeks  in  different  cases.  Post  mortem  examination  showed  the  presence 
in  the  intestines,  of  ulcers  which  had  a  tendency  to  perforate  the  wall.  Bacter- 
iological study  of  the  infected  parts  showed  exceptionally   large   numbers   of  a 


and  Laboratory  Methods.  1349 

bacillus  which  the  authors  regard  as  the  true  coll  bacillus.  Finding  this  bacillus 
in  such  large  numbers,  the  authors  were  led  to  experiment  with  it,  and  suc- 
ceeded in  demonstrating  that  cultures  of  the  bacillus  were  pathogenic  for  the 
horse.  By  the  use  of  these  cultures,  partly  with  food  and  partly  by  intravenous 
inoculation,  they  succeeded  in  reproducing  the  disease  in  experimental  animals. 
They  are  of  the  conclusion,  therefore,  that  the  widespread  coli  bacillus  is 
occasionally  the  cause  of  serious  and  fatal  epidemics  among  horses. 

H.  w.  c. 

Bienstock.     Du  role  des  Bacteries  de  I'intestin.      The    author    gives    a    very    suggestive 
Ann.  de  I'lnst.  Past.  XIV,  p.  yw,  igoo.  .,         -  .  ,    ,, 

■^  paper    upon    the    functions  of  the  or- 

dinary intestinal  bacteria.  He  has  previously  shown  in  the  intestine  of  animals 
the  presence  of  a  Bacillus  putrificus,  which  produces  a  putrefying  action  on 
proteids.  He  now  finds  that,  under  normal  conditions,  such  putrefaction  of  the 
contents  of  the  intestine  does  not  occur.  This  fact  seems  surprising,  inasmuch 
as  B.  putrificus  is  constantly  present  in  the  intestine,  and  the  conditions  are 
apparently  proper  for  its  growth.  Bienstock  is  of  the  opinion  that  putrefactive 
action  is  checked  by  the  presence,  in  the  intestine,  of  certain  aerobic  bacteria, 
such  as  lactic  and  the  coli  bacilli.  Experiments  show  that  the  putrefaction 
produced  by  B.  putrificus  does  not  take  place  when  a  quantity  of  these  aerobic 
bacteria  are  present.  The  author  concludes,  therefore,  that  these  aerobic  bac- 
teria, which  are  uniformly  found  in  the  normal  intestine,  are  of  direct  value  to 
the  human  body  in  preventing  the  putrefaction  of  the  intestinal  contents.  He 
points  out  the  fact  that  sterilized,  and  even  pasteurized,  milk  is  not  so  readily 
digested  as  raw  milk,  especially  by  persons  with  intestinal  disturbances,  and 
this  he  attributes  to  the  fact  that  since  the  heat  has  destroyed  the  lactic  organ- 
isms, these  organisms  are  not  present  in  the  intestine  to  prevent  the  put?-ificus 
from  producing  putrefaction.  In  short,  the  author  concludes  that  the  reason 
why  ordinary  micro-organisms  are  needed  in  the  intestine  is  to  prevent  the 
putrefaction  which  would  otherwise  occur  in  the  intestinal  contents,  owing  to  the 
presence  of  certain  putrefying  micro-organisms  which  are  always  found. 

H.  W.  C. 

Reed,  Carroll  and  Agramonte.    The  Etiology      These  authors  have  presented  a  further 
of  Yellow  Fever.     Med.  Rec,  Feb.  i6,  1901.       ^^^^^^  ^^^^  ^j^gij.  conclusions  in  regard 

to  the  relation  of  yellow  fever  to  mosquitoes.  The  results  reached  are  of 
immense  importance  and  are  too  numerous  to  be  summarized.  The  most 
important  are,  that  the  disease  is  transmitted  from  yellow  fever  patients  to 
healthy  persons  by  the  bites  of  mosquitoes,  there  being  a  period  of  incubation 
from  41  hours  to  5  days.  They  have  repeatedly  succeeded  in  reproducing 
the  disease  by  allowing  mosquitoes  (culcx  fasciatus)  to  bite  patients  and,  subse- 
quently, healthy  individuals.  They  find  that  an  attack  of  yellow  fever  conveyed 
by  a  mosquito  bite  confers  immunity  against  disease.  A  house  is  only  infested 
with  yellow  fever  when  containing  no  mosquitoes.  Yellow  fever  is  not  dis- 
tributed by  soiled  articles  of  clothing  or  bedding,  as  has  been  supposed.  The 
spread  of  yellow  fever  may  be  most  effectually  prevented  by  protecting  the 
patient  from  mosquito  bites.     Tiiese  conclusions,  which  represent  only  a  few  of 


1350  Journal  of  Applied  Microscopy 

the  important  results  of  the  work  of  these  investigators,  are  clearly  of  the 
utmost  importance  in  the  future  study  of  this  serious  disease.  h.  w.  c. 

„,.,.,    ^^  ,  .  The  author  studies  the  bacteria  present 

Hilsuiii.    Baktenologische  Untersuchung  eines  '^ 

Schwimmbades  in  Bezug  auf  Selbstreinig-      in  a  swimming  bath  which  was  in  con- 
ung.    Cent.  f.  Bak.  u.  Par  I,  2:  66i,  1901.         ^^^^^  ^ge.     He  finds  that  the  number 

of  bacteria  increased  regularly  during  the  first  day,  after  being  newly  filled  with 
water,  and  then  constantly  decreased.  This  decrease  in  the  number  of  bacteria, 
he  points  out,  could  be  due  neither  to  the  action  of  light  nor  to  sedimentation, 
since  the  number  of  bacteria  at  night  and  morning  was  essentially  the  same,  and 
since  the  water  was  in  constant  use,  a  condition  which  would  prevent  sedimen- 
dation.  Nor  does  he  believe  that  a  want  of  food  can  be  the  cause,  since  the 
water  filtered  through  a  pasteur  filter  is  an  excellent  culture  medium  for  bacteria. 
The  author  believes  that  the  matter  is  one  of  struggle  among  different  bacteria 
with  each  other,  resulting  in  a  destruction  of  many  individuals.  h.  w.  c. 


NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Viola,  C.  Ueber  das  "Glaukisiren"  verschied-  The  writer  proposes  the  term  "  Glau- 
ener  Feldspathe.  Zeit.  f.  Kryst.  34:  171-  kisiren  "  for  the  variety  of  schilleriza- 
195,  1901.  .  . 

tion  which  takes  place  m  moonstone 

— that  is,  when  the  inner  reflection  produces  a  silvery  or  bluish  light. 
Whether  or  no  a  convenient  English  form  of  this  term  will  be  made  remains  to 
be  seen,  but  Glaukisiren  may  be  translated  as  the  process  which  produces  the 
silvery  schiller  or  inner  reflection.  Hitherto  the  assumption  has  been  that  the 
process  was  one  of  internal  reflection  and  interference.  The  tests  made  by  Viola 
tend  to  prove  that  instead  of  interference  it  is  a  process  of  absorption.  The 
method  followed  in  examining  the  moonstone  of  Ceylon  was  as  follows : 

Ceylon  moonstone  consists  of  coarse  feldspar  crystals,  enclosing  and  inter- 
grown  with  quartz.  The  feldspar  is  not  absolutely  definite,  but  the  analyses 
indicate  orthoclase  with  slight  admixture  of  lime  and  soda.  It  is  usually  milk 
white  in  color,  and  the  cleavages  are  wave-like.  The  silvery  schiller  appears  to 
be  most  marked  parallel  to  the  face  201.  Sections  were  therefore  cut  parallel 
to  this  face,  and  about  1  mm.  thick.  These  were  mounted  in  an  ordinary 
Fuess  goniometer  with  201  vertical,  and  the  plane  of  reflection  of  the  schiller 
(found  by  experiment  to  be  approximately  (010))  horizontal.  Parallel  light 
from  the  collimator,  reflected  from  ^oi  as  parallel  light,  gave  a  sharp  signal  for 
the  plane  ;  the  section  was  then  revolved  until  the  sky-blue  schiller  signal  was 
obtained  ;  this  was  not  sharp,  but  diffused  through  four  to  five  degrees,  but  the 


and  Laboratory  Methods. 


1351 


brighter  center   could    be   determined    within   one 
degree. 

It  was  found  that  the  signal  was  obtained  for  all 
angles  of  incidence  precisely  as  if  it  was  due  to  a 
reflection  from  some  interior  surface,  and  it  might 
be  assumed  that  the  diffusion  was  due  to  this  sur- 
face not  being  perfectly  smooth.  In  order  to  find 
the  angle  between  the  supposed  interior  surface  of 
reflection  and  the  surface  of  the  section, 

Let  O  be  center  of  revolution, 

Let  C  be  collimator. 

Let  T  be  telescope. 

Let  N  be  normal  to  plate  when  yielding  ordi- 
nary signal, 

Let    N^    be    normal    to    plate    when    yielding 
schiller  signal. 

If  COTr=2  cp  then  CON^TON^^y  and,  see  Fig.,  a'=(^— e  and  b'=<7>+e. 
But  as  the  rays  from  C  and  to  T  must  have  been  refracted,  the  true  incident  and 
reflected  rays  for  the  internal  surface  cannot  have  been  these,  but  rather  such 
rays  as  C  O  and  T'  O,  making  angles  a  and  b  with  the  surface  normal  O  N',  in 

....             sin  a'        ...      sin  b'  ,    .        ,  .    ,         ^       ^ 

which  sm  a= and  sm  b= ,  //  being  the  mean  index  of  refraction 

for  the  moonstone.     The  normal  to  the  internal  surface  will  therefore  be  O  N", 
and  the  angle  between  the   two  surfaces  will  be  equal  to  N'O  N",  denoted  by  d, 


and,  from  the  figure,  b — d^a-f-d,  or  d= 


b— a 


For  Ceylon  moonstone  cut  parallel  ^oi  d=12°  5'  and  65°  19',  and  is 
essentially  independent  of  the  angle  of  incidence.  There  is  also  a  trans7nitted 
image  the  color  of  which  is  complementary,  that  is,  yellowish  to  reddish  orange. 
This  would  indicate  that  the  violet  and  blue  rays  were  diffused  and  reflected, 
while  the  red,  yellow,  etc.,  penetrate.  If  the  phenomenon  were  one  of  interfer- 
ence, then  if  for  a  certain  angle  of  incidence  the  reflected  color  is  blue,  it  must 
pass  into  red  for  a  larger  incident  angle,  and  this  it  does  not  do.  The  theory  of 
internal  reflection  and  absorption  seems  to  best  explain  the  phenomena.  Similar 
results  were  obtained  with  the  Amelia  Co.,  Va.,  albite  and  the  adular  of 
Zillerthal.  a.  j.  m. 

Thin  lamellae  with  bright  metallic  lustre. 

Color  on  cleavage,  silver  white  with 
normal  incidence,  reddish  brown  with  oblique  incidence.  G.  7.6  H.  1.5.  Streak 
lead  gray.     Ni2  TCg  with  traces  Pb,  Bi,  and  Au. 

Minerals  of  Japan.  Kotora  Jumbo,  in  Jour. 
Coll.  Sci.  Tokio,  11:  213-281,1899.  Nine 
page  Abs.  Zeit.  f.  Kryst.  34:  215,  igoi. 


Melonite.     From    Worturpa,  South  Australia. 
Trans.  Roy.  Soc.  S.  Australia,  23 :  211,  1899. 


According  to  Vogt  there  should  exist  a 
series   of   minerals,  belonging   to    this 
group,    low    in    silica    and    containing 
variable  amounts   of  calcium    and    aluminium,  varying   between  the    limits  of 


Fouque,  F.  Contribution  a  I'etude  des  miner- 
aux  de  group  de  la  melilite.  Bull.  Soc.  Min. 
23:  ID,  1900. 


^•>''">-  J(>iirn;>I   o['  Applied  Micrt^scopy 

j;c'hlrnit«>  .uul  .\kiM in.initr.  Author's  invostii;;i(ic)ns  on  artilioially  prcpaird 
nu>lilitos  i\o  not  piovo  the  oxistomo  of  this  sorios.  DilToroiU  arlilicial  inclilitos 
air  descMibi'il.  all  havinj;  in>sitivo  optical  ohatartor.  a>ul  some  sho\vii\g  spherolitic 
iorms  ami  70u,\\  stiuotm(\  Thev  aic  also  ('haiacteii/<>(l  hy  stioni;  [(ov  melilit(') 
ilonMt>  letiaetiou  (^.00^)     .t>0(iV  ,    m,  |   | 

Rojicra,  A.  F.  Sphaloiitc  nysi.ils  ol  .»  pc*  >ili;ii  Tlu'  iiystalsaie  i  tiUlish-hrown  in  voloi , 
lK»l>i(   aiul  willi  owe  lu'w  toin\,  \\om  (Jalcn.i,  ,        "  i     ■        i  i- 

K.>ns.«s.     Am.  loui.  S.i.  iv.Q:  i?.),  i.)oo,  ami  sijoitoned    in  the   direction  ot   one 

of    the    luMahedial    inteiaxes.        They 

ha\(>  a  hemimoiphic  aspect  dm>  to  the   pn'si-iue  ol    thi'  laces  ol    the  new  ioiin,  a 

positive  hemi  tetiai;onal    ti  isoctalu^lion  o  (^S;>.'!V  ti  umatiiij;    hall    ol    tlu-  dodeca 

hiulial  Climbs,  tlu>  doilccaliedion  ,;'  ^1  It*"!  luMiii;  the   chiet  loim.      Twins  aii'  nu>n> 

ciMUiUvHi    than    sunple    vivstals.      MiMsuienuMits    ueie   made   only    with    t'ontait 

jjoniomi'tii.  i    m.  i   i 

I'roslon.  H.  L     lllmoislJuloh  M.-u>ou.<-,     A,u.        ,-,,,iii(,>  niesent  in  very  small  quantity. 

lion  shows  iu>  etihiui;  lii;ures.      Rhah- 
dite  crystals  probably  present.     \\  cMj;hl       '.2.115;')  grains.    (\      7.7.     .Analysis^ivcn. 

1..  Mil.  1., 

Suh«i.l(c.  ..  now  minoval.  i;.  A.  ^\.^^c^  ,\  copper  -  sulpho-yanadile,'"  :U\i..S  . 
}o\\\.  I  \\('\\\.  .^oo.,  77:   ie>).).  looo.  '  '  '  ■ 

\  ,jS   ,  from   "near  the  buna,  in  St)u(h 

.Australia."  associated  with  mal.ichiti\  .i/nrite.  ijuart/,  yanadium  ochre,  gypsum, 

and  calcile.      I'iist  reciMded  inst.iuie  ol  a  sulphide  mineral  cont.iinin<;  yanadium 

as  one  ot  its  piiucip.d  constituents.       Two  .in.il\s(-s  i;.i\e   tlu>  loUowini;  ; 

C\i  \.  S. 

.\.  .....        51.57  i:?.b>  :m.5^7 

iv  ....     xl.\)(\  i;{.7o  :u.0'2 

riu-oiciual  tor  :u'u„s  •  \".,s  ,.  -    51.50  i;>.ss  :?4.(>'2 

Some  ot  the  physic. d  pioperties  .ire  :  massive,  luster  metallic  to  sub-metallic, 
lolor  bion/e  yellow,  streak  nearly  black.  H  .'5.5  (>  ■{.  No  crxstalline  form 
vK'tecled.  .\.  I",  u. 

Rlchnnls.  Joseph  \V.,  and  I'owcll.  N«rm«n  S.  p,,^,  experiments  were  undertaken  to 
r.iihi>ii.»u\s.  Knii.  AiuriuMu.  .^vv.  22:  1 1 ;,  lind  a  satisfactory  substitute  for  hydro- 
Nt.iuli,  looo.  ehloric  acid  for  testing  carbonates  in 

the  tield.  The  action  of  -0  per  cent,  solutions  oi  potassium  acid  sulphate,  citric 
acid,  tartaric  acid,  and  U>  per  cent,  solution  of  oxalic  acid,  on  sixteen  of  the 
nuire  common  c.vrbonate  minerals,  in  lump  aiul  powder,  are  recorded  in  tabular 
form.  Tartaric  acid  is  regarded  as  the  best  reagent,  with  citric  acid  as  a  close 
second.  Some  sulphides  give  olT  hydrogen  sulphide  with  the  reagents.  The 
authors  seem  \o  have  been  utterly  unaware  o\  Holton's  previous  work  along  this 
line.  .Annals  N.  \.  .\cad.  Sci..  I:  I.  l>7i>;  rhemical  News,  .?0:  •J45»  ;  37:  11, 
*J»,  t>5,  S(),  «)S  ;    4.1:   :51.  ;5'.>.  A.  K.  K. 

Nichols,  Henry  W.  A  Now  'l'<-si  loi  (."hloiiuo  'Potest  a  substance  for  chlorine  it  is 
toi  I'so  with   tlio  lUowpipo.       .\nuM.  C'lioiu.  ,  i      ■  i  •  •  ^        ^    \     . 

1o«i.  2.S:  ;,i  s.  Apiil.  ukm.  powdered  with  potassium  acid  sulphate 

and  placed  in  a  closed  tube.  .\  frag 
n\ent  ol  tiltei  papei,  moistened  with  cobalt  nitrate  solution,  is  put  into  the  mouth 
of  the  tube,  and  the  mixture  fused.  If  chlorides  are  present  the  paper  will  turn 
a  bright  blue,  and  if  bromides  or  iodides  are  present  the  color  will  be  green.  In 
the  case  of  minute  quantities  it  mav  be  necessary  to  dry  the  paper  before  the 
color  appears.  V^ther  details  of  manipulation  are  given.  Specimens  of  sodalite 
(CI.  7.1>  per  cent.),  /unyite  (^Cl.  -.5>  per  cent.\  and    apatite  gave  good  reactions. 

A.   K.   R. 


and  Laboratory  Methods.  1353 


MEDICAL   NOTES. 

Methods  for  the  Detection  of  Sugar  in  Urine — Haine's  Test. — This  is 
considered  the  best  of  the  copper  tests  for  sugar,  and  is  made  with  the  following 
solution : 

Copper  sulphate,         -         -         -         -         -     0  grs. 

Water,  distilled, 3  c.  c. 

Dissolve  the  CuSO^  thoroughly  in  the  water  and  add, 

Glycerin,  pure,  -         -         -         -         -     3  c.  c. 

which  should  be  thoroughly  mixed,  after  which  add, 

Liquor  potassse,  ....        30  c.  c. 

To  make  a  test  for  sugar  in  a  sample  of  urine,  boil  1  dram  (3.7  c.  c.)  of  the 
solution  in  a  test  tube,  and  add  2  or  3  drops  of  the  urine ;  continue  to  boil  and 
if,  after  a  few  seconds,  no  reaction  occur,  add  '1  or  3  drops  more,  and  so  on  until 
8  drops  are  added,  after  which  no  more  urine  should  be  added.  It  is  best  to 
use  the  least  possible  amount  of  urine  that  will  produce  the  reaction.  The  solu- 
tion should  not  be  allowed  to  boil  more  than  one-half  minute.  If  sugar  is  present 
in  the  urine,  a  yellow  or  yellowish-red  precipitate  forms. 

This  test  is  simple  in  application,  and  is  sufficiently  reliable  to  be  depended 
upon  in  general  practice.  The  solution  is  perfectly  stable,  and  may  be  kept 
indefinitely  without  deteriorating. 

Phenyl-Hydrazi7i  Test. — This  is  an  exceedingly  delicate  test,  and  is  very  desir- 
able when  the  routine  test,  above,  leaves  any  doubt  as  to  the  presence  of  sugar 
in  the  urine.  The  test  is  performed  by  adding  to  50  c.  c.  of  the  suspected  urine, 
'1  gms.  of  phenyl-hydrazin  hydrochloride,  1.5  gm.  of  sodium  acetate,  and  20  c.  c. 
of  distilled  water.  This  solution  should  be  heated  moderately  in  a  water  bath 
for  an  hour,  after  which,  when  cooled,  if  the  smallest  amount  of  sugar  be  present, 
a  yellowish  crystalline  precipitate  is  deposited. 

With  the  above  methods  the  presence  or  absence  of  sugar  in  the  urine  may 
be  readily  and  conclusively  ascertained.  When  sugar  is  detected,  it  is  very  im- 
portant to  determine  the  amount  present.  This  may  be  accomplished  with 
accuracy  by  pursuing  the  following  method  : 

Purdy^s  Method  for  the  quantitative  determination  of  sugar  in  urine. — Dissolve, 
with  gentle  heat,  .5  gm.  pure  cupric  sulphate,  and  3.8  c.  c.  glycerol  in  20  c.  c. 
distilled  water.  With  this  mix  a  solution  of  2.4  gms.  potassium  hydroxide  in 
20  c.  c.  distilled  water,  and  add  35  c.  c.  strong  ammonia.  Make  up  to  100  c.  c.  with 
distilled  water. 

Place  exactly  35  c.  c.  of  this  solution  in  a  flask,  dilute  with  an  equal  amount  of 
distilled  water,  and  bring  to  boil.  To  this  add  slowly,  drop  by  drop,  the  urine 
to  be  tested  until  the  solution  loses  its  blue  color  and  becomes  perfectly  colorless. 
The  amount  of  urine  required  contains  exactly  .02  gm.  of  sugar.  If  it  takes 
1  c.  c.  of  urine,  there  is  2  per  cent,  of  sugar  ;  if  it  takes  \  c.  c.  of  urine,  there  is  8 
per  cent,  of  sugar.  c.  w.  j. 


1354  Journal  of  Applied  Microscopy 


NEWS  AND  NOTES. 

The  article  "  The  University  of  Montana  Biological  Station,  "  which  appeared 
in  the  May  number  of  the  Journal,  elicited  a  number  of  questions,  in  answer  to 
which  the  author,  Prof.  Morton  J.  Elrod,  has  added  the  following: 

The  microscopical  equipment  during  the  past  summer  consisted  of  four 
large  compound  microscopes,  with  two-thirds  and  one-sixth  objectives  each  ;  one 
small  microscope  with  three  objectives ,  several  additional  objectives  of  higher 
and  lower  powers ;  a  dozen  or  more  hand  lenses,  doublets ;  an  abundance  of 
glass  slips  and  covers  ;  an  assortment  of  common  stains  and  chemicals  ;  glassware 
necessary  to  carry  on  the  work,  such  as  watch  glasses,  small  beakers,  pipettes, 
staining  dishes,  etc.  A  centrifugal  apparatus  was  used  to  determine  the  quantity 
of  the  plankton.  The  camera  had  a  Zeiss  anastigmat  lens.  Series  IV,  telephoto 
attachment,  and  ray  filter.  The  vials  for  containing  microscopic  life,  plankton, 
were  of  three  different  lengths,  with  the  same  diameter,  making  it  necessary  to 
carry  corks  of  one  size  only.  This  was  found  to  be  a  great  convenience,  especi- 
ally as  several  gross  of  vials  were  carried.  The  vials  were  straight  shells,  without 
neck.  As  a  preservative  formaldehyde  was  used.  The  concentrated  or  forty 
per  cent,  solution  was  carried  in  small  bottles,  so  that  if  one  should  be  broken  but 
a  portion  of  the  supply  would  be  lost.  The  concentrated  solution  was  diluted  as 
used.  No  alcohol  could  be  carried  while  collecting,  the  laws  preventing  alcohol 
from  being  taken  into  an  Indian  reservation. 

During  the  past  summer  seventeen  students  took  advantage  of  the  facilities 
for  work  offered  by  the  station.  The  microscopical  work  was  largely  elementary. 
Much  use  of  the  instrument  was  made  in  the  study  of  Entomostraca  from  the  lakes. 
Animal  and  plant  structures  were  examined,  several  students  using  the  micro- 
scope for  the  first  time.  Simple  mounts  were  made,  and  the  method  of  using 
stains  was  made  known  through  practice.  Four  of  those  attending  took  regular 
courses  in  either  botany  or  general  zoology,  with  daily  use  of  microscope  and 
microscopic  material.  One  microscope  was  in  constant  use  for  two  months  in 
study  of  Entomostraca.  Four  students  devoted  most  of  the  summer  to  orni- 
thology.    One  worked  on  fishes,  one  on  butterflies. 

In  the  light  of  past  experience  the  following  conclusions  have  been  reached. 
Although  the  state  is  large  and  the  population  small,  there  is  much  more  interest 
shown  in  the  station  work  than  was  at  first  anticipated.  The  obstacles  in  the  way 
are  not  so  great  as  would  naturally  be  expected.  The  chief  difficulty  is  in  get- 
ting over  the  country,  but  if  one  is  not  crowded  for  time  this  makes  little  differ- 
ence. The  lakes  in  the  mountains,  though  containing  cold  water,  have  many 
very  interesting  forms  of  life.  Very  few  of  the  lakes  have  been  touched.  Flat- 
head lake,  with  its  inlets  and  outlet,  has  sufficient  territory  for  a  large  working 
force,  and  sufficient  material  for  wide  range  of  study  and  experiment.  Natural- 
ists from  the  East  who  have  a  month  to  spare  in  the  summer  and  who  want  to 
see  the  West,  and  at  the  same  time  wish  to  do  some  work,  may  find  the  station  of 
advantage,  and  will  be  able  to  get  into  the  field  and  into  the  hills  without  wast- 
ing most  of  the  time  in  learning  how  and  where  to  go.     The  field  is  rich  enough  to 


and  Laboratory  Methods.  1355 

warrant  greater  expenditure  in  apparatus  and  material.  A  house  boat  would  be  a 
great  convenience  and  of  great  utility.  The  great  needs  of  the  station,  in  order 
to  secure  the  best  results,  are  more  extensive  working  material  and  a  longer 
working  period.  For  many  years  the  station  will  be  a  place  for  investigation 
rather  than  a  summer  school  for  students.  Its  best  work  will  be  done  by  mak- 
ing provision  for  both.  There  is  excellent  opportunity  to  establish  a  station  on 
a  larger  scale,  in  a  region  offering  great  variety  of  life,  from  Alpine  to  that  at 
3000  feet  altitude,  and  from  swamp  to  barren  hill.  It  is  hoped  the  income  of 
the  university  will  warrant  the  increased  expenditure  at  an  early  date.  While 
there  is  no  comparison  between  the  life  of  the  region  and  that  of  a  favorable 
ocean  locality,  the  problems  offered  are  of  a  different  nature,  fully  as  interesting, 
and  quite  as  important.  Nowhere  is  there  better  opportunity  to  study 
variation  and  its  effects  than  in  mountain  regions. 

The  work  of  the  coming  season  will  be  better  than  in  preceding  years.  In 
addition  to  the  work  of  the  director.  Principal  P.  M.  Silloway,  of  the  Fergus 
county,  Montana,  Free  High  School,  will  have  charge  of  work  in  ornithology, 
which  he  did  so  ably  the  past  season.  Maurice  Ricker,  principal  of  the  Burling- 
ton, Iowa,  High  School,  will  give  the  instruction  in  nature  study  and  physiography. 
The  New  York  Botanical  Garden  will  cooperate  in  the  botanical  work  of  the  sta- 
tion. Dr.  D.  T.  MacDougal,  director  of  the  laboratories  in  that  institution,  will 
join  the  party  in  the  field  for  the  purpose  of  making  collections  and  pursuing 
some  investigations ^upon  the  results  of  climate  and  vegetation,  and  will  continue 
both  lines  of  work  at  the  station.  The  botanical  work  during  the  session  will  be 
under  his  guidance.  Attention  will  be  given  to  general  botany,  and  to  the  spec- 
ial features  of  the  flora  of  Montana.  Mr.  R.  S.  Williams,  of  the  same  institution, 
will  spend  the  month  of  June  making  collections  in  the  northwestern  part  of 
the  state,  and  will  be  present  during  a  part  of  the  session,  giving  special  atten- 
tion to  mosses  and  ferns. 

During  the  five  or  six  weeks  previous  to  the  opening  of  the  station  the 
instructors  will  devote  their  time  to  collecting  in  the  immediate  region.  An 
outfit  has  been  provided  which  will  make  the  trip  quite  comfortable,  and  the 
entire  time  will  be  devoted  to  collecting  in  different  fields,  from  snowy  mountain 
summits  to  the  marshes  of  the  lakes  and  rivers.  During  this  trip  additional  col- 
lections will  be  made  in  lakes  not  yet  visited,  and  in  regions  where  collectors 
have  not  yet  been.  After  the  collecting  trip  the  party  will  proceed  to  the  station, 
and  will  take  care  of  the  students  attending,  at  the  same  time  continuing  the 
collecting  and  making  additional  observations. 

The  collecting  trip  is  made  possible  through  a  contribution  from  Hon.  Wm. 
A.  Clark,  who  has  contributed  annually  for  this  purpose,  and  to  whom  the  sta- 
tion is  greatly  indebted.  The  purchase  of  the  boats,  erection  of  building,  and 
expense  of  the  instructors  during  the  past  two  years  has  been  met  by  contribu- 
tions from  friends,  the  principal  contributors  being  E.  L.  Bonner,  H.  W.  Ham- 
mond, A.  B.  Hammond,  Dr.  W.  P.  Mills,  W.  P.  Murphy,  and  W.  A.  Clark. 
The  expense  for  the  coming  summer,  in  addition  to  the  contribution  by  Senator 
Clark,  will  be  met  by  university  appropriation. 


1356 


Journal  of  Applied  Microscopy 


Demonstration  of  Reticu- 
late Vessels. — The  teacher  of 
plant  histology  is  usually  seek- 
ing for  the  best  possible  mater- 
ials to  illustrate  the  several 
kinds  of  cells  that  are  to  be 
examined  by  his  students.  For 
well  defined  reticulate  vessels,  I 
have  seen  nothing  to  equal  those 
found  in  the  thickened  roots  of 
Arenaria  striata,  which  grew  in 
our  botanic  garden.  They  are 
somewhat  different  from  those 
found  in  the  stems  of  Impaficfis. 
W.  J.  Beal. 


In  the  cuts  of  starchy  gran- 
ules of  the  pea,  they  are  repre- 
sented as  having  cracks,  or  checks  in  the  middle.  I  wonder  if  it  is  generally 
known  that  the  checks  seldom  appear  if  the  peas  are  placed  in  alcohol  or  glycerin 
before  drying  ?  W.  J.  Beal. 


QUESTION   BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

5.  Will  you  kindly  refer  me  to  a  good  method  of  how  to  make  a  biologic 
aquarium  ?  In  the  Journal  of  Applied  Microscopy  are  a  few  notes  on  Culti- 
vation of  Algae,  but  not  sufficient  for  an  amateur.  What  should  be  the  soil- 
gravel,  sand,  or  mud  from  a  pond  ?  How  proceed  to  stock  it  and  with  what  ?  Is 
the  evaporation  to  be  supplied  from  a  pond  or  hydrant  ?  If  glass  cover  on,  can 
there  be  enough  air  to  pass  between  for  living  ?  I  am  desirous  of  having  a 
number  of  jars  for  general  elementary  biology  work  ;  to  supply  Amoeba,  Hydra 
Chara,  Vorticella,  Spirogyra,  Vaucheria,  Nitella,  Vallisneria,  etc. — v.  a.  l. 

6.  What  is  Scott's  method  for  the  examination  of  blood  ? — t.  g.  s. 

7.  Can  tinted  paper  be  used  for  the  haemoglobin  test  ?^ — t.  g.  s. 

REPLY  TO  QUESTIONS  3  AND  4  IN  THE  MAY  NUMBER. 

The  Welsbach  light  is  practically  worthless  for  high-power  microscopy,  either 
visual  or  photographic,  for  with  condenser  focused,  as  it  should  be,  an  image  of 
the  fabric  of  the  mantle  is  projected  into  field  of  view.  Have  found  no  difficulty 
in  making  photographs  up  to  Xl'-^OO,  using  H.  I.  objectives  and  Huyghenian 
oculars,  with  oil  lamp,  half-inch  wick.  Render  rays  approximately  parallel  with 
bulls-eye,  or  better,  a  large  size,  short  focus  photographic  lens  placed  at  its  focal 
distance  from  lamp  flame,  and  converge  on  object  with  substage  condenser 
sharply  focused,  achromatic  condenser  decidedly  preferable,  but  Abbe  con- 
denser will  answer  fairly  if  nothing  better  is  available.  No  noticeable  advantage 
derived  from  the  use  of  achromatic,  periscopic,  orthoscopic,  or  compensation 
oculars  over  the  Huyghenian,  when  used  with  achromatic  objectives. — f.  j.  k. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 

VOLUME  IV.  JULY,  1901.  Number  7 


The  Value  of  Methylen  Blue  as  an  Intravitam  Stain  in 

the  Tunicata. 

While  working  for  special  results  on  the  tunicate  nervous  system,  with 
methylen  blue,  I  found  that  this  anilin  could  be  made  of  much  value  as  a  gene- 
ral stain  where  living  material  was  obtainable.  Small  species,  such  as  Amorxcium, 
Botryllus,  and  Pcrophora,  as  well  as  young  Molguh^,  left  in  sea  water  containing  just 
enough  methylen  blue  to  color  the  water  a  lively  blue  (about  1  part  to  500U)  for  half 
an  hour,  will  give  almost  diagrammatically  the  branchial  basket  and  its  organs,  as 
well  as  the  free  mesenchyme  cells  of  the  body  cavity,  leucocytes  and  phagocytes. 
This  method  is  especially  favorable  for  cilia;  the  demonstration  of  cilia  in 
motion,  the  arrangement  of  cilia  in  rows  on  the  surface  of  the  cell,  and  the  pecu- 
liar thickened  basal  portion  of  the  tunicate  cilium  can  all  be  well  shown.  Long, 
whip-like  flagellae,  which  are  found  in  the  endostyle  and  ciliated  funnel,  also  take 
the  blue  and  stand  out  with  wonderful  distinctness.  The  above  named  cells  are 
usually  the  first  to  stain.  The  sensory  or  peripheral  portion  of  the  nervous  sys- 
tem stains  relatively  early  (from  1  to  1^  hours  after  immersion),  while  the 
deeper  lying  nerve  cells  and  motor  fibers  stain  later.  Cells  of  the  central  nervous 
system  are  sometimes  found  colored  blue  as  much  as  five  hours  after  immersion. 
But  all  of  this  so-called  staining  of  the  different  tissues  is  transitory,  sometimes 
lasting  only  a  few  minutes — as  in  the  case  of  the  very  delicate  neurofibrils — or 
for  several  hours,  or  even  days,  in  the  case  of  the  mesenchyme  cells. 

Special  Methods. — The  best  results  for  the  staining  of  the  nervous  system 
were  obtained  by  the  two  following  methods  :  Molgulse  were  placed  in  a  weak 
solution  (1-5000)  of  Meyer's  BX  methylen  blue  in  sea  water,  and  allowed  to  remain 
from  one  to  five  hours,  according  to  the  size  of  the  animal  and  the  tissue  to  be 
stained.  For  staining  by  immersion  small  specimens  were  used.  It  was  found 
necessary  to  have  the  animals  absolutely  fresh,  or  satisfactory  results  could  not 
be  depended  upon.  Molgulae  which  had  remained  in  aquaria  for  so  short  a 
period  as  two  to  three  days,  frequently  refuse  to  take  the  stain  ;  or  give  a  diffuse 
staining.  The  exact  intensity  of  the  blue  in  solution  does  not  seem  to  be  as 
important  a  factor  as  the  length  of  immersion.     Just  before  taking  out  the  animals 

(1357) 


1358  Journal  of  Applied  Microscopy 

for  examination  they  were  removed  to  a  dish  of  running  sea  water  aerated  by  a 
pipette  nozzle.  Specimens  thus  treated  gave  uniformly  good  results,  while  those 
in  which  this  process  was  omitted  did  not.  Whether  the  former  result  was  due 
to  the  revival  of  the  animal  or  the  oxygenation  of  the  tissues,  is  hard  to  say.  But 
the  physical  condition  of  the  animal  seems  to  play  an  important  part. 

The  other  method  successfully  used  was  to  inject  a  fairly  strong  solution 
(1  to  4  per  cent.)  of  methylen  blue  into  the  ovarian  vein  of  Molgula;.  From 
thence  the  fluid  reached  the  heart  and  was  pumped  to  all  parts  of  the  body.  A 
small  amount  of  fluid  should  be  used  and  great  care  taken  not  to  allow  too  much 
of  the  body  fluid  to  escape  through  the  hole  made  by  the  needle.  The  per- 
centage of  successful  staining  by  this  method  is  small ;  but  the  results  are 
valuable,  because  of  the  use  of  large  specimens.  The  peripheral  system  seems 
to  come  out  especially  well  by  this  method.  Animals  which  die  as  a  result  of  the 
injecting  process  give  a  diffuse  staining  of  tissues  that  is  of  no  neurological 
value.  This  diffuse  staining  can  be  recognized  at  once,  by  the  fact  that  the  whole 
animal  stains  a  pale  greenish  blue  color  ;  while  in  a  perfectly  successful  stain  the 
color  is  deep  blue. 

The  use  of  ammonia  is  advocated  by  Apathy  in  bringing  out  the  stain.  He 
believes  that  in  exposure  to  the  air  just  before  examination,  the  specimen  takes 
up  some  ammonia  from  the  atmosphere.  Molgulai  exposed  to  the  fumes  of 
ammonia  gave  negative  results.  The  same  may  be  said  regarding  the  exposure 
of  stained  tissue  to  the  air.  It  could  not  be  proven  that  oxygen  was  a  factor  in 
the  bringing  out  of  the  stain. 

The  fixation  of  the  stain  for  permanent  preservation. — Most  of  the  methods 
used  by  investigators  {I'ide  Arnstein,  Apathy,  Bethe,  Dogeil,  Huber.  Meyer, 
Peabody,  Retzius,  and  others)  agree  in  the  use  of  picrate  ammonia  as  a  fixing 
agent.  Aqueous  solutions  are  usually  employed,  and  the  material  is  allowed  to 
remain  in  a  saturated  solution  of  ammonium  picrate  from  a  few  minutes  to  several 
hours,  or  even  days,  according  to  the  size  and  permeability  of  the  material.  The 
macerating  effect  of  the  fluid  is  avoided  in  some  cases  by  the  addition  of  one 
part  to  100  of  a  1  per  cent,  osmic  acid  solution.  Such  material  may  then  be 
either  mounted  permanently  in  a  solution  of  saturated  ammonium  picrate  and 
glycerin  in  equal  parts  (rvV/f  Meyer,  Retzius)  ;  or  in  chemically  pure  glycerin 
without  washing  out  {inde  Dogeil) ;  or  by  the  somewhat  complicated  method  of 
Apathy,  when,  after  fixation  in  ammonium  picrate  plus  a  little  (;")  drops  to  100  c.  c.) 
concentrated  ammonia,  the  material  is  passed  through  glycerin  ;  glycerin  and 
gum  arable  ;  and  finally  mounted  in  a  solution  of  gum  arable,  cane  sugar,  and 
water,  in  equal  quantities.  I  may  say,  in  passing,  that  with  tunicates  I  found 
the  last  named  method  the  least  useful. 

By  far  the  best  of  my  results  were  obtained  by  the  following  modification  of 
the  methods  of  iJethe  and  Apathy  : 

Cut  out  the  part  to  be  used,  and  after  examination  under  a  fairly  high  power 
of  the  microscope,  remove  the  successfully  stained  material  to  a  saturated  solu- 
tion of  ammonium  picrate  in  sea  water.  The  pieces  of  tissue  are  then  immediately 
removed  to  a  slide,  or  small  dish,  where  they  are  left  for  a  few  minutes  (10  to  20, 
according  to  the  size  of  piece),  in  the  following  solution  : 


and  Laboratory  Methods. 


1359 


Sea  water  (or  normal  salt),         -         -         .         -         50  c.  c. 
Cone.  c.  p.  glycerin,     -         -         -         -         -         -         50  c.  c. 

Cone,  ammonia,  .-.-..  i  drop. 

Add  more  glycerin,  and  finally  mount  in  glycerin  containing  just  enough  of 
the  ammonium  picrate  to  color  it  slightly.  Specimens  put  up  in  this  way  have 
kept  their  color  now  for  over  three  years,  no  noticeable  change  having  taken  place 
during  that  time. 

The  above  method  was  frequently  modified.  I  did  not  find,  as  some  later 
writers  have  done,  that  better  results  were  obtained  by  a  bath  of  longer  duration 
in  ammonium  picrate.  A  short  bath  of  from  y^  to  \  minute  suffices  for  sensory 
nerves  and  peripheral  sense  organs,  and  somewhat  longer  for  deeper  lying  nerves. 
I  am  inclined  to  take  Apathy's  view  regarding  the  successful  staining  of  the 
nerve  fibers.  He  believes  that  he  gets  a  true  stain  and  not  an  impregnation. 
In  a  few  very  successful  cases  I  have  succeeded  in  following  to  the  nerve  cell  a 
bundle  of  fibers,  which  I  believe  to  correspond  to  primitive  neurofibrils. 

More  frequently,  however,  an  impregnation  probably  takes  place,  the  whole 
interfibrillar  space  taking  the  blue.  This  can  best  be  shown  by  following  the 
successive  changes  which  take  place  in  the  tunicate  nerve  fiber  after  death,  or 
during  the  diffuse  staining  which  takes  place  in  the  later  stages  of  every  suc- 
cessful nerve  staining  with  methylen  blue.  The  successful  stain  of  a  fiber  shows 
an  almost  unbroken  wavy  blue  line,  or  series  of  interlacing  fibrils,  around  which 
can  be  seen  very  faintly  the  sheath.  This  fiber  has  almost  no  knobs  or  granula- 
tions in  its  course  except  at  the  true  ending.  At  a  point  of  branching  a  triangular 
blue  area,  probably  caused  by  the  stretching  of  the  sheath,  can  be  seen.     As  the 

tissue  dies,  however,  a  change 
takes  place.  The  nerve  fiber 
begins  to  bead,  at  first  hardly 
noticably,  but  later  these  head- 
ings become  so  large  as  to  dis- 
tort the  whole  fiber  and  com- 
pletely change  its  appearance. 
Ultimately  all  trace  of  the  fiber 
disappears  except  a  line  of 
irregularly  placed  blue  globules. 
(See  figure.) 

This  same  beading  of  the 
fiber  is  frequently  induced  by 
fixation  with  ammonium  picrate, 
and  it  is  only  in  rare  cases 
when  we  have  the  fiber  fixed  so  that  it  shows  the  individual  fibrils.  Frequently 
there  appears  to  be  a  vacuolization  of  the  nerve.  Large  vacuoles  appear,  at 
one  side  of  which  there  is  a  heavier  deposit  of  blue.  This  is,  perhaps,  a 
pathological  condition  induced  by  the  injection  of  the  methylen  blue. 

Fixation  for  imbedding  and  sectioning. — Parker's  sublimate  and  alcohol  method 
was  tried  with  no  success.  Bethe's  ammonium  molybdate  method  (Arch.  f.  Mik. 
Anat.  xliv.   '94)  yielded  poor   results.     His  later  method  (Anat.  Anz.  xii.  '96) 


Four  stages  in  the  degeneration  of  a  nerve  fibre : 
I.  A  successful  impregnation. 
II.  Ten  minutes  later;  beading  commencing. 

III.  Twenty  minutes  later;  much  beading. 

IV.  Two  hours  later. 


1360  Journal  of  Applied  Microscopy 

proved  of  more  value.  After  staining  tissues  in  a  concentrated  solution  of 
ammonium  picrate  (which  I  used  in  sea  water)  the  material  is  brought  into  the 
following  solution  : 

Ammonium  molybdate,     -         -  1  gr. 

H^O,       ;         .         -         -         -         "g'-loriUccH^O. 
oper  cent,  osmic,      -         -         -         lU  gr.  )  ^ 

Peroxide  of  hydrogen,       -         -  1  gr. 

or  (with  somewhat  better  results  for  tunicates),  phosphomolybdate  of  soda  may  be 
substituted  in  the  above  formula  for  the  ammonium  molybdate. 

After  3^  to  1  hour  in  above  solution  (or  4  to  I'J  hours  in  the  osmic  solution), 
we  wash  in  water,  rapidly  pass  through  the  alcohol,  xylol,  and  imbed  in  paraffin. 
Results  from  this  method  have  not  been  uniformly  successful. 
De  Witt  Clinton  High  School,  N.  Y.  C.  GeorGE  William  Hunter,  Jr. 


Spermatozoa  of  Man,  Domestic  Animals,  and  Rodents. 

The  male  cell,  or  spermatozoon,  is  of  minute  size,  and  in  its  locomotor 
energy  and  vitality  resembles  a  flagellate  monad.  Anatomically  it  is  a  true  cell, 
consisting  of  the  "  head,"  composed  mainly  of  nucleus,  and  the  motile  "  tail," 
which  may  be  fibrillated,  and  a  small  central  portion  between  the  head  and  tail, 
which  is  sometimes  regarded  as  the  "  centrosome." 

In  studying  the  spermatozoon  of  the  mastifif,  I  noticed  the  striking  resem- 
blance it  bore  to  that  of  man  ;  finding  that  if  the  spermatic  fluids  were  allowed  to 
stand  for  a  time,  even  staining  did  not  furnish  sufficiently  satisfactory  evidence 
to  enable  one  to  distinguish,  beyond  question,  the  spermatozoon  of  man  from  that 
of  the  dog,  except  where  careful  measurements  were  employed  ;  and  a  fact  ever 
to  be  borne  in  mind  is,  that  these  measurements  may  vary  slightly  in  different 
persons  and  animals,  and  even  in  the  same  specimen. 

Through  the  courtesy  of  Dr.  James  Johnston  I  secured  testicles,  preserving 
as  much  as  possible  of  the  vasdeferens,  from  animals  slaughtered  at  the  Phila- 
delphia abattoir,  to  which  collection  was  added  those  from  certain  animals 
employed  for  experimental  work  in  the  laboratory.  The  spermatic  fluids  studied 
were  taken  from  the  vasdeferens,  and  from  an  incision  of  the  testicle.  In  man, 
the  fluid  ejaculated  at  intercourse  was  also  studied  and  the  findings  compared 
with  those  where  the  fluid  was  taken  from  the  vasdeferens  and  from  the  paren- 
chyma of  the  testicle. 

Examinations  were  made  immediately  after  the  testes  had  been  removed,  and 
on  the  first,  second,  third,  fourth,  and  fifth  days  after  their  removal.  Spermatic 
fluids  thus  collected  were  placed  in  cold  water,  after  which  it  was  found  that  the 
tails  became  coiled,  and  were  soon  detached  from  the  heads.  In  no  case  were 
the  spermatozoa  found  to  possess  individual  movement  twenty-four  hours  after 
the  testicle  had  been  removed,  or  after  the  death  of  the  animal ;  nor  were  they 
ever  found  motile  in  man  even  a  few  hours  after  death.  These  findings  differ 
from  the  statements  often  made,  that  spermatozoa  remain  active  for  a  long  time 
after  the  death  of  the  animal.  To  determine  this  point  one  testicle  was  kept  in 
a  cool  room,  and  the   other  at  a  temperature  of  about  75°  Fahr.,  when  certain 


and  Laboratory  Methods. 


1361 


other  changes  were  also  observed.  After  twenty-four  hours  it  was  common  to 
find  several  free  heads  and  tails,  their  number  increasing  daily,  until  by  the  fourth 
day  it  was  often  difficult  to  find  a  perfect  spermatozoon  ;  yet  these,  when  present, 
showed  evidence  of  marked  degeneration,  and  their  reaction  to  staining  was  not 
constant. 

Staining  was  accomplished  by  the  various  anilin  dyes,  of  which  carbol- 
fuchsin  was  found  to  be  of  most  value,  therefore  the  accompanying  illustrations 
were  sketched  from  specimen  slides  stained  by  carbol-fuchsin.  In  examining 
specimens  stained  in  1899,  I  find  that  the  tail  is  the  first  to  give  up  its  stain, 
and  from  one-fourth  to  one-seventh  of  the  tails  of  the  spermatozoa  of  the  sheep 
and  rats  show  no  stain,  and  are  seen  with  difficulty.  Fading  was  noted  to  take 
place  earlier  where  methylen  blue  was  employed.  In  but  one  instance,  that  of 
the  mouse,  was  it  necessary  to  apply  heat  in  order  to  stain  the  spermatozoa. 

Measurements  were  made  by  the  use  of  both  the  stage  and  eye-piece  microm- 
eters, always  measuring  the  entire  length,  dimensions  of  head,  and  length  of 
tail ;  the  latter  being  markedly  altered  whenever  the  staining  was  imperfect.  It 
was  thought  to  be  of  possible  service  to  have  all  measurements  recorded  both  in 
millimeters  and  in  inches.  All  measurements  and  sketches  were  made  with  a 
1-6  objective  and  2  eye-piece.  In  the  sketching  no  attempt  was  made  to  preserve 
the  original  size  of  the  cells.  The  total  lengths  given  were  obtained  by  the 
measuring  of  complete  cells,  recording  the  greatest  and  smallest  measurements 
only;  while  the  measurements  of  the  heads  and  tails  were  often  taken  after  these 
parts  had  separated.  Therefore  the  total  length  is  not  always  equivalent  to 
the  sum  of  the  lengths  of  the  head  and  tail. 


Fig.  1.— Man. 


Fig.  2.— Dog  (Mastiff). 


The  spermatozoa  of  man  (Fig.  1)  were  found  to  stain  deeply  with  carbol 
fuchsin ;  the  heads  and  tails  were  equally  stained,  and  appeared  more  distinct 
and  uniform  than  did  those  from  any  other  member  of  this  series.  The  tail  was 
seldom  found  to  be  coiled  or  twisted,  except  where  water  had  been  previously 


1362 


Journal  of  Applied  Microscopy 


0.051  to  0.058  mm., 
0.004  to  0.006  mm., 
0.003  to  0.004  mm., 
0.041   to  O.053  mm., 


or  0.00-2     to  0.002-2  in. 

or  0.0001  to  0.000-2  in. 

or  0.0001  to  0.0001  in. 

or  0.0016  to  0.002    in. 


added  to  the  spermatic  fluid.     Measurements  showed  the  spermatozoon  of  man 
to  be  the  smallest  member  of  this  series. 

Total  length, 
Length,  head, 
Width,  head. 
Length,  tail. 

The  spermatozoa  of  the  dog  (Fig.  2)  presented  many  features  which  would 
readily  distinguish  them  from  those  of  man  in  the  fresh  and  well  prepared  speci- 
men ;  but  if  both  were  subjected  to  the  action  of  certain  secretions  and  fluids,  or 
allowed  to  dry  before  properly  smeared  on  the  cover-glass,  great  question  would 
doubtless  arise  as  to  the  identity  of  either  of  these  cells.  A  small  portion  of  the 
head,  located  at  the  junction  of  the  tail  and  head,  stained  deeply,  while  the 
remainder  of  the  head  appeared  as  a  homogeneous  structure  bounded  by  a 
rather  distinct  margin.  The  tail,  too,  showed  but  moderate  affinity  for  stains. 
The  head  and  tail  of  an  individual  cell  were  occasionally  seen  to  unite  at  right 
angles,  and  a  few  specimens  were  observed  having  two  distinct,  well  formed 
heads  projecting  from  a  single  tail.  The  dog  furnishing  the  specimen  for  this 
series  weighed  105  pounds,  and  it  was  the  intention  to  compare  the  following 
measurements  with  those  obtained  by  the  study  of  spermatozoa  taken  from  a 
terrier,  but  such  opportunity  did  not  offer  itself. 

Total  length,  .        0.007  to  0.074  mm.,  or  0.0026  to 

Length,  head,  .     0.004  to  0.008  mm.,  or  0.0002  to 

Width,  head,  .     .   0.003  to  0.002  mm.   or  0.0001  to 

Length,  tail,  .  .      0.050  to  0.067  mm.,  or  0.0023  to 


0.0028  in. 
0.0003  in. 
0.0001  in. 
0.0026  in. 


Fig.  H.— Rabbit 


Horse. 


The  spermatozoa  of  the  rabbit  (Fig.  3)  possess  many  features  common  to 
those  of  the  dog ;  the  head,  in  addition  to  being  narrower,  presents  a  deeply 
stained  area  at  its  junction  with  the  tail,  and  a  less  marked  deepening  of  the 
stain  was  seen  at  the  other  extremity,  occupying  nearly  one-third  of  the  whole 
head.     Between    these    two   stained   portions   a  lighter   zone  was  seen.     It    is 


and  Laboratory  Methods. 


1363 


characteristic  of  the  tail  to  form  coils,  which  were  often  seen  to  surround  the  head, 
and  if  the  smear  be  at  all  thick  these  coils  render  it  impossible  to  outline  the 
individual  cells.  An  abrupt  bend,  at  right  angle,  which  extends  for  but  a  short 
distance  and  then  forms  another  equally  abrupt  angle  to  assume  the  course  pre- 
viously taken,  was  a  common  finding.  This  irregularity  in  the  course  of  the  tail 
may  be  seen  near  the  head,  but  more  commonly  at  the  junction  of  the  first  and 

second  thirds. 

Total  length,       .     .     0.051  to  0.066  mm.,  or  0.002    to  0.0025  in. 

Length,  head,  .     .        0.006  to  0.009  mm.,  or  0.0002  to  0.0003  in. 

Width,  head,       .     .     0.003  to  0.004  mm.,  or  0.0001  to  0.0001  in. 

Length,  tail,      .     .       0.045  to  0.058  mm.,  or  0.0017  to  0.0022  in. 

In  studying  the  spermatozoa  of  the  horse  (Fig.  4)  it  was  observed  that  their 

general  characteristics  and  reaction  to  stain  were  similar  to  those  of  man,  except 

that  after  the  fluid  had  been  kept  for  a  few  days  the  tails  of  certain  cells  appeared 

to  be  fibrillated.     The  measurements  of  equine  spermatozoa  were  found  to  be  : 

Total  length,      .     .     0.064  to  0.067  mm.,  or  0.0025  to  0.0026  in. 

Length,  head,  .     .        0.0(»6  to  0.008  mm.,  or  0.0002  to  0.0003  in. 

Width,  head,       .     .     0.003  to  0.004  mm.,  or  0.0001  to  0.0001  in. 

Length,  tail,      .     .        0.054  to  0.060  mm.,  or  0.(H»21  to  0.0022  in. 


Fig.  .5. — Bull.  Fig.  6. — Mouse. 

The  spermatozoa  of  the  bull  (Fig.  5)  were  always  accompanied  by  many  free 
heads,  and  comparatively  few  free  tails.  The  head  of  each  spermatozoon  stained 
feebly  except  for  a  small  portion  at  its  junction  with  the  tail  (centrosome),  which 
stained  deeply.  The  tail  was  always  found  to  be  well  stained ;  its  course  rather 
irregular ;  but  never  was  it  seen  to  change  abruptly,  as  was  commonly  observed 
in  the  rabbit,  nor  did  it  ever  extend  in  a  direct  course  from  the  head,  as  is  the 
rule  in  man  and  in  the  horse.      The  measurements  of  bovine  spermatozoa  were 

found  to  be  : 

Total  length,  .     .     0.087  to  0.093  mm.,  or  0.0033  to  0.0036  in. 

Length,  head,  .  .     .  0.009  to  0.009  mm.,  or  0.0003  to  0.0003  in. 

Width,  head,  .     .     0.006  to  0.006  mm.,  or  0.0002  to  0.0002  in. 

Length,  tail,     .  .     .   0.077  to  0.083  mm.,  or  0.003    to  0.0032  in. 


1364 


Journal  of  Applied  Microscopy 


The  spermatozoa  of  the  mouse  (Fig.  6)  presented  many  features  in  striking 
contrast  with  other  members  of  this  series.  The  head  stained  deeply  and  pre- 
sented a  sUghtly  curved  spine  at  one  extremity,  while  surrounding  the  head  was 
a  clear  zone  (apparent  capsule).  From  the  portion  of  this  capsule  correspond- 
ing to  the  larger  extremity  of  the  head,  a  delicate,  faintly  stained  tail  was  seen 
to  emerge.  This  tail  was  rendered  more  distinct  by  the  application  of  heat 
while  staining.     Measurements  of  these  cells  were  found  to  be  as  follows  : 

Total  length,       .     .     0.12    to  0.158  mm.,  or  0.0046  to  0.0061  in. 

Length,  head,  .     .     .  0.008  to  0.009  mm.,  or  0.0003  to  0.0003  in. 

Width,  head,        .     .     0.0O3  to  0.004  mm.,  or  O.OOOl  to  0.0001  in. 

Length,  tail,    .     .     .    0.112  to  0.138  mm.,  or  0.0043  to  0.0057  in. 


Fig.  7. — Sheep. 


Fig.  8.— Cat, 


The  spermatozoa  of  the  sheep  (Fig.  7)  stained  evenly  throughout  except  for  a 
small  central  portion  at  the  union  of  the  head  and  tail.  Its  measurements  were 
found  to  be  : 

Total  length,     ....    0.083  mm.,  or  0.0032  in. 

Length,  head,       .     .     .       0.009  mm.,  or  0.0003  in. 

Width,   head,     ....     0.006  mm.,  or  0.0002  in. 

Length,  tail,    ....       0.074  mm.,  or  0.0028  in. 

Spermatic  fluid  from  the  cat  (Fig.  8)  was  found  difficult  to  study,  as  but  few 
spermatozoa  were  present.  Each  spermatozoon  presented  a  tibrillated  tail,  and 
at  times  this  fibrillation  was  seen  to  surround  the  head.  The  head  stained 
deeply,  and  at  times  a  centrosome  was  distinct.  Decided  irregularity  was  noted 
in  the  size  and  form  of  the  heads,  which  partially  explains  the  variations  found 
in  the  measurements  of  these  cells. 

Total  length,       .     .     0.058  to  0.074  mm.,  or  0.0022  to  0.002s  in. 

Length,  head,  .     .     .  0.004  to  0.007  mm.,  or  0.001    to  0.0002  in. 

Width,  head,       .     .     0.003  to  0.003  mm.,  or  0.0001  to  O.OOOl  in. 

Length,  tail,     .     .     .  0.053  to  0.066  mm.,  or  0.002    to  0.0025  in. 


and  Laboratory  Methods. 


1365 


At  first  sight  the  spermatozoon  of 
the  rat  (Fig.  9)  reminds  one  of  the 
immature  male  cell  of  the  bird,  and 
it  is  not  impossible  that  these  cells 
may  undergo  further  development. 
The  spermatic  fluids  of  both  white 
and  gray  rats  were  studied,  and 
no  marked  difference  was  found  to 
exist  between  these  cells.  The 
head  and  tail  of  this  cell  stains 
well,  the  concave  border  of  the 
head  staining  slightly  deeper  than 
its  body.  Measurements  were 
found  to  be  : 


Fig.  9.— White  Rats. 


Total  length, 
Length,  head, 
Length,  tail. 


0.225  to  U.288  mm.,  or  0.0087  to  0.0092  in. 
0.012  to  0.016  mm.,  or  0.0004  to  0.0006  in. 
0.209  to  0.222  mm.,  or  0.0081  to  0.0086  in. 


The  spermatozoon  of  the  guinea 
pig  (Fig.  10)  differs  widely  from 
any  other  member  of  the  series. 
Its  head  is  nearly  spherical,  and  a 
minute,  deeply  stained  portion  was 
noted  at  the  junction  of  the  tail. 
Each  head  was  provided  with  a 
neatly  fitting,  semi-lunar  cap,  which 
was  also  well  stained.  At  times 
these  caps  were  seen  detached,  and 
at  others  deformed,  giving  that 
portion  of  the  head  either  a  con- 
cave or  a  pointed  appearance. 
That  portion  of  the  tail  nearest 
the  head  was  always  deeply  stained, 
and  the  course  of  the  tail  was 
never  found  to  be  tortuous.     The 


Fig.  10.— Guinea  Pig. 


following  measurements  were  obtained  for  these  cells  : 

Total  length,  .  .  0.113  to  0.138  mm.,  or  0.0053  to  0.0057  in. 
Length,  head,  .  .  .  0.006  to  0.012  mm,,  or  0.0006  to  0.0004  in. 
Width,  head,  .  .  0.007  to  0.011  mm.,  or  0.0004  to  0.0004  in. 
Length,  tail,     .     .     .  0.125  to  0.132  mm.,  or  0.0048  to  0.0051  in. 

L.  Napoleon  Boston,  M.  D. 
Philadelphia  Hospital. 


1366  Journal  of  Applied  Microscopy 

LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 

AN  IMPROVED  PHOTO-MICROGRAPHIC  APPARATUS. 

I  have  recently  had  constructed  for  Cornell  Medical  College  a  photo-micro- 
graphic  apparatus  by  B.  &  L.,  a  description  of  which  may  be  of  interest  to  the 
readers  of  the  Journal,  since  there  are  certain  departures  from  the  regular  type 
of  outfit.  Some  of  these  departures  were  suggested  by  me,  but  they  were  worked 
out  in  detail  and  put  into  practical  shape  by  the  makers.  To  begin  with,  I  will 
point  out  the  various  alterations,  all  of  which,  I  think,  are  improvements  over 
the  old  style,  and  afterwards  proceed  to  describe  the  apparatus  as  a  whole. 

In  the  first  place,  then,  on  the  old  plan  the  optical  bench  and  camera  stand 
being  practically  all  in  one  piece,  the  only  way  of  finding  the  desired  field  for 
photographing  is  to  push  the  camera  back  in  its  bed  and  bend  over  to  look 
through  the  eye-piece.  In  this  uncomfortable  and  back-breaking  position  it  is 
impossible  to  manipulate  the  slide  except  by  means  of  one  of  those  clumsy 
mechanical  stages  which,  though  all  very  well  for  special  purposes  such  as  blood 
cell  counting,  are  not  to  be  compared  with  one's  fingers,  which,  after  all,  were 
made  at  a  much  earlier  date,  for  rapid  manipulation. 

In  a  Zeiss  camera  stand  which  I  had  previously  used,  the  steel  rods  on  which 
the  camera  moved  could  themselves  be  pushed  back  so  that  one  could  sit  on  a 
stool  between  the  optical  bench  and  camera  stand  in  order  to  find  the  field.  The 
chief  objections  to  this  plan  are  that  the  rods  are  apt  to  sag,  and  that  the  bench 
and  stand  cannot  be  connected  into  one  solid  piece,  making  it  difficult  to  preserve 
the  exact  optical  axis,  or  arrange  a  satisfactory  method  for  mechanical  focusing. 

Two  years  ago  I  saw  at  the  Jenner  Institute  in  London  a  photo-micrographic 
outfit  with  a  revolving  optical  bench,  but  had  no  opportunity  of  examining  the 
details  or  of  finding  out  if  the  arrangement  worked  satisfactorily  or  not,  since  it 
was  entirely  new.  On  mentioning  the  idea  to  llausch  &  Lomb,  they  suggested  a 
modification  of  one  of  their  revolving  microscopical  tables.  The  details  will  be 
described  further  on  ;  sufficient  to  say  here,  that  by  this  arrangement  I  can  sit 
comfortably  at  one  side  of  the  apparatus  and  with  my  fingers  manipulate  the  slide 
on  the  microscope  stage  as  easily  as  if  I  were  sitting  at  an  ordinary  table. 

The  camera  stand  has  a  connecting  rod  between  its  two  cast  iron  supports, 
just  above  their  feet,  and  I  then  suggested  that  the  rod  be  continued  on  to  the 
single  upright  of  the  revolving  bench  in  order  to  give  more  rigidity,  but  was  told 
that  so  low  down  it  would  be  of  no  use.  After  much  discussion,  it  was  decided 
to  have  a  rod  connecting  the  upper  parts  of  the  uprights,  and  so  made  that  it 
could  be  put  up  and  clamped  in  at  the  time  of  setting  up  the  apparatus.  This 
seems  a  very  satisfactory  arrangement,  undoubtedly  adding  to  the  rigidity  and 
helping  to  keep  the  optical  bench  and  camera  stand  in  optical  axis. 

The  next  difficulty  was  how  to  manage  for  mechanical  focusing.  When  the 
bench  and  stand  are  fixed  there  is,  of  course,  no  trouble,  since  the  focusing  rod 
can  be  prolonged  under  the  fine  adjustment  of  the  microscope,  but  where  the 
bench  revolves  this  prolongation  of  the  rod  must  be  got  out  of  the  way  tempora- 


and  Laboratory  Methods. 


1367 


1368 


Journal  of  Applied  Microscopy 


HYPO 


rily  while  the  field  is  being  found.  Mr.  Bausch  asked  me  how  I  proposed  to 
effect  this,  and  I  was  forced  to  reply  that  I  had  not  the  least  idea,  but  must  leave 
it  to  him.  Mr.  Patterson,  who  had  charge  of  the  work,  came  to  the  rescue  with 
a  very  ingenious  device,  which  will  be  described  further  on. 

It  is,  however,  always  best  to  avoid  mechanical  focusing,  and  be  able  to  reach 
the  fine  adjustment  with  one's  fingers,  if  possible,  especially  for  the  higher  powers. 
In  the  B.  &  L.  stands  that  I  had  hitherto  seen,  this  appeared  to  be  an  impossi- 
bilitv,  except  within  a  very  limited  range,  since  the  arrangement  of  the  bellows 
was  such  that  a  shorter  distance  than  fourteen  inches  could  not  be  obtained 
between  the  eye-piece  of  the  microscope  and  the  ground  glass  of  the  camera. 

Besides  this,  the  front  of  the  camera  was  always  made  the  same  size  as  the 
back,  i.  e.,  11  inches  square  for  a  i}}2  x8i4  plate,  so  that  the  wrist  had  to  be 
bent  round  over  the  front  to  reach  the  fine  adjustment,  thus  losing  three  or  four 
inches  of  distance  for  hand  focusing. 

Not  only  is  the  hand  focusing  superior  in  accuray,  but  it  is  essential  for 
another  reason  to  be  able  to  get  the  ground  glass  close  up  to  the  front,  since  the 
up-to-date  apochromatic  lenses  will  stand  very  high  eye-piecing,  with  consequent 
shortening  of  focus.  The  higher  the  eye-piecing  the  shorter  the  focus  for  a 
o-iven  enlargement,  as  is  illustrated  by  the  accompanying  photographs,  all  of 
which  were  made  with  this  apparatus. 

It  will  be  noticed  from  the 
description  and  the  accompany- 
ing cut  that  the  disadvantages 
mentioned  have  been  over- 
come. With  the  new  apparatus 
I  can  bring  the  ground  glass 
as  near  as  seven  inches  from 
the  eye-piece,  and  can  focus  by 
hand  without  any  fiexing  of  the 
wrist,  the  total  range  for  hand 
focusing  being  about  sixteen 
inches  with  a  Zeiss  projection 
ocular.  The  choice  of  a  micro- 
scope stand  was  narrowed  down 
to  a  Zeiss  photographic  and  a 
Bausch  &  Lomb  DD,  the  latter 
of  which  was  finally  decided 
upon  since  the  former  does  not 
appear  to  offer  any  very  special 
advantages,  and  the  fine  adjust 
ment  is  situated  so  far  forward 
that  two  inches  in  length  would 
be  lost  for  hand  focusing. 

Another  improvement  sug- 


5  I  N  K 


D  E  V 


LOAD 


DARK 


ROOM 


0  PT/ 


O 


BE 


STOOL 


CAM 


STA 


NCH 


E  RA 


ND 


Fig.  2. —  Plan  of  Room. 


gested  was  in  making  the  camera  stand  entirely  of  metal.     The  wooden  bed  of 
the  old  style  is  liable  to  warp,  and  thus  throw  the  rods  out  of  true. 


and  Laboratory  Methods. 


1369 


It  would  have  been  an  advantage  to  have  had  one  of  the  rods  marked  off  in 
quarter  inches,  so  that  once  the  length  of  bellows  for  a  certain  enlargement  had 
been  accurately  measured,  the  back  of  the  camera  could  at  any  time  be  brought 
to  the  same  position  without  any  delay  in  measuring.  This  point  was  overlooked, 
but  I  have  had  (]uarter-inch  spaces  marked  in  white  paint  on  the  flat  surface  of 
the  upper  connecting  iron  casting,  and  this  answers  the  purpose  just  as  well. 
(See  plan  of  room,  Fig.  2.) 

The  optical  bench  carrying  lamp,  microscope,  and  accessories  is  4  feet  by 
15  inches  and  is  a  turntable,  revolving  upon  the  supporting  column,  so  that  it 
can  be  turned  partially  round  ;  the  operator  sitting  on  a  low  stool  to  find  the  field 
and  rough  focus. 

Since  the  dark  room  is  on  the  left  of  the  apparatus,  looking  towards  the 
source  of  illumination,  the  table  has  been  constructed  to  turn  to  the  left,  and 
everything  else  is  so  arranged  that  it  can  be  manipulated  from  the  left,  thus 
obviating  any  necessity  for  walking  around  the  apparatus.  ^ 

The  accompanying  diagram  illustrates  the  plan  of  the  room,  11  xl5  feet,  the 
dotted  lines  showing  the  optical  bench  in  position  for  finding  the  field.  Of  the 
camera  stand  only  the  steel  rods  on  which  the  camera  runs,  and  the  central  beam 
with  some  of  the  markings,  are  indicated.  The  lamp  and  accessories  run  on  steel 
rods,  which  are  supported  on  a  wooden  table,  supported  in  its  turn  on  leveling 
screws,  which  run  in  a  metal  groove  on  the  surface  of  the  optical  bench,  so  that 
the  table  can  be  moved  backward  or  forward  as  required. 


Fig.  3 — Epithelioma    invading  lympli   node,     x  70.  Lens,  Powell  & 

Leland   i    in.  apochromatic ;    ocular,    none;  exposure,    '^ 

sec. ;    distance    of   plate    from    hood,    70    inches. 

The   rheostat   is  a  Colt  adjustable,  and  stands  on  the  floor  just  below  the 

illuminating  end  of  the  optical  bench.     I  had  previously  used  an  automatic  arc 


1370 


Journal  of  Applied  Microscopy 


lamp,  but  finding  that  as  a  rule  I  had  to  be  my  own  automaton,  decided  that  a 
hand  feed  lamp  would  do  better,  since  less  liable  to  get  out  of  order,  and  so  far 
have  no  reason  to  regret  the  change. 

The  condenser  and  accessories  are  the  regular  Bausch  &  Lomb,  so  need  no 
special  description.  In  the  figure  the  order  from  the  lamp  is  :  (1)  condenser, 
(2)  iris  diaphragm,  (3)  paralleliser,  (4)  ray  filter,  (5)  water  tank,  (0)  iris  dia- 
phragm shutter.  In  practice  so  far,  however,  I  have  dispensed  with  the  iris 
diaphragm,  paralleliser,  and  ray  filter,  putting  the  water  tank  next  to  the  con- 
denser, and  between  tank  and  shutter  using  a  flat  tray  on  which  ground  or  colored 
glasses,  or  a  glass  trough  containing  Zeltnow's  solution,  can  be  placed,  so  that 
the  ray  filter  can  be  changed  at  a  moment's  notice.  The  lamp  and  condenser 
are  then  arranged  so  that  the  latter  focuses  directly  on  the  substage  condenser 


Fig.    4. — Same  as   Fig.    3.  X70,  Lens,  B.  &  L.   i  in.  apochromatic  : 

ocular,    B.  &   L.   i  in.  compensation ;  exposure,   i  sec ; 

distance  of  plate  from  hood,   12  inches. 

or  on  the  slide  if  no  substage  condenser  is  used.  By  this  means  a  tremendous 
flood  of  light  is  thrown  on  the  object,  and  exposures  can  be  cut  down  to  a  fraction 
of  a  second  without,  so  far  as  I  can  judge,  affecting  the  results  for  the  worse. 
On  the  day  of  writing  this,  with  an  exposure  of  one-half  second,  I  photographed 
a  small  round  cell  sarcoma  25<)  diameters  on  a  Cramer  slow  isochromatic. 

The  microscope  stand  rests  on  a  fixed  wooden  block  to  which  its  horseshoe 
foot  can  be  clamped. 

The  camera  stand,  as  seen  by  the  cut,  consists  of  two  cast  iron  uprights, 
connected  above  by  a  solid  cast  iron  beam  on  the  top  of  which  are  the  already 
mentioned  one-quarter  inch  spaces  marked  in  white  paint.  This  beam  supports 
the  steel  rods  upon  which  the  camera  runs. 


and  Laboratory  Methods. 


1371 


The  front  of  the  camera  is  five  inches  square,  and  has  vertical  and  horizon- 
tal movements.  The  bellows  is  divided  into  two,  being  so  constructed  that  the 
back  part  can  be  taken  off  the  central  wooden  frame  when  a  short  focus  is 
required,  and  pushed  back  out  of  the  way.  The  ground  glass  and  plate-holder 
can  then  be  fitted  into  the  central  frame.  Both  the  back  and  central  frames  have 
vertical  and  horizontal  movements,  and  are  precisely  alike  in  every  particular,  so 
that  either  can  be  used  for  focusing  and  exposing. 


Fig.  5. — Same  as  Figs.  3  and  4.     x  100.  Lens  B.  &  L.  i  in.  apochromatic ; 

ocular,  y^,  inch ;  exposure,  2  sec. ;  distance  of  plate 

from  hood,   7  inches. 

With  regard  to  the  arrangements  for  locking  the  optical  bench,  and  mechan- 
ical focusing,  I  append  a  short  description. 

In  making  the  mechanical  attachment  of  a  photo-micrographic  camera  for  the 
purpose  of  operating  the  fine  adjustment  of  the  microscope,  two  points  must  be 
considered,  viz.  : 

First.  An  arrangement  whereby  the  attachment  can  be  operated  from  any 
position,  and 

Second.  The  operation  of  the  microscope  fine  adjustment  without  lost  motion 
or  back  lash. 

In  the  camera  described  above,  a  third  problem  is  presented,  in  that  the 
optical  bench  carrying  the  microscope  and  illuminating  apparatus  is  arranged  to 
rotate  upon  the  supporting  column,  enabling  the  operator  to  adjust  the  specimen, 
illumination,  and  preliminary  focus  before  connecting  the  microscope  with  the 
camera. 

It  will  be  seen  that  this  arrangement  necessitates  the  detaching  of  the  focus- 


1372 


Journal  of  Applied  Microscopy 


ing  rod  of  the  camera 
from  the  microscope. 
To  avoid  the  displac- 
ing and  replacing  of 
the  belt  connecting 
the  mechanism,  the 
following  arrange- 
ment is  adopted  : 

On  the  table  of 
the  optical  bench, 
directly  beneath  the 
fine  adjustment  head 
of  the  microscope,  is 
situated  a  milled 
wheel  on  suitable 
standard.  A  belt 
extends  from  this 
wheel  to  the  fine 
adjustment  head  of 
the  microscope. 
Through  the  axis  of 
the  wheel  is  located 
a  rod  carrying  at 
the  end    toward   the 


Fig.    6. — Anthrax,  impression  preparation    from    edge    of    colony  on 
gelatin.      xyoo.     Lens,    j'^    oil  immersion ;    ocular  i  in.  compensa- 
tion;    substage  condenser,  oil  immersion  N.  A.   1.40. 

camera  a   clutch  which  can    be  quickly  connected  with  the 

focusing  rod  of  the 
camera  by  sliding 
adjustment,  oper- 
ated by  a  milled 
head.  The  optical 
bench  must  of 
course  be  brought 
to  its  proper  posi- 
tion with  relation 
to  the  camera  in 
order  to  make  this 
connection,  and, 
that  this  position 
may  be  quickly  and 
accurately  located, 
an  automatic  catch 
is  provided,  which 
catch  can  be  re- 
leased by  a  lever, 
shown  in  the  illus- 
tration. 


Malarial  Parasite  in  Human  Blood.     Cresentic  form.     X1200. 


B.  H.  Buxton. 

Cornell   Medical  Col- 
lege. 


and  Laboratory  Methods.  1373 

MICRO-CHEMICAL  ANALYSIS. 

XV. 

Magnesium  Group — ,G1,  Mg,  Zn,  Cd. 

GLUCINUM. 

This  element,  doubtless,  should  not  be  included  in  the  present  series  of 
articles  introductory  to  the  methods  of  micro-chemical  analysis,  since  it  is  rare 
that  the  analyst  is  called  upon  to  search  for  it. 

The  element  glucinum  being,  however,  of  much  interest  from  the  standpoint 
of  pure  chemistry,  the  writer  has  not  been  able  to  resist  the  temptation  to  include 
it  among  the  few  elements  to  be  considered. 

Glucinum  resembles  members  of  Group  I  in  the  crystallizing  power  of  its 
chlorplatinate,  this  salt  being  analogous  to  that  of  sodium  as  regards  its  solubility 
and  general  appearance,  but  differs  from  the  latter  in  that  it  crystallizes  with 
more  water  of  crystallization  and  in  the  tetragonal  system. 

Glucinum  resembles  aluminum  and  other  trivalent  metals  in  the  gelatinous 
character  of  its  hydroxide  precipitated  by  ammonium  hydroxide,  but  differs  from 
them  in  that  this  hydroxide  is  soluble  in  solutions  of  ammonium  carbonate. 

Like  magnesium,  its  salts  unite  to  form  double  salts  with  ammonium  ;  and  its 
chloride,  when  evaporated  to  dryness  from  aqueous  solution,  is  decomposed. 

Like  zinc,  it  is  soluble  in  sodium  or  potassium  hydroxide,  the  compound 
formed  being  a  glucinate  of  the  formula  Gl(OM)o. 

It  has  already  been  seen  that  glucinum  can  replace  magnesium,  zinc,  or 
cadmium  in  the  triple  acetate  of  sodium,  magnesium,  and  uranyl. 

It  is  thus  obvious  that  in  the  progress  of  a  micro-chemical  analysis,  glucinum, 
if  present,  may  appear  when  testing  for  Group  I,  Group  II,  and,  perhaps. 
Group  III. 

There  are  only  three  reagents  which  can  be  considered  as  giving  satisfactory 
crystals  for  the  micro-chemical  detection  of  glucinum.     These  are  : 
I.  Chlorplatinic  Acid. 
II.  Normal  Potassium  Oxalate. 
III.  Uranyl  Acetate  and  Sodium  Acetate. 

Of  the  three,  the  best  undoubtedly  is  normal  potassium  oxalate.  The  other 
two  are  subject  to  too  many  disturbing  conditions  and  sources  of  error. 

/.   Glucinum  unites  with  Chlorplatinic  Acid  to  form  Glucinum    Chlorplatinate. 

GISO^  +  H2PtCl6  =  GlPtClg  .  8H2O  +  H2SO4. 
Method. — Evaporate  to  dryness  a  drop  or  two  of  the  solution  to  be  tested,  so 
as  to  obtain  a  thin,  uniform  film  of  residue.  Place  a  drop  of  the  reagent  next  to 
the  dry  residue,  and  carefully  draw  it  across  the  latter.  If  the  glucinum  is 
present  in  considerable  amount,  there  will  appear  neat,  transparent,  square,  and 
rectangular  plates  and  prisms  of  a  faint  yellow  color  (Fig,  64). 


1374 


Journal  of  Applied  Microscopy 


Remarks. — If  it  is  desired  to  hasten  the  separation  of  the  glucinum  salt,  tip 
^   ,  ^  .,     ^  up  the  slide  and  add  a  drop  of  alcohol  to  the  test  drop 

>4^/iv^~^  ^^ter  the   reagent  has  been  drawn   across.     Generally 

^r       ^f  r^\    ^^  the  addition   of  the   alcohol  is  essential  in  order  that 

any  crystals  of  the  glucinum  chlorplatinate  be  obtained. 

"^^A^^^  rl^-A^      \1  ^        ^^^   ^^^^   ^^  satisfactory  only   when  the   air   of  the 
-      '  ^^  laboratory  is   quite  dry.      In   a   moist   atmosphere  the 

glucinum  chlorplatinate  is  deliquescent,  hence  the  test 
fails.  In  such  an  event  it  is  necessary  to  add  abso- 
lute alcohol,  or  place  the  preparation  in  a  desiccator, 
or  cover  it  with  a  watch-glass  carrying  a  drop  of  con- 
centrated sulphuric  acid. 

In  case  the  quantity  of  glucinum  present  is  small, 
and  that  of  the  members  of  Group  I  great,  it  is  essential  that  sufficient  reagent 
be  added  to  unite  with  all.  If,  therefore,  on  examination  of  the  preparation 
after  the  addition  of  the  chlorplatinic  acid,  it  is  seen  that  members  of  the 
potassium  group  are  present,  it  is  wise  to  make  a  second  addition  of  the  reagent, 
and  follow  it  with  alcohol. 

When  sodium  is  present  in  considerable  amount,  it  is  often  difficult  to  dis- 
tinguish the  glucinum  salt  from  the  sodium  chlorplatinate  ;  if,  however,  the 
preparation  be  examined  between  crossed  nicols,  the  problem  is  simplified,  since 
the  chlorplatinate  of  sodium  exhibits  oblique  extinction  (triclinic)  and  a  brilliant 
play  of  colors,  while  the  glucinum  compound  gives  parallel  extinction  (tetragonal) 
and  but  faint  colors  (usually  none).  The  chlorplatinates  of  the  potassium  group 
are  isometric. 

If  solutions  containing  glucinum  in  the  form  of  sulphate  are  employed,  care 
must  be  taken  to  avoid  confusing  this  salt  with  the  chlorplatinate,  since  the 
glucinum  sulphate,  GISO4  •  4H2O,  which  is  also  to  be  referred  to  the  tetragonal 
system,  sometimes  separates  in  thin,  six-sided  plates. 

//.   Normal  Potassium   Oxalate  added  to  solutions  of  salts  of  Glucinum  causes 
the  separation  of  a  difficultly  soluble  Double   Oxalate  of  Potassium  and  Glucinum. 
2  K2C2O4  +  GISO4  =  K2C2O4  .  GIC2O4  +  K2S04- 

Method. — To  the  moderately  concentrated  solu- 
tion add  a  little  acetic  acid,  then  a  fragment  of  the 
reagent  about  twice  as  large  as  is  usually  the  case 
in  micro-chemical  work.  Almost  immediately  large, 
clear,  colorless,  highly  refractive  prisms  of  the 
monoclinic  systems  are  obtained.  These  prisms 
unite  to  form  twins  and  radiating  masses  (Fig.  (»5). 

Remarks. — The  appearance  of  the  crystals 
varies  greatly,  according  to  the  amount  of  the 
reagent  present,  as  compared  with  that  of  glucinum. 
Too  little  potassium  oxalate  will  yield  only  a  pre- 
cipitate of  tiny  crystals  which  probably  consist  of  ^'^ 
the   normal    oxalate    of   glucinum.     Too  much  re- 

■=»  Fig.  65. 


and  Laboratory  Methods.  1375 

agent,   on   the  other    hand,  gives    rise  to   skeleton   crystals  and  to   masses   of 
thin  lenticular  plates. 

During  the  disintegration  of  the  fragment  of  potassium  oxalate  while  passing 
into  solution  (particularly  in  concentrated  solutions),  crystals  of  the  reagent 
appear  mof/w/itari/y,  which  bear  a  striking  resemblance  to  some  of  the  forms 
assumed  by  the  double  glucinum  potassium  oxalate.  In  testing  unknown  solu- 
tions the  worker  must  be  on  his  guard  lest  he  fall  into  error  by  deciding  too 
hastily. 

The  double  oxalate  of  glucinum  and  potassium  can  be  readily  recrystallized  from 
water  by  gently  warming  the  preparation  and  allowing  it  to  cool  slowly.  The 
salt  is  also  soluble  in  solutions  of  ammonium  carbonate,  a  property  which  can 
be  utilized  when  there  is  doubt  as  to  the  nature  of  the  precipitate  obtained  in  the 
course  of  an  analysis. 

The  addition  of  a  little  mercuric  chloride  will  induce  the  production  of  long 
prisms  and  twins,  and  hence  is  useful  when  good  crystals  cannot  otherwise  be 
obtained. 

Neither  primary  potassium  oxalate,  sodium,  nor  ammonium  oxalates  can  be 
substituted  for  the  normal  oxalate  of  potassium. 

With  zinc,  the  reagent  gives  tiny  double  globulites  and  pseudo-octahedra  of 
normal  zinc  oxalate,  and  later,  as  the  test  drop  concentrates  by  evaporation, 
neat  hexagonal  plates  appear,  which  are  probably  due  to  a  double  oxalate  of 
potassium  and  zinc  (?).  Mercuric  chloride  seems  to  favor  the  formation  of  the 
hexagonal  plates. 

Cadmium  treated  in  like  manner  yields,  apparently,  only  crystals  of  normal 
cadmium  oxalate  (g.  v.).     No  double  salt  seems  to  separate. 

The  reagent  gives  nothing  with  magnesium,  providing  the  test  drop  is  not  too 
concentrated  and  does  not  contain  an  excessive  amount  of  free  acetic  acid. 

When  zinc  or  cadmium  is  also  present,  the  crystal  form  of  the  glucinum 
potassium  oxalate  is  changed.  It  then  becomes  difficult  to  decide  whether  or 
not  glucinum  is  present. 

Magnesium,  aluminum,  and  iron,  on  the  other  hand,  have  practically  no 
influence,  unless  present  in  relatively  large  amount.  But  the  double  oxalate  of 
glucinum  and  potassium  crystallizing  from  such  solutions  will  always  occlude  an 
appreciable  quantity  of  the  potassium  double  oxalate  of  these  elements. 

Calcium,  strontium,  and  barium  may  mask  the  reaction. 

Ammonium  salts,  if  present,  must  first  be  removed  by  gentle  ignition  before 
testing  with  potassium  oxalate. 

Free  mineral  acids  must  be  absent. 

Stannous  salts  may  at  times  give,  with  potassium  oxalate,  crystals  of  stannous 
oxalate  which  may  be  mistaken  by  an  inexperienced  worker  for  the  glucinum 
double  salt.*  After  the  tin  salt  has  been  allowed  to  grow  for  a  short  time, 
there  is  little  danger  of  confusing  the  two.  If  still  in  doubt,  recrystallize  from 
warm  water,  treat  with  ammonium  carbonate,  or  apply  tests  for  tin. 

When  the  solution  to  be  tested  contains  copper,  cobalt,  or  nickel,  it  is  gener- 


*  This  error  cannot  arise  in  the  course  of  a  systematic  analysis. 


1376  Journal  of  Applied  Microscopy 

ally  best  to  avoid  testing  it  directly  for  glucinum,  but  to  first  practice  a  separation 
where  the  former  are  removed  from  the  latter. 

Exercises  for  P7-acticc. 

To  a  drop  of  a  solution  of  a  pure  salt  of  Gl  add  KoC^O^  in  the  manner 
directed  above.  Try  the  experiment  several  times,  varying  the  amount  of  the 
reagent. 

Try  again  under  as  nearly  like  conditions  as  possible,  but  this  time  having 
first  introduced  a  little  HgClg. 

Try  the  action  of  HKCjO^  ;  (NH4)2C204  ;  NaoCgO^. 

Try  reagent  on  salts  of  Mg;  Zn  ;  Cd  ;  Cu;  Co;  Ni. 

Then  try  mixtures,  as  for  example,  Gl  and  NH^  ;  Gl  and  Mg  ;  Gl  and  Al ; 
Gl  and  Fe ;  Gl  and  Zn  ;  Gl  and  Ca,  etc.,  and  also  more  complicated  mixtures. 

///.    With  Urhnyl  Acetate  and  Sodium  Acetate. 

This  reaction  of  glucinum  salts  has  already  been  alluded  to  under  Sodium, 
Method  II.*  The  equation  for  the  reaction  is  the  same  as  that  there  indicated, 
save  that  glucinum  replaces  the  magnesium. 

To  the  material  to  be  tested  a  little  sodium  acetate  is  added  (unless  it  is 
known  that  sodium  is  present).  The  drop  is  then  evaporated  to  dryness,  and 
the  solution  of  the  reagent  is  drawn  across  the  film  of  dry  residue.  Skeleton 
crystals,  long,  imperfect  prisms,  and  almost  colorless  tetrahedra  result. 

There  is,  at  times,  some  difficulty  in  clearly  distinguishing  between  the  triple 
acetate  of  glucinum,  sodium,  and  uranyl,  the  double  acetate  of  sodium  and 
uranyl,  and  the  crystals  due  to  a  separation  of  uranyl  acetate.  It  has  been  the 
experience  of  the  writer  that  students  trying  the  method  for  the  first  time  are 
invariably  in  doubt  as  to  the  nature  of  the  crystals  obtained. 

The  tetrahedra  of  the  triple  acetate  differ  only  in  size  and  color  from  those 
of  the  double  acetate,  the  former  attaining  a  greater  size  than  the  latter,  and 
being  only  very  faintly  yellow  instead  of  exhibiting  a  distinct  yellow  tint. 

The  amount  of  sodium  present  must  be  small,  otherwise  only  the  double 
acetate  will  appear. 

Salts  of  ammonium,  potassium,  and  of  the  calcium  group  will  generally 
interfere,  if  present  in  excessive  quantity. 

Phosphates  and  other  compounds  precipitating  uranium  should  be  absent. 

Free  mineral  acids  must  be  removed  by  evaporation  to  dryness,  as  has  been 
suggested. 

It  is  obvious  that  this  method  cannot  be  employed  for  the  detection  of  gluci- 
num save  in  the  absence  of  magnesium,  zinc,  cadmium,  cobalt,  nickel,  iron, 
manganese. 

MAGNESIUM. 

The  micro-chemical  detection  of  magnesium  in  complex  mixtures  is  usually  a 
matter  of  not  a  little  difficulty,  since  this  element  is  commonly  associated  with 
others  closely  related,  which  are  prone  to  interfere  with  or  prevent  the  formation 
of  typical  crystals  with  the  reagents  employed  for  its  recognition. 


*  Jour.  App.  Micros.  Ill,  1900,  985. 


and  Laboratory  Methods. 


137^ 


Tests  applied  to  pure  salts  and  simple  mixtures  are  quite  satisfactory,  and 
would  scarcely  lead  the  worker  to  anticipate  the  annoyances  and  difficulties 
which  may  beset  him  in  other  cases. 

In  ordinary  practice  three  reagents  will  be  found  useful : 

I.  Secondary  Sodium  Phosphate  in  Ammoniacal  solution. 
II.  Potassium  Antimonate, 
III.  Uranyl  Acetate  with  Sodium  Acetate. 


/.  Jhc  addition  of  Scconda?y  Sodium  Phosphate  to  Ammo7iiacal  solutionis  am- 
tainiug  Magnesium  pj'ecipitatcs  Ammoniiim  Magnesium  Phosphate. 

MgSO^  +  HNa2P04  +  NH^OH  =  NH4MgP04  •  GH2O  +  Na2S04  +  HgO. 

Method. — Two  methods  are  available  ;  the  choice  of  procedure  depending 
upon  the  nature  of  the  salts  present  in  the  drop  to  be  tested.  In  all  cases  where 
there  is  a  doubt  as  to  the  probable  composition  of  the  material  to  be  examined, 
it  is  best  to  have  recourse  at  once  to  the  modification  B.* 

A.  To  the  solution  of  the  material  to  be  tested,  which  must  not  be  too  con- 
centrated, add  several  fragments  of  ammonium  chloride ;  stir,  then  a  very  slight 
excess  of  ammonium  hydroxide,  and  warm  the  preparation.  (If  a  precipitate 
results  it  is  best  to  draw  off  the  clear  solution.)  To  the  warm  solution  add  a 
small  crystal  of  secondary  sodium  phosphate.  Crystals  of  ammonium  magnesium 
phosphate  soon  appear. 

B.  To  the  solution  to  be  tested  add  a  fragment  or  two  of  citric  acid,  then  an 
excess  of  ammonium  hydroxide.  Evaporate  to  dryness.  To  the  residue  add 
dilute  ammonium  hydroxide.  Warm,  then  add  a  very  little  solid  secondary 
sodium  phosphate.     Crystals  of  ammonium  magnesium  phosphate  separate. 

The  crystals  of  the  ammonium  magnesium  phosphate  separate  as  skeletons 
and  hemimorphic  forms  of  the  orthorhombic  system  (see  Figs.  40  and  66). 

Remarks. — It  should  be  remembered  that  a  number  of  elements  are  pre- 
cipitated by  phosphates  in  alkaline  solution  ;  the  most 
frequently  met  with  in  the  course  of  micro-chemical 
/|.  analyses,  either  in  the  substance  to  be  tested,  or  present 
as  reagents  from  previous  tests,  are  doubtless,  lithium, 
members  of  the  calcium  and  magnesium  groups,  triva- 
lent  metals,  manganese,  nickel,  cobalt,  tin,  lead,  silver, 
copper,  uranium. t  Of  these  elements,  lithium,  iron, 
manganese,  cobalt,  and  nickel  form,  with  ammonium 
and  phosphoric  acid,  salts  of  similar  composition  to 
and  isomorphous  with  the  magnesium  salt. 

The    ammonium    glucinum    phosphate,    ammonium 
zinc   phosphate,  and   ammonium  cadmium  phosphate 
are   not  precipitated   in   crystal  form. 


Fig.  60. 


*  Romijn,  Zeit.  anal.  Chem.  37,  300. 

t  Most  of  these  elements  will  have  been  removed  in  the  progress  of  the  analysis  before  the 
addition  of  the  sodium  phosphate. 


1378  Journal  of  Applied  Microscopy 

In  A  the  reaction  sometimes  fails  for  lack  of  sufficient  ammonium  chloride, 
magnesium  hydroxide  being  precipitated.  A  slight  excess  of  this  salt  will  do  no 
harm. 

Both  modifications  fail  if  there  is  an  insufficiency  of  ammonium  hydroxide, 
for  it  should  be  remembered  that  there  must  be  not  only  enough  ammonium 
present  to  unite  to  form  the  proper  compound,  but  that  this  salt  will  not  separate 
save  in  alkaline  solution. 

The  advantage  of  employing  modification  7?  lies  in  the  fact  that  owing  to  the 
presence  of  ammonium  citrate,  there  is  little  danger  of  the  interference  of  the 
elements  listed  above.  If,  in  following  this  method,  the  residue  after  evapora- 
tion is  not  completely  soluble  in  the  ammonium  hydroxide  solution,  it  is  best, 
though  not  essential,  to  draw  off  the  clear  liquid  before  adding  to  it  the  sodium 
phosphate. 

Reactions  A  and  B  work  equally  well  in  the  cold,  but  are  then  a  trifle  slower. 
Generally,  an  amorphous  precipitate  is  at  first  produced,  which  begins  to  crystal- 
lize in  a  few  seconds.  The  formation  of  merely  an  amorphous  precipitate  must 
never  be  taken  as  evidence  of  the  presence  of  magnesium. 

It  must  also  be  borne  in  mind  that  the  use  of  too  strong  ammonium  hydroxide 
in  excess  so  reduces  the  solubility  of  many  salts  as  to  cause  their  separation, 
hence  it  is  necessary  to  beware,  in  reactions  of  this  character,  of  deciding  too 
hastily  as  to  the  result  of  a  test. 

See  remarks  made  under  Ammonium,  Method  II  (Journal,  p.  1190),  and 
Calcium,  Method  V  (Journal,  p.  1247). 

In  the  presence  of  phosphates  the  detection  of  magnesium  becomes  quite 
difficult,  particularly  if  other  elements  are  present  which  form  phosphates  insol- 
uble in  ammonium  hydroxide.  If  arsenates  are  also  present,  a  still  further  com 
plication  arises,  for,  as  we  have  already  seen,  double  ammonium  arsenates  of 
calcium,  zinc,  etc.,  are  formed,  which  are  isomorphous  with  ammonium  magnes- 
ium phosphate. 

Of  course  it  may  happen  that  in  some  cases  the  mere  addition  of  ammonium 
hydroxide  will  cause  the  separation  of  characteristic  crystals  of  ammonium  mag- 
nesium phosphate.  Generally,  however,  it  is  first  necessary  to  remove  the 
phosphoric  acid.  This  can  be  accomplished  by  tin  and  nitric  acid,  or  by  means 
of  ammonium  tungstate  and  nitric  acid.     Details  will  be  given  later. 

Exercises  for  Practice. 

Try  method  I A  on  z.  solution  of  MgSO^,  then  try  it  on  salts  of  Fe,  Mn,  Co, 
Ni,  Al,  Zn,  Cd.  Repeat  the  experiments,  this  time  adding  the  HNa2P04  before 
the  NH4OH. 

Try  IB  in  like  manner. 

Make  mixtures,  trying  various  combinations  of  the  above  with  members  of 
Groups  I  and  II. 

Consult  notebook  on  the  results  obtained  with  the  experiments  tried  under 
Ammonium  II  and  Calcium  V. 


and  Laboratory  Methods.  1379 

//.  Potassium  Atitwionate  added  to  solutions  containing  Magnesium  causes  the 
separation  of  Magnesium  Pyro-antimonate. 

MgSO^  +  HaKaSbgO-  =  HgMgSb.O^  .  9H2O  +  K2SO4.* 

Method. — First  prepare  an  almost  saturated  solution  of  the  reagent  by  heat- 
ing a  fragment  with  water.  A  drop  of  this  solution  is  placed  next  the  test  drop, 
and  the  two  caused  to  unite.  A  dense  amorphous  precipitate  is  usually  imme- 
diately produced.  After  a  time,  crystals  of  magnesium  pyro-antimonate  appear, 
generally  near  the  circumference.  The  forms  most  frequently  obtained  are  thin, 
colorless,  transparent  hexagonal  plates,  and  spherical  masses  more  or  less 
crystalline  in  appearance  (Fig.  67).    Less  often,  short  hexagonal  prisms  are  seen. 

Re7narks. — The  solution  to  be  tested  must  be  dilute  and  neutral.  Free  acid 
not  only  interferes  with  the  formation  of  characteristic  ^ 

crystals,  but  also  causes  the  reagent  itself  to  yield  an  /— \   Q    ■=■ 

amorphous  precipitate.  '^ C\  O    ^  C^l 

The  development  of  the  crystals  of  magnesium  pyro-  ^^ 

antimonate  is  quite  slow,  and  eventually  they  may  attain   yr^  Q     sSto    ^  ^ 
a  size  of   double    or  even    triple  that    of   those  shown  in  \     j     ^  ^^7\    r\ 

Fig.  67.  0^(fi^ 

Alcohol  can  be  employed  to  hasten  crystallization,  but  ^^ 

it  is  better  to  allow  the  preparation  to  take  all  the  time  it  \p^;;^!otw«,. 

needs.  fig.  tiv. 

Lithium  sometimes  yields  crystals  not  to  be  distinguished  from  those  of 
magnesium,  more  often  circular  disks  and  sperulites. 

Sodium  (q.  v.)  gives  fusiform  crystals. 

Members  of  the  calcium  group  are  precipitated  in  an  amorphous  form,  and 
interfere  with  the  test  for  magnesium. 

Ammonium  salts  should  be  absent. 

r 

Exercises  for  Practice. 

Try  reaction  on  salts  of  Mg. 

Repeat  the  experiment  in  the  presence  of  salts  of  NH^. 

Make  a  mixture  containing  Na  and  Mg  ;  test  as  above. 

Test  a  salt  of  Li.     Try  the  effect  of  the  reagent  on  salts  of  Zn  and  of  Cd. 

Test  a  mixture  of  Mg  and  Zn. 

///.    With  Uranyl  Acetate  and  Sodium  Acetate. 

The  method  of  applying  the  test  has  been  described  in  Method  III  of  Gluci- 
num ;  there,  and  under  Sodium,  Method  II,  the  properties  of  the  triple  acetates 
have  been  discussed  in  detail. 

The  formula  and  appearance  of  the  triple  acetate  of  sodium  magnesium  and 
uranyl  will  be  found  on  page  9X5  of   Vol.  Ill,  of  this  Journal.     To  this  article, 
and  to  that  on  Glucinum,  the  reader  is  referred  for  details  as  to  methods  of  pro- 
cedure, sources  of  error,  etc.  E.  M.  Chamot. 
Cornell  University. 

*  See  foot  note,  Sodium,  Method  V,  Jour.  App.  Micros.  Ill,  1900.  p.  1048. 


1380  Journal  of  Applied  Microscopy 

Journal    of  Among  the  many  questions  clamoring 

for     decision,    that    of    the    standard 

AppllCCl    iVllCrOSCOpy  of  equipment  in  the  various  classes  of 

""■^  laboratories   has    received    very    little 

Laboratory   Methods.  organized   consideration,   and  yet  it  is 

Edited  by  L.  B.  ELLIOTT.  °"^  °^  ^'  ^'"^^  practical  value  as  any 

■  connected   with  science  teaching,  and 


Issued  Monthly  from  the  Publication  Department  one  which    WOuld    seem    tO    admit    of    a 

of  the  Bausch  &  Lomb  Optical  Co., 

Rochester,  N.  Y. very  easy  and  practical  settlement.     It 


■SUBSCRIPTIONS:  is  not  our  purpose  here  to  make  any 

One  Dollar  per  Year.     To  Foreign  Countries,  $1.25       suggestions    as    tO    what    might     COnsti- 
per  Year,  in  Advance.  °°  ° 

tute  a  standard  equipment,  but  to  point 


The  majority  of  our  subscribers  dislike  to  have  their  .,i              i         .         j           ,•           i-        .•..• 

files  broken  in  case  they  fail  to  remit  at  the  expiration  OUt  the  ValUC  tO  CducatlOUal  inStltUtlOUS 

of  their  paid  subscription.     We  therefore  assume  that  no  r      j        i-                  +       A       A 

iiiterruption   in   the   series  is  desired,  unless  notice  to  01  adopting  3.  Standara. 

discontinue  is  sent.  rr-n                   •                 ,      r                 i        i              r 

1  he  requirements  for  each  class  of 

laboratory,  so  far  as  the  most  important 
apparatus  goes,  are  practically  the  same.  The  biological  laboratory  requires  a 
microscope  having  powers  ranging  from  a  minimum  to  a  maximum.  The  histo- 
logical, bacteriological,  chemical,  high  school,  and  each  other  class  is  likewise 
limited.  There  is,  however,  no  unity  of  opinion  as  to  the  kind  of  stand  these 
powers  of  lenses  are  to  be  used  on,  or  the  accessories,  such  as  nosepiece,  con- 
densers, etc.,  which  are  to  be  used  with  them.  The  same  is  true  in  a  general 
way  in  regard  to  microtomes,  incubators,  and  the  unit  equipment,  for  each 
student,  of  glassware,  stains,  and  reagents.  Each  laboratory  director  is  a  law 
unto  himself,  and  an  inspection  of  the  purchases  made  for  the  various  labora- 
tories of  the  country  for  the  year  would  seem  to  indicate  that  each  had  done  his 
best  to  be  original  in  the  make-up  of  his  equipment. 

This  is  all  well  enough  from  the  standpoint  of  the  individual  who  is  equip- 
ping the  laboratory,  but  the  practice  costs  the  institutions  of  the  country  an 
immense  sum  of  money,  far  greater  than  any  one  not  fully  conversant  with  the 
conditions  can  realize,  and  the  cause  is  obvious. 

The  cost  of  any  article  is  dependent  very  largely  on  the  number  consumed. 
Where  the  number  is  small  the  cost  of  production  is  high,  because  it  does  not 
pay  the  manufacturer  to  build  expensive  machinery  and  make  up  a  large  quantity 
to  be  held  a  long  period,  and  in  addition  run  the  chance  of  his  stock  becoming 
antiquated  through  the  development  of  more  suitable  models. 

So  with  this  laboratory  apparatus ;  the  ceaseless  demand  for  variations  from 
existing  models,  the  selection  of  every  grade  of  apparatus  for  the  same  kind  of 
work,  makes  it  necessary  for  the  maker  to  build  an  endless  variety  of  perfectly 
useless  instruments,  and  to  charge  an  average  advance  on  all  to  compensate  him 
for  the  extra  cost  to  him  of  doing  his  work  piecemeal. 

If  the  subject  of  laboratory  equipment  could  be  taken  up  by  a  committee 
from  each  of  the  organizations  of  laboratory  men  interested  in  this  work,  and  a 
joint  recommendation  made,  there  is  no  question  but  that  the  majority  of  labora- 
tories would  accept  the  findings,  and  that  the  uniform  demand  thus  established 
would  result  not  only  in  better  apparatus,  but  at  a  decreased  cost. 


and  Laboratory  Methods.  1381 

CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Murbeck,  S.     Ueber  das  Verhalten  des  Pollen-  Since  Other  species  of  Alchemilla  have 

schlauches  bei  Alchemilla  arvensis  und  das  been  found  by  the  same  writer  to  be 

Wesen  der   Chalazogamie.     Lunds  Univer-  ^,  ..      •.   .„     r  •    .„ ^4.  <_„  „„4.^ 

sitets  Arsskrift.  36:  T-19,  pis.  1-2,  .901.  parthenogenetic,  it  is  of  interest  to  note 

that  A.  arvensis  has  a  pollen  tube  at 
all.  In  the  development  of  the  ovule  a  micropyle  is  formed,  but  long  before 
pollination  occurs  the  continued  growth  of  the  integument  entirely  closes  the 
micropyle.  The  pollen  tube  grows  down  through  the  style  and  enters  the  ovule 
at  the  chalazal  end,  then  traverses  the  entire  length  of  the  integument,  growing 
through  the  tissue,  and  enters  the  micropylar  region  of  the  sac.  Although  the 
act  of  fertilization  was  not  observed,  it  may  be  reasonably  assumed  to  take  place 
in  spite  of  the  fact  that  other  species  of  Alchemilla  are  parthenogenetic.  In  this 
case  the  pollen  seems  perfectly  normal.  The  author  does  not  regard  this  as  a 
case  of  chalazogamy  such  as  is  found  in  Casuarijia,  Corylus,  Carpimis,  Betiila, 
Alnus,  and  /iigla?is,  but  rather  as  a  type  intermediate  between  genuine  chalazo- 
gamy and  the  condition  found  in  Ulmus.  Chalazogamy  is  regarded  as  a  derived 
condition,  and  as  a  physiological  phenomenon  of  no  phylogenetic  significance. 

c.  J.  c. 

Brand,  F.  Bemerkungen  Uber  Grenzzellen  und  The  heterocysts  of  the  NastOCacea^ 
liber  spontanrothe  Inhaltskrirper  der  Cyano-       have  been    described    as    cells   poor    in 

phycecc.     Ber.   d.  deutsch.    bot.    Gesell.  19:  ^      ^  -^i         1  ^  ^      ^ 

j^j.        Qj  contents,  or  with  only  watery  contents, 

and  they   have   been   supposed   to    be 

concerned  only  in  false  branching  and  in  breaking  filaments  up  into  hormogonia. 

The  present  writer,  in  investigating  Nostoc  convnniie,  finds  that  in  addition  to  the 

empty  heterocysts  there  are  also  heterocysts  with  contents,  which  are  not  watery 

but  elastic,  and  of  considerable  consistency.     By  pressure  on  the  cover-glass,  the 

walls  of  these  heterocysts  may  be  broken   and  dissociated  from   the   contents 

which  retain  their  spherical   form.     It  was   found  that  the  contents  divide  like 

ordinary  vegetative  cells,  and  give  rise  to  filaments.     It  was  also  found  that  in 

some  cases  the  contents  of  the  heterocyst  pass  over  into  the  neighboring  cells, 

and  may  induce  in  them  a  renewed  activity.     The  writer  believes  that  the  red 

granules  of  the  Wasserbliithe,  forming  members  of  the   Cyanophyceae,  are  rot 

due  to  gases.  c.  j.  c. 

Ernst,  A.  Beitrage  zur  Kentniss  der  Entwicke-  This  paper  treats  in  considerable  detail 
lung  des  Embryo-sackes  und  des  Embryo  the  life  history  of  Tlilipa  Gesneriaua, 
(Polyembrvonie)  von  Tulipa  Gesneriana  L.        ,  .  ,  ^,  ,  .    , 

Flora.  88:  37-77,  pis.  4-8,  190 1.  ^""^^  ^^^  appearance  of  the  archesponal 

cell  in  the  nucellus  of  the  ovule  up  to 
the  ripe  seed.  Besides  the  original  work,  there  is  a  very  convenient  summary  of 
the  literature  of  polyembryony.     A  few  of  the  points  noted  are  the  following : 


138:2  Journal  of  Applied  Microscopy 

The  first  division  of  the  nucleus  of  the  embryo-sac  in  which  the  reduction  in 
the  number  of  chromosomes  is  effected,  takes  place  after  the  opening  of  the 
flower.  The  number  of  chromosomes  in  the  gametophyte  is  twelve,  but  in  one 
case  six  were  counted.  The  antipodal  nuclei  become  fragmented  into  a  varying 
number  of  pieces.  The  generative  cell  of  the  pollen  grain  often  occupies  the 
greater  portion  of  the  space  within  the  spore,  and  has  an  unusually  thick  mem- 
brane. The  vegetative  nucleus  remains  in  the  end  of  the  tube  after  the  two 
male  nuclei  have  been  discharged  into  the  sac.  One  of  the  male  nuclei  conju- 
gates with  the  nucleus  of  the  egg,  and  the  other  becomes  applied  to  the  upper 
polar  nucleus,  so  that  the  definitive  nucleus  results  from  the  fusion  of  three 
nuclei,  the  two  polar  nuclei  and  one  of  the  male  nuclei. 

After  fertilization,  the  egg  gives  rise  to  an  irregular  mass  of  cells,  in  which 
the  beginnings  of  several  embryos  may  be  distinguished.  The  polyembryony  of 
Tulipa  is  very  much  like  that  described  by  Jeffrey  for  Erythronium. 

c.  J.  c. 

CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole,  Cornel]  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Doflein,  F.     Cell  Division  in  Protozoa.  Zool.       Dr.    F.    Doflein    has    Studied    Noctiliua 

Jahrb.    14 :    1-16,    1900.    E.xtracts  from      miliar  is  with  especial  reference  to  the 

Royal  Mic.  Jour.  April,   iqoo.  ,              ,                                           .                , 

■'                        f    '     ^  nuclear    changes     accompanying    cell 

division.  The  life  cycle  is  as  follows  :  Afterdividingrepeatedly  the  adult  comes 
to  rest,  copulation  of  two  individuals  occurs,  followed  by  rapid  budding.  The 
liberated  buds  are  at  first  similar  to  Dinoflagellata,  but  ultimately  become  con 
verted  into  adults.  Division  occurs,  a  sphere  appears  near  the  nucleus,  and 
a  process  takes  place  believed  by  the  author  to  have  a  superficial  resem- 
blance to  metazoan  karyokinesis.  The  division  of  the  nucleus  appears  to  be 
in  some  degree  independent  of  the  division  of  the  sphere,  the  division  of  the 
latter  being  closely  associated  with  plasmic  division;  the  author  believes  that  the 
former  structure  is  but  a  concentration  of  the  plasma  itself,  hence  the  close 
relation.  The  budding  after  copulation  consists  of  a  rapid  cell  division  during 
which  the  division  products  remain  united  by  a  common  stroma.  It  is  uncertain 
whether  this  stroma  indicates  a  reduction  process  or  not.  A  discussion  on  the 
structure  of  protoplasm  and  the  causation  of  its  movement  is  also  given  in  the 
paper.  a.  m.  c. 

The   authors  use  the  following  simple 
Stephens,  J.  W.  W.  and   Christopher,  R.  S.  R.  .,     j  r  1    4^  •    •      c, 

Technique  for  Malaria  Blood.  Roy.  Soc.  method  for  preparmg  and  stammg  films 
Report  to  Malaria  Comm.  od  Series,  1900.  of  malaria  blood  :  The  finger  is  pricked 
Ext.   from   Royal   Mic.  Jour.,  April,   igoo.  ■,  .    ■  ,  ■      ,  ,, 

■'  J       1     f    ■>    J  ^\m     a     triangular     surgical     needle 

and  a  clean  glass  slide  touched  to  the  exuding  blood.     The  drop  thus  obtained 


and  Laboratory  Methods.  1383 

on  the  slide  is  spread  by  the  shaft  of  the  needle  in  a  broad,  even  streak,  a  little 
time  being  allowed  for  the  drop  to  run  along  the  needle  by  capillarity.  The 
most  perfect  films  are  thus  obtained.  The  slides  are  then  placed  in  absolute 
alcohol  for  5  minutes,  after  which  the  films  are  stained  with  saturated  alcoholic 
solution  of  haematein.  To  every  10  c.  c.  of  this  solution  is  added  50  c.  c.  of  alum 
solution  (alum  50  grams,  water  1000  c.  c).  In  this  solution  the  slides  are  left 
5  to  '20  minutes  or  even  hours.  Oil  is  applied  directly  to  the  slide  without  a  cover 
and  the  specimen  examined.  A  permanent  mount  can  be  made  by  washing  off 
the  oil  with  xylol  and  mounting  the  preparation  in  balsam.  If  placed  in  a  clean 
box  and  wrapped  in  paper,  the  slides  will  keep  a  year  unmounted.        a.  m.  c. 


Qurwitsch,  A.     Die  Vorstufen  der   Flimmer-      ^^  the  eighteenth  number  of  this  peri- 
zellen  unci  ihre  Beziehungen  zu  Schleimzel-       odical     a     work     on     M.     Heidenhain 

len.     Anat.  Anz.  19:  44-48,  looi.  j      u-    u       •  ^      -^ 

^  appeared  which  gives   an   opportunity 

for  a  few  remarks.     Heidenhain's  work  came  to  the  author  so  shortly  before  the 

appearance  of  his  previous  communication  on  this  subject  that  it  was  impossible 

to  discuss  in  the  same  issue   Heidenhain's  opinions   of  the  author's  statements. 

These  are  here  set  forth.     The  peculiarly  shaped  epithelial  cells  in  the  mouth  and 

pharynx  of  salamander  larvae  is  the  subject  under  special  discussion. 

The  author  has  presented,  beginning  with  the  earliest  stages,  the  develop- 
ment and  its  modifications  of  the  peculiar  superficial  border  of  the  cells  ;  first 
appearing  as  an  apparently  homogeneous  "  crest "  not  sharply  separated,  the 
cell  border  is  next  clearly  foam-like  ;  in  the  course  of  the  further  development 
the  foam-like  structure  is  efifaced  to  make  a  "  felt  work."  (Arch.  f.  Mikros. 
Anat.  57:  209,  Fig.  16-18.) 

As  an  end  product  of  development  there  comes  a  clearly  formed,  sharply 
isolated  border  of  small  rods,  of  which  the  separate  little  hairs  correspond  in 
their  height  exactly  to  the  cilia  of  the  mature  ciliated  cells  and  provisionally 
remain  covered  with  a  thin  but  very  sharply  apparent,  net-like  film.  At  this 
stage  of  development  direct  observation  ceased,  as  the  oldest  of  the  remaining 
larvae  showed  no  more  continuance  of  the  process.  Based  on  this  observation 
the  author  felt  drawn  to  the  conclusion  that  these  cells  possessed  of  rods  were 
the  early  stages  of  ciliated  cells,  and  also  to  infer  that  in  one  kind  of  cells  at 
least  the  cilia  are  formed  before  the  basal  bodies. 

This  interpretation  of  these  questioned  cells  is  now  doubted  by  M.  Heiden- 
hain. He  regrets  that  the  "  last  step  of  development,  the  peculiar  transforma- 
tion into  the  true,  mature,  free  cilia,  was  not  observed."  The  reason  for  this 
deficiency  was  given  thus  :  "  I  sought  to  remedy  the  lack  somewhat  by  figuring 
in  my  detailed  work  (Fig.  21)  a  mature  ciliated  epithelial  cell  next  to  two  inter- 
mediate ones  from  the  transition  of  esophagus  to  pharynx."  Heidenhain  is  much 
more  disposed  to  consider  the  pharyngeal  cells  as  forerunners  of  mucous  cells. 
It  seems  that  a  misunderstanding  arises  each  time.  Although  in  both  articles 
the  authors  speak  only  of  pharyngeal,  not  of  esophageal  cells,  and  although  Heid- 
enhain mentions  each  time  that  the  pharyngeal  epithelium  of  the  salamander 
was  the   object  treated  of,   he   suggests  as  a  possible   cause  for  confusion  on 


1384  Journal  of  Applied  Microscopy 

Gurwitsch's  part  the  circumstance  that  "  the  epithelium  of  the  esophagus  and 
stomach  in  the  transition  region  may  not  be  sharply  distinct  from  each  other,  so 
that  ciliated  regions  could  be  found  in  the  surface  epithelium."  This  is  impos- 
sible ;  since  the  part  of  the  epithelium  which  was  used  for  investigation  lay  above 
the  esophagus,  and  since  the  whole  region  was  covered  with  a  similar  unbroken 
coat  of  these  questioned  cells,  just  as  in  later  life  the  ciliated  coat  is  entirely 
unbroken,  the  author's  conclusion  is  again  justified  and  Heidenhain's  assump- 
tion would  only  be  right  if  there  were  ground  for  the  belief  that  the  whole 
pharyngeal  epithelium  was  changed  to  a  mucous  condition  and  that  later  ciliated 
epithelium  arose  dc  novo  from  some  unknown  source.  No  evidence  exists  for 
this  and  such  a  process  could  not  easily  escape  notice.  Moreover,  if  Heiden- 
hain's explanation  were  applied  to  all  these  questioned  cells,  then  this  rodded 
border,  which  occurs  in  so  many  forms  and  kinds  of  cells,  must  be  declared  the 
forerunner  of  mucoid  formation  on  the  ground  alone  that  it  seems  to  be  the  case 
in  this  one  kind  of  cell. 

It  cannot  be  doubted  that  identical  tissues  in  two  nearly  related  species  of 
animals  have  in  similar  developmental  stages  an 'entirely  different  appearance. 
Therefore  it  is  no  objection  to  the  writer's  hypothesis  that  the  methods  of 
development  of  ciliated  cells  in  salamanders  may  differ  in  various  cases.  The 
only  apparent  question  existing  is  whether  these  steps  are  those  of  ciliary  or 
mucoid  formation  in  the  cells. 

The  facts  important  for  histogenesis  in  general  must  satisfy  us  that  similar 
structures  may  owe  their  origin  to  different  methods  of  development,  and  that 
the  histogenetic  processes  which  are  found  in  one  species  may  not  be  applied 
to  a  nearly  related  one.  This  is  also  true  in  other  lines,  as  for  example,  the  his- 
togenesis of  crystalline  lens  in  different  animals  and  the  ectodermal  origin  of 
cartilage  in  Petromyzon. 

The  author  adds  a  few  words  on  the  beaker  cells  in  the  same  epithelium. 
This  in  the  salamandar  larva  has  always  two  layers,  the  one  with  the  peculiar  rod- 
cells  is  set  on  a  layer  of  cubical  or  more  or  less  sloping  cells.  In  not  a  single 
case  was  the  first  or  rodded  layer  found  in  contact  with  the  basilar  membranes. 
On  the  other  hand,  all  the  mucous  cells,  independent  of  their  condition  of  func- 
tion, were  placed  directly  on  the  basilar  membrane.  This  immediately  suggested 
the  latter  to  be  the  mother  cells  of  the  beaker  cells.  While  this  could  not  be 
maintained  by  direct  evidence,  not  sufficient  transitional  stages  being  examined, 
it  is  enough  for  this  argument  to  state  the  facts  that  there  is  a  good  criterion  for 
the  differentiation  of  the  immature  mucous  cell  from  the  rod  cell.  "  Thus  the 
beaker  or  goblet  mucous  cells  of  the  pharyngeal  epithelium  point  clearly  to  a  layer 
underlying  the  latter  ciliated  cells.)  This  relation  holds  true  in  all  cases  and  all 
stages  in  development.  e.  j.  c. 


and  Laboratory  Methods.  1385 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Bergh,  R.  S.  Kleinere  histologische  Mittheil-  The  author  commends  the  use  of 
ungen,  Zeitsch.  f.  Wiss.  Zool.  69 :  444-456.  maceration  methods  in  the  study  of 
Taf.  32,  33,  1 90 1.  ^^g    histology    and   organology   of  the 

larva  of  the  leech  Aulastoma.  Very  dilute  acetic  acid  or  a  mixture  of  three  or 
four  parts  of  30  per  cent,  alcohol  with  one  part  of  '1  per  cent,  acetic  acid  was 
employed  with  good  results,  both  for  maceration  and  as  a  fluid  for  examination. 
For  the  demonstration  of  cell  boundaries  the  silver  method  of  Fischel  was  used, 
though  the  finest  results  were  secured  with  a  mixture  of  equal  parts  of  1  per 
cent,  nitric  acid  and  1  per  cent,  silver  nitrate  allowed  to  act  for  a  longer  time 
than  that  usually  employed  in  silver  impregnation.  Reduction  was  accomplished 
by  sunlight  or  by  a  weak  solution  of  formic  acid  in  alcohol.  By  this 
method  very  fine  demonstrations  of  the  cell  boundaries  in  the  nephridia  of  the 
Lumbricidce  can  be  secured,  the  cell  limits  being  defined  even  in  the  intra- 
cellular lumen.  Bergh  confirms  his  earlier  thesis  of  the  presence  of  a  larval 
epidermis  in  Aulastoma  consisting  of  about  thirty  large  multinucleate  cells, 
whose  nuclei  multiply  by  amitotic  division.  The  structure  of  the  nephridium  of 
Lumhricus  hcnuleus  is  considerably  elucidated  by  the  silver  method.  The  cells 
in  the  margin  of  the  funnel  alone  have  the  usual  straight  cell  walls.  Within  the 
funnel  and  in  the  straight,  ciliated,  intracellular  lumen  of  the  adjacent  section  of 
the  nephridium  the  cell  walls  are  very  tortuous.  In  the  narrower  regions  when 
the  lumen  is  intracellular  they  become  even  more  irregular  and  in  the  ampulla 
almost  labyrinthine,  though  everywhere  transverse  in  general  direction,  each 
cell  forming  a  short  transverse  section  of  the  nephridial  tube.  Attempts  to 
demonstrate  cell  boundaries  in  the  nephridia  of  the  aquatic  oligochaites  by  the 
silver  method  failed  entirely.  c.  a.  k. 

Coe,W.R.  Papersfrom  the  Harriman  Alaska  ^r.  Coe  reports  thirty-twO  species,  of 
Expedition,    XX.     The  Nemerteans,    Proc.       which    all    but    tWO    are     new     tO    the 

Wash.  Acad.  Sci.  3:  i-i  10,  pi.  1-13,  iqoi .  t-.      -c  •  1  ^         ^ 

^         ^    ^  Pacmc  region  and  twenty-seven  new  to 

science.     The    methods  employed  with  this   refractory   group    are    of  general 

interest.     The  worms  die  well  extended  if  a  few  drops  of  formalin  are  added  to 

the  sea  water  in  which  they  are  placed,  and  if  handled  with   care  they   do   not 

always   break  up  into    fragments.     Material    was    hardened   in    2    to    5    per 

cent,  solution  of  formalin  in  sea  water  and  eventually  transferred  to  alcohol. 

Formalin  gives  good  results  for  anatomical  work  or  for  the  histology  of  epithelial 

structure,    but    it    is    disastrous  to  the  connective  and    nervous  tissues.     For 

supplementary  work,  strong  alcohol,   sublimate-acetic,  Gilson's  fluid,  and — for 

nervous  system — Flemming's  fluid  were  used.     Iron   hematoxylin  followed  by 

orange  G  was  the  most  effective  stain  for  sections.  c.  a.  k. 


1386  Journal  of  Applied  Microscopy 

Burckhard,  Q.     Die   Implantation   des   Ei  der       It  was  the  purpose  of  this  investigation 
Mans  in  die  Uterusschleinihaut  und  die  Um-  ,  ,,  ■  i  ■         i 

bildungderselben  zur  Decidua.Arch.f.  Mik.      to    toUow    the    changes   in    the    uterus 
Anat.  u.  Entwick,  57:  528-569,  Taf.  26-28,      from  the  time  the   egg  enters   it   from 

the  oviduct  until  the  embryo  is  fully 
encapsuled  in  its  walls  in  the  so-called  decidua  rejfcxa.  About  the  beginning  of 
the  fifth  day  after  impregnation  in  the  mouse,  the  ova  are  clustered  at  the  lower 
end  of  the  oviduct  in  an  advanced  stage  of  cleavage  with  a  small  cleavage  cavity 
appearing.  About  this  time  they  enter  the  uterus  and  are  immediately  distrib- 
uted, probably  by  movements  of  the  uterine  walls,  at  somewhat  regular  intervals 
throughout  the  uterine  lumen,  lying  in  crypt-like  depressions  on  the  antimeso- 
metrial  side  of  the  lumen.  The  process  of  implantation  is  completed  by  the 
eighth  day  and,  owing  to  the  rapidity  with  which  it  takes  place,  has  been  over- 
looked in  large  part  by  all  previous  investigators  of  the  subject.  At  the  end  of 
the  eighth  day  the  embryo  is  separated  from  the  lumen  and  embedded  in  the 
decidua,  composed  entirely  of  mucosa  cells  from  which  all  traces  of  the  uterine 
epithelium  have  disappeared.  Other  investigators  have  suggested  that  the  embryo 
sinks  beneath  the  epithelium  and  develops  in  the  mucosa,  but  Burckhard's 
results  show  conclusively  that  this  position  of  the  embryo  is  brought  about  by  a 
degeneration  of  the  uterine  epithelium  of  the  walls  adjacent  to  the  embryo.  By 
the  middle  of  the  fifth  day  the  epithelium  near,  but  not  as  yet  in  contact  with,  the 
ovum,  shows  traces  of  flattening  and  the  cells  of  the  subjacent  mucosa  exhibit 
nuclear  activity.  Eosinophilous  leucocytes  invade  this  territory  and  the  capil- 
laries branch  and  spread  toward  the  region  of  the  ovum,  while  the  uterine  glands 
close  and  degenerate  from  the  uterine  lumen  toward  the  musculature.  By 
the  middle  of  the  sixth  day  the  epithelium  near  the  ovum  (lining  of  the  decidual 
cavity)  is  much  flattened  and  the  walls  on  the  mesometrial  side  meet  above  the 
ovum  uniting  with  its  ectoplacental  one  (Trager),  thus  completely  separating  the 
decidual  cavity  from  the  uterine  tract.  By  this  time  the  epithelium  near  the 
ovum  has  disappeared  entirely  either  by  degeneration  or  retraction,  while  the 
remainder  of  the  lining  of  the  decidual  cavity  degenerates  by  evident  desquama- 
tion and  karyolysis. 

The  ovum  now  lies  in  the  uterine  mucosa  beneath  the  epithelium  on  the 
antimesometrial  side  of  the  uterus.  Multiplication  of  the  decidual  cells  and 
their  subsequent  growth,  combined  with  increased  vascular  supply,  result  in 
further  closure  of  the  uterine  walls  above  the  ovum  until  only  a  small  lumen 
remains.  The  thickened  vascular  region  above  the  ovum  becomes  the  placenta, 
while  the  persistent  lumen,  now  on  the  mesometrial  side  of  the  ovum,  comes  at  a 
later  stage  to  lie  on  the  antimesometrial  side.  The  manner  in  which  the  change 
is  accomplished  is  not  at  present  known.  The  probability  of  a  method  of  implan- 
tation of  the  ovum  in  the  human  uterus  similar  to  that  found  in  the  mouse  is 
suggested  and  discussed  at  length.  The  material  examined  was  secured  entirely 
from  white  mice.  The  uteri  were  removed  immediately  after  death  to  picro-subli- 
mate,  Zenker's,  Flemming's  or  Hermann's  fluids  for  twenty-four  hours.  Safranin 
iron-hsmatoxylin  was  used  after  the  last  two  fluids,  borax  carmin  or 
haematoxylin-eosin  after  the  other  fluids.  These  last  preparations  gave 
the  best  results  ;  since  the  eosin  differentiates  the  blood  corpuscles  and  also 
gives  the  epithelium  of  the  uterus  and  its  glands  a  characteristic  tint.  The 
figures  illustrating  this  paper  are  very  fine,  being  based  upon  micro  photographs 
prepared  by  Sobotta's  method.  c.  a.  k. 


and  Laboratory  Methods.  1387 

NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Lewy.  Die  Beziehungen  der  Charcot-Leydenn'-  Lewy  shows  that  where  eosinophihc 
schen  Krystalle  zu  den  eosinophilen  Zellen.  ^^^^  abound,  there  Charcot-Levden 
Zeitschr.  f.  khn.  Med.  40:  59,  1900.  .     .  -^ 

crystals    appear.     This    association  is 

found  in  all  the  tissues  in  leuksemia,  in  the  sputum  in  various  diseases  of  the 
respiratory  organs,  in  nasal  polypi,  in  tumors,  in  the  faeces  in  helminthiasis,  and 
in  the  normal  bone  marrow.  When  the  crystals  are  not  present  in  the  fresh 
preparations,  they  form  quickly  if  the  blood,  pus,  bits  of  tissue,  or  other  material 
are  preserved  and  kept  from  drying.  The  crystals  can  also  be  produced  by  the 
action  of  various  metallic  salts  upon  the  eosinophilic  cells. 

The  crystals  can  arise  within  the  eosinphilic  cells,  and  those  that  form  out- 
side the  cells  probably  take  their  origin  from  eosinphilic  granules,  which  lie 
free  in  the  tissue-spaces.  It  is  not  to  be  assumed  that  the  eosinophilic  granules 
are  directly  transformed  into  crystals  or  simply  supply  the  material  by  a 
chemical  change.  Lewy  advances  the  hypothesis  that  the  mother-substance  of 
the  crystals  is  formed  physiologically  in  different  tissues  and  is  destroyed  by  the 
round  cells  attracted  thither  by  chemotaxis.  Under  certain  conditions  the  mother- 
substance  is  formed  in  such  a  large  amount  that  a  residue  remains  from  which 
the  crystals  arise.  The  eosinophilic  granules  are  formed  from  this  mother-sub- 
stance by  the  action  of  the  round  cells,  which  become  transformed  into 
eosinophiles.  Lewy  himself,  however,  brings  forward  objections  to  this 
hypothesis.  j.  h.  p. 

Glinski,  L.   K.    Zur    Kenntniss    des   Neben-      A  firm,  oval,  reddish  gray,  sharply  cir- 

pankreas       und       verwandter       Zustande.       cumscribed    body,  4.5    cm.     long,     was 
Virchow  s  Archiv,  lo4 :   132-145,  1901.  _  _  •'  ° 

discovered  in  the  wall  of  the   stomach 

near  the  pyloric  end.     It  produced  a  bulging  of  the  overlying  mucous  membrane 

into  the  cavity  of  the  stomach.     On  microscopical  examination  the  structure  was 

recognized  as  an  accessory  pancreas.     It  was  embedded  in  the  muscular  coat. 

In  all  the  other  recorded  cases  the  accessory  pancreas  has  been  situated  in  the 

submucosa  or  between  the  serosa  and  the  muscular  layer.     The  author  collected 

from  the  literature  thirteen  cases  in  which  an  accessory  pancreas  has  been  found. 

Three  times  it  was  located  in  the  stomach  wall;  ten  times  in  the  wall   of  the 

intestine.     The  pancreas  in  some  of  the  lower  vertebrates  is  situated  in  the  wall 

of  the  stomach  or   intestine,  hence  the   accessory  pancreas   in    man  is   regarded 

as  a  reversion  to  the  original  type.  j.  h.  p. 

Warthin,  A.  S.    A  Contribution  to  the  Normal      Haemolymph    nodes  differ  from   ordi- 

Histology   and    Pathology  of   the    Haemo-  ,  ,  j       •     ^1     ^  ^1 

lymph    Glands.     Journal     of    the     Boston       "^ry  lymph  nodes  m  that  they  contam 

Society    of    Medical    Sciences,  5:  416-436,       blood-sinuses    in    place    of   the    lymph- 

^    *  sinuses.    These  bodies  were  discovered 

by  Gibbes  in  1889.     Six  years  later  they  were  described  in  more  detail  by  Robert- 


1388  Journal  of  Applied  Microscopy 

son,  who  gave  them  the  name  of  ha^molymph  glands.  Clarkson  and  others  have 
studied  the  occurrence  and  minute  anatomy  of  these  organs  in  the  lower  animals, 
but  little  attention  has  been  paid  to  the  haemolymph  nodes  of  man.  The  author 
bases  his  report  on  the  investigation  of  these  structures  in  autopsies  on  eighty 
subjects.  Haemolymph  nodes  occur  in  greatest  number  in  the  prevertebral 
retroperitoneal  region  near  the  great  vessels,  near  the  adrenal  and  renal  vessels, 
along  the  brim  of  the  pelvis,  and  in  the  root  of  the  mesentery.  The  differ  as  to 
location,  number,  and  size  in  different  individuals.  They  undergo  atrophy  in  old 
age.  They  usually  lie  embedded  in  fat,  and  as  a  rule  very  near  to  the  wall  of 
some  large  vessel.  An  interesting  and  suggestive  feature  is  the  richness  of  their 
blood  supply.  The  haemolymph  nodes  cannot  be  definitely  distinguished  from 
the  ordinary  lymph  nodes  by  naked-eye  examination.  This  is  owing  to  the  fact 
that  the  blood  sinuses  are  usually  empty  and  collapsed  after  death.  When  the 
sinuses  are  filled  with  blood  the  bodies  are  deep  red  or  bluish,  and  the  smaller 
ones  are  easily  mistaken  for  blood  clots. 

Two  types  of  haemolymph  nodes  exist,  to  which  Warthin  has  given  the  names 
splenolymph  gland  and  marrowlymph  gland,  as  indicating  their  structure  and 
probable  function.  Between  these  types  are  transition  forms,  and  also  between 
these  bodies  and  the  spleen  on  the  one  hand  and  ordinary  lymph  nodes  on  the 
other. 

The  splenolymph  node  is  the  more  frequent  form.  It  possesses  a  relatively 
thick  capsule.  Trabeculae  pass  from  this  into  the  organ  dividing  it  into  irregu- 
lar lobules.  Branches  of  a  peripheral  blood  sinus  accompany  the  trabeculae, 
increasing  in  size  as  they  approach  the  center.  Between  the  sinuses  lies  the 
lymphoid  tissue.  Round  collections  of  lymphoid  cells,  suggesting  splenic  follicles, 
are  common.  Next  to  the  small  lymphocyte  the  large  mononuclear  cell  is  the 
most  common  form  in  the  lymphoid  tissue.  Red  blood  corpuscles  lie  free  in  the 
meshes  of  the  reticulum,  and  there  is  a  varying  amount  of  blood  pigment. 
Mononuclear  phagcoytes  containing  red  blood  corpuscles  and  blood  pigment 
are  found  in  the  recticulum  and  in  the  central  blood  sinuses.  Scattered  areas 
of  a  hyaline  substance  which  stains  blue  with  Mallory's  connective  tissue  stain 
occur  in  the  lymphoid  tissue.  Fuchsinophile  bodies,  probably  the  product  of 
the  destruction  of  the  red  blood  corpuscles,  are  seen  in  the  reticular  meshes  and 
also  in  the  mononuclear  phagocytes.  In  the  marrowlymph  node  there  is  a 
greater  variety  of  cells  than  in  the  splenolymph  node.  Mononuclear  eosinophiles 
are  more  numerous,  and  multinuclear  cells  and  large  mononuclear  forms  with 
deeply  staining  knobbed  nuclei  occur. 

Warthin  believes  that  under  normal  conditions  the  haemolymph  nodes  are 
probably  concerned  chiefly  in  haemolysis  and  leucocyte  formation  and  play  but 
little  part,  if  any,  in  the  production  of  red  blood  corpuscles.  Under  pathologi- 
cal conditions  of  the  blood  these  bodies  may  assume  a  blood-forming  function. 
The  haemolymph  nodes  take  on  the  structure  of  either  spleen  or  bone-marrow 
and  compensate  for  these  organs  when  their  functional  power  is  diminished  by 
disease.  J-  h.  p. 


and  Laboratory  Methods.  1389 

GENERAL   PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

,     „  ,  ,  The  alternation  in  the  direction  of  the 

Scbultze,    L.    S.       Untersuchungen    iiber    den  .  . 

Herzschlag  der  Salpen.  Jenaische  Zeitschr.  heart  beat  m  the  tunicates  IS  a  phe- 
f.  Naturwiss.  xxxv,  N.  F.  xxviii,  pp.  221-  nomenon  which  has  frequently  been 
328.     Taf.  ix-xi,  1901.  ,  ,         ,    ,  .,      ,    ,  , 

observed  and  described,  but  the  present 

paper  is  by  far  the  most  comprehensive  and  detailed  study  of  the  subject  which 
has  ever  appeared.  The  work  was  done  on  several  different  species  of  the 
transparent  pelagic  tunicate  Sa/pa.  A  complete  period  in  the  activity  of  the  nor- 
mal heart  of  Salpa  includes  a  succession  of  series  of  advisceral  and  abvisceral 
pulsations,  with  an  intervening  short  pause  after  each  series.  The  different 
phases  of  the  activity  of  the  heart  vary  so  widely,  both  absolutely  and  relatively, 
that  it  is  impossible  to  give  a  normal  value  for  any  one  of  them.  As  an  example 
illustrative  of  this  great  variability  may  be  taken  the  relative  number  of  pulsations 
in  the  advisceral  and  abvisceral  series  in  the  case  of  a  specimen  of  Salpa  demo- 
cratica-nuuronata.  The  advisceral  pulsations  were  to  the  abvisceral  as  lOU  is  to 
115  in  one  individual  of  the  colony,  while  in  another  individual  of  the  same  size 
in  the  same  colony  the  two  series  were  related  as  100  is  to  45.  The  rate  of  the 
abvisceral  and  advisceral  pulsations  is  in  general  the  same.  The  condition  of 
the  water  has  a  very  decided  influence  on  the  activity  of  the  heart.  Stale  water 
produces  an  increase  in  the  number  of  pulsations  and  an  acceleration  in  their 
rate.  The  author  gives  a  detailed  account  of  the  phenomena  observed  in  an 
animal  slowly  dying  in  foul  water.  The  most  significant  appearance  under  these 
conditions  is  the  loss  of  coordination  in  the  heart  beat.  For  example,  abvisceral 
and  advisceral  pulsations  may  start  from  opposite  ends  of  the  heart  at  the  same 
time  and  meet  in  the  middle,  neutralizing  each  other  and  disappearing.  An 
advisceral  series  may  be  extended  to  a  great  length  ;  in  some  cases  to  as  many 
as  241  single  pulsations  without  any  pause.  A  section  on  the  effects  of  poisons 
is  mainly  devoted  to  an  account  of  the  action  of  two  drugs,  nicotine  and  hellebore. 
Nicotine  decreases  the  number  of  advisceral  pulsations,  while  hellebore  increases  it. 
Experiments  were  performed  to  discover  the  source  of  the  cardiac  stimulus. 
It  was  found  that  a  heart  completely  isolated  from  the  body  beats  in  the  normal 
manner,  thus  showing  that  the  cause  of  the  pulsation  is  not  peripheral.  To  test 
the  effect  of  the  central  nervous  system  on  the  heart  beat,  stimulation  and  extir- 
pation experiments  were  performed.  Electrical  stimulation  of  the  ganglion  had 
no  effect  either  on  the  number  or  the  rate  of  the  pulsations.  Extirpation  of  the 
ganglion  causes  a  decrease  in  the  number  of  pulsations  in  a  series,  but  it  is  shown 
that  this  is  not  a  specific  effect  of  the  removal  of  the  nervous  system,  but,  instead, 
is  a  result  of  the  loss  of  a  certain  amount  of  body  substance.  Experiments  in 
which  the  heart  was  cut  transversely  in  pieces  gave  the  result  that  these  pieces 
will,  after  a  time,  begin  to  beat  rhythmically  whether  they  are  from  the  ends  or  the 
middle  region  of  the  heart.     Emptying  the  heart  of  all  blood  did  not  affect  the 


1390  Journal  of  Applied  Microscopy 

normal  beat.  The  motor  stimulus  which  causes  the  rhythmical  pulsation  must 
arise  in  the  muscles  of  the  heart  itself,  since  a  very  careful  search  with  a  great 
variety  of  histological  methods  failed  to  reveal  either  ganglion  cells  or  nerve  fibers 
in  this  organ. 

The  next  general  subject  considered,  is  the  cause  of  the  periodical  change 
in  the  direction  of  the  blood  flow  and  the  heart  beat.  After  a  critical  review  of 
the  theories  which  have  been  advanced  by  previous  workers,  Schultze  proceeds 
to  an  account  of  his  own  explanation.  By  isolating  one  end  of  the  heart  he 
found  that  its  activity  showed  a  marked  periodicity.  There  were  periods  of  maxi- 
mal activity,  in  which  the  pulsations  were  strong  and  rapid,  followed  by  periods 
of  minmal  activity  during  which  the  beats  were  weakened  and  nearly  dis- 
appeared. Both  ends  of  the  heart,  under  normal  circumstances,  would,  of  course, 
show  this  periodicity.  From  the  fact  that  any  single  muscle  fiber  of  the  heart 
cannot  be  made  by  extra  stimulation  to  further  react  when  already  contracted  or 
nearly  contracted,  together  with  the  fact  that  the  stimulus  is  conducted  in  the 
muscle  fibers  themselves,  it  is  shown  that  the  end  of  the  heart  which  is  beating 
more  rapidly  and  strongly  will  determine  the  beat  of  the  whole.  Taking  this  in 
connection  with  the  periodicity  in  the  activity  of  either  end  of  the  heart,  the 
result  is  that  first  one  and  then  the  other  end  will  determine  the  beat  of  the  whole 
heart.  When,  for  example,  the  abvisceral  end  is  in  its  period  of  maximal  activity 
it  will  set  the  whole  organ  to  beating  synchronously  with  it,  outweighing  and 
obscuring  the  weaker  pulsations  of  the  advisceral  end.  After  a  time,  however, 
its  period  of  maximal  activity  ends  and  that  of  the  opposite  end  begins  and  con- 
trols in  turn  the  beat  of  the  whole.  The  continuation  of  this  process  results  in 
the  observed  alternation  in  the  direction  of  the  heart  beat  and  blood  flow. 

R.  p. 

RadI,  Em.    Ueber  den   Phototropism  einiger      This  paper  gives  an  account  of  the  effect 
Arthropoden.       Biol.  Centralbl.  21 :  75-S6,       of   light  on   the  movements  of  the  eyes 

of  various  Cladocera,  and  the  relation 
of  these  eye  movements  to  the  phototaxis  of  the  organisms.  It  was  found  that 
sudden  shading  of  a  Daphnia  caused  an  immediate  drawing  in  of  the  eye  stalks. 
Careful  study  showed  that  under  all  conditions  the  eye  was  directed  towards  the 
source  of  greatest  illumination.  If,  for  example,  a  Daphnia  on  a  slide  on  the 
microscope  stage  be  shaded  by  the  hand  from  above,  the  eye  stalk  will  be  rotated 
so  as  to  point  its  tip  towards  the  opening  in  the  diaphragm  ;  while,  on  the  other 
hand,  if  the  light  be  diminished  from  below,  the  eye  will  turn  up  towards  the 
light  now  coming  from  above.  When  the  organism  (Daphnia)  is  oriented  with 
its  back  towards  the  light  the  eye  is  in  its  normal  position,  with  all  the  muscles 
of  the  stalk  in  a  state  of  equal  tension.  If  now  the  preparation  is  turned  so  as 
to  bring  the  animal  out  of  its  position  of  orientation,  it  is  found  that  the  eyes 
maintain  their  orientation  while  the  body  turns  about  them  as  a  fixed  point  until 
it  is  again  in  a  position  such  that  the  eye  muscles  are  in  a  state  of  equal  tension. 
These  reactions  of  the  eyes  do  not  usually  appear  in  strong,  direct  sunlight,  there 
being  apparently  an  upper  limit  of  intensity  beyond  which  the  normal  phenomena 
do  not  appear. 

Observations  were  made  on  the  method  of  orientation  to  light  of  specimens 


and  Laboratory  Methods.  1391 

of  Simocephahts  swimming  freely  in  the  water.  They  always  keep  the  back 
towards  the  source  of  light  even  though  this  necessitates  an  entire  reversal  of  the 
usual  position  with  reference  to  the  force  of  gravity.  Furthermore,  to  a  sudden 
shading  the  animals  react  by  a  strong  spring  towards  one  side  or  the  other. 
This  results  in  getting  all  the  individuals  out  of  a  shaded  area  in  a  short  time. 

The  author  considers  the  eyes  of  the  Cladocera  as  physiologically  comparable 
with  the  statocyst  of  the  decapod  Crustacea.  The  eye  orients  itself  along  the 
"  lines  of  force  "  of  the  light  rays,  and  thus  effects  differences  of  muscle  tonus. 
On  the  other  hand,  the  statolith  moves  along  lines  of  the  force  of  gravity  to  the 
lowest  point  of  the  statocyst  and,  through  the  sense  hairs,  causes  differences  in 
the  tonus  of  the  body  muscles.  The  general  biological  significance  of  the  photo- 
tactic  reaction  is  discussed,  and  the  orientation  of  swarms  of  Ciilicidce  to 
surrounding  objects  is  explained  as  such  a  reaction.  R.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Karlinski  has  made  a  study  of  the  nasal 

Karlinski.     Zur  Kenntnis  der  saurefesten  Bak-       uj^o  ^t  ^,,u^  -,  l^r-rr^  mimKor-  ^f  ;nrI5 

.  „    ^  ,  „  ,       „         in.  ,^,  .„^.       cavities  or  quite  a  large  number  ot  inai- 

tenen.     Cent.  f.  Bak.  u.  Par.  i,  29:  521,  1901.  ^  ° 

M  ....  ^  -A      ■  .       viduals,  originally    for  the  purpose  of 

Murray.     A  preliminary  report  on  acid  resist-  >         &  J  r      1 

ing  bacilli,  with  special  reference  to  their      determining  whether  the  lepra  bacillus 

occurrences  in  the  lower  animals.     Jour,  of  j^    ^      j      ^^    -^    ^^ese    cavities    in 

Exp.  Med.,  p.  205,  1900. 

people  not  suffering  from  leprosy.     In 

the  course  of  this  study  he  has  discovered,  in  the  nose  in  19  cases  out  of  235,  a 
very  characteristic  bacillus,  which  holds  its  stains  against  the  action  of  acids  in 
the  sarne  manner  as  the  tuberculosis  bacillus.  This  bacillus  is  larger  than  the 
tubercle  bacillus  or  the  lepra  bacillus,  and,  indeed,  when  compared  with  the  vari- 
ous other  "sauerfest"  bacilli,  proved  to  be  quite  different  from  any  of  them.  It 
appears  to  be  the  nearest  to  the  organism  discovered  by  Rabinovitsch,  although, 
in  some  respects,  it  is  different  from  that  variety.  The  author  thinks  it  is  a  new 
type  of  bacillus  holding  stains  against  the  action  of  acids.  The  organism  does 
not  appear  to  be  pathogenic  for  animals  or  for  man  when  simply  placed  in  the 
nasal  cavities,  although  it  is  commonly  found  in  individuals  showing  certain 
ulcers  in  the  nose. 

The  second  author  studies  the  bacilli  from  the  genital  organs  of  dogs,  horses, 
cows,  cats,  guinea  pigs,  rabbits,  and  white  rats.  He  finds  in  all  cases,  except  in 
those  of  cats  and  rabbits,  acid  resisting  bacilli,  resembling  the  smegma  bacillus. 
They  are  not  all  alike,  and  the  author  thinks  they  form  a  group  of  closely  allied 
but  variable  bacteria.  h.  w.  c. 

Reichenbach.  Ueber  Verzweigung  bei  Spir-  During  recent  years  many  questions 
illen.  Cent.  f.  Bak.  u.  Par.  i,  29:  553  have  been  raised  in  regard  to  the  rela- 
''^°''  tions   of  bacteria  to   other  fungi,  and 

there  are  many  who  have  a  strong  suspicion,  amounting  to  a  belief,  that  they  are 


1392  Journal  of  Applied  Microscopy 

to  be  regarded  as  stages  in  the  development  of  the  higher  fungi.  This  conclus- 
ion is  no  new  one,  inasmuch  as  it  was  advanced  in  the  early  days  of  bacteriological 
study ;  but  it  has  been  revived  in  recent  years,  because  of  evidence  based  upon  quite 
new  data.  The  most  important  fact  pointing  in  this  direction  has  been  the  dis- 
covery among  some  bacteria,  for  example  the  tubercle  bacillus  and  the  diphtheria 
bacillus,  of  undoubted  branching  forms.  Branching  is  not  supposed  to  be  char- 
acteristic of  typical  bacilli,  and  wherever  it  occurs  has  suggested  a  relation  to 
some  of  the  higher  fungi.  Our  author  believes  that  the  evidence  for  the  branch- 
ing of  bacilli  hitherto  advanced  is  not  quite  conclusive,  being  of  the  opinion  that 
many  or  most  of  the  facts  may  possibly  be  explained  upon  the  ground  that  the 
branching  forms  are  degenerate  types.  He  conceives  that  the  spirilli  are  the 
most  promising  organisms  for  the  proper  solution  of  the  question,  and  makes, 
therefore,  a  careful  study  of  Spirilhim  riibrum.  Under  proper  culture  media  he 
is  able  to  obtain  undoubted  instances  of  branching  forms  of  this  spirillum,  seve- 
ral excellent  photographs  of  which  are  given.  Whether  these  branching  forms 
are  to  be  regarded  as  normal  or  as  degenerate  types,  he  is  unable  positively  to 
ascertain,  inasmuch  as  the  various  branches  do  not  all  show  the  typical  spiral 
coiling,  and  he  concludes  that  if  the  branching  is  a  normal  feature,  every  branch 
should  become  spirally  coiled  and  should,  perhaps,  subsequently  show  branching 
in  turn.  These  characters  he  does  not  find,  and  while,  therefore,  he  is  confident 
that  these  spirilli  have  a  true  branching,  he  is  unable  to  determine  positively 
whether  it  can  be  regarded  as  a  normal  or  abnormal  character.  The  question  of 
the  relation  of  bacilli  to  the  higher  moulds  is  therefore  left  still  uncertain. 

H.  w.  c. 

Some  investigators  of  cholera  epidemics 
Bliesener.     Beitrag    zur    Lehre   von    Sporen-       ,  i     j  .u  i      ■         r 

bildung.    Zeit.i  Hyg.  32:  71,  1901.  ^'^^'^  x^^&y^A  the  conclusion,  from  van- 

ous  facts  connected  with  the  distribution 
of  the  disease,  such  as  the  breaking  out  of  the  disease  anew  in  the  spring  after 
a  season  of  winter  weather,  or  its  sudden  appearance  in  localities  after  having 
disappeared  for  a  long  time,  that  this  bacillus  must,  under  certain  circumstances, 
produce  spores  or,  at  least,  resisting  forms  capable  of  lying  dormant  for  a  long 
time.  The  author  has  obtained  direct  evidence  that  something  of  this  kind 
occurs.  A  quantity  of  water  contaminated  with  a  large  amount  of  organic  mate- 
rial was  filtered,  and  subsequently  steriHzed  by  discontinuous  heat ;  into  this  a 
small  amount  of  cholera  bacillus  was  inoculated  and,  after  some  time,  the  author 
finds  in  the  precipitate,  which  appears  at  the  bottom,  a  number  of  oval,  highly 
refractive  bodies,  which,  in  appearance  and  their  relation  to  stains  resemble 
spores.  Experimental  evidence  which  followed  showed  him  that  these  were 
forms  of  the  cholera  bacillus,  since  they  develop  into  the  typical  forms.  He  is- 
inclined  to  believe,  therefore,  that  they  are  spores  of  the  cholera  bacillus,  but 
recognizes  that  the  conclusion  is  not  very  satisfactory,  inasmuch  as  a  half  hour 
heating  at  a  temperature  of  50°  C  is  sufficient  to  destroy  the  vitality  of  these 
bodies,  whereas  true  spores  resist  a  much  higher  heat.  In  spite  of  this,  the 
author  is  inclined  to  believe  that  he  has  discovered  a  spore  formation  in  the 
cholera  bacillus.  h.  w.  c. 


and  Laboratory  Methods.  1393 

NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Clark,  F.  W.,  and   Steiger,  G.    The  Action  of  The  data  given  yield  further  proof  of  the 

Ammonium  Chloride  upon  Natrolite,  Scole-  availability  of  the  method  for  investiga- 

cite,   Prehnite,    and    Pectolite.     Am.   Jour.  .        ■    ^      ^,          i         •      i             ,-^    ^-            r 

Sci  iv  9:  ^41;  igoo.  ^^°"  ^"^°   ^"^  chemical  constitution  of 

silicates. 
Natrolite  and   Scokcite  proved   to   have   no   constitutional   water,  but  similar 
chemical  formulae,  which,  however,  are  not  those  of  ortho-silicates  : 
Natrolite,  NasAUSigO^o  .2H2O. 
Scolecite,  Ca  A^Si^Oio  -SHjO. 
Prehnite  may  be  regarded  as  having  constitutional  water   and  ortho-silicate 
formula. 

Pectolite  differs  widely  from  these  other  minerals  as  regards  the  ammonium 
chloride  reaction ;  but  experiments  suggest  the  probable  existence  of  a  hydrous 
ammonium  pectolite.  l.  Mci.l. 

Clark,  F.  W.  and  Steiger,  0.     The  Action  of      Authors  show  that   both   analcite  and 
Ammonium    Chloride   upon    Analcite    and       ,         .,  ,         ,       ^    j  ^      ocao      -^i     ..i  • 

Leucite.     Am.  Jour.  Sci.  iv,  9:  117,  19C0.  leucite,  when  heated  to   350°  with  this 

reagent  in  a  sealed  tube,  yielded  the 
same  definite  compound,  ammonium  leucite  NH^Al  SioOy.  This  reaction  was 
fairly  general  in  character,  and  analagous  results  were  obtained  with  other 
species. 

By  substituting  in  many  minerals  a  volatile  base  for  fixed  bases,  silicates  are 
obtained  which  split  upon  ignition  in  such  a  way  as  to  shed  light  upon  their 
molecular  constitution. 

The  conclusion  was  reached  that  analcite  and  leucite  were  not  true  meta- 
silicates,  but  pseudo  mixtures.  l.  Mci.  l. 

Parsons,  Charles  L.    The  Use  of  Metalhc  Sod-      Contrary  to  the  statement  of  Hempel, 
ium  in  Blowpipe  Analysis.    Jour.  Am.  Chem.  i       c     ^  j  i.u  r        j- 

Soc.23:  159,  Mar.  1901.  ^^o  first  proposed  the  use  of  sodium 

in  blowpipe  analysis,  the  author  finds 
that  the  reduction  of  metallic  compounds  by  means  of  sodium  takes  place  with 
the  greatest  ease  on  charcoal.  The  following  procedure  is  recommended  :  A 
piece  of  metallic  sodium  3  or  4  mm.  in  diameter  is  hammered  flat.  The  powdered 
substance  to  be  tested  is  spread  upon  and  pressed  into  the  metal,  and  the  whole 
turned  into  a  little  ball.  It  is  placed  in  a  slight  depression  of  a  piece  of  char- 
■  coal,  and  ignited  with  a  match  or  Bunsen  flame.  The  residue  is  heated  before 
the  blowpipe,  and  fusible  metallic  particles  readily  collect  into  a  globule ;  at  the 
same  time  coatings  of  the  volatile  metals  are  obtained.  Treated  in  this  way, 
garnierite  gives  magnetic  nickel,  chrysocolla  a  copper  button,  and  cassiterite  a  tin 


1394  Journal  of  Applied  Microscopy 

button,  as  readily  as  a  lead  button  is  obtained  from  cerussite.  Large  quantities 
of  sodium  are  to  be  avoided,  and  care  must  be  taken  not  to  touch  the  metal  with 
the  fingers,  as  the  reaction  sometimes  begins  spontaneously.  a,  f.  r. 

Morgan,  Leonard  P.,  and  Smith,  E.  F.  Experi-  Weighed  portions  of  chalcopyrite  were 
ments  on  Chalcopvrite.     Jour.  Amer.  Chem.  i    .       .1  •  r     1         ,      ^ 

Soc.  22:  107,  Feb.  1 90 1.  exposed  to    the   action   of  dry   hydro- 

chloric acid  gas  in  a  heated  combustion 
tube.  The  liquid  in  the  tube  was  titrated  with  potassium  permanganate,  and 
gave  an  iron  content  of  from  30.56  to  30.72  per  cent.,  theory  requiring  for 
chalcopyrite  30.5  per  cent.  Results  indicate  complete  decomposition,  and  show 
that  all  the  iron  is  in  a  ferrous  state.  The  results  obtained  by  heating  the 
mineral  in  a  closed  tube  with  a  solution  of  copper  sulphate  confirm  this. 

A.   F.  R. 

Atkinson,  Elizabetli  Allen.  Indium  in  Tungsten  From  150  to  300  grams  of  wolframite. 
Minerals.     Jour.  Amer.  Chem.  Soc.  20:  811,       ,  ...  -^  j        1       i-^       r  1 

jg  g  hubnerite,  and  scheelite,  from   several 

localities,  were  carefully  examined  for 
indium,  but  only  in  the  wolframite  from  Zinnwald  was  any  found.  Author  comes 
to  the  conclusion  that  indium  cannot  be  regarded  as  an  associate  of  tungsten, 
and  that  Hoppe-Seyler's  suspicion  as  to  blende  being  its  origin  is  probably  cor- 
rect, for  only  in  the  Zinnwald  mineral  was  it  found,  zinc  also  being  present  only 
there.  a.  f.  r. 

Vater,  Heinrich.  Ueber  den  Einfluss  der  Losun-  This  paper  discusses  the  action  of  the 
genossenauf  die  Krystallisationdes  Calcium-  solution  upon  gypsum  and  anhydrite, 
carbonates.       Theil  viii,  Zeit.  f.    Kryst.,  31 :  ,  1    j        ^1     ^      ^   i 

-^g_._g  jg„Q  -^  and  concludes  that  at  lower  tempera- 

tures, below  30°C.,  calcite  alone  results, 
and  that  the  only  known  cause  by  which  pure  calcium  carbonate  separates  as 
aragonite  is  a  temperature  exceeding  80°C.  a.  j.  i\i. 

Qoldschmidt,  V.  Ueber  Trogerit  und  kiinst-  Concludes  trogerite  is  tetragonal,  not 
lechen  Uranospinit.     Zeit.  f.  Kryst.  31 :  460-  ...  j  ^ 

478  iSqq  j  -'  ^        monoclmic,   and  ventures   supposition 

that  all  other  uranium  micas  :  autunite, 

uranospinite,  torbernite,  zeunerite,  and  phosphuranylite  are  also  tetragonal. 

A.  J.   M. 

Viola,  C.     Zur    Kentniss    des    Anorthits    vom       Crystallographic  Study. 
Vesuv.     Zeit.  f.  Kryst.  31:  484-498,  1899.  j  b     f  j 

Ward,  H.  L.    Notice  of  an  Aerolite  that  recently       The  Stone    is    light    ash-gray   in    color, 

fell  at   Allegan,  Mich.      Am.   Jour.    Sci.  iv.  r  •    ui  j  1       -^i.       ui      1 

g.  jg  °    '  very  friable,  and  covered  with  a  black 

crust,  which  is  1-2  mm.  thick,  and  has 

a  smooth  or  wavy   surface.     The   structure   is   chondritic,   and    the  following 

minerals  are  present :    enstatite,  chrysolite,  feldspar,  troilite  and  iron,  the  two 

latter  being  granular  and  evidently  scattered.     G.^3.558.  l.  mci.  t,. 

Qrimsley,  G.  P.,  and  Bailey,  E.  H.  S.  Report 
on  Gypsum  and  Gypsum  Cement.  Vol.  v, 
Univers.  Geol.  Sur.  of  Kansas. 


and  Laboratory  Methods.  1395 

.      MEDICAL  NOTES. 

METHODS  FOR  STAINING  TUBERCLE  BACILLI. 

Ehrlich-Weigert  Anilin-Methyl-Violet  Method.  —  Place  a  dried 
cover-glass  preparation,  film  down,  in  the  following  soliation,  and  heat  gently 
until  steam  rises,  then  allow  to  stand  for  2  to  5  minutes  : 

Methyl  violet,  sat.  ale.  sol.,  ...  1.1  part. 

Alcohol,  absol.,      ......     1.     part. 

Anilin  water,     .  .         .  .  .  .10.     parts. 

Anilin  water  is  made  by  using  1  part  anilin  oil  with  20  parts  of  distilled 
water,  and  after  standing  a  short  time  and  becoming  thoroughly  saturated,  filter- 
ing the  mixture.  Decolorize  for  a  few  seconds  in  1  part  nitric  acid  and  3  parts 
water.  Wash  one  or  two  seconds  in  60  per  cent,  alcohol,  then  in  water.  If  it 
is  desired  to  counterstain  the  specimen,  allow  a  few  drops  of  saturated  aqueous 
solution  of  vesuvin  to  cover  the  specimen  for  about  five  minutes.  When  the 
staining  is  complete  the  preparation  is  washed,  dried,  and  mounted  in  balsam. 
Gabbett's  Method. — This  method  is  simple  and  rapid,  and  is  considered 
by  many  to  be  a  most  excellent  method  for  routine  work.  The  cover-glass  pre- 
paration is  stained  for  2  to  5  minutes  in  Ziehl's  carbol  fuchsin  solution,  after  the 
formula : 

Fuchsin,       .......  1  part. 

Alcohol,  .......  10  parts. 

Carbolic  acid  (5  per  cent,  sol.),     .  .  .     100  parts. 

The  fuchsin  should  be  dissolved  in  the  alcohol  before  the  acid  is  added. 
After  this  solution  is  allowed  to  act  for  the  desired  length  of  time,  the  prepara- 
tion is  placed  for  1  to  2  minutes  in  Babbett's  methylen-blue  sulphuric  acid  solu- 
tion, consisting  of : 

Methylen  blue,         ......      1  part. 

Sulphuric  acid  (25  per  cent,  sol.),         .  .         50  parts. 

The  specimen  is  then  washed  in  water,  dried,  and  mounted  in  balsam. 
Tubercle  bacilli  are  stained  red,  while  other  elements  of  the  mount  are  blue. 

Ziehl-Neelson  Method. — By  this  method  tubercle  bacilli  are  stained  with 
the  following  solution  : 

Fuchsin,  1  part,  dissolved  in  10  parts  alcohol,  to  which  is  added  lOO  parts 
5  per  cent,  solution  of  carbolic  acid.  The  cover-glass  preparation  is  floated,  film 
down,  on  the  solution,  to  which  gentle  heat  is  applied  until  steam  rises.  The 
specimen  is  then  washed  in  water,  and  decolorized  in  25  per  cent,  nitric  or  sul- 
phuric acid,  then  in  60  per  cent,  alcohol  for  a  very  short  time,  after  which,  with 
thorough  washing  in  water,  it  is  mounted  in  balsam.  c.  w.  j. 

NEWS  AND  NOTES. 

The  New  Haven  Microscopical  Society  has  just  issued  a  very  neat  booklet 
containing  the  constitution  and  by-laws  of  the  society,  as  well  as  a  list  of  the 
members  with  the  address  of  each.  The  officers  of  the  society  are  :  President, 
Robert  Brown,  Observatory  place.  New  Haven  ;  Secretary  and  Treasurer,  J.  F. 
Malone,  25  Wooster  place,  New  Haven. 


1396  Journal  of  Applied  Microscopy 

The  annual  meeting  of  the  American  Microscopical  Society  will  be  held  in 
Denver,  Col.,  August  "29  to  •>!.  Efforts  are  being  made'to  secure  exceptionally 
low  rates,  and  an  enthusiastic  and  profitable  meeting  is  assured. 


The  Hopkins  Seaside  Laboratory,  of  Leland  Stanford  Junior  University, 
began  its  tenth  session  at  Pacific  Grove  on  Monterey  Bay,  Monday,  June  10, 
1901.  The  regular  courses  of  instruction  continue  six  weeks,  closing  July  'JOth. 
Provision  is  made  at  the  laboratory  to  accommodate  three  classes  of  students  : 
(1)  teachers  and  students  who  wish  to  pursue  laboratory  courses  in  botany  and 
zoology;  (2)  advanced  students  in  zoology,  physiology,  and  botany;  and  (o) 
investigators  who  are  prepared  to  carry  on  researches  in  morphology  and  physi- 
ology.    The  regular  courses,  with  the  instructors  in  charge,  are  as  follows  : 

1.  General    Zoology — Professor   G.  C.  Price,  Leland   Stanford  Jr.  Univ. 

2.  Elementary  Botany — Professor  S.  C.  Price. 

3.  Advanced  course  on  Structure  and  Physiology  of  Algae. — -Professor  G. 

J.  Peirce,  Leland  Stanford  Jr.  Univ. 

4.  Embryology — Professor  G.  J.  Peirce. 

5.  Comparative  Morphology  and  Histology  of  the  Nervous  System  and 

Sense  Organs. — Professor   F.   M.  MacFarland,   Leland   Stanford 
Junior  University. 

6.  Advanced  course  in  Zoology. — Professor  F.  M.  MacFarland. 

7.  General  Ornithology. 


We  have  received  the  announcement  of  the  third  session  of  the  Rhode  Island 
Summer  School  for  nature-study  to  be  held  at  the  Rhode  Island  College  of  Agri- 
culture and  Mechanic  Arts,  Kingston,  R.  I.,  July  5  to  20,  1901.  The  instruction 
is  to  be  almost  wholly  in  the  nature  of  field  work,  the  schedule  being  made  up  of 
excursions,  led  by  competent  men  in  every  branch  of  natural  science.  The  time 
given  to  class-room  exercises  will  be  just  enough  to  indicate  what  is  to  be 
observed  and  done  in  the  field.  Special  evening  lectures  will  be  given.  Com- 
munications should  be  addressed  to  "  Summer  School,"  Kingston,  R.  I. 

QUESTION   BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

8.  Where  can  "  Stabilite  "  insulating  material  be  bought  ?  Is  it  used  much 
in  American  laboratories  ?  l.  h. 

9.  What  is  the  best  method  of  drying  and  mounting  microscopic  fungi  ?  Can 
you  refer  me  to  a  good  book  dealing  with  the  subject.  m.  r. 

REPLIES  TO  QUESTIONS. 

"  What  is  meant  by  the  growing  tip  in  allium  ?  "     (Question  No.  1.) 

The  growing  part  of  a  root  {a),  or  the  "  growing  tip,"  is  a  short  space  about 

v..>^;^= -.  -  ^.:v.:;:^K-;Jv;- •,:;!;     oue-sixth  of   an  inch  back  from  the  extreme  end. 

^^^  In  this  part  of  the  root  the  cells  divide  rapidly, 

■*K<'/  ^"^^  ^^^  length  is  thereby  increased.     This  is  the 

/  only   part    of    the  root   in   which   growth  takes 

4         —  -'""  'v;^:^v       place.     The    end  of  the  root  is  usually  covered 

by  a  protecting  coat  of  dead  cells  {b),  derived  from  the  living  zone  just  back  of  it. 

These  dead  cells  constitute  what  is  known  as  the  root  cap. 

C.  A.  Whiting. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 

VOLUME   IV.  AUGUST,    1901.  NUMBER  8 


Table  of  Specific  Gravities  of  Saturated  Solutions  and 
Solubilities  of  Anilin  Stains. 

In  the  following  tables  will  be  found  in  the  first  two  columns  the  specific 
gravity  of  saturated  solutions  of  the  various  anilin  stains,  made  in  the  cold  and 
by  boiling.  Those  made  in  the  cold  were  allowed  to  stand  with  an  excess  of 
the  stain,  the  bottle  being  vigorously  shaken  daily,  for  two  weeks. 

The  solutions  made  with  hot  water  were  kept  at  the  boiling  point  for  twenty 
minutes  and  well  stirred.  Both  solutions  were  carefully  filtered  and  the  s.  g. 
ascertained  by  means  of  the  specific  gravity  bottle.  Correction  was  made  for 
temperature  and  the  average  of  five  weighings  was  taken  in  each  case. 

The  third  column  contains  the  increase  in  weight  of  95  per  cent,  alcohol 
when  various  stains  are  dissolved  in  it  to  saturation.  The  s.  g.  in  this  case  may 
be  obtained  by  adding  the  weight  of  the  alcohol  in  use  to  the  figures  given  in 
the  table.  The  s.  g.  of  the  solution  is  not  given  for  the  reason  that  the  various 
samples  of  alcohol  obtained  commercially  as  95  per  cent,  vary  considerably  in 
actual  percentage  and  hence  in  weight.  The  solvent  power  varies  practically 
not  at  all,  so  that  by  adding  the  weight  of  the  sample  at  hand  to  the  figures  in 
the  table  the  s.  g.  may  be  obtained  sufficiently  accurate  for  most  working 
purposes. 

The  various  formulae  which  one  encounters  constantly  call  for  saturated 
solutions  of  the  various  stains,  and  it  is  usually  the  custom  to  add  three  or  four 
times  as  much  of  the  dry  stain  as  is  really  necessary.  This  can  be  obviated  by 
a  glance  at  the  table.  While  it  is  true  that  all  solutions  will  change  slightly  in 
volume  when  they  have  substances  dissolved  in  them  to  the  point  of  saturation, 
still  the  change  is  usually  small,  and  we  may  also  take  the  table  as  a  guide  to 
the  solubilities  of  the  stains  in  making  them  up,  thereby  avoiding  waste. 

In  the  first  column  the  approximate  solubilities  in  grams  per  hundred  c.c.  of 
water  may  be  obtained  by  simply  subtracting  100  from  the  s.  g. 

In  the  last  column  the  increase  in  weight  is  given  and  may  be  taken  directly 
as  the  approximate  solubility. 

A  slight  error  will  be  found  in  several  of  the  stains,  which  seems  to  be  caused 
by  a  chemical  change  when  the  stains  are  boiled.     This  is  notably  the  case  with 

(1397) 


1398  Journal  of  Applied  Microscopy 

gentian  violet,  which  upon  boiling  yields  a  solution  which  is  lighter  than  the 
water  was  at  first  (s.  g.  99.928.) 

All  the  stains  used  in  this  work  were  of  the  first  quality  for  histologic  pur- 
poses, not  the  ordinary  commercial  dyes. 

I  desire  to  acknowledge  my  indebtedness  to  R.  H.  Hough,  for  his  assistance 
in  this  work. 

Anilin  Blue  Black 100.280 100.792 0.44 

"       Green 100.530 100.740 1.71 

"       Red 100.052 100.100 1.95 

"       Violet 100.172 100.076 2.76 

"       Black 100.478 100.264 0.54 

Benzoazurin 100.504 101.120 0.99 

Benzo-Purpurin 100.394 100.424 0.61 

Biebrich  Scarlet 101.972 101.054 0.27 

Bismark  Brown 100.440 100.752 0.55 

Blue  Lumiere Ins.        Ins 0.64 

Chrysoidin   100.543 100.608 2.13 

Congo  Red 100.464 100.768   0.22 

Corralin 100.926 101.012 2.74 

Dahlia 100.294 100.180 2.12 

Delta  Purpurin 100.674.  ,    100.528 0.64 

Eosin  (Bluish) 102.496 101.872 2.43 

Erythrosin 100.800 101.732 1.99 

Fuchsin 100.018 100.072 1.54 

Fuchsin  Acid 101.200 101.452 0.72 

Gentian  Violet 100.068 99.928 3.65 

Gold  Orange 100.366 100.528 0.02 

Indulin 100.606 101.024 0.79 

Magenta 100.072   100.072 2.98 

Malachite  Green 100.232 100.372 3.56 

Metanyl  Yellow 100.420 100.580 1.12 

Methyl  Blue 100.446 100.672 0.28 

Methyl  Green 100.744 101.425 2.69 

Methyl  Violet 100.072 100.260 2.29 

*Methylene  Blue 100.060 100.580 0.28 

Orange  B.  Naphthol 100.210 100.836 0.28 

Orange  G 100.944 101.172 trace 

Orange  II 100.292 100.520 0.56 

Orseille  G 100.456 100.812 0.40 

Rocellin 100.372 100.312 0.33 

Rosanilin  Hydrochlorate 100.048 100.080 1.79 

Rosein 100.053 100.076 1.48 

Rose  Bengal 101.416 101.856 2.12 

Rubin  G 100.046 100.072 2.53 

Rubin  S 102.464 102.364 0.15 

Saffranin 101.526 100.596 2.36 

Solferino 100.014 100.052 2.543 

Solid  Green 100.564 100.844 3.00 

Tropeolin  000  No.  2 100.448 100.660 0.58 

Vesuvin 100.412 100.944 0.81 

Victoria  Blue 100.102 100.160 1.77 

Violet  B 100.286 100.440 3.84 

Vanderbilt  University.  LOUIS    LerOY. 

*The  sample  of  Methylene  Blue  here  tested  was  but  very  slightly  soluble,  and  stained  very  poorly. 


and  Laboratory  Methods. 


1399 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 

PHOTOMICROGRAPHY. 

Introductory. 

Photomicrography,  as  it  may  be  practiced  to-day,  is  of  prime  importance  in 
several  different  kinds  of  work.  Nothing  can  take  its  place  as  a  means  of  illus- 
tration in  popular  lectures.  In  class  lectures,  for  review,  a  term's  work  can  be 
summarized  more  quickly  and  correctly  by  means  of  the  lantern  slides  than  in 
any  other  way.  It  has  the  advantage  over  microscopic  observation,  of  directing 
everyone's  attention,  at  the  same  time,  to  the  same  thing.  In  all  our  colleges 
and  universities,  students  of  history,  sociology,  psychology,  etc.,  are  calling 
for  lectures  on  the  laws  of  growth,  on  what  is  known  about  heredity,  on  the 
principles  of  kinship,  etc. — students  who  have  neither  the  time  nor  the  skill  to 


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Fig.  I. — Four  day  chick,     x  15. 

get  the  foundation  facts  in  the  laboratory.  Wherever  illustration  in  histology  is 
desirable,  photomicrography  has  advantages  that  will  not  longer  permit  it  to  be 
neglected. 

There  is  no  class  work  that  must  be  seen  through  the  microscope  that  the 
half-tone  and  the  lantern  slide  cannot  faithfully  present.  It  is  the  purpose  of 
the  accompanying  cuts  to  sustain  this  proposition.  Five  of  the  microscopic  slides 
photographed  were  made  by  students  in  the  regular  work  of  the  class-room. 
The  negative  for  Fig.  1  was  made  with  a  70  mm.  apochromatic  objective  without 
eye-piece,  and  with  a  camera  extension  of  three  feet.  The  low-power  objective 
gives  penetration,  and  the  extension  gives  the  necessary  amplification  ;  by  just 


1400 


Journal  of  Applied  Microscopy 


this  means  the  depth  of  focus  necessary  in  any  case  can  be  obtained.  The 
lowest  objective  that  will  resolve  the  details  wanted  is  selected,  and  then  the 
requisite  power  is  obtained  by  camera  extension ;  this  preserves  at  all  times  the 
relative  depth  of  focus.  It  is  true  that  a  histological  section  can  be  so  thick  or 
prepared  so  poorly  that  the  photography  of  it  is  difficult.  A  better  section 
should  be  made  (and  this  is  one  reason  for  the  use  of  photomicrography — it  will 
at  once  lead  to  the  preparation  of  better  microscopic  slides).  Any  section  can, 
however,  be  so  photographed  as  to  show  all  that  can  be  seen  at  any  one  look, 
and  by  repeated  exposures  all  can  be  shown.  Fig.  2  was  made  with  an  8  mm. 
objective  and  a  No.  4  projection  eye-piece,  and  a  camera  extension  of  four  feet, 


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Fig.  2. — Frog's  blood  ;  hJEmatoxylin-eosin  stain,     x  400. 

and  shows  the  same  things  in  flatness  of  held  and  depth  of  focus  for  its  power, 
400  diameters,  that  Fig.  1  does  for  its. 

Fig.  3  is  a  section  of  onion  root  with  chromosomes  lying  in  several  different 
planes,  and  shows  the  same  for  1500  diameters.  It  was  made  with  a  2  mm.  oil 
immersion,  apochromatic  objective,  and  a  No.  4  projection  eye-piece,  with  a 
camera  extension  of  thirty-seven  inches.  Fig.  4,  Fig.  5,  and  Fig.  G,  were  made 
with  the  same  combination. 

Fig.  6  is  a  blastula  of  ascaris  ;  it  was  photographed  from  an  unsectioned 
blastula ;  the  instrument  was  focused  on  the  midsection  of  the  ball  of  cells  ;  the 
light  had  to  pass  through  the  lower  cells  before,  and  the  upper  cells  after  passing 
the  points  shown ;  this  is  a  means  of  sectioning  with  the  microscope.  Fig.  1 
represents  low-power  work.  Fig.  2  medium-power  work,  and  the  remaining  figures 


and  Laboratory  Methods. 


1401 


high-power  work,  which  for  depth  of  focus,  magnification,  and  extent  of  field, 
cannot  be  reproduced  with  cheap  or  improvised  apparatus. 

If,  for  instance,  an  ordinary  microscope  is  used  with  an  ordinary  camera, 
none  of  the  figures  here  shown  can  be  duplicated,  no  matter  how  good  the  lenses 
may  be,  for  to  produce  any  power  here  given  a  higher  objective,  with  a  narrower 
field  and  less  deep  focus,  would  be  indispensable,  and  this  would  sacrifice  part 
of  the  field  entirely,  and  the  focus  over  the  part  retained. 

One  reason  why  photomicrography  has  not  hitherto  succeeded  better,  is  that 
cheap  apparatus,  scraped  together  from  a  microscopic  and  a  photographic  outfit, 
has  been  recommended.     This  cheap  apparatus  was  always  the  most  expensive 


Fig.  3. — Cells  from  onion  rootlet ;  iron-haematoxylin  stain,     x  1500. 


to  be  had,  for  the  reason  that  the  time  consumed  in  getting  ready  for  and  making 
a  successful  exposure  costs,  in  the  end,  more  than  the  investment  for  a  correct 
outfit. 

In  the  second  place,  the  results,  for  reasons  above  given,  were  never  valuable 
except  in  the  case  of  slides  so  perfectly  prepared  that  they  had  to  be  the  best  of 
an  expert  microscopist's  work.  I  again  and  again  concluded,  while  using  these 
makeshifts,  that  histological  slides  could  not  be  successfully  photographed.  I 
thought  photomicrography  was  an  art,  the  usefulness  of  which  was  confined  to 
the  resolving  of  lines  on  diatoms,  and  reproducing  the  silhouettes  of  bacteria  so 
prepared  that  the  contrast  was  sharp  and  the  field  flat. 

The  cheap  way  to  make  successful  photomicrographs  is  to  have  a  complete 


1402 


Journal  of  Applied  Microscopy 


Fig.  4. — Early  telophase  of  mitosis  in  Ascaris  megalocephala  var.  bivalens; 

polar  bodies  at  i ;  egg-cell  wall  at  2.     Centrosome  divided 

just  below  polar  bodies,     x  1500. 

apparatus;  microscope  stand,  lenses,  camera,  and  illuminating  appliances,  dedi- 
cated to  this  one  work  ;  mounted  to  stay,  on  tables  adapted  to  the  purpose, 
resting  on  a  floor  that  cannot  be  jarred,  with  a  fully  equipped  dark  room  imme- 
diately at  hand.  The  essentials  of  such  an  apparatus  will  be  fully  described  in  a 
succeeding  paper. 


Fig.  5. — Mitotic  figure  from  pollen  mother-cell  of  Lillium  candidum. 
Slide  by  Prof.  David  M.  Mottier.     x  1 500. 


and  Laboratory  Methods. 


1403 


With  such  an  apparatus,  ten  first  quality  negatives,  of  any  diameters  from 
4000  down,  can  easily  be  made  in  a  couple  of  hours.  They  can  be  more  easily 
and  certainly  made  than  the  same  number  of  fair  to  middling  negatives  of  from 
40  to  100  diameters  on  any  improvised  outfit. 


.^^^- 

Fig.  6. —  Unsectioned  blastula  of  ascaris.     x  1500. 

This  makes  photomicrographs  of  all  powers  really  usable.  Previous  to 
1899,  experts  were  happy  with  ten  failures  to  one  success ;  one  correct  negative 
had  to  pay  for  an  entire  evening's  work.  This  made  them  expensive,  too 
expensive  for  any  use,  however  cheap  the  apparatus. 

Earlham  College.  D.  W.  DennIS. 

Notes  on  Testing  for  B.  Coli  in  Water 

Test  for  B.  Coli  in  Large  Volumes. 

In  1898  we  published  a  description  of  the  methods  employed  at  the  Lawrence 
Experiment  Station  in  the  routine  examination  of  water  for  B.  coli.  These  tests 
were  all  made  in  one  cubic  centimeter. 

The  description  of  the  preliminary  test  in  large  volumes,  usually  100  cubic 
centimeters,  was  omitted  at  this  time,  as  the  test  was  then  in  the  experimental 
stage.  This  test,  as  it  is  now  made,  has  been  in  use  in  this  laboratory  for  about 
two  years,  and  has  proved  very  satisfactory.  The  water  to  be  tested  is  collected 
in  clear,  glass-stoppered  bottles,  of  the  tall  Blake  pattern,  holding  about  eight 
ounces,  with  a  graduation  mark  for  100  cubic-centimeters.  To  100  c.  c.  of  the 
water,  in  the  bottle,  we  add  10  c.  c.  of  a  strong  carbolated  dextrose  broth,  which 
is  prepared  as  follows:  100  grams  of  dextrose  and  50  grams  of  peptone  are 
dissolved  in  1  liter  of  boiling  water.  After  cooling,  and  filtering  through  paper 
till  clear,  50  c.  c.  of  a  5  per  cent,  aqueous  solution  of  phenol  is  added,  or  approxi- 
mately 0.25  per  cent,  of  phenol.  The  solution  is  now  ready  for  use,  and 
requires  no  sterilizing.  There  is  enough  phenol  present  to  prevent  the  spoiling 
of  the  solution,  especially  if  kept  on  ice,  as  we  always  do. 


1404 


Journal  of  Applied  Microscopy 


The  dilution  in  the  test  reduces  the  amount  considerably  below  that  neces- 
sary to  retard  the  growth  of  B.  coli  and  B.  typhosus,  but  is  still  sufificient  to  pre- 
vent the  growth  of  many  species  which  would  ferment  the  broth  and  interfere 
with  the  test.  After  inoculation,  the  bottles  are  placed  in  an  incubator  at  38°C. 
for  twenty  hours.  At  the  end  of  this  time,  if  B.  coli  be  present,  there  should  be 
a  slight  bead  on  the  surface.  Immediately  on  removing  from  the  incubator, 
give  the  bottle  one  quick,  hard  shake,  and  set  it  up  in  front  of  a  window.  The 
gas,  if  B.  coli  be  present,  will  now  separate  from  the  liquid  and  rise  slowly  to  the 
top,  giving  the  same  appearance  as  when  a  bottle  of  highly  carbonated  water  is 
opened.  This  appearance  is  quite  characteristic.  The  pressure  of  the  liberated 
gas  is  frequently  sufficient  to  blow  the  stopper  out  of  the  bottle.  We  always 
plate  out  a  sample  showing  positive  indications  and  test  to  confirm  the  diagnosis, 
and  in  over  75  per  cent,  of  such  samples  we  have  found  B.  coli  to  be  the 
organism  responsible  for  the  fermentation.  On  the  other  hand,  we  have  plated 
out  many  hundred  negative  tests,  and  have  yet  to  find  one  in  which  B.  coli  was 
present. 

A  Fermentation  Tube  Adapted  to  Rapid  Handlino  in  Routine  Work. — 
In  1897,  when  we  first  began  making  routine  tests  for  B.  coli,  we  found  that  the 
usual  type  of  fermentation  tube,  with  a  base,  a  large  bulb,  and  a  constriction  at 
the  bend,  was  not  suited  to  rapid  work,  accordingly  I  had  some  tubes  made 
omitting  these  features. 

These  tubes  have  given  such  general  satisfaction 
in  our  laboratory,  that  I  hope  the  following  descrip- 
tion and  illustration  will  be  of  some  value  to  others 
in  the  same  line  of  work.  The  shape  and  dimen- 
sions of  the  tube  are  shown  in  the  sketch.  The 
tubes,  when  filled  about  as  in  the  sketch,  are  set  in 
a  wire  basket  in  rows,  cotton  wool  being  placed  in 
the  bottom  for  them  to  rest  on.  We  use  baskets 
3x5  inches,  and  5  inches  deep  ;  these  will  hold  ten 
tubes  in  a  row,  and  by  putting  in  layers  of  cotton 
wool,  three  tiers  can  be  placed  in  a  basket.  In  this 
way,  thirty  tubes  occupy  about  as  much  space  as 
five  or  six  of  the  ordinary  style  of  tube  would 
occupy,  and  can  be  handled  as  a  unit  in  steriliza- 
tion, etc.  The  open  arm  of  the  tube  is  sufficiently 
long  to  hold  all  of  the  liquid  when  it  is  forced  out 
of  the  closed  arm,  without  wetting  the  cotton  plug. 
Determinations  of  the  volume  and  composition  of  the  gas  are  made  as  easily  and 
accurately  as  in  the  old  style  of  tube.  We  have  all  of  our  tubes,  test  tubes 
included,  made  without  lips,  as  we  think  it  tends  to  decrease  the  breakage,  and 
make  the  tubes  pack  better  in  baskets.  The  abolishment  of  the  bulb  and  con- 
striction at  the  neck  also  makes  cleaning  somewhat  less  of  a  grind. 

Lawrence  Experiment  Station.  "  STEPHEN  DeM.  Gage. 


and  Laboratory  Methods. 


1405 


The  Cone  Net. 

This  net  was  originally  described  in  the  Transactions  of  the  Wisconsin 
Academy  of  Sciences,  Arts,  and  Letters,  Volume  VIII,  page  397,  1892.  Since 
that  time  I  have  had  numerous  inquiries  for  directions  for  the  construction  of 
the  net,  and  in  response  to  these  I  give  the  following  account  of  the  net  as  I 
now  use  it.  The  apparatus  is  still  very  crude  and  improvements  can  easily  be 
suggested.  It  has  met  my  purposes,  however,  and  therefore  for  some  years  I 
have  given  no  attention  to  improving  its  details  of  construction.  The  metal 
parts  of  the  apparatus  are  illustrated  in  the  annexed  figures.  The  diameter  of 
the  base  of  the  cone  top  in  the  net  which  I  now  use  is  three  inches.  From  this 
the  scale  of  the  drawings  can  easily  be  computed.     The  cone  top  is  represented 


in  Fig.  1.  It  consists  of  a  rim  of  stout  tin  (A)  three  inches  in  diameter,  one 
inch  in  height,  with  a  stout  wire  turned  into  the  lower  edge.  To  the  inside  of 
this  is  soldered  a  wire  loop  (B),  which  lies  under  the  cone  and  projects  through 
its  apex  in  a  loop  to  which  the  line  is  attached.  The  top  of  the  net  is  formed 
by  a  cone  of  brass  wire  netting  (C),  one-twentieth  of  an  inch  mesh,  with  a  slant 
height  of  four  inches  soldered  to  the  rim  and  to  the  wire  loop  at  the  top.  The 
bottom  of  the  net  (Fig.  3)  is  formed  by  the  screw  top  of  a  kerosene  can,  to 
which  is  soldered  a  cylinder  of  tin  about  one  inch  in  height  and  one  and  one- 
fourth  inch  in  inside  diameter,  into  whose  upper  edge  a  stout  wire  is  turned. 
The  dredge  net  is  fastened  to  these  two  metal  parts,  being  firmly  tied  both  at 
top  and  bottom.  The  tying  must  be  firm,  or  if  the  net  is  thrown  when  contain- 
ing water  it  may  be  pulled  off  from  the  cone  top.  I  have  found  it  a  good 
method  of  tying  to  use  a  dry  string  drawn  tightly,  of  a  kind  which  will  shrink 
when  wet,  and  to  tie  the  net  twice,  turning  the  edge  down  over  the  string  of  the 
first  tying  before  fastening  it  the  second  time.     For  a  net  I   use  a  fine  cotton 


1406  Journal  of  Applied  Microscopy 

cloth  of  the  sort  known  to  the  trade  as  •'  India  linen."  This  is  faced  with 
muslin  at  the  top  and  bottom. 

In  my  first  nets  I  made  the  cone  top  removable,  and  this,  of  course,  can  easily 
be  done.  I  discovered,  however,  that  in  working  among  weeds  I  practically  always 
used  the  netting  and  that  it  was  easier  to  carry  a  second  net  of  the  small  size 
for  work  in  open  water  than  to  have  the  more  complicated  and  heavier  arrange- 
ment necessitated  by  making  the  net  removable. 

The  collecting  funnel  (Fig.  2)  is  the  part  of  my  apparatus  which  seems  to 
have  been  least  used,  but  which  I  regard  as  even  more  important  than  the  cone 
dredge.  The  funnel  is  two  inches  in  diameter  at  the  top  and  has  a  cylindrical 
spout  about  three  inches  in  length.  The  bottom  of  this  is  formed  by  a  cylinder 
of  tin  (A)  one-half  inch  in  height,  connected  to  the  body  of  the  funnel  by  two 
strips  of  folded  tin  (B).  Outside  of  this  is  soldered  a  cylinder  of  fine  brass 
wire  netting  (C).  I  have  used  a  fine  milk  strainer,  which  is  about  one-fiftieth  of 
an  inch  mesh,  and  also  a  finer  netting,  one-hundredth  of  an  inch  mesh,  for  this 
cylinder.  For  general  use  the  coarser  mesh  is  sufficiently  fine  and  is  decidedly 
more  convenient.  The  diameter  of  the  spout  is  about  half  an  inch.  It  is  made 
of  such  a  size  as  to  slip  into  the  opening  of  an  eight  dram  homeopathic  vial, 
short  form.  This  funnel  is  used  in  connection  with  a  tin  cup  in  collecting 
material,  especially  from  among  weeds.  The  material  collected  by  the  net  is 
washed  into  the  cup,  which  is  then  filled  with  water  and  allowed  to  stand  for  a 
short  time.  The  vegetable  debris  settles  to  the  bottom  and  most  of  the  animals 
remain  in  the  water  above  it,  together  with  some  of  the  lighter  parts  of  the  veg- 
etation. The  water  is  then  poured  through  the  funnel,  the  lower  end  of  which 
may  be  stopped  with  a  cork,  or,  as  I  find,  more  conveniently  by  the  finger,  and 
when  the  water  is  drained  off,  the  spout  of  the  funnel  is  thrust  into  a  homeo- 
pathic vial  filled  with  alcohol,  and  its  contents  washed  out.  In  this  way  the 
greater  part  of  the  animals  can  be  separated  from  most  of  the  accompanying 
vegetable  debris ;  thus  greatly  facilitating  the  subsequent  study  of  the  collection. 
Still  further,  the  results  of  a  large  number  of  hauls  of  the  net  can  be  con- 
centrated and  preserved  in  a  single  bottle.  In  all  cases,  however,  some  of  the 
material  which  settles  to  the  bottom  of  the  cup  should  be  preserved,  since  it 
always  happens  that  part  of  the  animal  life  seeks  refuge  at  once  in  this.  The 
cup  which  I  use  is  made  of  such  a  size  as  to  contain  the  funnel,  and  the  dredge 
net  with  the  bottom  part  easily  finds  place  in  the  interior  of  the  cone. 

The  net  can  be  used  as  a  tow  net,  or  can  be  thrown  either  from  the  shore  or 
from  a  boat.  In  working  among  weeds,  I  prefer,  if  possible,  to  use  it  as  a  tow 
net,  keeping  it  close  to  the  boat  and  working  it  by  the  line  in  and  out  among  the 
weeds.  There  is  very  little  use  in  putting  out  a  long  line  and  allowing  it  to 
drag  through  the  weeds,  as  the  vegetation  collects  and  very  quickly  entirely 
covers  the  top  of  the  dredge.  In  hauling  the  net,  it  is  better  to  use  a  violent 
jerking  motion  than  to  pull  steadily,  and  the  line  and  net  should  be  so 
strong  as  not  to  suffer  by  this  method.  In  throwing  the  net,  it  is  often 
found  difficult  to  make  it  sink  after  the  cloth  has  once  been  wet.  If  the  throw 
is  not  to  a  great  distance  the  net  can  often  be  made  to  fall  with  bottom  down- 
ward by  a  little  manipulation  of  the  line.     The  sinking  can  also  be  secured  by 


and  Laboratory  Methods. 


1407 


a  weight  at  the  apex  of  the  cone,  either  formed  by  melted  solder  poured  in,  or 
by  a  lead  weight  which  may  be  tied  to  the  interior  of  the  cone  or  to  the  screw- 
cap.  In  dragging  the  net  along  the  bottom,  I  prefer  to  have  the  cone  and  net 
without  weights  and  to  attach  a  weight  to  a  line  so  long  that  it  will  drag  a  little 
way  behind  the  end  of  the  dredge.  In  this  way  the  net  draws  over  the  bottom 
instead  of  digging  into  it,  as  it  is  apt  to  do  if  weighted  at  the  apex. 

The  materials  and  construction  of  the  apparatus  are  such  that  it  can  be  con- 
structed by  any  tinman,  and  nets  can  be  made  of  any  size  for  various  purposes. 
I  have  used  them  up  to  six  inches  in  diameter  and  with  a  cone  provided  with  a 
quarter-inch  mesh.  Such  a  net  was  used  for  collecting  in  salt  water,  to  prevent 
the  entrance  of  floating  algae.  In  nets  so  large  it  is  well  to  make  the  cone  top 
in  two  pieces  so  that  the  net  can  be  easily  removed  from  the  cone. 

In  collecting  on  expeditions  where  it  was  not  convenient  to  carry  a  large 
number  of  small  bottles,  I  have  furnished  little  bags  of  such  a  size  as  to  slip 
over  the  spout  of  the  funnel.  The  material  collected  is  washed  out  into  these 
bags,  which  are  tied,  numbered  and  labeled  and  placed  in  a  large  bottle  or  can 
of  preserving  fluid.  E.  A.  Birge. 

University  of  Wisconsin. 


A  Modification  of  the  Birge  Collecting  Net. 

To  collectors  of  the  smaller  aquatic  organisms,  especially  of  such  as  Crusta- 
cea, Hydrachnidae,  etc.,  the  Birge  collecting  net,  or  some  similar  apparatus,  is  an 
indispensable  part  of  their  equipment.  The 
writer  has  used  it  with  general  satisfaction 
for  several  years  in  the  collecting  of  water-mites. 

There  are  occasions,  however,  when  its  use 
in  the  ordinary  form  becomes  awkward  or  im- 
possible owing  to  the  place  or  conditions  about 
the  place  where  it  is  desired  to  make  collections. 
Frequently  it  is  desirable  to  secure  material 
from  narrow,  tortuous  streams  in  which  a  straight 
haul  for  any  distance  is  impossible,  or  from  small 
springs  or  pools,  or  from  the  shore  when,  owing 
to  the  uncertain  footing  or  to  the  interposition 
of  branches  or  other  objects,  casting  is  extremely 
difficult,  or  in  water  so  choked  with  vegetation 
that  only  small  open  spaces  are  left  here  and 
there  amongst  the  weeds.  To  meet  such  condi- 
tions the  author  devised  and  has  put  to  successful 
use  the  following  modification,  which  he  ventures 
to  suggest  to  others  who  may  have  felt  a  similar 
want. 

Briefly  stated,  it  is  as  follows :  A  groove 
(Fig.     1)    is    passed    about    the     tin    cylinder. 


1408 


Journal  of  Applied  Microscopy 


"^ 


which    is    fastened    to   the  back  end   of  the  cone  of   metal  gauze,  and    a  ring, 
affixed  to  the  end  of  a  handle   of  about  the   size   of  an   ordinary  walking  stick 

(Fig.  2),  fits  into  this  groove  and  is  firmly 
clamped  in  this  position.  The  collector 
then  has  perfect  control  of  his  net  and  can 
dip  into  a  contracted  space,  scrape  the 
margin  of  a  pool,  reach  beneath  a  dock  or 
other  obstruction,  or  while  wading  sweep 
the  bed  of  vegetation  which  carpets  the 
bottom  by  passing  the  net  back  and  forth 
from  right  to  left  and  left  to  right  before  him 
as  he  walks.  The  figures  given  will  illus- 
trate the  method  of  construction  more 
clearly  than  can  be  done  by  means  of  a 
description,  and  to  them  the  reader  is 
referred  for  further  particulars. 

Reference  may  also  be  made  to  certain 
other  details  of  construction  which  exper- 
ience has  shown  to  be  of  advantage.  First,  it  is  found  that  if  the  wire 
which  forms  the  ring  (r)  to  which  the  cord  is  attached  be  carried  down  inside 
the  gauze  cone  to  its  base  and  there  firmly  soldered  to 
another  ring  which  forms  the  margin  of  the  base  of  the  cone, 
the  stiffness  thus  gained  is  of  great  advantage.  Second,  if 
the  tip  of  the  cone  be  filled  in  with  lead  for  a  short  distance 
the  weight  serves  to  hold  the  "  nose  "  of  the  net  down,  and 
it  draws  through  the  weeds  to  better  advantage.  The  attach- 
ing of  a  "  sinker  "  to  the  cord  attached  to  the  net  has  been 
tried,  but  the  "  sinker  "  is  liable  to  be  caught  and  the  "  nose  " 
of  the  net  is,  also,  not  carried  down  into  a  bed  of  Chara,  as 
is  desirable  in  the  work  of  the  writer.  Third,  it  is  suggested 
that  the  free  margin  of  the  tin  cylinder  (t)  be  made  to  project  in  the  form  of  a 
flange,  and  that  by  "  running  "  a  cord  through  the  overlapped  border  of  the 
cloth  net  (n)  this  latter  can  be  easily  adjusted  for  use  and  as  readily  removed. 
Fourth,  the  writer  has  found  that  if  the  cup  at  the  end  of  the  cloth  net  which 
receives  the  collection  be  heavy  by  being  made  of  a  piece  of  lead  pipe  (Fig.  4) 
the  net  is  less  liable  to  "  foul  "  itself  in  casting,  and  also  sinks  more  quickly,  as 
this  end  of  the  net  is  carried  downward  the  more  rapidly.  For  his  own  nets  the 
writer  uses  a  fine  cheese-cloth,  and  lines  this  on  the  inside  in  the  lower  two-thirds 
with  a  fine  quality  of  China  silk,  which  costs  about  seventy-five  cents  a  yard,  and 
is  thus  much  cheaper  than  bolting  cloth  while  serving  equally  well. 

The  author  is  in  the  habit,  during  collecting  trips  taken  to  different  portions 
of  the  state,  of  gathering  material  of  all  kinds,  having  in  view  the  use  of  it  in 
the  interest  of  a  complete  faunal  survey  of  the  state,  which  it  is  hoped  before 
many  years  may  become  possible.  He  has  found  it  very  useful  to  standardize 
his  nets  and  thus  to  make  them  all  capable  of  insertion  into  the  same  ferrule, 
whereby  it  becomes  possible,  by  carrying  one  handle,  to  affix  it  as  desired  to  any 


I 


<^^.4. 


and  Laboratory  Methods.  1409 

particular  form  of  net,  whether  it  be   one  serving  for  the   collecting  of  insects, 
.minute  or  large  aquatic  animals,  and  either  Birge  net,  dip-net,  sieve  or  scoop. 

Explanation  of  Figures. 

Fig.  1. — The  upper  part  of  the  net  with  the  upper  margin  of  the  cloth  "  leg." 
Fig.  2. — The  ring  which  clamps  it  to  the  handle. 
Fig.  3. — Method  of  clamping  suggested. 
•  Fig.  4. — The  collecting  cup  at  the  lower  end  of  the  cloth  "  leg." 


r,     Ring  of  wire  for  attaciiment  of  cord  if  net  is  to  be  used  in  casting. 

rt,    Same  wire  carried  down  and  soldered  to  the  wire  ring 

w,    whicii  forms  the  rim  at  the  base  of  the  cone  of  gauze  netting,  c. 

a,    Mass  of  lead  at  the  tip  of  the  cone  to  add  weight. 

t,     Short  tin  cyHnder  soldered  to  the  base  of  the  cone,  with  a  groove  at  the  upper  end  (g) 

and  a  flange  at  the  lower  (f),  over  which  is  fastened  the  end  of  the  cloth  "leg  "  of 

the  net. 
n,    The  cloth  "  leg,"  which  is  about  eighteen  inches  long,  tapers  toward  the  lower  end,  and 

is  made,  as  before  indicated,  of  cheese-cloth  and  China  silk, 
cl.    The  clamping  ring,  the  end  of  which  (cle)  is  soldered  to  a  block   (b),  which  in  turn  is 

capable  of  being  fastened  firmly  to  a  base  (ba)  by  a  screw  (s)  fitting  into  a  hole  (sh). 

In  the  writer's  nets  these  parts  are  all  of  brass. 
1,     Cup  at  lower  end  of  the  "leg"  made  of  lead  pipe  flared  at  the  upper  rim  and  grooved 

near  this  rim  for  tying  of  the  "  leg." 
st,  Screw  top  such  as  is  used  for  kerosene  cans.     The  material  is  accumulated  in  the  cup 

and  then  by  unscrewing  of  the  cap  allowed  to  run  into  a  bottle,  can,  or  vial  as  may 

be  desired. 
Zoological  Laboratory,  University  of  Nebraska.  ROBT.    H.    WOLCOTT. 


A  Method  of  Determining  the  Comparative  Gravity  of  Alcohol 
when  Dehydrating  by  Osmosis. 

In  dehydrating  by  osmosis  it  is  not  always  easy  to  tell  when  the  two  fluids 
have  reached  an  exact  balance.  The  following  simple  method  is  very  accurate, 
and  practically  no  trouble  at  all.  When  it  is  thought  that  a  balance  has  been 
reached,  take  a  couple  of  drops  of  the  dehydrated  fluid  in  a  dropping  pipette, 
and  carefully  drop  it  into  the  dehydrating  fluid.  If  there  is  any  difference  in  the 
gravity  of  the  two  fluids  the  drops  will  descend  to  the  bottom  with  the  peculiar 
oily  appearance  always  seen  upon  the  mixture  of  two  grades  of  alcohol.  If  the 
grades  are  equal,  however,  the  drops  will  not  be  seen  after  touching  the  surface. 
Chicago.  R-  P-  Woodford. 


A  simple  plan  of  preparing  permanent  specimens  to  demonstrate  any  desired 
structures  in  the  earth-worm,  is  to  place  the  specimen,  after  careful  dissection, 
into  glass  .tubes  of  suitable  size,  one  end  of  which  has  been  sealed  with  a  flame 
before  the  specimen  is  inserted,  the  other  corked  and  sealed  with  sealing  wax 
after  the  tube  containing  the  specimen  has  been  filled  with  3  per  cent,  formalin. 


1410  Journal  of  Applied  Microscopy 

Journal   of  Perhaps  there  is  no  time  throughout 

A  1  •      1     li  yi .  ^^^  year   when    laboratory    apparatus,. 

Applied    Microscopy  especially  in  high  school  laboratories, 

^"'^  suffers  so  much  through  lack  of  atten- 

Laboratory   Methods.  tion  as  during  the  summer  months.     In 


Edited  b     I     R    FI I  lOTT  ^^^  majority  of  school  laboratories  the 

apparatus  lies  untouched  and  forgotten 


Issued  Month^^ from  the^PubiicaHon^  Department       throughout  the  entire  summer  vacation. 
Rochester,  N.  Y. This  no  doubt  does   very   well  where 


SUBSCRIPTIONS:  the  necessary  precautions  are  taken  at 

One  Dollar  per  Year.     To  Foreign  Countries,  $1.25       the  close    of    the    SChool    year  tO    place 
per  Year,  in  Advance.  •'  ^ 

the  equipment  in  such  condition  as  will 


The  majority  of  our  subscribers  dislike  to  have  their  .,            •            ■        i           ^m   •>_     •        i 

files  broken  in  case  they  fail  to  remit  at  the  expiration  preSCrVC   it     Unimpaired     Until   it     IS    de- 

of  their  paid  subscription.     We  therefore  assume  that  no  .        ,      /•                                •               tt                          ^l 

interruption   in   the   series  is  desired,  unless  notice  to  Sired     lOr    USC     again.         HowevCr,     the 
discontinue  is  sent. 


closing  days  of  the  school  year  bring 

so  many  things  to  take  the  attention, 
that  the  condition  in  which  the  working  equipment  of  the  laboratory  is  to  be  left 
is  likely  not  thought  of,  or  receives  only  unsatisfactory  care.  One  has  but  to 
visit  a  few  laboratories  at  this  time  of  year  to  be  convinced  that  the  primary 
reason  why  many  teachers  have  to  work  with  scanty  equipment  is  that  either 
they  or  their  predecessors  have  neglected  to  keep  in  perfect  working  order  what 
has  been  provided  for  them.  To  accumulate  a  satisfactory  equipment,  the  pieces 
as  they  are  obtained  must  not  be  allowed  to  become  worthless  long  before  they 
have  served  their  possible  period  of  usefulness.  School  boards  should  not  rely 
wholly  on  their  teachers  to  care  for  the  public  property,  for  the  care  and  preser- 
vation of  which  they  are  even  more  responsible  than  the  teachers.  If  they  have 
delegated  those  duties  to  the  teacher,  it  is  still  for  them  to  know  that  the  work 
has  been  properly  done. 

The  summer  vacation  affords,  really,  the  only  time  when  apparatus  can  be 
spared  from  the  laboratory  for  repairs.  It  is  the  best  time  to  go  over  the  equip- 
ment carefully  and  make  sure  that  every  available  piece  is  in  the  best  possible 
shape  for  the  coming  year's  work,  and  in  case  repairs  are  needed,  such  may  be 
made  with  no  inconvenience  to  teacher  or  pupils.  Chemical  apparatus  should 
be  cleaned,  and  thus  saved  from  the  action  of  destructive  chemicals.  Physical 
and  optical  instruments  should  be  thoroughly  cleaned,  and  protected  from  dust 
and  moisture.  A  few  hours'  work  now  may  save  an  endless  amount  of  trouble, 
delay,  and  expense,  when  the  apparatus  is  brought  out  for  use  at  the  beginning 
of  another  school  year. 

*  * 

Owing  to  a  short  leave  of  absence  of  the  author,  for  the  purpose  of  securing 
much  needed  rest,  the  series  of  articles  on  Micro-chemical  Analysis,  by  Prof.  E. 
M.  Chamot,  Cornell  University,  is  not  continued  in  this  number,  but  will  appear 
again  next  month. 


and  Laboratory  Methods.  1411 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Guignard,   L.     La   Double  Fertilization    dans       Double  fertilization  in  Zea  mays,  which 
leMais.    Jour.duBot.    15:  1-14, 1901-  j^^g    ^een    suspected    for    some    time, 

and  which  is  believed  to  be  the  cause  of  the  phenomenon  known  as  xenia,  is 
described  in  a  recent  paper  by  Guignard. 

The  mature  pollen  grain  contains,  besides  the  vegetative  nucleus,  two  very 
small  elongated  generative  cells,  each  in  the  form  of  a  slender  rod,  curved  or 
straight.  The  ends  are  often  pointed.  The  protoplasm  of  these  cells  is  much 
reduced  and  difficult  to  distinguish.     Their  nuclei  appear  almost  homogeneous. 

The  two  synergids  and  the  oosphere  are  large.  The  pyramidal  synergids 
occupy  the  summit  of  the  sac,  and  in  many  cases  have  a  large  vacuole  in  the 
base.  They  show  near  the  tip  a  conspicuous  longitudinal  striation,  especially  in 
material  fixed  in  absolute  alcohol.  Their  nuclei  do  not  stain  readily  at  the 
time  of  fertilization. 

The  nucleus  of  the  oosphere  is  very  large  and  contains  much  chromatin. 
The  protoplasm  surrounding  the  nucleus  is,  ordinarily,  highly  granular  and 
much  massed  together  at  the  time  of  fertilization. 

Near  the  oosphere,  sometimes  in  the  median  line  of  the  embryo-sac,  some- 
times at  one  side,  are  the  two  polar  nuclei.  These  nuclei  do  not  fuse  before 
fertilization.  They  have  each  a  relatively  large  nucleus  and  small  amount  of 
chromatin. 

As  many  as  a  dozen  multi-nuclfeate  cells  may  be  found  in  the  much  narrowed 
antipodal  end  of  the  embryo-sac. 

The  pollen  tube,  after  penetrating  the  embryo-sac,  usually  seems  to  dis- 
charge its  contents  into  one  of  the  synergids.  In  one  instance  the  two  elongated 
male  nuclei  were  observed  resting  against  the  base  of  a  synergid  ;  under  high 
magnification  their  chromatin  was  distinct.  One  of  the  male  cells  unites  with 
the  oosphere,  the  other  with  the  polar  nuclei.  These  nuclei  are  bound  together 
by  the  last  male  cell.  Fertilization  proceeds  with  such  great  rapidity  that  it 
could  be  observed  in  very  few  preparations.  In  general,  the  ovules  at  the  base 
of  the  ear  are  first  fertilized. 

In  hybrids  many  ovules  are  not  fertilized. 

After  fertilization,  one  of  the  synergids  usually  persists  for  a  time,  with  its 
contents  finely  granular  and  refractive.  Division  of  the  fertilized  polar  nuclei 
proceeds  with  such  rapidity  that  the  course  of  cell-division  could  not  be  fol- 
lowed. The  first  two  nuclei  of  the  endosperm  are  large,  each  one  having  an 
enormous  nucleolus  and  many  smaller  nucleoli. 

It  is  to  be  regretted  no  figures  are  given.  W.  J.  G.  Land. 

Chicago. 


1412  Journal  of  Applied  Microscopy 

Richards,    H.    M.     Ceramothamnion    codii,    a       Recent    collections    of    Codiicm    tomoi- 
new    Rhodophyceous    alea.     Bull,    Torrey,       ^  i      ■      ti  i     i_      tvj^      h-    l 

Bot.  Club.  28:  257-265^  pis.  21-22,  1901.  f'''"»'  "^^de  in  Bermuda  by  Mr.  Rich- 
ards reveal  an  addition  to  our  list  of 
Rhodophyccie.  The  form  discovered  is  epiphytic  on  Codiiim,  scarcely  visible 
to  the  naked  eye,  appearing  only  as  a  slight  reddening  on  the  host  plant.  The 
new  plant  seems  to  combine  in  itself  the  characters  of  four  other  algae.  In 
habit  it  is  like  Rhodochorton,  a  prostrate  filament  sending  up  erect  filaments 
and  sending  down  rhizoids  into  the  host.  In  structure  it  resembles  Callitham- 
nio/i,  and  Ccramiian,  having  monosiphorous  internodal  cells  and  a  node  of  three 
or  four  rows  of  closely  packed  smaller  cells.  Alternate  nodal  and  internodal 
cells  are  cut  off  by  the  apical  cell  of  each  filament ;  the  nodal  cell  divides  longi- 
tudinally and  transversely  to  form  the  rows  ;  the  internodal  cell  merely  enlarges 
in  all  directions.     Elongated  hairs  may  arise  from  the  nodal  cells. 

Reproduction  is  by  tetraspores  and  polyspores.  The  tetrasporangium  arises 
from  an  upper  cell  in  a  young  node,  often  enclosed  later  by  a  bract-like  growth 
of  other  adjoining  nodal  cells.  After  the  maturity  and  discharge  of  the  cruciate 
spores,  proliferation  of  the  basal  cell  of  the  sporangium  occurs  and  another 
tetrasporangium  is  formed  within  the  old  wall.  Mr.  Richards  reports  finding 
sometimes  four  or  five  older  walls  surrounding  a  developing  sporangium.  The 
antheridia  also  arise  from  nodal  cells,  which  by  their  activity,  spreading  up  and 
down,  completely  envelop  the  internode.  Usually  the  antheridial  plants  are 
separate.  The  polyspores  resemble  quite  strongly  the  favellae  of  CaUithamnioti, 
but  entirely  lack  the  functioning  trichogyne.  They  occur  in  the  axils  of  special 
branches  near  the  tip.  After  careful  investigation  the  author  is  emphatic  in 
declaring  that  polysporic  development  is  purely  asexual  and  that  where  a  hair  is 
present,  it  in  no  way  acts  as  a  trichogyne. 

For  the  reason  that  this  alga  resembles  Rodochortoti  in  habit,  CanUhamnion 
in  cell   and   chromatophore  structure,  Ceramiiim  in  filamentous  form,  Ptilota  in 
polyspores  and  has  besides  proliferation  of  the  sporangium,  the  author  makes  it 
a  new  genus,  Ccramothamuion.     The  paper  is  accompanied  by  two  plates. 
Chicago.  Philip  G.  Wrightson. 

Chamberlain,  Charles  J.,  A.  M.,  Ph.  D.    In-      The  series  of  articles  which  appeared 

structor    in    Botany    in     the    University    of       under  the  above  title  in  The   JOURNAL 
Chicago.      Methods     in    Plant    Histology.  .  ,,  ,  , 

Price  ^i  qo  *^^^    APPLIED    MICROSCOPY    has    been 

thoroughly  revised  and  enlarged  by 
about  one-third  and  is  now  published  in  book  form  by  the  University  of 
Chicago  Press.  Directions  are  given  for  collecting  and  preparing  plant  mate 
rial  of  all  groups  for  microscopic  investigation.  The  various  processes  of  fixing, 
embedding,  sectioning,  staining  and  mounting  are  treated  in  detail.  In  the 
later  chapters  specific  directions  are  given  for  making  those  mounts  which  are 
needed  by  classes  studying  the  development  of  the  plant  kingdom.  It  is 
intended  to  meet  the  requirements  not  only  of  the  student  who  has  the  assist- 
ance of  an  instructor  in  a  well  equipped  laboratory,  but  also  of  the  student  who 
must  work  by  himself  and  with  limited  apparatus.  Formulae  are  given  for 
stains  and  other  reagents. 


and  Laboratory  Methods.  1413 

Lawson,  A.  A.    Origin   of  the   Cones  of  the      Investigations  have  shown  that  multi- 
Multipolar  Spindle  in  Gladiolus.     Bot.  Gaz.  i  •     n  r  i 

30:  145-153,  pi.  12,  1900.  Po'ar  spindles  are  of  very  general  oc- 

currence in  higher  plants,  at  least  in 
the  mother  cells  of  spores.  Since  this  type  of  spindle  formation  does  not 
require  a  centrosome,  and  since  most  investigators  do  not  believe  that  centro- 
somes  are  present,  some  other  explanation  must  be  sought  for  the  ultimate  bipo- 
larity  of  the  spindle.  Those  who  have  investigated  the  multipolar  spindle 
agree  that  it  arises  from  a  weft  of  kinoplasmic  fibers,  but  they  have  not  studied 
the  earliest  stages.  In  1898,  the  present  writer  in  studying  the  pollen  mother 
cells  of  Cobea  scandens  found  that  the  weft  of  kinoplasmic  fibers  arises  from  a 
granular  zone  which  he  designated  as  the  perikaryoplasm.  In  Gladiolus,  as  in 
Cobca,  there  is  a  granular  zone  about  the  nucleus  and  it  is  probably  from  this 
that  the  felted  zone  of  fibers  arises.  The  felted  network  about  the  nucleus 
does  not  grow  uniformly,  but  some  portions  grow  more  rapidly  than  others  and 
appear  as  projections  which  become  the  poles  of  the  multipolar  spindle.  The 
fibers  of  the  spindle  are  formed  by  the  elongating  meshes  of  the  network.  The 
nuclear  membrane,  the  nucleolus  and  the  linin  take  no  part  in  spindle  forma- 
tion.    The  cones  fuse  into  two  groups  to  form  the  bipolar  spindle. 

c.  J.  c. 

Longo,   B.     La    mesogamia    nella    commune  When  Treub,    in    1891,   found    that   in 

Zucca   (Cucurbita    Pepo   Lin.)     Rendiconti  Casuarina   the   pollen   tube   enters   the 

della  R.   Accademia    dei    lincei.     10:    i68-  ,                    ,                    r     i         ■     i           i 

172,  I  go  I.  embryo-sac  by  way  of  the  chalaza,  he 

gave  the  name  chalazogamy  to  this 
peculiar  phenomenon  and  designated  as  porogams  those  plants  in  which  the 
pollen  tube  reaches  the  embryo-sac  by  the  usual  route  of  the  micropyle.  Chal- 
azogamy has  since  been  observed  in  several  other  members  of  the  Amentiferce. 
and  in  Uhniis  a  condition  somewhat  intermediate  between  chalazogamy  and 
porogamy  has  been  described.  According  to  Dr.  Longo  the  pollen  tube  in 
Cucurbita  traverses  the  tissues  of  the  funiculus  and  outer  integument  before 
entering  the  micropyle.     He  proposes  the  name  mcsogamy  for  this  phenomenon. 

c.  J.  c. 

Bessey,  Chas.  E.    The  modern  conception  of  Prof.    Bessey    accepts    Miiller's    view 

the  structure  and  classification  of  Diatoms,  ,  i        ,-,  i-  •         •        i 

^vith  a  revision  of  the  tribes  and  a  rearrange-  that    the    filamentous   condition    IS   the 

ment  of  the  North  American  genera.  Trans.  primitive  one,  and  that  diatoms  should 

of  the   American    Mic.   Soc.  21:   61-8=;.  pi.  ,  ,     ,  •      n       <-, 

r  igoo_  be    regarded    as  typically    filamentous 

rather  than  as  unicellular  forms.  They 
should  then  be  classed  between  the  Peridinialus  on  the  one  hand  and  the  Des- 
midiaceae  and  Zygnemaceae  on  the  other.  The  Zygnemaceae  are  regarded  as 
the  most  primitive  of  the  Conjugatae,  while  the  Desmids  and  Diatoms  are 
believed  to  represent  two  similar  and  somewhat  parallel  genetic  lines  in  which 
the  filaments  tend  to  break  up  rather  early  into  independent  cells.  The  larger 
part  of  the  paper  is  occupied  by  a  key  to  the  tribes  and  genera  of  the  American 
forms.  c.  J.  c. 


1414  Journal  of  Applied  Microscopy 


CYTOLOGY,   EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole,  Cornell  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Rothig.  P.  "Ueber  einen  neuen  Farbstoff  Kresofuchsin  is  an  amorphous  powder 
Namens  Kresofuchsin."  Arch.  f.  Mikrosk.  of  grey  blue  color,  it  is  easily  soluble 
Anat.  Bd.  s6,  igoo.  .  ^••11.1  i-i 

in  acetic  acid  and  aceton,  less  readily 

but  quite  soluble  in  alcohol,  only  very  slightly  so  in  water.  It  is  insoluble  in 
benzol.  The  alcoholic  solution  appears  blue,  the  aqueous  red.  The  former 
stains  elastic  tissue  deep  blue,  mucous,  cartilaginous  and  horny  substances  red- 
dish ;  while  the  latter  does  not  stain  elastic  tissue  at  all,  but  mucous,  cartilage, 
keratin,  as  well  as  nuclei,  are  colored  deep  red.  The  author  suggests  that  the 
stain  contains  two  components  of  which  one  has  an  affinity  for  chromatin,  mucin, 
chondrin,  and  keratin,  while  the  other  takes  to  elastin.  To  determine  the  stain- 
ing of  tissues  by  this  new  agent,  the  investigation  was  made  on  material  fixed 
24  hours  in  a  weak  alcoholic  sublimate  solution  (  9  parts  of  concentrated 
aqueous  sublimate  solution  to  1  part  05  per  cent,  alcohol),  then  washed  with 
alcohols  of  increasing  strength  through  the  iodized  alcohol  till  free  from  subli- 
mate, and  finally  embedded  in  xylol-paraffin.  The  sections  were  fastened  to 
slides  by  weak  alcohol  and  placed  successively  in  xylol,  absolute  alcohol,'  and 
then  the  stain.  A  simple  alcoholic  solution  gives  unsatisfactory  results :  the 
addition  of  hydrochloric  acid  gives  better  differentiation.  Results  are  still  better  if 
a  small  quantity  of  picric  acid  is  added  as  well  as  the  hydrochloric  acid.  These 
substances  are  combined  as  follows  :  0.5  gram  of  kresofuchsin  in  1UI>  c.c.  of  95 
per  cent,  alcohol,  and  o  c.c.  hydrochloric  acid.  With  40  c.c.  of  this  solution,  in 
which  the  particles  of  stain  are  not  dissolved,  are  mixed  32  drops  of  a  picric 
acid  solution,  consisting  of  1  part  concentrated  picric  acid  solu- 
tion and  2  parts  water.  The  sections  remain  2  hours  or  longer  in  the  stain  ;  24 
hours  do  no  harm.  Then  into  95  per  cent,  alcohol,  in  which  they  must  stay  for 
a  long  time.  From  that  into  absolute  alcohol  until  the  remaining  color  is 
removed  and  the  sections  are  dehydrated,  thence  into  xylol  and  balsam.  The 
best  counterstain  is  orange  G,  For  a  final  nuclear  stain  carmin  is  used  ;  if 
haematoxylin  is  used  it  is  necessary  to  overstain  with  it  owing  to  the  picric  acid 
constituent  of  the  kresofuchsin  solution.  After  staining  for  an  hour  the  sections 
are  washed  with  water,  treated  with  alcohol  of  increasing  concentration  and 
stained  several  hours  with  kresofuchsin.  The  violet  color  of  the  nuclei  passes 
into  a  dark  brown  to  brown  red.  The  author  has  also  sought  to  stain  unfixed 
material  with  the  new  stain.  He  obtained  the  same  results  as  in  sublimate  fixed 
tissues.  E.  J.  c. 


and  Laboratory  Methods.  1415 

The  author  has  carried  on   investiga- 
Reuter.  K.     Zur   Frage  der   Darmsresorption.       ^-^^^^   ^^    ^j  ^^^   obstetrcans,    in    regard 
Anat.  Anz.  19:  190-203,  1901.  -^  _  >  o 

to  the    morphological  changes   in   the 

intestinal  epithelium  during  its  periods  of  secretion  and  absorption. 

A  previous  paper  on  the  intestinal  spiral  of  AlytesX^d  to  a  number  of  experi- 
ments on  vertebrates — mouse,  rat,  guinea  pig — accompanied  by  histological  stud- 
ies. Mingazzini  worked  on  the  absorption  processes  in  the  intestine  of  the  fowl,  and 
finds  that  there  are  distinct  changes  in  the  epithelium  corresponding  to  the  dif- 
ferent stages  of  absorption.  The  absorption  of  albumen  is  especially  worthy  of 
mention,  while  fat  absorption  proved  to  be  of  much  less  value  in  the  investiga- 
tion. Mingazzini  finds  the  first  indication  of  absorption  in  the  cells  to  be  a 
granular  clouding  at  the  base  ;  as  the  process  continues  the  cell  contents  become 
watery.     The  whole  process  appears  in  some  sort  as  an  internal  secretion. 

The  evidence  of  these  researches  goes  to  show  that  the  so-called  Gruen- 
hagen's  spaces  are  no  artifact,  but  that  they  are  truly  the  morphological 
expression  of  an  internal  secretion  formed  from  absorbed  nutritive  material. 
By  careless  fixation  and  hardening  on  these  absorption  studies  artifacts  can  easily 
be  made.  Careful  work  has  made  the  following  points  fairly  certain.  The 
whole  absorptive  process,  the  passage  from  alimentary  tract  to  blood  and  lymph, 
takes  place  in  two  chief  stages  :  First,  from  the  lumen  of  intestine  through  the 
epithelium  into  adenoid  tissue ;  second,  the  passage  from  the  adenoid  tissue 
into  the  blood  and  lymph  capillaries.  In  the  first  part  of  the  course,  albumens 
and  fats  pass  readily.  In  this  process  two  stages  are  visible  :  the  taking  up  of 
material  from  the  free  ends  of  the  epithelial  cells,  and,  second,  the  passage  into 
the  basal  part  of  the  cell  below  the  nucleus.  This  is  doubtless  due  to  the 
activity  of  the  living  cell.  In  all  previous  investigations  absorption  has  been 
considered  a  purely  physical  process,  due  to  osmosis,  no  account  has  been 
made  of  the  individuality  of  different  cells.  Quite  notable  variations  in  absorp- 
tion are  due  to  this  factor.  Thanhoffer  thought  that  the  cells  absorbed  through 
pseudopodic  processes,  but  the  author,  repeating  his  experiments  most  carefully, 
could  distinguish  no  changes  ;  the  cell  margins  were  uniform  and  at  rest,  the  fat 
drops  showing  brownian  movement,  but  no  evidence  existed  of  a  mechanical 
taking  up  of  these.  Hardened  material  also  showed  no  change  in  these  margins 
that  could  be  counted  as  any  morphological  change  during  absorption.  In  the 
author's  opinion  the  border  of  the  cylindrical  epithelium  acts  as  an  osmotic 
membrane  through  which  the  soluble  substances  diffuse  into  the  epithelial  cells. 
The  fat  absorbed  in  small  granules  can  be  followed  by  the  use  of  osmic  acid, 
while  the  absorbed  albuminous  substances  lying  above  the  nucleus  can  only  be 
traced  by  the  protoplasmic  conditions.  The  process  of  separation  from  the 
cylinder  cells  is  in  both  substances  entirely  unlike.  That  of  albuminous  sub- 
stances is  intracellular ;  the  whole  process  closely  resembles  mucous  secretion 
with  this  difference,  that  the  excretion  product  cannot  be  fixed  and  stained,  this 
can  be  done  only  for  the  protoplasmic  network  that  contains  it.  The  fat  on  the 
contrary,  is  passed  into  the  intercellular  spaces  between  the  separate  cylinder 
cells.  The  contents  of  these  spaces  passes  into  the  spaces  of  the  adenoid  tissue, 
where  they  probably    undergo    further    changes  by  the  agency  of  leucocytes, 


1416  Journal  of  Applied  Microscopy 

followed  by  their  final  passage  into  the  blood  and  chyle.  The  fat,  appears  to 
pass  into  the  adenoid  meshes  in  comparatively  large  particles  and  here 
disappears  as  if  in  solution.  It  is  frequently  taken  up  by  leucocytes,  in  the 
central  chyle  vessels  which  have  been  filled  by  absorption  abundant  fat  occurs 
in  the  form  of  a  very  fine  emulsion.  The  albuminous  substances  cannot  be 
demonstrated  by  fixatives  and  stains,  but  the  protoplasmic  network  surrounding 
these  substances  can  be  demonstrated.  Till  they  reach  the  central  chyle  vessel 
each  drop  is  undoubted  albumin.  a.  u.  c. 

Benda,  C.    Eine    Makro-und  Mikrochemishe  The  author  applied  Weigert's  method 

Reaction    der     Fettegewebsnecrose.      Vir-  for    neuroglia    demonstration    tO    Other 

chow's  Arch.  Bd.  i6i,  iqoo.  i  •   ^   i      •      i       l-      ^         rr^i 

^  histological  subjects.      The  tissues  are 

hardened  in  a  10  per  cent,  formalin  solution,  then  after  one  or  several  days  put 
into  Weigert's  Neuroglia  mordant,  a  mixture  of  copper  acetate,  chrom-alum 
and  acetic  acid.  This  impregnation  is  best  accomplished  in  an  incubator, 
with  the  exception  of  the  bony  substance,  which  is  a  deep  blue  from  the  copper 
salt,  all  the  other  organs  after  a  week's  treatment  in  this  mordant  take  a  pale 
green  grey  color,  somewhat  bleached  by  washing  with  water.  The  necrotic  fat 
tissue  (  omentum  with  a  little  of  the  pancreas  )  appears  after  24  hours'  treatment 
in  the  incubator  covered  with  green  flakes  or  rust,  both  on  the  surface  and  deep 
in  the  tissue. 

It  was  easily  ascertained  that  it  was  the  copper  solution  that  caused  this 
color  and  that  it  was  the  necrotic  tissue  exclusively  that  had  taken  it.  This  color 
was  so  sharp  that  it  was  possible  to  distinguish  areas  so  small  as  to  be  otherwise 
indistinguishable  macroscopically.  Microscopically  from  preparations  or  from 
frozen  sections  (  prepared  from  the  pancreas  treated  with  the  copper  solution  ) 
it  was  ascertained  that  the  normal  fat  cells  contained  no  trace  of  blue,  while  the 
necrotic  areas  were  clearly  blue  green.  The  most  intense  color  was  in  the 
needle-shaped  fatty  acid  crystals.  Before  embedding  a  counterstain  may  be  used, 
either  alum  or  copper  haematoxylin.  The  latter  stain  brings  out  some  more 
points.  A  section  of  the  copper  treated  tissue  is  stained  with  an  aqueous  solu- 
tion of  crystalized  haematoxylin  and  there  comes,  as  in  the  Weigert  process,  a 
blue  black  color,  while  the  copper  salt  taken  up  goes  over  into  a  copper 
haematoxylin. 

The  fatty  acid  crystals,  however,  retain  their  blue  green  color.  The  author 
thinks  in  consequence  that  the  copper  salt  in  the  crystals  cannot  be  merely 
absorbed,  but  is  present  in  some  different  chemical  substance  too  firmly  to  be 
displaced  by  ha;matoxylin.  This  must  be  a  copper  salt  of  the  fatty  acid.  The 
acids  that  are  here  concerned  are  stearic,  palmitic,  and  oleic.  The  inner  part 
of  the  necrotic  fat  cell  contains  stearic  and  palmitic  acids,  while  the  outer 
part  of  the  cells  contains  a  considerable  amount  of  oleic  acid.  This  new 
reaction  has  several  advantages  for  histological  investigation.  The  other  fat 
staining  methods,  osmic  acid  and  sudan-red,  stain  fats  and  fatty  acids  equally. 
The  sudan-red  stain  is  extraordinarily  adapted  to  show  the  parenchymatous 
inflammation  of  the  pancreas  cells.  But  through  the  intensity  of  the  staining  of 
the  fat,  the  difference  between  the  normal  and  necrotic  cells  disappears,  since 
the  fatty  acids,  which  are  always  surrounded  by  a  fatty  detritus,  can  scarcely  be 


and  Laboratory  Methods.  1417 

recognized.  On  the  contrary,  the  copper  treatment  brings  out  the  blue  green 
crystals  from  the  amorphous  stained  material  and  the  entirely  unstained  normal 
and  abnormal  neutral  fat  drops  are  very  sharp.  The  smallest  indication  of  dis- 
ease can  thus  be  detected  microscopically.  The  author  has  found  in  a  but 
slightly  affected  pancreas  entirely  isolated  necrotic  fat  cells,  which  were  not  to 
be  detected  by  any  other  method.  e.  j.  c. 

Noeoske,  H.    EosinophileZellenund  Knochen-  The      author     considers     the    Staining 

mark,    insbesondere    bei    chirurgischen    In-  technique   for   eosinophile   cells    to    be 
fectionskrankheiten       und      Geschwiilsten.  .  ,       ,  ,  ,i       r   n       • 

Deutsche  Zeitsch.  fiir  Chir.,  Bd.  55,  1900.  very  important  and  uses  the  followmg 

method  :  The  organ  to  be  investigated 
is  fixed  12  to  24  hours  in  4  per  cent,  formol  solution  at  body  temperature,  hard- 
ened in  alcohol  and  embedded  in  paraffin.  Celloidin  was  not  used.  Fixation 
in  Mueller's  fluid  was  less  satisfactory ;  better  results  came  from  5  per  cent,  sub- 
limate solution  and  Altmann's  nitrous  acid,  fixing  from  a  few  minutes  to  2 
hours.  The  sections  ;  from  3  to  6  f.i  thick,  were  stained  with  a  1  per  cent, 
aqueous  solution  of  Gruebler's  eosin  for  2  to  o  minutes,  rinsed  with  water  and 
counterstained  with  the  following  alkaline  methylen  blue :  lithium  carbonate, 
cone.  aq.  sol.  5  pts.,  distilled  water  80  pts.,  alcohol  10  pts.,  methylen  blue 
(cone.  ale.  sol.)  2  pts.  This  staining  solution  is  poured  abundantly  over  the 
eosin  stained  sections ;  it  remains  for  a  half  minute  or  longer,  according  to  the 
method  of  fixing  or  thickness  of  the  section,  washed  off  with  absolute  alcohol, 
cleared  in  xylol  and  mounted  in  balsam.  In  a  section  thus  stained  the  radiating 
structures  about  the  tubercle  bacilli,  the  granules  in  their  immediate  neighbor- 
hood, the  eosinophile  cells,  and  in  part  the  red  blood  cells,  are  all  bright  red, 
while  the  rest  of  the  tissue  is  bluish.  The  alkaline  condition  of  the  methylen 
blue  solution  is  necessary  for  this  differentiation  by  the  eosin.  This  seems  im- 
portant since  weak  neutral  alcoholic  or  aqueous  blue  solutions  remove  the  eosin 
entirely  from  the  section,  which  is  not  bound  firmly  as  a  tissue  element.  Lyons 
blue  has  also  been  used,  made  as  follows  :  20  parts  of  a  1  per  cent,  aqueous 
solution  of  Lyons  blue  with  one  drop  of  ofticinal  solution  of  caustic  potash  boiled 
about  5  minutes  and  diluted  with  20  pts.  of  alcohol.  In  the  same  way  20  parts  of  a 
Bismark  brown  solution  mixed  with  a  drop  of  caustic  potash  solution  boiled  about 
5  minutes  and  diluted  with  20  pts.  alcohol.  Thirty  parts  of  the  first  standard 
solution  are  mixed  with  5  parts  of  No.  2,  while  shaking  ;  to  this  mixture  are 
added  25  pts.  of  alcohol  and  filled  to  100  parts  with  distilled  water.  This 
brownish-violet  stain  is  used  on  the  section  with  cautious  warming,  allowing  steam 
to  form  and  then  cooling  slowly.  The  result  of  this  is  to  give  the  sections  a 
brownish  yellow  tone,  then  the  stain  is  washed  off  with  acid  alcohol  (HCl.) 
whereby  the  brownish  color  changes  to  a  faint  blue.  This  is  followed  by  a  care- 
ful short  wash  with  a  mixture  of  equal  parts  of  pure  anilin,  alcohol  and  distilled 
water.  The  latter  differentiating  fluid  should  act  only  until  the  sections 
appear  a  light  brown;  this  at  most  takes  but  a  few  seconds.  Washing  in  alcohol 
follows  ;  clearing  in  xylol  and  mounting  in  balsam  complete  the  process.  Excel- 
lent plates  illustrate  the  granules  of  the  eosinophile  cells  stained  in  this  manner. 

E.  J.   c. 


1418  Journal  of  Applied  Microscopy 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Sand,    Rene.      Etude   Monographique  sur   le       Since     the     publication     of    Blitschli's 

Groupedes  Infusoires  tentaculiferes.  441  pp.       monograph   of   the    Protozoa    no    more 

24  pis.,  Bru.xelles,  1901.  Extrait  des  Ann.  „„.„.,.:„„„  fli<;p„ccinn  of  these  interest- 
de  la  Soc.  Belg.  de  Micros.  T :  24,  25  et  26.      pretentious  discussion  ot  tnese  interest 

ing  forms  than  this  of  Dr.  Sand  has 
been  attempted.  The  morphology,  taxonomy,  and  especially  the  ecology  of  the 
group,  are  treated  at  length.  Full  synoptic  keys  for  the  determination  of  all 
known  species  are  provided,  and  a  synonymic  bibliography  is  given  for  all 
recognized  forms.  The  author  rejects  the  idea  that  the  Siidoria  have  been 
derived  from  the  Ciliata,  and  seeks  their  ancestors  among  the  Heliozoa.  The 
ciliated  embryos  of  the  Siictoria  are  mere  adaptive  phenomena  .without  phylo- 
genetic  significance,  and  still  further,  according  to  our  author,  the  fundamental 
biogenetic  law  is  not  applicable  to  the  Protozoa.  The  Suctoria  are  found  in 
fresh,  brackish,  and  salt  waters,  most  abundant  in  the  last,  on  Bryozoa  and 
hydroids  at  depths  of  10  to  20  meters.  In  fresh  water  they  are  most  abundant  in 
stagnant  pools.  In  fresh  water  aquarium  cultures  they  appear  after  the  carnivor- 
ous Ciliata,  which  succeed  the  herbivorous  forms.  A  very  large  number  of  kill- 
ing fluids  and  stains  were  tried,  but  best  results  were  obtained  by  the  following 
simple  method  :  Kill  in  2  per  cent,  osmic  acid,  wash  in  water  to  which  a  trace 
of  ammonia  has  been  added,  and  mount  under  a  cover-glass  in  a  drop  of  acetic 
methyl  green  prepared  in  these  proportions : 

Distilled  water, 80  c.  c. 

Alcohol,  0.4°, lOc.c. 

Concentrated  glycerin,  .         .         .  10  c.  c. 

Methyl  green, 0.5  gm. 

Glacial  acetic  acid,        .  .  .  .  2  c.  c. 

As  the  fluid  evaporates  about  the  preparation  10  per  cent,  glycerin  is  added. 
Bordeaux  red,  borax  carmin,  and  a  mixture  of  safranin,  methyl  violet,  and  eosin 
gave  good  results.  The  peduncle  was  stained  by  chrysoidin.  Dehydration  and 
mounting  in  balsam  usually  deforms  the  Suctoria.  c.  a.  k. 

Chapman,  F.  M.  Bird  Life;  a  Guide  to  the  Study  Anew  edition  of  this  well  known  work 

of  our  Common  Birds.     With  75  colored  adds  colored   figures  of  one  hundred 

plates    after    drawings     by    Ernest     Seton  ,      .    j  1  •    1       fir    o^-^^v,  XT,.v,-fV,    a  .,-..^,-; 

Thompson.     Populaf  colored  edition,  8  vo.  selected  birds  of  Eastern  North  Amen- 

pp.  xii  and  195,  with  25  illustrations  and  an  ca.  Mr.  Thompson's  lifelike  and 
appendix  for  use  of  teachers.  Pp.  85,  190 1.  _  •  -.^j  Hrawinfrs  have  been  colored 
D.  Appleton  &  Co.,  New  York.    52.00.  spirited  drawings    nave    oeen    coiorea 

under  the  direction  of  Mr.  Chapman, 
and  are  here  reproduced,  thus  greatly  enhancing  the  value  of  the  book  to  amateur 
students  of  birds  and  to  teachers  of  nature-study  and  biology  in  the  secondary 
schools.  The  teacher's  appendix  contains  many  suggestions  for  utilizing  the 
book  in  the  class-room,  and  especially  in  the  field.  Fortunately  there  is  no  hint 
of  taxidermy  or  birds-nesting  in  its  pages.  c.  a.  k. 


and  Laboratory  Methods.  1419 

Herrick,  F.  H.    The  Home  Life  of  Wild  Birds,      g    means  of  a  portable  tent  and  camera, 
A  New  Method  of   the    Study  and  Photog-  ^  ^ 

raphy  of  Birds.  With  141  original  illustra-  Professor  Herrick  has  solved  the  prob- 
tions  from  nature.,  pp.  xix,  148,  4to,  1901.  ^^^  ^^  ^.^e  successful  Study  of  the  life 
G.  p.  Putnam's  Sons.      $2.50.  .  .    ,,        ,      . 

of  the  nesting  bird,  especially  during 

the  period  between  hatching  and  the  lirst  flight  of  the  fledglings.  The  nest, 
usually  with  the  nesting  bough,  is  moved,  if  necessary,  to  a  place  of  easy  access 
near  the  original  site,  and  the  observation  tent  is  set  up  close  at  hand.  Con- 
cealed within  it  the  observer  can  study  at  close  quarters  the  behavior  of  both 
parents  and  young,  and  can  record  with  the  camera  the  various  phases  of  the 
domestic  life  of  birds.  The  author  gives  very  full  directions  for  the  use  of  his 
method,  and  offers  a  number  of  suggestions  for  its  wider  application  by  others 
who  would  follow  this  fascinating  sport.  The  apparatus  used  is  in  the  main 
very  simple,  and  can  be  easily  managed  by  any  one  familiar  with  photography 
and  possessed  of  the  naturalist's  patience  and  the  ornithologist's  enthusiasm.  A 
brief  review  can  give  but  little  idea  of  the  originality  and  freshness  of  these 
pages  (for  this  is  no  ordinary  "  bird  book  "),  which  bring  within  sight  and  touch 
of  the  reader  the  secrets  of  the  home  life  of  our  native  birds.  The  abundant 
illustrations  secured  under  these  ideal  conditions  record  what  no  naturalist  has 
before  seen,  and  "  what  no  artist  could  hope  to  portray."  Professor  Herrick 
details  in  this  book  the  results  of  his  study  by  this  method  of  fifteen  of  the  land 
birds  of  New  England,  and  his  pages  will  prove  to  be  a  rich  mine  of  suggestive 
information  for  teachers  of  nature-study  and  the  "  new  "  natural  history. 
Though  strictly  scientific  in  method  and  treatment,  the  book  is  well  adapted  to 
the  general  library.  c.  a.  k. 

Zoologisches  Addressbuch     Namen  und  Ad-  Zoologists  everywhere  will  welcome  this 

dressen  der  lebenden  Zoologen,  Anatomen,  °                  ■' 

Physiologen,  und  Zoopalseontologen  sowie  supplement  to  the  admirable  directory 

der  Kunstlerischen  und  technischen  Hiilfs-  j^gued  by  this  firm  in  1895.      The  pres- 

kraften.    Theil.  II,  8vo,  pp.  517,  1901,  Preis  ■'                                                      ^ 

,     M.  6.        Herausgegeben  im  Auftrage  der  ent  issue  is    practically  a  new  edition 

deutschen  zoologischen  Gesellschaft  von  with  the  names,  academic  positions,  ad- 
R.  Fnedlander  &  Sohn,  Berlin. 

dresses,  and  specialties  of  over  seven 

thousand  zoologists  and  collectors.  There  are  full  indices  of  places  and  names, 
and  the  latter  are  also  conveniently  grouped  according  to  the  specialties  given. 
A  list  of  deceased  (since  1895)  zoologists,  and  an  appendix  or  two,  bring  the 
directory  up  to  date.  This  directory  is  indispensable  to  all  publishing  zoologists. 
The  American  section  needs  revision  sadly  in  places,  and  to  this  end  it  is  to  be 
hoped  that  our  zoologists  will  respond  to  the  firm's  request  for  corrections  and 
additions  for  the  next  edition.  These  should  be  sent  to  the  publishers  at  Berlin, 
N.  W.,  Carlstrasse  11.  c.  a.  k. 

Peter,  K.     Mitthdlungen  zur  Entwicklungs-      ^^g  author  gives  a  very  full  account  of 
geschichte  der  tidechse.     II,  Die  Schlund-  °  •' 

spalten  in  ihrer  Anlage,  Ausbildung  und  the  origin,  growth,  disappearance,  and 
Bedeutung.  Arch  f.  Mik.  Anat.  und  Ent-  derivations  df  the  gill-clefts  in  the 
wick.     57:  705-756.     Taf.  30-40,  1901.  .  ° 

lizard.     The  process  was  followed  from 

embryos  of  4  somites  (length  of  embryo  1.8  mm.)  to  those  whose  length  was 
6.3  mm.      The  greater  part  of  this  period  is  passed  before  the  eggs  are  laid. 


1420  Journal  of  Applied  Microscopy 

Reconstructions  were  made  in  wax  from  serial  sections,  and  very  instructive 
figures  of  these  models  are  given  in  the  plates.  The  author  concludes  that  the 
entoderm  only  is  active  in  the  formation  of  the  gill-clefts.  It  is  marked  by 
abundant  mitoses,  and  by  a  thickening  of  the  epithelium,  and  moves  out  to  form 
a  junction  with  the  ectoderm.  The  outer  gill  pockets  appear  only  on  first,  fourth, 
and  fifth  pairs,  and  are  wholly  of  a  secondary  character,  the  ectoderm  being 
passively  drawn  in  by  the  withdrawing  entoderm  of  the  throat  pocket.  The 
closing  membrane  of  clefts  I  to  IV  separates,  but  the  openings  are  later  closed 
and  the  entoderm  retreats  from  the  epidermis.  The  aortic  arches  appear  after 
the  formation  of  the  gill  pockets,  arising  as  outgrowths  from  the  dorsal,  and  later 
the  ventral  aortae.  The  gill-clefts  are  thus  in  no  way  conditioned  upon  the 
appearance  of  aortic  arches.  Six  aortic  arches  appear,  though  in  one  case  a 
seventh  vessel  was  found  and  in  another  instance  the  pulmonary  (VI)  was  miss- 
ing in  the  sixth  gill-arch,  and  was  found  posterior  to  the  sixth  cleft.  The  deriva- 
tives of  the  clefts  are  as  follows :  The  first  cleft,  as  usual,  gives  rise  to  the 
middle  ear  and  the  eustachian  tube.  Epithelial  buds  from  the  dorsal  ends  of 
clefts  I-III  form  the  thymus,  while  the  fourth  disappears  entirely  save  for  some 
ephemeral  epithelial  corpuscles.  The  third  cleft  also  gives  rise  to  some  similar 
elements.  The  fifth  cleft  leaves  no  trace,  but  the  sixth  (on  the  left  side  only) 
gives  rise  to  the  supra-pericardial  body.  All  of  these  organs  are  of  purely 
entoder??ial  origin,  thus  our  author  regards  this  last  pair  of  throat  pockets  as 
belonging  to  the  branchial  category,  and  concludes  that  the  Laccrtilia  have  six 
gill-clefts.  The  utility  of  these  ephemeral  embryonic  organs  is  found  (1)  in 
their  relation  to  permanent  organs  (ear,  thymus,  supra-pericardial  body)  ;  (2)  in 
their  relation  to  the  formation  of  the  ganglia  of  the  seventh,  ninth,  and  tenth 
cranial  nerves.  The  fact  that  open  clefts  are  found  only  in  those  amnota  with 
yolk-laden  eggs  (reptilia,  aves,  echidna)  leads  to  the  further  suggestion  that  these 
open  clefts  facilitate  the  passage  of  nutrient  fluids  from  the  yolk.  c.  a.  k. 

Richardson,  Harriet.     Key  to  the  Isopods  of  'i^^ese     papers    of  Miss    Richardson's 

I  hhe  Atlantic  Coast  of  North  America,  with  dealing    with     the  taxonomy     of     this 

■    descriptions  of  new  and  little  known  species.  .,    ,        i-   .    -i     ,     ,                      r          •       i 

^Proc.U.  S.Nat.  Mus.  23:  493-579,  1901.  widely   distributed  group   of    animals, 

„.  .     ^  u     .  ..      r.  r    ivT     ,       are  welcome  aids  for  all  who  wish  to 

Richardson,      Harriet.      Synopses     of    North 

American  Invertebrates,  VIII,  The  Isopoda.  Study  Isopods.  The  first  paper  is 
Pt.  I,  Am.  Naturalist,  34:  207-230;  Pt.  II,  concerned  only  with  species  of  the 
ibid,  295-309,  1900.  .  -^ 

Atlantic  Ocean  and  of  our  contiguous 

coasts.  Synonomy,  bibliography,  geographical  and  bathymetrical  distribution  of 
all  the  known  species  in  this  habitat  are  given,  and  fourteen  new  forms  described. 
The  second  paper  includes  keys  to  the  fresh-water  forms  and  the  marine  species 
of  all  North  America,  with  only  the  most  general  statements  as  to  their 
geographical  distribution.  c.  a.  k. 

Reighard,  J.,  and  Jennings,  H.  S.    Anatomy  of  A     laboratory   manual    of    mammalian 

the  Cat.    pp.  XX,   498.     With  173  original  anatomy,      complete,     of      convenient 

figures  drawn  by  Louise  Burridge   Jennings.  .                        •        .•        ,1                     1 

Henry  Holt  &  Co.,  1901.  Size,   approximating  the  general   usage 

in    terminology,    and    containing  both 

description  text  systematically  arranged  and  brief  directions  for  preparations  of 


and  Laboratory  Methods.  1421 

material  and  for  dissection,  has  long  been  a  desideratum.  This  need  is  most 
adequately  met  by  Reighard  and  Jennings'  work.  The  book  is  based  upon  a 
decade  of  practical  experience  in  anatomical  instruction  in  the  University  of 
Michigan  and  elsewhere,  and  is  thus  well  adapted  to  the  demands  of  university 
courses,  while  at  the  same  time  affording  a  valuable  guide  to  the  private  student 
of  anatomy  and  a  desirable  adjunct  to  the  high  school  library.  In  symmetry  of 
treatment,  in  freedom  from  extraneous  matter,  in  clearness,  comprehensiveness, 
and  at  the  same  time  brevity  of  statement,  and  in  technical  execution,  the  book 
may  well  serve  as  a  model.  Mrs.  Jennings'  accurate  figures  supplement  the 
text  in  all  important  subjects. 

The  book  contains  adequate  directions  for  the  preparation  of  the  various  sys- 
tems of  organs  and  advocates  the  use  of  formalin  as  a  preservative.  A  5  per 
cent,  solution  of  commercial  formalin  is  injected  into  the  femoral  artery  to  the 
amount  of  300  to  400  cubic  centimeters.  Specimens  thus  injected  may  be  pre- 
served thereafter  by  immersion  in  1  per  cent,  formalin.  Color  and  pliability 
are  better  preserved  by  the  following  method  :  Inject  5  per  cent  formalin  to 
which  has  been  added  one-sixth  of  its  volume  of  glycerin  and  keep  the 
specimen  in  a  tight  box,  wrapping  all  exposed  parts  in  cloths  wet  in  the  injecting 
fluid.  The  suggestion  is  made  that  the  addition  of  fungicides  to  the  injecting 
fluid  might  prevent  the  growth  of  molds  which  soon  attack  exposed  surfaces. 

c.   A.  K. 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

,.,,,,        ,      ,.    ,  This  is  a  consideration  of  the  general 

Pappenheim,  A.     Ueber   das  Vorkommen  ein-  ° 

kerniger  Zellen  ein  gonorrhoischen  Ure-  histology  of  inflammation,  and  it  pre- 
thralsecret.  Virchow's  Archiv  fur  path.  ^^^^^  ^^  admirable  survey  of  recent 
Anat.,  164:  72-119,  1901.  .  .     ,  . 

work  in  this  field.     The  author  dissents 

from  the  view  of  the  Cohnheim  school  and  adopts  in  modified  form  the  original 

view  of  Virchow  that  the  fixed  cells  of  the  part,  as  well  as  the  extravasated  cells 

from  the  blood,  participate  in  the  formation  of  pus.     The  polynuclear  leucocytes 

are  haematogenous  in  origin,  but  the  lymphocytes  and  mononuclear  leucocytes 

he  regards  as  formed  locally  from  the  cells  of  the  connective  tissue  group. 

Mononuclear  cells  occur  in  the  gonorrhoeal  discharge  in   all   stages   of  the 

disease  and  they  are  always  present  in  larger  proportion,  in  relation  to  the  poly- 

nuclears,  in  the  exudate  than  in  the  blood.     The  lymphocytes  are  not  simply  the 

small  typical  lymphocytes  of  the  blood ;  many  are  of  larger  size,  and  forms  as 

large  as  the  great  lymphocytes  of  acute  lymphsemia  are  not  uncommon.     The 

mononuclear  cells  are  found   in  the  pus  in  clumps.     As  the  process  becomes 

more  chronic  there  is  a  considerable  increase  of  mononuclear  leucocytes  and 

particularly  of  lymphocytes.     These  facts  favor  the   histogenetic  origin  of  the 


1422  Journal  of  Applied  Microscopy 

cells.  If  this  theory  be  true  we  have  a  measure  of  the  degree  of  productive  cell 
activity  in  the  appearance  and  still  more  in  the  number  of  the  mononuclear 
leucocytes  and  lymphocytes  in  the  pus,  and  hence  a  point  of  diagnostic  value  in 
determining  the  stage  of  the  inflammation.  Especially  in  gonorrhtea  the  finding 
of  many  gonococci,  eosinophiles  and  few  lymphocytes  would  speak  for  an  acute 
infection,  while  the  finding  of  few  cocci  and  abundant  mononuclear  cells  would 
indicate  an  exacerbation  of  a  chronic  process.  j.  h.  p. 

„.„„._       »;r    .     .     ir  o    •  •  This  method  is  based  on  the  fact  that 

Harris,   H.  r.     A    rsew    Method   of    htaining 

Elastic  Tissue.  Proceedings  of  the  Patho-  haematin  solutions  have  a  remarkable 
logical  Society  of  Philadelphia,  4:  167-16S,  affinity  for  elastin.  Harris  directs  that 
1 90 1 .  \ 

the  stain  shall  be  prepared  as  follows  : 

Haematoxylin,  0.2  gm. ;  aluminum  chloride,  U.l  gm.  ;  .50  per  cent,  alcohol,  100 
c.c.  Dissolve  the  haematoxylin  and  aluminum  chloride,  and  then  carefully  heat 
the  solution  to  the  boiling  point ;  0.6  gm.  of  mercuric  acid  is  now  slowly  added, 
and  as  soon  as  the  mixture  assumes  a  dark  purple  color  it  is  removed  from  the 
flame  and  cooled  rapidly.  The  stain  is  filtered  and  one  drop  of  hydrochloric 
acid  is  added.  The  stain  requires  several  weeks  to  ripen.  It  appears  to  keep 
indefinitely. 

Sections  are  stained  from  five  to  ten  minutes  and  are  then  washed  for  about 
a  minute  in  a  1  per  cent,  solution  of  nitric  acid  in  alcohol ;  the  acid  alcohol  is 
then  thoroughly  removed  with  pure  alcohol,  and  the  sections  are  cleared  and 
mounted.  j.  H.  p. 

Howard,  W.  T.  Observations  on  the  Character  In  1898,  Councilman  published  his 
of  the  Cells  in  the  Exudation  in  Acute  Inter-  ,         ,  ,        .    ,        .-,•    1  1     •^' 

stitial    Nephritis,  with  Special  Reference  to  ^tudy    of     acute     mterstltial    nephritis, 

the    Presence  of   Cells  with    Eosinophilic  He  showed  that  the  disease  is  charac- 

granulations.       American    Journal    of    the  ,     •      j  i_  1         11        1  •    ci^     ^- 

Medical  Sciences,  121 :  1 51-163,  1901.  terized  by  general  and  local  infiltration 

of  the  interstitial  tissue  of  the  kidney 
with  Unna's  plasma  cells.  He  agreed  with  Marschalko  that  these  cells  are 
derived  from  lymphocytes,  and  stated  that  they  are  carried  to  the  kidney  in  the 
blood  current.  In  addition  to  the  plasma  cells  Councilman  found  a  variable 
number  of  lymphocytes  and  polynuclear  leucocytes  in  the  exudation. 

Howard  has  confirmed  Councilman's  observations,  and  in  addition  found 
large  numbers  of  typical  eosinophilic  leucocytes  in  the  interstitial  exudation  and 
in  the  blood  vessels  in  the  three  cases  of  the  disease  which  he  has  studied.  The 
eosinophilic  leucocytes  are  for  the  most  part  brought  to  the  kidney  by  the  blood- 
vessels and  reach  the  interstitial  tissue  by  emigration,  but  they  may  be  formed 
locally  from  plasma  cells.  j.  h.  p. 

Fuchs,  E.  Beitrage  zur  Kenntniss  der  Entste-  F"chs  holds  that  the  eosinophilic  cells 
hung,  des  Vorkommens  und  der  Bedeutung  have  no  single  mode  of  origin.  Eosin- 
•' eosinophiler "     Zellen,      mit      besonderer  ,  ...  ,  ,       ,  j         ..     r 

Berucksichtigung  des  Sputums.     Deutsches       oP^iliC  granules   can   be  formed  out  of 
Archiv  fiir  klinische  Medicin,  63:  427-443,      neutrophilic    granules,    or    from    frag- 
^^'  ments  of  broken  down  red  blood  cor- 

puscles, which  when  ingested  by  cells  transform  the  cells  into  eosinophiles. 
This  probably  explains  the  increase  of  eosinophiles  in  various  haemorrhagic  pro- 
cesses.    Eosinophiles  can  be  formed  in   all   the   tissues.     They  are  especially 


and  Laboratory  Methods.  1423 

numerous  in  those  organs  and  tissues  that  are  exposed  to  bacterial  invasion. 
The  eosinophilic  cells  of  the  sputum  probably  arise  in  the  respiratory  tract. 
They  occur  in  varying  number  in  all  diseases  of  the  respiratory  tract  which  are 
not  associated  with  fever.  In  febrile  conditions  they  ordinarily  do  not  appear 
until  the  temperature  has  returned  to  the  normal. 

For  the  study  of  the  sputum  Fuchs  highly  recommends  a  modification  of 
Teichmiiller's  method.  A  thin  layer  of  sputum  is  spread  upon  cover  slips,  and 
the  preparations  are  fixed  by  drawing  them  three  times  through  the  flame.  They 
are  stained  for  two  minutes  in  a  0.5  per  cent,  alcoholic  solution  of  eosin,  and 
then  decolorized  in  50  per  cent,  alcohol.  Everything  is  decolorized  except  the 
red  blood  corpuscles,  which  retain  the  stain  partially,  and  the  eosinophilic  granu- 
lations.    Counterstain  with  methylen  blue.  j.  h.  p. 

Meek,  E.  R.    Method  of  Staining  the  Elastic      The    writer   has    devised    a    stain    for 

Fibers  of  the  Skin      Boston   Medical    and       gj^Stic      tissue      which     she     considers 
Surgical  Journal,  143:  23-24,  1900. 

superior  to  Weigert's  method,  as  it  is 

less  complicated  and  requires  less  time.       The  sections  are  taken  out  of  strong 

alcohol  and  immersed  in  the  following  solution  : 

Orcein,  ....  3.0 

Absolute  alcohol,  .         .         100.0 

Hydrogen  peroxide,        .  .  40.0 

If  the  sections  are  thin,  three  minutes  suffice  for  staining.  For  differentiation 
the  same  solution  in  which  the  orcein  was  dissolved  is  used  : 

Absolute  alcohol,  .         .  KM). (J 

Hydrogen  peroxide,       .  .  40.0 

For  thin  sections  one  minute  suffices;  the  elastic  fibers  are  then  shown  very 
clearly,  while  the  rest  of  the  tissue  is  lightly  stained.  j.  h.  p. 


GENERAL   PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

„,      „,       J.     „, .    .    ,     or.        J  /-       u        During    recent    years  there    has    been 
Oker=Bloin,  M.     Thiensche  Safte  und  Gewebe  _  °  ■' 

in  physikalisch-chemischer    Beziehung.  manifest  a  growing  tendency  to   apply 

I.  Die  elektrische  Leitfahigkeit  des  Blutes.  ^j^e  methods  and  laws  of  physical 
Arch.  f.  d.  ges.  Physiol.  79:   111-145,    1900- 

II.  Die  Abhangigkeit  der  elektrischen  chemistry  tO  the  Study  of  physiological 
Leitfahigkeit  des  Blutes  von  den  Blut-  problems.  This  series  of  papers  by 
korperchen.  Beitrag  zur  Lehre  von  der  ^  r  r  j 
Leitfahigkeit  der   Suspensionen.    Ibid.    79 :  Oker-Blom    forms    the    most   extensive 

5'°7533-.  i9°o-  ,,...,.,         ,       ^^,  and    detailed    contribution    along  this 

III.  Die  Durchlassigkeit  der  rothen  Blut-  ° 
korperchen  fiir    verschiedene   Stoffe,  beur-  line    which     has    yet     appeared.      The 
theilt  nach  der  elektrischen  Leitfahigkeit.  fi^st    paper     of     the     series     dealing 
Ibid.     81:   167-221,   1900.  ^   ^                                                             ** 

IV.  DieelektromotorischenErscheinungen       with  the   electrical   conductivity  of  the 

am  ruhenden    Froschmuskel.      Ibid.     84:      ^lood  is  introduced   by    an    excellent 
191-259,  I90I.  •' 

brief  summary   of  the   present  knowl- 
edge of  solutions,  from  the  physico-chemical  standpoint.     In  this  study  the  resist- 


1424  Journal  of  Applied  Microscopy 

ance  of  a  certain  quantity  of  blood  in  a  standard  cell  was  compared  by  the 
"  telephone  method  "  with  a  known  resistance  on  a  rheostat.  The  results  are  for 
convenience  expressed  10,000  times  greater  than  their  absolute  value  in 
Kohlrausch  units.  It  was  found  that  the  conductivity  of  defibrinated  ox  blood 
was  from  52.50  to  70.89,  while  an  equal  amount  of  serum  alone  showed  a  con- 
ductivity of  from  114.40  to  131.08.  Furthermore  the  conductivity  of  a  .7  per 
cent,  solution  of  NaCl  under  similar  conditions  was  124.10,  very  nearly  the  mean 
value  for  the  serum  determinations  ;  thus  indicating  in  another  way  that  such  a 
solution  of  NaCl  is  a  "  physiological  normal  "  solution.  The  degree  of  dissoci- 
ation of  the  serum  electrolytes  was  determined  to  be  about  .65  to  .76.  It  was 
found  that  between  20°  and  40°  C.  the  conductivity  rises  with  the  temperature. 
There  is  no  difference  in  the  conductivity  between  arterial  and  venous  blood. 
The  electrolytes  of  the  blood  corpuscles  contribute  very  little  to  the  carrying  of 
the  current  while  they  are  in  the  corpuscles,  but  as  soon  as  they  diffuse  out  into 
the  serum  they  become  active.  From  this  it  follows  that  by  measuring  the  con- 
ductivity of  both  serum  and  corpuscles  a  method  is  given  whereby  it  may  be 
determined  what  quantity  of  an  electrolyte  added  to  the  blood  has  entered  into 
the  corpuscles  and  how  much  has  remained  in  the  serum.  The  conductivity  of 
the  blood  is  not  simply  proportional  to  the  serum  content,  but  is  considerably 
influenced  in  some  way  by  the  resistance  of  the  corpuscles  present. 

To  determine  precisely  this  effect  of  the  suspended  corpuscles  on  the  con- 
ductivity of  the  whole  blood  is  the  purpose  of  the  second  paper  in  the  series.  A 
large  number  of  experiments  were  performed  to  test  the  effect  on  the  conduct- 
ivity of  solutions  of  different  electrolytes  of  the- presence  of  suspended  particles 
of  some  non-conductor,  as  for  example  sand  grains.  The  solutions  of  the  elec- 
trolytes were  made  in  gelatin,  which  was  allowed  to  harden  and  thus  hold  the 
sand  grains  in  any  desired  arrangement.  Essentially  the  same  methods  of 
measuring  the  resistance  were  used  in  these  experiments  as  in  the  preceding  in- 
vestigation. The  results  show  that  the  electrical  conductivity  of  a  solution  is 
mechanically  influenced  by  the  presence  of  non-conducting,  suspended  particles, 
and  that  this  influence  is,  within  certain  limits,  independent  of  the  size  of  the 
particles  and  the  conductivity  of  the  solution,  but  is  markedly  affected  by  the 
amount  and  arrangement  of  the  non-conducting  bodies.  Formulas  are  given 
by  means  of  which  an  absolute  value  for  this  effect  can  be  determined.  From 
parallel  experiments  it  appears  that  the  blood,  in  so  far  as  its  conductivity  is 
concerned,  behaves  as  an  electrolyte  in  which  the  corpuscles  play  the  part  of 
suspended  non-conducting  bodies. 

The  third  paper  in  the  series  discusses  the  permeability  of  the  red  blood 
corpuscles  for  different  substances.  The  method  used  was  that  which  has  been 
indicated  above,  namely  the  measurement  of  the  electrical  conductivity  of  the 
blood  after  the  addition  of  the  substance  to  be  tested.  Solutions  were  made  in 
both  water  and  serum.  When  potassium  chloride,  potassium  sulphate,  or  mag- 
nesium sulphate  are  dissolved  in  serum  and  mixed  with  defibrinated  ox  blood, 
by  which  process  the  osmotic  pressure  of  the  serum  is  of  course  raised,  they 
only  enter  the  corpuscles  to  a  very  slight  degree.  On  the  other  hand,  under 
similar  conditions  ammonium  sulphate  and  ammonium  chloride  are  taken  up  by 


and  Laboratory  Methods.  1425 

the  corpuscles  to  a  much  greater  extent.  When  these  same  substances  in  the 
form  of  water  solutions  are  mixed  with  the  blood  it  is  found  that  potassium 
chloride  and  potassium  sulphate  are  only  taken  up  by  the  corpuscles  when  the 
osmotic  pressure  of  their  solutions  is  higher  than  that  of  the  serum.  Magnesium 
sulphate  when  in  solution  of  lower  osmotic  pressure  than  the  serum  does  not 
enter  the  corpuscles.  Ammonium  chloride  and  sulphate  in  water  solutions  are 
taken  up  by  the  corpuscles  whether  the  solutions  are  hypertonic  or  hypotonic. 
This  method  of  the  electrical  conductivity  gives  excellent  results  in  the  measure- 
ment of  the  permeability  of  the  corpuscles  for  electrolytes,  but  is  not  so  well 
adapted  for  the  treatment  of  the  resorption  of  non-conductors,  as  for  example,  urea. 
The  last  paper  deals  with  the  so  called  "  demarcation  current  "  of  a  resting 
muscle  which  has  been  injured  in  some  way.  The  sartorius  muscle  of  the  frog 
was  used  and  this  was  injured,  either  by  wounding  with  a  knife  or  by  the  appli- 
cation of  chemicals  to  its  surface,  or  by  both  methods  in  combination.  The 
arrangement  of  the  experiments  was  as  follows  :  on  the  ends  of  two  "normal," 
unpolarisable  electrodes  filled  with  .In  KCl  were  placed  secondary  electrodes, 
one  of  which  was  filled  with  an  indifferent  fluid  -  .In  NaCl  -  and  the  other  with 
the  substance  \vhose  effect  on  the  muscle  was  to  be  tested.  On  the  ends  of 
these  secondary  electrodes  the  muscle  from  a  curarised  frog  was  laid.  Before 
beginning  each  experiment  the  muscle  was  tested  and  found  to  give  no  current. 
From  the  primary  electrodes  wires  were  led  to  a  suitable  apparatus  for  measuring 
the  current.  With  such  an  arrangement  it  was  found  that  distilled  water  in  con- 
tact with  the  surface  of  the  muscle  produces  a  negative  electrical  condition  at 
that  point.  After  a  short  time  (  fifteen  to  thirty  minutes),  however,  this  affected 
area  becomes  positive  with  reference  to  the  rest  of  the  muscle,  and  later  again 
changes  to  negative.  If  very  dilute  solutions  of  KCl  are  brought  in  contact  with 
the  muscle,  analogous  phenomena  appear,  the  only  difference  being  that  in  this 
case  the  negative  and  positive  phases  of  the  current  are  of  shorter  duration  than 
when  water  is  used.  The  sheath  of  the  muscle  fibril  has  an  important  influence 
on  the  electromotive  force  of  the  muscle,  the  current  becoming  weaker  as  this 
surface  layer  becomes  more  and  more  injured.  The  author  believes  that  all 
these  phenomena  can  be  brought  into  agreement  with  the  laws  of  physical  chem- 
istry. It  appears  that  the  contractile  substance  and  the  sheath  are  affected  sep- 
arately by  the  irritating  agent.  In  case  of  each  there  arise  decomposition 
products,  whose  positive  ions  move  faster  than  their  associated  negative  ions. 
The  diffusion  of  these  ions  is  the  primary  cause  of  the  electrical  phenomena,  and 
the  changes  in  the  direction  of  the  current  are  the  results  of  changes  in  the 
permeability  of  the  sheath.  This  paper  forms  an  important  step  towards  the 
bringing  of  animal  electricity,  one  of  the  most  peculiar  of  physiological 
phenomena,  under  physical  and  chemical  laws.  r.  p. 

Courtade,     D.     L'Irritabilite    dans    la      Serie       These  two  numbers  in  the   "  Scientia  " 
animale.     Paris  (Carre  &  Naud).     Pp.  86,       ^^^-^^    ^^^  ^^^j^j    jj^^j^    hand-books   on 

Bonnier,    P.    L'Orientation.    Paris  (Carre  &      their  respective  subjects.     Their  pur- 

p.  90,  1900.  pose  is  not  the  publication  of  new  facts 

or  theories,  but  rather  to  give   a  clear,  concise,  and  more  or  less  elementary 


1426  Journal  of  Applied  Microscopy 

discussion  of  the  different  phases  of  the  subjects  treated.     Considering  the  space 
at  the  authors'  disposal,  this  end  is  very  well  attained. 

The  first  of  the  two  books  is  a  discussion  of  all  the  so-called  irritable  phe- 
nomena displayed  by  the  living  organism.  After  a  brief  but  excellent  intro- 
ductory historical  chapter,  the  morphology  and  chemical  composition  of  living 
matter  are  treated  in  a  very  general  way.  A  chapter  is  then  given  to  the 
discussion  of  the  external  conditions,  such  as  oxygen  supply,  heat,  nutriment,  etc., 
necessary  for  the  performance  of  vital  functions.  Under  the  caption 
"Nutritive  Irritability  "  the  process  of  digestion  is  described,  particular  attention 
being  given  to  the  action  of  the  different  ferments.  Under  "Functional  Irritability" 
are  treated  the  subjects  of  animal  heat,  the  phenomena  of  movement,  including 
the  various  "  taxes  "  and  "  tropisms,''  animal  electricity  and  phosphorescence. 
Considerable  space  is  devoted  to  the  functions  and  activities  of  the  nervous 
system,  the  subject  being  introduced  by  a  somewhat  doubtful  comparison  of  the 
nucleus  of  the  cell  to  the  central  nervous  system.  A  section  is  given  to  the 
phylogenetic  development  of  the  nervous  system.  The  final  chapter  takes  up 
briefly  the  nature  of  irritability  in  general,  from  the  standpoint  of  the  chemical 
relations  of  protoplasm. 

The  other  book,  "L'Orientation,"  is  psychological,  both  in  point  of  view  and 
treatment.  Its  purpose  is  to  show  how  an  organism's  notions  of  its  relations  in 
space  are  developed.  The  term  "  orientation  "  is  used  throughout  in  this  general 
sense  of  "  space  relation,''  rather  than  in  the  more  ordinary  restricted  physiolog- 
ical sense  of  a  particular  sort  of  a  reaction  to  certain  stimuli.  Such  subjects  as 
the  muscle  sense,  the  senses  of  active  and  passive  movement,  equilibrium,  and 
tactile,  visual  and  auditory  space  localization  are  discussed,  among  others.  A 
chapter  is  devoted  to  the  migratory  and  homing  instincts  of  birds.  These  the 
author  explains  (?)  as  due  to  heredity  and  a  well  developed  sense  of  direction. 

R.  p. 

Levene,  P.  A.  This   series   of    papers,   while   dealing 

I  On  the  Nucleoproteids  of  the  Brain,  ^^j^i^  technical  matters  in  the  subject  of 
Arch.    Neurol,    and  Psychopathol.     V.  II,  p. 

1-14,  1899.  physiological  chemistry,  are,  on  account 

II.  Iodine  Compounds  in  the  Tissues  after  of  ^heir  breadth  of  view  and  clear 
Administration  of  Potassium  Iodide.     Ibid,  p. 

15-20,  1899.  method  of  presentation,  of  considerable 

III.  On  the  Absorption  of  Proteids  (with  interest  to  the  biological  reader.  The 
I.  Levin).     Ibid,  p.  551-556,  1899.  ° 

IV.  Embryochemical  Studies.  Ibid,  p.  557-      first  paper  of  the  series  discusses  the 

5^5>  'S99.  .    ,  „  ,    .      , .      .  ^  „  . ,       chemical   nature   of  the  nucleoproteid 

V.  1  he  Chemical  Relationship  of  Colloid, 

Mucoid  and  Amyloid  Substances.  Ibid,  p.  571-      or  chromatin  of  the  brain.      Extracts  of 

573.  1899.  ^Y^Q  brains  from   freshlv   killed   calves 

(Reprints  dated  1900.) 

were  used.  The  nucleocompound  ob- 
tained by  this  extraction  was  found  to  be  a  true  nucleoproteid,  differing  from 
other  nucleoproteids  by  its  low  percentage  of  phosphorous,  by  the  nature  of  its 
xanthin  bases,  and  by  the  large  amount  of  proteids  bound  to  its  nuclein.  There 
does  not  seem  to  be  any  evidence  of  more  than  one  nucleoproteid  in  the  brain, 
but  on  the  contrary  it  seems  probable  that  the  chromatin  of  the  cytoplasm  (in 
the  Nissl's  granules  of  the  ganglion  cells)  does  not  differ  chemically  from  that  of 
the  nucleus. 


and  Laboratory  Methods.  14'i7 

The  second  paper  deals  with  the  question  of  how  a  drug  acts  on  the  cell,  that 
is,  whether  by  merely  changing  the  physical  condition  of  cell  or  tissue,  or  by 
forming  new  chemical  compounds  with  the  cell  constituents.  The  method  of 
attacking  the  problem  was  to  examine  the  eggs,  and  finally  the  tissues  of  hens 
that  had  been  given  regularly  certain  doses  of  potassium  iodide,  in  order  to  deter- 
mine whether  any  actual  combination  of  iodine  with  the  cell  substance  had  taken 
place.  In  the  analysis  of  the  eggs  only  iodides  were  found  ;  that  is,  no  iodopro- 
teids  had  been  formed.  Analysis  of  all  the  principal  tissues  of  the  body  gave 
the  same  result,  so  that  it  appears  that  the  drug  does  not  act  by  forming  new 
chemical  compounds. 

The  third  paper  in  the  series  deals  with  the  question  of  whether  the  proteid 
material  is  taken  up  from  the  digestive  tract  by  the  blood  or  by  the  lymph.  The 
method  employed  was  to  inject  into  a  ligated  portion  of  the  alimentary  tract  a 
certain  amount  of  an  artificially  prepared  iodoproteid.  After  some  time  the 
lymph  of  the  animal  was  collected  and  tests  for  iodoproteid  were  made.  In  the 
eight  experiments  no  iodoproteid  compounds  were  found  in  the  lymph,  the  work 
thus  tending  to  confirm  the  old  view  that  the  proteids  are  absorbed  by  the  blood. 

The  fourth  paper  describes  some  of  the  chemical  changes  which  take  place 
in  the  developing  egg,  the  point  of  view  being  that,  since  in  the  process  of  devel- 
opment assimilation  is  greatly  in  excess  of  dissimilation,  a  chemical  study  of  the 
egg  at  different  stages  ought  to  furnish  a  good  opportunity  for  the  working  out 
of  the  synthetic  processes  in  the  metabolism  of  the  organism.  As  material,  cod- 
fish eggs  were  used.  The  results  of  analyses  of  eggs  at  different  stages  of  devel- 
opment seem  to  indicate  that  the  process  of  synthesis  is  preceded  by  decomposi- 
tion, since  immediately  after  fertilization  the  proteids  decrease  in  quantity  and 
basic  nitrogenous  substances  are  formed.  Later  the  proteids  grow  in  quantity  and 
complexity.     The  amount  of  mineral  salts  in  the  egg  increases  with  development. 

The  last  paper  in  the  series  is  a  preliminary  communication  in  regard  to  a 
technical  point  with  reference  to  the  relation  or  possible  identity  of  colloid, 
mucoid  and  amyloid  substances.  r.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Buchner.    Immunitat.   Hyg.   Rund.  11:    301,      The  dispute  between  the  chemical  and 

biological  theory  of  immunity  has  been, 
in  the  last  few  years,  carried  on  largely  by  the  two  leaders,  Buchner  and 
Metschnikoff,  the  former  holding  to  the  chemical  theory  of  immunity,  and  the 
latter  being  the  leading  exponent  of  the  biological  theory.  It  has  been  evident 
that  the  two  schools  have,  in  recent  years,  been  rapidly  coming  together,  each  of 
them  admitting  certain  conclusions  of  the  opposing  school.  In  an  address 
recently  given  by  Buchner  this  harmony  of  the  views  is  distinctly   stated,  and 


1428  Journal  of  Applied  Microscopy 

Buchner  admits  that,  though  in  earlier  years  he  was  inclined  to  place  no  weight 
upon  the  phagocytosis  theory,  he  has  in  recent  years  come  to  believe  that  this 
theory  of  Metschnikoff  represents  a  truth.  The  phagocytes  are  active  agents  in 
immunity,  and  produce  their  effect  from  the  fact  that  they  eliminate  certain 
poisonous  products  which  are  the  direct  cause  of  the  repressing  action  upon  the 
invading  bacteria.  With  this  admission,  the  French  and  German  schools  are 
very  close  together.  Buchner  makes  a  further  classification  of  these  poisonous 
products  produced  in  the  animal  body,  showing  that  they  are  to  be  divided  into 
two  different  types.  One  class  is  destroyed  by  a  heat  of  (30°  C,  and  the  other  is 
not  destroyed  by  this  temperature.  He  thinks  that  these  two  should  be  separ- 
ated from  each  other  and  would  call  the  first  group,  which  can  resist  the  temper- 
ature of  60°,  by  the  name  of  alexines,  while  the  latter  group,  which  cannot  resist 
this  temperature,  he  calls  "  anti-bodies,"  (antikorper ),  for  example  anti-toxin, 
anti-haematin,  etc.  h.  w.  c. 

Sternberg,  Carl.    Zur  Kentniss   des    Aktino-      The     author    makes   a   study  of   three 
mycespilzes.  Hyg.  Rund.  II  :  207,  iqoi.  r  •  ■      •  r 

cases  of   actmomycosis   m  man,  from 

which  he  succeeds  in  isolating  three  different  cultures  of  Actinomyses.     These 

cultures,    when    inoculated    into    guinea    pigs    and    rabbits,    produced    typical 

abscesses  in  which  great  quantities  of  the  fungus  were   found   in  form   of   rods. 

The  author  concludes  that  the  species  which  he  has  described  are  identical  with 

the    Wolf-Israel  Actinomyses   although   the   results  of   animal  inoculations  are 

somewhat  different.     He  believes  that  the  Actinomyses  species,  liable  to  attack 

the  human  body,  are  two  in  number.     One  of  these  he  has  described,  while  the 

other  is  that  described  by  Bostroem.  h.  w.  c. 

Miscellaneous  Studies  of  Cancer.  The  whole  of  number  52  of  \\\^  Journal 

of  the  Boston  Society  of  Medical  Science  issued  October,  23,  1900,  is  taken  up 
with  a  report  of  the  series  of  studies  on  cancer  made  at  the  Harvard  Medical 
School.  There  are  several  distinct  papers  devoted  to  various  aspects  of  the 
problem.  The  problems  considered  include,  statistics  of  cancer  ;  the  etiolog}'  of 
cancer  ;  a  report  on  the  presence  of  "  Plimmer's  Bodies  "  in  cancer  ;  the  study 
of  tumors  and  sporozoa  in  fishes  ;  a  paper,  with  a  figure,  on  a  reconstruction  of 
a  cancer  nodule,  and  a  series  of  reports  on  culture  experiments  made  with 
carcinomatous  tissue.  The  papers  are  useful  as  giving  an  outline  of  various 
facts  known,  but  they  reach  no  positive  conclusion  as  to  the  cause  of  this 
mysterious  disease.  h.  w.  c. 

^    ...    .         TT  u     J     ,0  ,    ,  By  improving  his  method  of  technique 

u.  a  Arrigo.     Ueber  die  Gegenwart  und  uber  ./         r-  o  -1 

die  Phasen  des  Kochschen  Bacillus  in  den  the  author  has  made  a  careful  study  of 
sogenannten  skrophulosen  Lymphdrussen.  scrofulous  glands  for  the  purpose  of 
Hyg.  Rund.  II  :   292,  1901.  ^  r      r 

determining  to  what  extent  they  are 
tuberculous  in  origin.  His  general  conclusion  is  that  all  so-called  scrofulous 
glands  are  tubercle  lesions.  He  further  finds  that  in  ages  from  four  to  twelve 
years  the  cervical  and  sub-axillary  glands  are  most  liable  to  be  affected,  while  in 
later  life  the  axillary  glands  are  more  commonly  attacked.  The  author  further 
finds  that  the  bacilli  in  scrofulous  glands  have  certain  morphological 
peculiarities.  h.  w.  c. 


and  Laboratory  Methods.  1^29 

Busquet.  Transmission  de  la  tuberculose  par  The  author  has  described  a  new  source 
les  timbrespost.  Hyg.  Rund.  11:  289,  of  distribution  of  tuberculosis  by  pos- 
tage stamps  which  the  collector  of 
postage  stamps  moistens  with  his  tongue  for  the  purpose  of  sticking  them  into 
stamp  albums.  A  case  of  tuberculosis  having  such  a  source  was  brought  to  the 
author's  attention,  the  patient  being  a  soldier,  who  was  a  stamp  collector.  The 
author,  thinking  that  the  stamps  were  possibly  the  source  of  the  trouble,  made 
careful  studies  of  these  stamps,  inoculating  guinea  pigs  with  a  watery  solution 
made  from  them,  and  in  every  case  the  animals  showed  tuberculosis.  The 
author  thinks  that  this  is  a  new  source  of  distribution  which  should  be  guarded 
against.  h.  w.  c. 

,     ,  .    u        J  »;  11'       T-.  J  J        After  alluding;  to  the  close  biological 

Leclainche   and  Vallee.     Etude   comparee   du       -^i^v-i    anu^i  ^  t, 

vibrion  septique  et  delabacteriesdu  charbon       relations  of  the   bacterium    of    sympto- 
SQo^i'g^o^"'^'''"     ^""-  '^^  '■'"''■  ^^''■'^'      matic  anthrax    and  the    septic  vibrio, 

the  authors  state  that  it  is  possible  to 
distinguish  the  two  microbes  for,  while  the  septic  vibrio  produces  long  forms 
both  in  the  serum  of  the  specific  oedema  and  in  the  peritoneal  sac  of  guinea 
pigs,  these  are  constantly  absent  in  the  case  of  symptomatic  anthrax.  The  same 
methods  for  immunising  against  anthrax  are  applicable  to  the  vibrio,  and  the 
immunising  serums  are  in  both  cases  rigorously  specific.  The  same  holds  good 
for  agglutination  by  these  serums.  Animals  vaccinated  against  anthrax  are  not 
immunised  against  the  vibrio;  and,  reciprocally,  vaccination  against  the 
septicaemia  does  not  protect  against  anthrax.  h.  w.  c. 

Funck.  A  Preliminary  Note  on  the  Etiological  The  author  claims  to  have  finally  solved 
Agent  in  Vaccinia  and  Variola.  Brit.  Med.  (.}^g  problem  of  the  specific  agent  in 
Jour.,  p.  448,  1900.  .  .  ,  .    ,  .  J. 

vaccme  virus  and  variola.     According 

to  his  views  this  is  not  a  bacterium,  but  a  protozoon,  belonging  to  the  group  of 
Sporozoa,  which  the  author  has  named  Sporidium  vaccinale.  This  protozoon 
he  finds  uniformly  present  in  the  vaccine  pustules,  as  well  as  those  of  variola. 
He  finds  that  vaccine  material,  treated  in  such  a  way  as  to  render  it  impossible 
for  bacteria  to  live,  still  contains  these  protozoa.  The  most  significant  part  of 
his  work  consisted  in  separating  the  organisms  in  question,  in  what  seems  to  be 
a  pure  culture.  His  method  is  as  follows :  In  the  pustules  he  finds  that  the 
protozoon  produces  sporocysts  which  are  of  tolerably  good  size.  These  sporo- 
cysts  are  large  enough  for  him  to  fish  out  successfully  with  a  platinum  needle. 
He  then  places  them  in  a  small  amount  of  sterilized  agar  and  makes  an  emul- 
sion with  a  sterilized  liquid.  Such  an  emulsion  he  found  capable  of  reproducing 
the  disease,  and  he  is  convinced,  consequently,  that  this  protozoon  is  the  long 
sought  cause  of  variola  and,  probably  therefore,  of  small  pox.  If  these  conclus- 
ions are  correct,  they  will  doubtless  inaugurate  an  new  era  in  the  study  of  small 
pox.  H.  w.  c. 

Fisher,  Alfred.  Die  Empfindlichkeit  der  Bak-  Fisher  has,  in  this  paper,  published  a 
terienzelle  und  das  baktericide  Serum.  ^^j.  pregnant  series  of  experiments 
Zeit.  f.  Hyg.,  35:   I,  1900.  ,      ^.         ^  ,  ,,  r\,        , 

bearing  upon  the  problem  of  the  alex- 

ines  in  the  blood.     Fisher  is  clearly  of  the  opinion  that  the  destruction  of  bacteria 


1430  Journal  of  Applied  Microscopy 

in  fresh  blood  is  to  be  explained  upon  some  other  ground  than  the  presence  of 
poisonous  substances,  which  Buchner  has  called  alexines.  Fisher's  experiments 
have  been  in  the  line  of  transferring  bacteria  from  one  culture  medium  to  another 
containing  a  larger  amount  of  salt,  and  in  all  cases  he  finds  that  the  change  from 
one  medium  to  another  is  followed  by  a  granulation  of  the  protoplasm  in  the 
bacteria  body,  and  a  greater  or  less  destruction  of  the  bacteria,  quite  similar  to 
that  which  has  been  described  under  the  influence  of  the  so-called  alexines. 
Fisher  experiments  with  a  large  number  of  micro-organisms,  stationary  and 
motile,  including  all  types.  He  finds  that  this  granulation,  which  he  calls  "plas- 
motyse,"  is  a  very  common  occurrence,  as  the  result  of  a  change  in  culture 
media.  He  believes  that  it  is  due  to  purely  physical  phenomena  affecting  the 
protoplasm  and  is,  therefore,  not  inclined  to  place  much  weight  upon  the  action 
of  alexines.  H.  w,  c. 

Gromakowsky.    Varieties  of  Pseuclodiplitiieria      The  author  states  that  there  are  three 
Bacilli.      Cent.  f.  Bac.  u.   Par.    i,   28:   136,       kinds      of      pseudodiphtheria      bacilli, 

which  are  distinguishable  by  their  cul- 
tural characters  and  by  their  growth  in  bouillon  :  (1)  A  relatively  thick  rodlet 
of  variable  length,  which  does  not  render  bouillon  turbid.  It  resembles  Loeffler's 
bacillus  in  staining  by  Neisser's  method,  and  in  the  acid  reaction  which  it 
develops  in  bouillon.  Its  distinguishing  characters  are  its  large  size  and  its  cul- 
tural appearances.  (2)  A  rodlet,  of  medium  thickness  and  length,  which  after 
25  hours  at  36°  C,  renders  bouillon  markedly  turbid  and  causes  a  copious 
deposit.  Morphologically  and  culturally,  it  closely  resembles  Loeffler's  bacillus. 
Its  distinguishing  feature  is  the  absence  of  acid  reaction  in  bouillon  and  a  nega- 
tive Neisser  staining,  (o)  A  short,  thin  rodlet  which  causes  only  slight  cloudi- 
ness in  the  medium  and  a  scanty  deposit.  It  has  some  resemblance  in  appearance 
to  Loeffler's  bacillus.  h.  w.  c. 

Clowes  and  Houston.   The  Bacterial  Treatment      ^^   a  report   on  the  general  subject  of 
of  London  Sewage.    Brit.  Med.  Jour.  p.  287,      London  sewage,  bacteriological  exami- 
^    ■  nation  is  made  to  determine  whether 

the  bacterial  method  of  treating  sewage,  which  is  now  coming  to  be  so  widely 
adopted,  is  efficient  in  removing  bacteria  as  well  as  chemical  products.  The  con- 
clusion reached  is  that,  although  the  water  may  be  chemically  purified,  it  is  hardly 
improved,  so  far  as  concerns  bacteria.  Crude  sewage,  which  contains  seven 
million  bacteria  before  treatment,  contains  about  five  millions  afterward,  a  reduc- 
tion of  only  thirty  per  cent.  The  reduction  in  number  of  coli  bacillus  is  about 
the  same  per  cent.,  a  fact  which  indicates,  of  course,  that  if  the  sewage  contains 
typhoid  bacilli,  the  treated  sewage  will  also  contain  them,  only  in  somewhat  less 
numbers.  In  other  words,  the  bacteria  treatment  of  sewage  has  practically  no 
influence  in  rendering  sewage  less  likely  to  distribute  sewage  borne  diseases. 

H.  w.   c. 


and  Laboratory  Methods.  1431 

NOTES  ON    RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

tenSiethoff,  E.  G.  A.     Eine  einfache  Construe-  The  accompanying      rough      tracing 

tiondersogenannte     Interferenzkreuzes  der  ^^.^^  ten  Siethoff's    diagram    will  iUus- 

zweiaxigen-Ivrystalle.     Centralblatt  fur  Min-  " 

eral.    Geol.  und  Palaent.    267,  1900.  trate  the     simple     method     taken     to 

make  clear  the  interference    cross  in 

biaxial  crystals.     As  is  well  known,  this  dark  cross  or  curve  obtained  between 

crossed  nicols  is  not  due  to  interference,  but  to  the  total  extinction  of  all  rays 

the  vibration  directions  of  which  are  par&llel  to  the  vibration  direction  of  either 

nicol.     In  the  diagram  the  little  crosses  represent  the  vibration  directions  (with 

X  K  ><  X      X  X  '"<  X  ^  ^  -'^-  V  4~h  i-^"A^>cX  \<  ^  ^  ^  ^  ^  y 

y.  y.  >c  %  y  X  X  x^  X  V  V  -^-+-4-+  -^^y-x  ^-X'x>cV>r;ir^  v 
i^-hi-i^  %%  v^x  X  X  v^  -v-f^-H-  i-i-i-x.  X  y  ^  )^  )^  ^  >^  ^  , 

+-4--h-h-H-^H--f  4-  4^-f-f  4-H--f-  +  -f#i-  H--h4-4--h-h+'^  "^ 

-V-V-V'V-v^-^\'^><X  T^f -f- f-+  \  :^  y  y^i-^^i-  ^i-"^-^  '^ 

^  A^^>r-Y>^x  .x^x:  %  ^  f  H-4-  -t-  A-  >-  X  x^x^  ><.  ^  "^  ^  y.  -a  ^ 
A  ^.^A- >rx  X  x>  >c  >cy  ~A-4--f-i--v  ^  ^  X  X    y^y^-^^^^^ 

\  k><X  K><  X^x-  xX/t^-H— f'  +  ^'V^^  X  xx^^  ^^"^^ 
\  X  X  xy  x^x^x  x-^"^  -7^  f-f— f  +  -vv^vx  xxx^^^  ^^ 
^  >c  X  K  ^XK^y.  y.  X  -A  -f  ^-f-4-  -I'-VA'V^^xxx^V^^^^^ 
xx-x-^^xVx  /.x^y^i^  +  +  4+-V  A-  V  ^xx  X^x'^xX  X  ^^^^ 

convergent  light)  of  the  different  rays  emerging  from  a  section  cut  normal  to  a 
bisectrix  ;  the  •  points  are  the  optic  axes.  As  the  plate  is  turned,  the  vibration 
directions  of  different  rays,  that  is,  different  series  of  little  crosses,  move  into 
parallelism  with  the  vibration  direction  of  the  nicols,  and  together  make  up  the 
hyperbola  or  the  cross.  By  turning  the  diagram,  while  resting  on  a  pad  the  edges 
of  which  may  represent  the  vibration  directions  of  the  nicols,  the  series  of  little 
crosses  parallel  to  the  edges  of  the  pad  at  any  moment  is  easily  observed.  The 
fact  that  with  an  optic  axis  in  the  field  the  rotation  of  the  plate  in  one  direction 


l-iS'i  Journal  of  Applied  Microscopy 

about  this  axis  is  accompanied  by  a  rotation  of  the  dark  brush  in  the  opposite 
directions,  is  shown  in  the  same  manner.  Minor  facts,  such  that  one  arm  of  the 
cross  is  broader  than  the  other,  and  that  the  hyperbola  branches  broaden  the 
further  we  go  from  optic  axis,  may  also  be  shown.  a.  j.  m. 

Schenck.  R     Ueber  die  Dynamik  der  Krystalle.       starting  with  the  proposition  that,  "with 
Central blatt  Min.   Geol.  Palaen.   313,  1900.  _  r     t-  ' 

crystallized   material  the  free  physical 

and  chemical  energy  is  dependent  on  the  direction,"  the  writer  assumes  that  all 
equations  involving  vapor  pressures  are  valid  for  solution  pressures  since  there 
is  perfect  analogy  between  these  pressures.  If,  then,  any  crystal  is  considered 
to  be  a  volatile  chemical  substance  with  different  crystal  faces,  and  if  all  but  two 
of  these  faces  are  coated  with  a  layer  which  hinders  vaporization,  then  on  these 
two  free  faces  there  will  be  different  pressures.  If  A  possesses  the  higher  pres- 
sure and  B  the  lower,  then  A  yields  vapor,  or  crystal  molecules,  until  the  surround- 
ing medium  is  saturated  therewith  but  the  medium  being  supersaturated  for  B  T, 
the  vapor,  or  crystal  molecules,  condense  on  B  T.  If  the  vapor  pressure  at  both 
faces  is  known,  the  work  done  in  transporting  one  gram-molecule  from  one  crystal 
face  to  another  can  be  calculated.  A  system  of  formula;  are  given  which  practi- 
cally apply  the  principles  of  thermodynamics  to  crystallographic  problems.  As 
a  practical  example,  crystals  of  potash  alum  were  coated  with  shellac,  but  on 
some  the  octahedral  faces  were  not  covered,  on  some  the  cube  faces,  and  on 
some  the  dodecahedral  faces.  The  velocity  of  weathering  at  constant  tempera- 
ture and  per  square  centimeter  surface  of  each  of  three  crystal  faces  was  deter- 
mined. The  relative  water  yield  per  square  c.  m.  for  octahedral,  cubic  and 
dodecahedral  faces  at  35°C  was  1.  :  1.27  :  1.60;  at  50°C  about  four  times  as 
much  water  was  yielded,  but  the  relation  for  octahedral  and  cubic  faces  remained 
nearly  the  same,  viz.,  1  to  1.25.  a.  j.  m. 

Qrunling,  Fr.     Ueber   die  Mineralvorkommen       Dr.  Griinling  was  sent  by  the  Tamnau- 
von    Ceylon.     Zeit.   f.  Kryst,  ll\    200-230,       „^..  r   -r.     i-  n        • 

inoo.  J  7    J7       btiftung  of  Berhn  on  a  collectmg  tour 

to  Ceylon  in  1896.     He  describes  the 

mineral  localities,  the  history  of  the  occurring  species,  and  the  native  method  of 

cutting,  and  gives    a    bibliography   and  map    and    statistics  of    production    of 

pearls,  rubies,  etc.     The  species  obtained  included  apatite,  phlogopite,  hydro- 

phlogopite,   serpentine,    pyrite,    spinel,  graphite,    ruby,    sapphire,    chrysoberyl, 

zircon,  tourmaline,  sillimanite,  moonstone,  garnet,  and  rutile.  a.  j.  m. 

Melczer,  G,    Ueber  einige  Mineralien  vorwie-      Determines  forms  and  axial  relations  of 
gend  von  Ceylon.     Zeit.  f.  Kryst.  33 :  240-       y-,      ,  ,  ,         ,  ,,  .       . 

262,1900.  Ceylon  chrysoberyl,  as  well  as  twmnmg 

law  and  optical  characters,  and  com- 
pares with  Brazilian  and  Siberian  chrysoberyls.  One  crystal  of  sillimanite  from 
Ceylon  was  exceptionally  fine.  It  was  a  transparent^  grayish  blue  prism,  with 
excellent  cleavage  in  one  direction.  H=6f  G=:3.249.  A  plate  was  cut 
from  the  center  perpendicular  to  the  length,  and  about  '1\  m.m.  thick,  which 
gave  a  beautiful  axial  figure  with  /J>z'.  In  the  Abbe  Refractometer  this 
yielded  : 


and  Laboratory  Methods.  1433 

2  V,^  from  a  ft  y 
31°48f 
31  111 
30  38| 


Y 

ft 

a 

Li      1.6730 

1.6542 

1.6527 

Na     1.0766 

1.6576 

1.6562 

Tl      1.6801 

1.6611 

1.6597 

By  measurement 

in  the 

Fuess  apparatus 
2  V, 

Li 

31.°  19 

Na 

30     57 

Tl 

30     35i 

The  pleochroism  was : 

For  vibration  parallel  c(=r)  deep  blue  with  feebly  violet  tone. 
"  "  "         a(=/' )  pale  yellow  to  brownish  yellow. 

"  "  "        b(=<!')  feeble  green  to  gray  green. 

The  transparent  blue  spinel  of  Ceylon  was  also  examined  optically  and  as  to 
form.  A.  J.  M. 

vonWorobleff,  V.    Krystallographische  Studien      ^f^g^  ^  historical    review  with  bibliog- 
iiber   Turmahn    von   Ceylon    una     einigen  ° 

anderen  Vorkommen.     Zeit.  f.  Kryst.  U :      raphy  the  author  proceeds  to  a  careful 
263-454,  1900.  examination  of  some  hundreds  of  fine 

crystals  obtained  by  Dr.  Griinling  in  Ceylon,  and  now  in  the  Berlin  museum,  and 
also  others  from  other  museums.  The  entire  recorded  series  of  forms  are  com- 
pared and  tabulated  by  zones,  and  with  respect  to  antilogous  and  analagous 
poles.  One  hundred  and  thirty-one  new  forms  are  recorded,  and  a  discussion  of 
the  relation  between  crystal  form  and  pyro-electrical  behavior  of  tourmaline  in 
general  is  given,  and  finally  the  conclusion  is  reached  that  the  mineral  belongs 
to  the  ditrigonal-pyramidal  class  of  symmetry.  a.  j.  m. 

Ooldschmidt,  v.,  and    Preiswerk,  H.      Chryso-      Good  illustrations  of  two-circle  measure- 
beryllzwilling    von    Ceylon.     Zeit.    f.  Kryst.  i      ,     . 

iZ:  455-467,  1900.  ment  and  calculation. 

Goldschmidt,  V.  Zur  Theorie  der  Zwillings-und 
Viellingsbildungen.  Zeit.  f.  Kryst.  li : 
468-476,  1900. 

Smith,  G.  F.  H.      A   Three-Circle  Goniometer.       ^^e   object   of  goniometrical   measure- 
Min.  Mag.  12:   175,  1899.  ■"  ^ 

ment  is  to   determine  in   the  simplest 

and  quickest  manner  the  geometrical  constants  of   crystals,  and  the  indices  of 

their  faces.     The  relative  advantages  and  disadvantages  of  the  one-  and  fev^-circle 

goniometers  are  pointed  out,  and  the  author  states  that  the  advantages  of  both 

instruments  may  be  combined  by  the   addition  of   a  third  circle.     A  detailed 

description    of   the    instrument    is  given   (with  plate)   and  the   method   of  use 

described,  some  actual  readings  being  recorded.  l.  mci.  l. 

Smith,  0.  P.  H.    Note  on  the  Identity  of  Para       The    oxychloride     of     lead    rafaelite 

laurioniteand  Rafaelite.    Min.  Mag.  12 :   183,        .,  -u     1  u        a  -n  j    i.       u 

jg  **  -^       (described  by   Arzruni)    proved  to    be 

identical  with  paralaurionite  by  com- 
parison of  angles,  made  by  twinning  and  optical  characters,  the  only  difference 
being  in  the  color,  P.  being  white  while  R.  is  violet-red,  and  shows  strong  pleo- 
chroism.    Axial  ratio  of  Y.^=a  :  b  :  r=2.7036  : 1  :  1.8019.  l.  mci.  l. 


1434 


Journal  of  Applied  Microscopy 


MEDICAL  NOTES. 


Robin,  A.     A  Contribution  to  the  Techinic  of 
the  Widal  Test.     Phila.  Med.  Jour.  7:  ii. 


Four   problems  present  themselves  to 
the  bacteriologist  who  attempts  to  per- 
form the  Widal   test  in  the  diagnosis  of  typhoid  fever,  viz.  :      1.  The  dilution. 

2.  The  best  way  of  obtaining   a   motile  culture   free   from   "  natural  "  clumps. 

3.  The  differentiation  between  a  true  and  a  pseudo-reaction.     4.  The  time  limit. 

To  these  problems  Dr.  Robin  offers  solutions  which  in  his  expe- 
rience have   proved   most  practical   and    satisfactory. 

1.  Accurate  dilutions  are  obtained  by  means  of  the  simple 
medicine  dropper  device  (Fig.  1)  described  in  Vol.  Ill,  No.  8,  p.  962 
of  the  Journal. 

2.  Motile  organisms  may  be  readily  obtained  for  the  test  by 
keeping  at  hand  pure  cultures  of  typhoid  bacilli  in  hermetically 
sealed  tubes.  When  a  test  is  to  be  made  a  fresh  agar  or  bouillon 
culture  is  made  from  the  stock  culture  and  kept  in  the  incubator  for 
eighteen  to  twenty-four  hours.  It  was  found  that  the  temperature  of 
a  fairly  warmed  room  produced  just  as  good  if  not  better  results 
than  the  incubator.  The  author  deems  the  bouillon  culture  unsatis- 
factory and  has  adopted  the  following  medium  :  An  agar  culture  is 
kept  in  the  incubator  or  at  room  temperature  for  twelve  to  eighteen 
hours,  when  two  or  three  loopfuls  are  transferred  into  bouillon  until 
a  marked  turpidity  results,  or  a  small  quantity  of  bouillon  is  added 
to  the  agar  culture  and  enough  of  the  growth  scraped  off  to  produce 
a  uniform  cloudiness.  The  latter  course  is  preferable  and  if  care- 
fully followed  the  "  natural "  clumps  so  frequently  observed  in 
bouillon  cultures    (Fig.    2  B)    are    entirely  avoided. 

3.  The  third  problem  is  met  by  using  a  slide  with  two  concavities    Fig.  i. 

(Fig.  3),  around  the  edges  of  each  of  which 
is  a  r^ng  of  vaseline.  On  each  of  two  clean 
cover-glasses  is  deposited  a  loopful  of  the 
culture  ;  to  one  a  loopful  of  the  blood,  diluted 
1:20  to  1:40,  is  added,  while  the  other  serves  as 
a  control.  The  behavior  of  the  bacilli  on  each 
cover  may  be  readily  observed.  If  the  reaction 
is  positive  the  bacilli  on  the  test 
cover  will  gather  in  clumps  of  two, 
three  or  a  dozen  and  will  soon  lose 
their  motility  (Fig.  2  D),  while  in  the 
pseudo-reaction  only  a  few  clumps 
will  form,  the  rest  of  the  bacilli  re- 
maining separated  (Fig.  2  C). 

4.  The  time  given  to  determine  whether  a  reaction  is  positive 
or  negative  varies  greatly  with  different  bacteriologists.  Dr. 
Robin  proposes  the  adoption  of  a  uniform  limit  and  offers  the 
following:  Dilution  1:10,  time  limit  5  to  15  minutes;  1:20,  15 
to  20  minutes;  1:40  to  1:100,  .'JO  to  (JO  minutes;  1:100  to  1:200, 
1  to  2  hours.  That  is,  if  within  the  specified  time  a  consider- 
able number  of  bacilli  are  found  actively  motile  or,  if  dead,  fail  to 
arrange  themselves  in  clumps,  the  reaction  is  negative,  irrespect- 
ive of  the  clumps  which  have  already  formed.  c.  w.  j. 


Fig. 


and  Laboratory  Methods.  1435 

NEWS  AND  NOTES. 

It  is  recommended  in  the  study  of  infusoria  to  use,  in  place  of  powdered 
carmin,  water-color  carmin.  The  infusoria  are  added  to  a  drop  of  water  in 
which  a  very  small  quantity  of  the  stain  has  been  dissolved. 


In  the  study  of  the  mouth  parts  of  the  crayfish  very  satisfactory  results  have 
been  obtained  by  sewing  the  parts,  in  their  proper  order,  to  a  piece  of  stiff  linen 
paper.  Thus  mounted  the  parts  may  be  preserved  in  vials  in  3  per  cent,  forma- 
lin, and  be  ready  at  any  time  for  study  in   connection   with  the  whole  specimen. 


A  Convenient  Table  for  Formalin  Solutions. 

100  %  formalin=:40%  formaldehyde,  usual  strength  of  commercial  solutions. 
50    "         "       =20 " 
25   "         "       ==10" 
121"         '<       =5" 
10""         "       =4" 


71"    .     "       ==3 " 
5"  "         "       =  2 " 

1^"  "       =0.4" 


:1  vol.  formalin -f-1  vol.  water.  Total=2  vols. 

— ^  <<  u  -1-3  "  "  "  =4     " 

=  1  "  "  +7  "  "  "  =8     " 

=  1  "  "  +9  "  "  "  =10  " 

=1  "  "  +121  "  "  =131" 

=1  "  "  +19"  "  "  =20  " 

=  1  "  "  +39"  "  "  =40  " 

=  1  "  "  +99"  "  "  =100" 


The  Biological  Department  of  Earlham  College  has  recently  issued  Bulletin 
No.  1,  containing  fifty-five  excellent  photo-micrographs  of  fertilization,  matura- 
tion, and  segmentation  stages  of  ascaris,  and  the  development  stages  of  the 
chick.  The  illustrations,  together  with  the  running  description  accompanying 
them,  briefly  and  concisely  summarize  the  steps  in  embryological  development 
from  the  unfertilized  egg  to  the  union  of  the  alantois  with  the  alimentary  canal. 


Referring  to  Mr.  H.  A.  Doty's  article  on  "  Conochilus  and  Vorticella  as 
Commensals  "  (J.  A.  M.  3 :  989),  Mr.  H.  D.  Thompson  of  Moline,  III,  notes 
the  following  observation  : 

"  For  some  years  I  have  been  wont,  when  other  methods  failed  me,  to 
obtain  Vorticellse  for  class  use  by  collecting  quantities  of  cyclops,  with  a  small 
muslin  net,  from  a  certain  spring  hole.  The  Vorticellae,  living  in  great  abun- 
dance on  these  cyclops  specimens,  are  invariably  provided  with  a  short  stalk, 
which  is  only  slightly  contractile,  and  incapable  of  assuming  the  usual  spiral 
posture." 


Pheln's  Method  of  Staining  the  Malaria  Parasite. — Wash  the  fixed 
specimens  for  three  or  four  minutes  in  absolute  alcohol,  after  which  they  are 
stained  for  five  or  six  minutes  in  the  following  solution  : 

Methylen  blue,  cone,  aq.  sol., 15 

Eosin,  Yz  per  cent.  sol.  in  alcohol,  75  per  cent.,       5 

Water,  dist., .      10 

Sodium  hydrate,  20  per  cent., 3  drops. 

After  thorough  washing  in  water,  mount  in  Canada  balsam. 


1436 


Journal  of  Applied  Microscopy 

A  Combined  Slide  and  Cover-glass  Forceps. 


Fig.  I. — The  forceps  with  the  slide  locked  in  position. 


Fig.  2. — As  a  cover-glass  forceps. 

The  forceps  illustrated  was  devised  by  Mr.  L.  Napoleon  Boston,  Philadelphia, 
and  combines  in  a  single  instrument  both  a  slide  and  cover-glass  holder.  It  is 
made  of  brass  wire.  A  slide  is  easily  picked  up  from  a  smooth  surface  and 
held  as  shown  in  Fig.  1. 


We  recently  received  a  copy  of  the  preliminary  announcement  of  the  fiftieth 
annual  meeting  of  the  American  Association  for  the  Advancement  of  Science,  to 
be  held  at  Denver,  August  24  to  31,  1901.  The  announcement  contains  lists  of 
officers  and  members  of  the  association,  and  a  general  programme  of  the  coming 
meeting.  Care  has  been  taken  in  the  preparation  of  a  guide  to  the  city  of 
Denver,  embracing  hotel  accommodations,  excursions  to  points  near  Denver, 
and  points  of  interest  within  the  city. 


QUESTION   BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

lU.  How  should  the  solution  of  gutta  percha  in  turpentine,  recommended  by 
V.  A.'  L.  for  cementing  liquid  mounts,  on  page  712  of  Journal  of  Applied 
Microscopy,  be  made? — i.  d. 

11.  For  what  kinds  of  vegetable  tissues  are  Amann's  media  (pp.  711-2  of 
Journal  of  Applied  Microscopy)  suitable  ?  Are  they  useful  for  hydrous 
tissues,  or  must  tissues  be  dehydrated  prior  to  mounting  in  them  ? — i.  d. 

12.  F.  M.  L.  wishes  to  secure  specimens  of  Selaghiella  lepidophylla  alive 
and  producing  spores.     Can  any  of  our  readers  supply  such  specimens  ? 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


VOLUME  IV.  SEPTEMBER,   1901.  NUMBER  9 


Laboratory  Courses  by  Correspondence. 

Not  every  one  who  desires  an  education  finds  it  possible  to  spend  three  or 
four  years  at  a  university.  Many  teachers  with  only  a  high  school  education 
hold  good  positions  which  they  would  not  feel  justified  in  resigning  for  an  ex- 
tended course  of  study.  Correspondence  courses  carefully  planned  by  compe- 
tent instructors  enable  such  teachers,  while  still  holding  their  positions,  to  devote 
some  time  to  a  systematic  study  of  branches  connected  with  their  work,  and  thus 
to  increase  their  own  knowledge  and  at  the  same  time  be  better  prepared  to  in- 
struct their  pupils.  Those  who  are  working  for  university  degrees,  but  are  com- 
pelled to  spend  the  shortest  possible  time  in  residence  at  the  university,  find  in 
the  correspondence  system  a  solution  of  the  problem.  Others  who  are  neither 
teachers  nor  university  students  are  deeply  interested  in  particular  subjects ; 
such  people,  even  when  relying  entirely  upon  their  own  resources,  will  advance 
along  the  chosen  line,  but  progress  is  more  rapid  and  satisfactory  when  efforts 
are  systematized  and  directed  by  those  who  have  often  traversed  the  ground 
before. 

It  has  for  some  time  been  recognized  that  many  university  courses  can  be 
pursued  successfully  by  correspondence.  The  favorable  results  secured  in  lan- 
guage, literature  and  history  suggested  that  an  attempt  be  made  to  conduct  lab- 
oratory courses  also. 

Several  years  ago  the  writer  was  asked  to  conduct  a  course  in  botany  by 
correspondence.  With  many  misgivings  as  to  the  success  of  any  laboratory  study 
by  this  method,  a  course  in  the  Morphology  of  Algae  and  Fungi  was  planned  and 
the  work  was  begun  with  a  single  pupil.  The  result  soon  showed  that  a  persist- 
ent student  could  do  the  work  thoroughly  in  spite  of  the  difficulties. 

Several  courses  were  then  announced,  each  course  being  the  full  equivalent 
of  the  same  course  as  conducted  at  the  university.  The  following  is  the  general 
plan  for  the  Algae  and  Fungi  and  the  other  two  morphological  courses  are  simi- 
lar :  Material,  selected  with  extreme  care,  is  sent  to  the  student  and  all  prepar- 
ations for  the  microscope  which  require  a  knowledge  of  technique  are  also 
included.  The  directions  for  study  are  in  the  form  of  twelve  lessons,  each  lesson 
covering  three  laboratory  exercises  as  conducted  at  the  university.  In  the  lab- 
oratory work  more  than  fifty  types  are  studied,  and  these  are  arranged  so  as  to 

(1437) 


1438  Journal  of  Applied  Microscopy 

give  a  view  of  the  structure,  development  and  relationship  of  all  the  great  groups 
of  Algae  and  Fungi.  The  lack  of  lectures  is  compensated  for  by  assigned  read- 
ings and  the  study  of  a  larger  number  of  types.  As  soon  as  a  lesson  is  com- 
pleted, it  is  sent  to  the  instructor,  who  returns  it  with  corrections  and  suggestions. 

Three  courses  in  botany.  (1)  General  Morphology  of  the  Algae  and  Fungi, 
(2)  General  Morphology  of  the  Ikyophytes  and  I^teridophytes.  and  (3)  General 
Morphology  of  the  Gymnosperms  and  Angiosperms,  have  been  thoroughly  tested 
by  the  writer,  nearly  a  hundred  students  having  taken  the  work.  The  results 
are  surprising.  Many  students  after  taking  one  or  more  of  these  courses  by  cor- 
respondence have  come  to  the  university  for  further  work,  and  have  not  only 
been  able  to  hold  their  own  in  classes  with  students  who  had  done  the  previous 
work  in  residence,  but,  on  the  whole,  have  shown  a  more  thorough  preparation. 

However,  it  must  not  be  inferred  that  correspondence  work  is  preferable  to 
residence  w^ork,  for  such  is  not  the  case.  The  explanation  is  to  be  sought  in  the 
fact  that  those  who  have  sufficient  interest  and  determination  to  carry  on  a  course 
by  correspondence  are  willing  to  devote  more  time  and  effort  than  can  be  required 
of  the  average  university  student.  It  is  particularly  noticeable  that  correspond- 
ence students,  when  they  come  for  resident  work,  are  more  independent  and  ask 
fewer  thoughtless  questions  than  those  who  have  always  had  an  instructor  at  the 
elbow.  Several  who  have  laid  the  foundation  for  morphological  work  by  corres- 
pondence have  subsequently  come  to  the  university  for  research  work,  and  have 
published  excellent  papers,  and  two  have  even  taken  the  doctor's  degree,  with 
botany  as  the  major  subject. 

After  the  success  of  these  courses  became  evident,  a  course  in  histological 
technique,  preeminently  a  laboratory  course,  was  offered  and  has  proved  a  suc- 
cess. Work  in  the  newer,  but  very  popular  field  of  Ecology,  is  also  being  con- 
ducted satisfactorily  by  correspondence. 

In  looking  over  the  list  of  those  who  have  studied  botany  by  correspondence, 
it  is  interesting  to  note  that,  aside  from  the  teachers  and  students  who  form  the 
great  majority,  there  are  also  lawyers,  business  men,  clerks  and  artisans,  who 
have  found  time  to  improve  themselves  in  their  chosen  subject. 

The  success  which    has  attended  the  correspondence  work  in  botany  sug- 
gests that  in  other  sciences  also  those  laboratory  courses  which  do  not  require 
very  expensive  apparatus  may  be  conducted  by  this  method. 
University  of  Chicago.  ClIARLES  J.  ChamI'.ERLAIN. 


Dr.  W.  IUjrck,  of  the  Royal  Academy  of  Sciences  of  Amsterdam,  has  re- 
cently published  some  observations  bearing  upon  the  subject  of  the  prevention 
of  hybridisation  in  plants.  His  experiments  showed  that  certain  chemical  sub- 
stances act  very  differently  on  the  pollen  of  different  plants.  Levulose  in  small 
quantities  greatly  accelerates  the  growth  of  pollen  tubes  in  some  plants,  while  in 
others  the  pollen  grains  are  caused  to  burst.  Saccharose  and  dextrose  produce 
different  effects  than  levulose.  According  to  the  author's  interpretation  these 
results  would  indicate  the  possibility  that  the  stigmatic  secretion  of  a  given 
species  contains  substances  which  promote  the  emission  of  pollen  tubes  in  that 
species,  but  prevent  the  growth  of  pollen  from  other  species. — Natiire^\\  1656. 


and  Laboratory  Methods. 


1439 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 
PHOTOGRAPHING   DIATOMS. 

In  photographing  diatoms  at  the  University  of  Iowa  the  apparatus  used  is  of 
the  simplest  character.  It  consists  primarily  of  a  "  Practical  "  photomicro- 
graphic  camera,  a  microscope  furnished  with  a  mechanical  stage,  apochromatic 
lenses,  and  with  compensating  and  projection  eyepieces.  The  remainder  of  the 
apparatus  is  mostly  home  made  and  consists  of  a  table,  condensing  lenses  suit- 
ably disposed,  an  acetylene  generator,  and  simple  lamp  or  burner. 

The  camera  is  hinged  so  that  it  may  be  used  in  either  a  vertical  or  horizontal 
position.  I  find  this  very  convenient,  as  the  bellows  may  be  quickly  raised  to 
allow  the  operator  to  make  a  direct  examination  of  the  object.  The  working 
lens  is  a  dry  apochromatic,  3mm.  of  .95  N.  A.  A  compensating  eyepiece  No.  S 
and  a  projection  eyepiece  No.  4  are  the  oculars  used.  The  table, 
which  serves  the  purpose  at  once  of  camera  table  and  optical 
bench,  is  about  three  feet  six  inches  long.  The  width  is  about 
sixteen  inches  and  the  height  so  adjusted  that  when  the  operator 
sits  in  a  chair  the  ocular  is  in  a  convenient  position  for  obser- 
vation. 

When  making  an  exposure  the  bellows  is  turned  down  and 
rests  on  the  leaf  of  the  table,  which  for  this  purpose  is  raised  to  a 
horizontal  position. 

The  condenser  is  composed  of  two  plano-convex  lenses  two 
and  one-half  inches  in  diameter,  an  achromatic  pair  two  and  one- 
fourth  inches  in  diameter,  and  a  one-inch  negative  to  effect  the 
parallelism  of  the  rays.  The  spherical  and  chromatic  aberration 
of  the  first  system  of  condensers  is  in  a  large  measure  corrected 
by  this  simple  device  ;  and,  although  it  is  conceded  that  every 
additional  lens  is  in  a  sense  an  added  obstacle,  nevertheless  the 
advantage  to  be  derived  from  the  introduction  of  the  negative  in 
the  series  at  this  point  will  quickly  become  apparent  to  anyone  who  chooses  to 
try  the  experiment. 

No  heat  filter  is  necessary  with  acetylene  gas  as  the  illuminant.  As  is  well 
known,  this  light  is  remarkably  cool.  The  substage  condenser  is  a  plain,  uncor- 
rected Abbe. 

A  small  acetylene  generator  and  a  round  flame  burner  complete  the  outfit. 
I  have  adopted  the  round  flame  burner  after  a  series  of  experiments  involving 
every  other  form  of  burner  offered  by  the  trade.  I  certainly  consider  it  the  most 
desirable  and  efficient. 

In  the  present  discussion  I  shall  assume  that  the  material  to  be  photographed 
is  properly  mounted  in  well  cleaned  styrax  on  cover-glasses  of  known  thickness. 
For  mounting  the  larger  species  a  mechanical  finger  will  be  found  convenient,  as 
such  species  should  be  mounted  singly.     In  photographing  from  spreads  I   pro- 


Fig. 


1440 


Journal  of  Applied  Microscopy 


ceed  as  follows  :  By  the  aid  of  the  mechanical  stage  I  carefully  review  the 
entire  slide,  noting  the  location  of  every  suitable  specimen  of  the  species 
desired,  of  all  species  not  yet  photographed,  and  their  degree  of  perfection. 
Such  specimens  should  lie  in  optical  contact  with  the  lower  side  of  the  cover- 
glass,  be  clearly  marked  and  free  from  foreign  matter.  These  observations  are 
recorded  with  the  readings  of  the  mechanical  stage,  and  by 
use  of  the  record  at  any  subsequent  time  the  desirable 
specimens  may  be  turned  to  readily  and  photographed  with- 
out loss  of  time.  This  search  work  may  be  done  by  day- 
light or,  what  I  consider  an  exceedingly  good  substitute,  by 
means  of  the  acetylene  lamp  with  a  rather  dense  ray  filter 
interposed.  The  filter,  which  has  given  me  excellent  results, 
is  simply  a  fiat  eight-ounce  bottle  filled  with  a  fluid  com- 
posed of  175  grams  of  copper  sulphate,  17  grams  potassium 
bichromate,  2  c.  c.  sulphuric  acid  and  500  c.  c.  of  water. 
The  color  is  restful  and  agreeable  to  the  eyes,  and  the 
density  is  not  sufiicient  to  interfere  in  any  serious  way  with 
accurate  vision  or  inspection. 

Focusing  and  adjusting  for  cover-glass  thickness  can  be 
learned  by  experience  only.  As  is  well  known,  however,  both 
arts  are  of  the  most  vital  importance.  In  this  paper  I  shall 
not  endeavor  to  give  any  advice  on  these  two  points  except 
merely  to  mention  a  little  matter  that  I  have  never  seen 
elsewhere  stated  and  which  has  been  of  great  service  to  me. 
In  my  experience  the  microscope  is  always  horizontal ; 
this  is  the  convenient  position. 

One  day  when  working  at  the  instrument  I  discovered 
that  when  I  placed  my  fingers  on  the  milled  head  of  the  fine 
adjustment  screw,  there  ensued  an  alteration  of  the  focus 
although  the  head  had  not  been  turned.  Further  investi- 
gation brought  out  the  fact  that  the  alteration  was  due  to  a 
springing  of  the  arm  induced  by  a  downward  pressure  on 
the  milled  head,  and  that  when  the  finger  was  removed  the 
object  came  again  into  perfect  focus.  I  also  found  that  a  slight  pressure 
upward  caused  the  object  to  pass  out  of  focus  in  the  opposite  direction. 

This  proved  to  be  an  exceedingly  delicate  test  of  the  correctness  of  the  focus. 
If  perfectly  focused  the  error  produced  by  this  slight  pressure  is  equal  in  both 
directions ;  but  if  not  perfectly  focused  the  error  will  be  more  evident  in  one 
direction  than  in  the  other.  This  apparently  commonplace  and  trifling  matter 
is  well  worth  the  attention  of  anyone  who  attempts  the  photography  of  these 
very  delicate  and  difficult  forms. 

The  time  of  exposure  will,  of  course,  vary  according  to  conditions.  I  use  two 
difTerent  amplifications,  660  diameters  and  lolJO  diameters.  All  my  photographs 
are  made  at  6(30  diameters  unless  the  objects  are  very  small  or  are  adorned  with 
very  fine  stria;.  When  the  forms  are  large  and  marked  with  fine  striae  two 
photographs  are  taken  ;  one  to  show  simply  the   outline,  and  the  other   at  the 


Fig.  2. 


and  Laboratory  Methods. 


1441 


higher  magnification  and  with  oblique  illumination    to   show  details,  as   in  Figs. 
1,  2  and  3. 

The  exposure  is  made  as  short  as  possible  without  sacrificing  detail;  then,  if 
the  plate  be  strongly  developed  the  requisite  contrast  will  be  secured.  I  find, 
too,  that  it  is  best  to  make  two  exposures  of  each  specimen. 

-On  removing  the  plate  from  the  holder  a  number  is  placed  on  one  corner  with 
a  soft  lead  pencil,  say  Dixon's  "  Ultimatum  "'  or  one 
similar  to  it.  This  number  is  also  placed  in  a  book 
kept  for  the  purpose  with  the  name  of  the  species, 
date,  magnification,  light,  number  of  slide,  and  loca- 
tion. With  these  data  it  is  an  easy  matter  to  re- 
photograph  any  particular  specimen  if  at  any  time 
the  negative  be  lost  or  broken,  or  if  for  any  reason 
it  prove  unsatisfactory. 

I  have  experimented  with  every  developer  to  be 
had  here,  and  have  tested  many  formulas,  but  none 
of  them  is  equal  to  the  one  known  as  Bromo- 
hydroquinon.  It  gives  the  requisite  amount  of  con- 
trast, a  thing  to  be  kept  constantly  in  mind  in 
photographing  objects  so  very  hyaline  as  are  diatoms. 

For  a  fixing  bath  plain  hypo  seems  to  give  better 
results  than  the  acid  alum  bath. 

Any  good  plate  will  answer  the  purpose  provid- 
ing it  is  heavily  coated ;  my  preference,  however,  is 
"  Cramer's  Instantaneous  Isochromatic."  Let  me 
say  again,  a  thin  plate  will  not  answer.  In  order 
to  economize,  I  get  the  4x5  plates  and  then  cut 
them  once  or  twice  as  the  size  of  the  diatom  de- 
mands ;  i.  e.,  the  plates  are  then  2^  x  2,  or  2;^  x  4. 
When  dry  they  are  put  in  appropriate  envelopes, 
filed  away  in  alphabetical  order,  and  a  full  record  of 
each  one  is  kept  in  a  card  index. 

Most  diatoms  lend  themselves  readily  to  photography,  the  side  of  the  valve 
which  is  most  important  usually  being  nearly  plane.  Some,  however,  are  more 
or  less  convex  or  concave.  Figure  5  represents  a  species  that  has  a  ridge  just 
inside  the  margin  and  a  depressed  center.  This  of  course  necessitates  a  com- 
promise, with  some  loss  of  detail.  Only  a  very  few  species  of  the  fresh  water 
forms,  however,  are  impossible  of  photography  as  here  described. 

As  is  well  known,  Navicula  is  the  typical  genus  of  the  Bacillariaceae,  with 
hundreds  of  species  ;  these  come  out  beautifully,  as  is  attested  by  Fig.  4. 

Previous  to  printing,  the  negative  is  placed  in  a  retouching  frame  and  the 
background  is  all  cut  out  by  the  application  on  the  back  of  a  heavy  coat  of 
"  Copelin's  Opaque."  This  cuts  out  everything  but  the  image  desired.  For 
printing,  all  sorts  of  paper  have  been  tried.  Among  those  that  I  have  used,  of 
the  developing  sort,  Velox,  and  of  the  printing  sort,  Solio,  seem  to  give  the  best 
results.     I  have  discarded  Solio,  however,  on  account  of  its  slowness.     In  print- 


Fig.  3. 


1442 


Journal  of  Applied  Microscopy 


ing  on  Velox  I  proceed  as  follows  :  Six  frames  are  prepared  and  arranged  on 
the  six  sides  of  a  hexagonal  wire  frame  on  which  are  stretched  two  thicknesses  of 
white  tissue  paper. 

This  screen  is  about  six  inches  in  diameter,  six  inches  deep,  and  open  at 
the  top.  The  printing  frames  are  placed  at  irreg- 
ular distances  from  the  screen,  according  to  the 
density  of  the  plate  in  each  case. 

A  bit  of  magnesium  ribbon  about  one  and  one- 
half  inches  long  is  then  ignited  in  an  alcohol  flame 
and  instantly  placed  within  the  screen  near  the 
center.  This  prints  all  six  pictures  at  once  and 
they  are  ready  to  be  developed.  The  screen  pre- 
vents the  edge  of  the  opaque  from  printing  up  as  a 
sharp  line.  The  use  of  the  magnesium  light  greatly 
increases  the  rapidity  with  which  the  prints  may 
be  produced  and  also  contributes  not  a  little  to  the 
sharpness  of  the  image. 

Of  course,  with  the  simple  appliances  here 
described,  the  highest  de- 
gree of  critical  photography 
may  hardly  be  attempted. 
Nevertheless,  it  may  be 
readily  seen,  from  the 
samples  herewith  submit- 
ted, that  illustrations  may 
easily  be  secured,  sufficient- 
ly accurate  for  practical 
purposes.  No  doubt  a  bet- 
ter apparatus  is  a  thing  to 
be  desired.  But,  if  the  matter  of  expense  must  be  taken  into  account  at  all, 
the  apparatus  which  we  have  here  described  and  successfully  used  will  com- 
mend itself  to  many  who  might  be  prevented  by  the  consideration  of  cost 
from  attempting  experiment  in  this  most  fascinating  field  of  work.  The  results 
of  our  labors  in  this  direction  will  form  the  subject  of  a  descriptive  paper  pres- 
ently to  appear  in  the  Bulletin  of  the   Laboratories   of   Natural  History  of  the 


Fig.  4. 


Fig. 


State  University  of  Iowa. 
University  of  Iowa. 


P.  C.  Myers. 


THE  5  mm.  APOCHROMAT,  AFTER    PROF.  CHARLES  S.  HASTINGS,  IN  THE 
PHOTOGRAPHY  OF  DIATOMS. 

We  are  in  receipt  of  a  very  interesting  series  of  photo-micrographs  of  diatoms 
from  Honorable  A.  A.  Adee,  Washington,  D.  C,  made  while  testing  a  5  mm. 
apochromatic  objective  after  the  formula  recently  computed  by  Prof.  Charles  S. 
Hastings,  Sheffield  Scientific  School,  Yale  University. 

The  following  brief  notations,  in  connection  with  data  which  show  subject, 


and  Laboratory  Methods. 


144.^ 


<T  i^','"  '>  '^  o  -^  "^ 


Orthoncis  splendida,  Grunow. 


ArachnoKdi&cus  indicus,  Ehf^ 


1444 


Journal  of  Applied  Microscopy 


Triceratium  tripolare,  Temp   Br. 


Cymbola  mexicana,  Ehr. 


and  Laboratory  Methods.  1445 

accessory  apparatus,  illuminant,  etc.,  used,  are  interesting  as  showing  advance 
made  by  American  opticians  in  constructive  optical  mathematics  and  the  possi- 
bilities of  the  application  of  theoretical  conclusions  in  the  production  of  an 
apochromatic  objective  system,  without  the  use  of  other  materials  than  the 
glasses  ordinarily  employed  : 

"  Although  I  cannot  claim  any  expert  knowledge  of  optical  science,  my  exper- 
ience during  the  past  six  years  in  difficult  photo-micrography  may  make  my  test 
of  this  glass  in  the  camera  of  some  worth  to  you.  I  find  it  superior  in  working 
quality  to  any  lens  of  apochromatic  focus  I  have  yet  tried  except  the  Zeiss 
apochromatic  of  4  mm.,  and  as  to  that  it  holds  its  own  for  photographing.  The 
correction  for  actinic  rays  is  surprisingly  good,  so  that  exquisite  definition  is 
obtainable,  even  with  a  projection  ocular  No.  4,  and  it  does  not  bring  it 
down  under  a  compensation  ocular  No.  8.  Notwithstanding  the  extremely 
wide  aperture,  the  field  is  perfectly  flat,  so  that  perfect  photographic  defini- 
tion is  obtained  to  the  edges  of  a  large  circle  on  the  focusing  screen.  It 
bears  more  light  than  any  others  I  have  tried,  and  I  can  open  the  condenser 
and  diaphragm  at  least  40  per  cent,  more  than  with  the  other  glasses,  and  still 
get  excellent  photographic  contrast. 

"  The  focus  of  this  lens  appears  to  be  a  trifle  less  than  5  mm.,  about  4.65  mm., 
as  nearly  as  I  can  estimate  it  by  comparison  of  the  negatives  with  it,  and  the 
Zeiss  4  mm."  l,  b.  e. 


The  New  Medical  Laboratories  of  the  University  of 

Pennsylvania. 

The  University  of  Pennsylvania  is  about  to  erect,  at  a  cost  of  more  than 
,000,  exclusive  of  grounds  and  equipment,  a  Medical  Laboratory  building 
which  will  be  unexcelled  in  every  respect.  The  trustees  are  also  contemplating 
the  erection  in  the  near  future  of  a  new  Medical  Hall,  Anatomical  Building,  and 
auxiliary  buildings,  which  will  adjoin  the  new  laboratory  about  to  be  erected, 
and  which  will  form  one  of  the  most  extensive  systems  of  buildings  devoted 
exclusively  to  the  teaching  of  medicine  in  Europe  or  America. 

The  new  Medical  Laboratory  building,  which  will  be  erected  at  once,  will  be 
quadrangular  in  shape,  and  will  be  located  on  the  south  side  of  Hamilton  walk, 
between  Thirty-sixth  and  Thirty-seventh  streets.  The  building  will  be  two 
stories  in  height  above  a  high  basement,  and  measure  340  feet  front  by  nearly 
200  feet  in  depth.  The  long  front  faces  north,  securing  a  maximum  amount  of 
the  best  light  for  laboratory  purposes.  All  along  the  front  are  arranged  small 
rooms  for  research,  rooms  for  professors  and  their  assistants,  a  library,  etc. ; 
these  open  into  a  private  corridor,  so  that  men  employed  in  these  rooms  may 
pursue  their  work  without  interruption  from  students  passing  through  the  main 
halls. 

Perfect  lighting  of  all  the  laboratories  has  been  obtained,  the  courts  being 
large  enough,  with  the  low  front  building,  to  furnish  good  north  light  to  the 


1446  Journal  of  Applied  Microscopy 

laboratory  of  pharmacodynamics  on  the  first  floor,  and  to  the  large  laboratories  on 
the  second  floor  devoted  to  pathology,  where  microscopical  work  is  done,  the 
north  front  of  these  rooms  facing  on  the  courtyard  being  made  almost  wholly  of 
glass,  and  extending  higher  than  the  front,  so  that  steady  north  light  will  be 
thrown  to  the  back  of  the  room. 

The  first  floor  of  the  new  laboratories  will  be  devoted  to  physiology  and 
pharmacodynamics. 

The  second  floor  will  be  devoted  exclusively  to  pathology.  An  examination 
of  the  commodious  plans  will  disclose  the  purpose  of  the  pathological  laboratory. 
After  providing  for  lectures  upon  general  topics  in  pathology,  the  chief  provision 
is  for  laboratory  instruction.  The  entire  north  front  of  the  building  is  devoted 
to  laboratories  for  advanced  students  in  pathology  and  pathological  bacteriology, 
and  to  the  special  research  and  assistants'  rooms.  Each  of  the  advanced  labora- 
tories measures  31  x  44  feet.  The  east  wing  accommodates  the  laboratory  of 
experimental  and  chemical  pathology,  while  the  west  wing  is  occupied  by  the 
museum  of  pathological  specimens.  This  latter,  which  measures  44  x  65  feet, 
adjoins  the  demonstration  hall  of  morbid  anatomy,  which  hall  communicates 
with  the  general  pathological-histological  laboratory.  The  last  laboratory,  the 
front  of  which  is  to  consist  almost  entirely  of  glass,  is  located  in  a  section  of  the 
building  looking  north  into  a  spacious  court.  This  room,  37  x  100  feet,  will  seat 
one  hundred  students,  and  will  be  devoted  entirely  to  microscopical  work,  for 
which,  on  account  of  the  excellent  lighting,  it  will  be  admirably  adapted.  In 
order  to  combine  in  one  harmonious  whole  the  study  of  the  microscopical 
features  of  diseased  organs  and  the  gross  alterations  in  them,  the  pathological- 
histological  laboratory,  the  laboratory  of  morbid  or  gross  pathological  anatomy, 
and  the  museum  of  pathology  are  made  closely  communicating  and  freely 
accessible  one  from  the  other.  Another  section  of  the  building,  of  equal  size 
with  the  first,  and  also  looking  north  into  the  court,  is  subdivided  into  three 
smaller  laboratories  for  the  instruction  in  comparative  (pathology  of  animal 
diseases),  neurological  (pathology  of  nervous  diseases),  and  surgical  pathology. 
The  same  method  of  lighting,  with  enormous  glass  windows,  is  to  be  carried  out 
in  this  group  of  laboratories.  Finally,  the  west  wing  of  the  building  will  also 
provide  for  photographic  and  micro-photographic  outfits. 

Besides  the  numerous  laboratories,  research  rooms,  etc.,  there  are  four  lecture 
rooms  in  the  building.  The  two  marked  "  Demonstration  Rooms  "  on  the  plan 
each  seats  184  students.  These  lecture  rooms  communicate  with  two  prepara- 
tion rooms  each.  At  the  rear  of  the  building  there  are  two  large  lecture  rooms, 
each  seating  400  students.  To  avoid  confusion  between  lectures,  the  corridors 
and  stairways  are  so  arranged  that  one  class  enters  the  large  lecture  room  from 
one  side  as  the  other  class  leaves  it  from  the  opposite  side.  Students  enter 
these  rooms  from  a  landing  at  the  main  stair,  midway  between  the  first  and 
second  floors.  The  floor  of  the  lecture  room  is  on  a  level  with  the  basement, 
and  the  lecturer  will  enter  directly  from  the  basement  level,  and  all  specimens 
needed  to  illustrate  the  lectures  will  be  brought  through  the  entrance,  thus  saving 
the  crossing  of  the  halls  through  which  classes  move. 

The  equipment  of  the  laboratory  will  be  adeq.uate  and  in  keeping  with  the 


and  Laboratory  Methods. 


1447 


1448  Journal  of  Applied  Microscopy 

advanced  ideas  of  the  times  regarding  laboratory  instruction.  That  of  the 
physiological  department  will  be  described  at  another  time.  The  outfits  for  the 
laboratories  of  pathology  will  include  modern  microscopes,  furnished  with  suit- 
able optical  parts  for  the  study  of  animal  tissues  and  bacteria,  complete  bacteri- 
ological outfits,  for  the  study  of  the  relation  of  bacteria  and  other  parasites  to 
pathological  formations,  new  and  complete  photographic,  micro-photographic, 
and  projection  apparatus,  and  a  special  outfit  consisting  of  kymographs,  respira- 
tory apparatus,  etc.,  for  the  study  of  subjects  in '  general  and  experimental 
pathology. 

The  assistants'  and  research  rooms  will  contain  individual  outfits  for  histo- 
logical and  bacteriological  study.  This  will  be  in  addition  to  those  provided  for 
the  use  of  undergraduates  and  advanced  (or  post-graduate)  students  in  the  gen- 
eral laboratories. 

It  is  intended  to  cultivate  and  promote  a  spirit  of  independent  and  research 
work,  both  in  respect  to  students  taking  the  course  in  medicine  and  graduates 
who  have  such  preliminary  training  as  to  adapt  them  to  this  work. 

A  feature  of  the  undergraduate  instruction  that  may  be  well  to  indicate 
especially,  is  the  close  union  between  the  laboratories  of  pathological-histology 
and  morbid  anatomy,  and  the  museum  of  pathology.  In  order  that  the  gross 
changes  in  and  appearances  of  organs  may  be  correlated  with  the  histological 
alterations  as  shown  by  the  microscope,  the  gross  specimens  will  be  exhibited  in 
the  laboratory  of  morbid  anatomy  during  the  exercises  on  pathological-histology. 
The  arrangement  of  rooms  and  seating  is  such  that  the  student  may  enter  one 
room  from  the  other  without  creating  disturbance,  or  interfering  with  the 
illumination  of  the  microscopes  in  his  rear. 

It  is  believed  that  the  use  of  enormous  glass  fronts  for  the  histological  labora- 
tories will  provide  such  abundance  of  north  light  as  to  make  all  the  seats  of 
equal  value  for  microscopical  work.  Simon  Flexner. 

University  of  Pennsylvania. 


Magnifiers. 

After  some  years'  experience  as  teacher  and  examiner  of  classes  requiring  in 
their  work  the  use  of  magnifying  lenses,  I  have  come  to  the  conclusion  that 
fewer  persons  know  how  to  make  good  use  of  simple  microscope  than  of  the 
compound  one. 

The  majority  of  students  whom  I  have  met  have  used  either  the  folding 
lens  or  the  tripod.  The  former  is  convenient  for  carrying  in  the  pocket,  but  has 
the  disadvantage  of  requiring  the  exclusive  use  of  a  hand,  leaving  only  one  free 
to  manipulate  or  dissect  the  object  under  examination.  Such  single-handed 
manipulation  is  tedious  and  frequently  gives  very  imperfect  and  unsatisfactory 
results.  With  wire  and  cork,  one  can  improvise  a  holder  for  the  folding  magnifier, 
but  so  mounted  it  is  less  satisfactory  than  the  tripod. 

Within  two  years  I  have  tried,  with  three  classes  of  nearly  one  hun- 
dred students  in  each,  the  magnifier  known  as  the  watchmaker's  glass  with  two 


and  Laboratory  Methods. 


1449 


lenses.  The  lens  on  the  tip  may  be  removed,  thereby  rendering  the  remaining  lens 
lighter  to  hold  in  the  eye,  while  at  the  same  time  giving  sufficient  amplification  for 
most  work.  The  great  advantage  of  this  magnifier  is  that  both  hands  are  free, 
and  the  object  can  be  placed  or  held  up  in  the  most  favorable  light.  The 
objection  to  its  use  is  that  a  considerable  portion  of  the  students,  despite  the 
most  careful  directions  and  praiseworthy  perseverance  on  their  part,  are  unable 
to  retain  the  magnifier  on  the  eye.  This  year  I  have  had  a  detachable  spring 
added  to  the  mounting.  This  is  a  heavy  watch  spring  which  goes  round  the 
head  and  when  properly  adjusted  holds  the  lens  comfortably  in  a  suitable  posi- 
tion. Even  those  who  can  hold  the  lens  on  without  the  spring  find  that  when 
the  protracted  use  of  the  instrument  is  necessary  fatigue  is  reduced  to  a  minimum 
or  eliminated  by  using  the  spring.  The  latter's  being  detachable  permits  the 
glass  to  be  carried  in  the  pocket  and  used  in  the  hand  for  simple  magnification 
as  conveniently  as  a  folding  lens.  The  spring  is  kept  with  the  kit  of  dissecting 
tools  and  attached  when  desirable.  Its  use  so  far  is  proving  highly  satisfactory. 
London  Normal  School,  London,  Canada.  J.  DearNESS. 


A  New  Thermo-Regulator. 

The  following  is  a  simple  and  extremely  efficacious  form  of  thermo-regulator 
which  was  shown  to  me  some  while  ago  by  one  of  my  ingenious  friends,  and 
who  kindly  undertook  to  provide  me  with  one.  I  have  been  trying  it  on  an  air 
sterilizer  (one  of  Jung's)  with  the 
most  satisfactory  results.  I  have 
also  got  one  for  low  temperatures, 
as  per  modifications  suggested  by 
myself :  i  e,  India  rubber  cork 
through  which  passes  the  tube  t, 
at  the  inner  end  of  which  is  a  coni- 
cal entrance,  c.  B  is  a  glass  float 
which  is  counterpoised,  and  which 
either  rises  so  as  to  obstruct  the 
gas  entry  (the  black  end  closing 
the  tube  t),  or  it  descends  into  the 
cup  A.  This  cup  is  provided  with 
a  tube  which  dips  into  the  mercury. 
The  apparatus  being  put  into  its 
place,  the  heat  causes  the  mercury 
to  rise  into  the  cup  A,  and  lifts  B, 
which  will  finally,  i  e.,  at  a  given 
temperature,  obstruct  the  gas  pas- 
sage so  as  to  limit  the  supply  of 
gas,  and  thereby  govern  the  tem- 
perature. 

The  form  shown  is,  of  course,  serviceable  for  only  one  temperature,  but 
by  interposing  a  metallic  cap  and  screw,  as  shown  in  the  left  figure,  acting  on  a 
leather  diaphragm,  the  apparatus  may  be  regulated  to  any  temperature. 

Thos.  Pal.mer. 


1450  Journal  of  Applied  Microscopy 

A  Rapid  Method  of  Making  Slides  of  Amoeba. 

If  a  small  film  of  detritus  which  contains  abundant  amoebae  be  placed  in  a 
solid  watch  glass  with  plenty  of  water  and  examined  with  a  magnification  of  Id 
to  '10  diameters,  amoeba;  can,  after  a  little  practice,  be  readily  seen  and  can  be 
picked  out  with  a  thin-walled  dipping  tube  such  as  any  one  can  readily  make  for 
himself.  The  medicine  droppers  which  one  buys  have  walls  too  thick  to  be 
available. 

The  drop  of  water  containing  the  amoeba  may  be  placed  on  a  coverslip  and 
with  a  little  care  any  fragments  of  dirt  taken  up  with  it  can  be  removed  to  a  dis- 
tance by  needles  and  then  taken  away  altogether  by  a  cloth  or  a  bit  of  filter 
paper.  With  a  little  experience  also  it  will  be  easy  so  to  manipulate  the  cur- 
rents as  to  bring  the  amoeba  to  the  center  of  the  slide.  As  much  water  should 
be  drawn  off  as  is  possible  without  incurring  the  risk  of  allowing  the  animal  to 
dry.  After  he  has  been  quiet  for  a  few  moments  and  has  begun  to  put  forth  his 
pseud opodia,  he  adheres  slightly  to  the  glass  and  it  is  now  possible  by  a  sudden 
move  to  drain  off  the  rest  of  the  water  and  to  replace  it  by  a  small  drop  of  picric 
alcohol  (saturated  solution  of  picric  acid  in  50  per  cent,  alcohol).  If  the  alcohol 
is  placed  directly  upon  him  and  is  not  allowed  to  fall  from  any  considerable 
height,  the  attachment  to  the  glass  will  not  be  loosened.  The  cover  may  now  be 
inclined  somewhat  and  a  gentle  current  of  50  per  cent,  alcohol  allowed  to  flow 
over  it  until  the  amoeba  appears  quite  colorless.  Dehydration  may  be  accom- 
plished by  allowing  two  or  three  c.c.  of  each  of  the  higher  grades  of  alcohol  to 
flow  over  it  in  the  same  way.  This  done,  the  animal  may  be  permanently  fixed 
to  the  cover,  as  Overton  suggests,  by  adding  a  small  drop  of  a  very  dilute  solution 
of  collodion,  which,  by  tilting  the  slip  in  various  directions,  may  be  spread  out 
into  the  thinnest  possible  film.  As  soon  as  the  collodion  ceases  to  flow  it  may 
be  completely  hardened  by  dropping  the  cover,  amoeba-side  up  of  course,  into 
80  per  cent,  alcohol. 

In  this  alcohol  the  preparation  may  be  left  as  long  as  convenient,  or  it  may 
at  once  be  stained  with  any  suitable  stain,  such  as  borax  carmin  or  haematoxylin. 
I  am  accustomed  to  use  Syracuse  watch-glasses  for  manipulation  of  such  covers, 
and  the  only  precaution  necessary  is  to  incline  the  cover  somewhat  as  it  is  put 
into  a  fluid,  since  if  attention  is  not  given  to  this  point  the  entire  film  with  the 
specimen  may  float  away. 

The  collodion,  like  the  amoeba,  will  of  course  be  colored,  but  if  the  film  was 
not  too  thick  it  may  be  entirely  decolorized  before  the  color  is  withdrawn  from 
the  specimen.  In  dehydrating,  amylic  alcohol,  which  does  not  dissolve  collo- 
dion, should  be  substituted  for  the  ordinary  absolute  ethyl  alcohol.  If  the  speci- 
men is  so  large  that  supports  are  needed  for  the  cover,  two  slips  of  paper  pre- 
viously soaked  in  xylol  may  be  used  instead  of  wax  feet.  The  final  step  is  of 
course  to  place  a  drop  of  balsam  upon  a  slide  and  invert  the  cover  upon  it. 

Although  the  process  may  sound  somewhat  tedious,  it  is  really  a  rapid  one.  I 
have  repe<itedly  put  away  a  completed  specimen  in  my  cabinet  less  than  half  an 
hour  from  the  time  when  the  amoeba  was  crawling  about  in  his  home.  I  should 
add  that  I  have  not  made  any  cytological  study  of  specimens  prepared  in  this 
way.     For  purposes  of  demonstration,  however,  they  are  exceedingly  satisfactory. 

Wellesley  College.  M.  A.  WiLLCOX. 


and  Laboratory  Methods. 


1451 


MICRO-CHEMICAL  ANALYSIS. 
XVI. 

ZINC. 

Although  zinc,  from  its  position  in  the  periodic  system,  closely  resembles 
magnesium  in  general,  in  its  chemical  behavior,  the  majority  of  the  micro- 
chemical  reactions  of  the  two  elements  are  quite  different.  We  have  already 
seen,  however,  that  with  uranyl  acetate  and  sodium  acetate,  and  with  arsenic 
acid,  magnesium,  zinc  and  cadmium  give  identical  reactions. 

A  number  of  reagents  have  been  proposed  for  the  detection  of  zinc,  but  of 
these  only  the  folio  vving  need  to  receive  our  attention  : 

I.  Double  Sulphocyanate  of  Mercury  and  Ammonium. 
II.  Oxalic  Acid. 
III.  Primary  Sodium  Carbonate. 
Of  these,  the  third  is  the  most  sensitive  and  most  characteristic,  but  is  not 
so  simple,  convenient  nor  so  easily  applied  as  is  the  first.  The  second  reagent- 
oxalic  acid  —  is  unsatisfactory  and  of  comparatively  little  value. 


/.   Ammonintfi  Mercuric  Sulphocyanate  added  to  neutral  or  slightly  acid  solu- 
tions containing  Zinc,  precipitates  a  Double  Sulphocyanate  of  Zinc  and  Mercury. 


ZnSO^    +    [^(NH^CNS) 


Hg(CNS)2]=   [Zn(CNS)2 
(NH,),SO, 


Hg(CNS)2]   + 


Method. — The  reagent  is  prepared  by  adding  to  an  almost  saturated  solution 
of  mercuric  chloride  a  saturated  solution  of  ammonium  sulphocyanate  in  slight 
excess  of  the  amount  required  by  theory  to  form  the  double  salt  of  the  formula 
given  above.  The  solution  thus  prepared  is  employed  as  the  reagent.  It  suffers 
no  deterioration  on  keeping. 

Next,  to  a  small  drop  of  the  solution  to  be  tested,  place  a  tiny  drop  of  the 
reagent  and  cause  the  latter  to  flow  into  the  test 
drop  by  means  of  a  glass  rod,  at  the  same  time  in- 
clining the  slide.  Almost  immediately,  pure  white 
feathery  crosses  and  branching  feathery  aggregates 
separate  (Fig.  68).  These  skeleton  crystals,  when 
thick,  appear  black  by  transmitted  light,  snow 
white  by  reflected  light.  The  normal  crystal  of 
the  double  sulphocyanate  of  zinc  and  mercury  is 
said  to  be  a  right-angled  prism  of  the  orthorhombic 
system,  but  under  the  conditions  which  obtain  in 
ordinary  practice,  only  skeleton  and  dentritic  forms 
will  be  seen. 

Remarks. — Employ  dilute  solutions  only.  Mitigate  the  action  of  free  min- 
eral acids  by  the  addition  of  ammonium  or  sodium  acetate. 


Fig.  OS. 


1-1'52  Journal  of  Applied  Microscopy 

Avoid  adding  too  much  reagent.  This,  however,  is  a  matter  of  little  import- 
ance when  zinc  alone  is  present,  but  it  is  quite  necessary  when  dealing  with 
mixtures. 

Neither  magnesium  nor  aluminum  interfere  with  this  test,  save  that  when 
magnesium  is  present  in  large  amount  the  separation  of  the  zinc  salt  is  retarded, 
and  that  aluminum  under  similar  conditions  renders  the  skeleton  crystals  of 
the  zinc  salt  somewhat  less  feathery. 

The  reagent  gives  reactions  with  zinc,  cadmium,  copper,  cobalt  and  indium. 
These  reactions  are  among  the  most  interesting  and  elegant  of  micro-chemistry 
and  leave  little  to  be  desired. 

When  zinc  alone  is  present  the  crystals,  as  has  been  stated  above,  are  snow 
white  and  of  the  form  shown  in  Fig.  68  ;  but  if  copper  is  present  in  minute 
amount,  the  crystals  of  the  zinc  salt  are  colored  chocolate  brown  without  under- 
going any  change  of  form.  These  brown  crystals  begin  to  appear  after  the 
white  ones  have  separated.  More  copper  than  sufficient  to  yield  the  brown  tint 
produces  black  crystals  of  modified  form  ;  still  a  greater  proportion  of  copper 
completely  changes  the  appearance  of  the  crystals,  and  jet  black  spheres  and 
botryoidal  masses  result.  Finally  a  point  is  reached  where  crystals  of  copper 
mercuric  sulphocyanate  predominate,  accompanied  by  the  black  crystals  just 
mentioned. 

This  change  in  color  of  the  zinc  salt  brought  about  by  the  presence  of  copper 
is  a  most  interesting  one.  The  zinc  compound  —  Zn(CNS)  2  •  Hg(CNS)2  —  con- 
tains no  water  of  crystallization,  while  the  copper  salt  normally  separates  as — 
Cu(CNS)2  •  Hg(CNS)2  '  ^2^  —  ^nd  being  hydrated  is  greenish  in  color.  The 
presence  of  water  of  crystallization  in  salts  of  copper  seems  to  determine  their  color. 
The  removal  of  the  water  leads  to  the  production  of  a  brown  or  almost  color- 
less body.  The  nature  of  this  change  is  not  yet  thoroughly  understood.  It 
seems  probable  that  in  the  case  of  the  brown  and  black  copper-zinc-mercury 
sulphocyanates  we  have  to  deal  with  a  case  of  solid  solution,  although  it  is  also 
conceivable  that  an  anhydrous  copper-mercury  double  salt  may  exist  in  the  pres- 
ence of  the  zinc  compound,  isomorphous  with  the  latter,  yet  incapable  of  exist- 
ing alone. 

In  the  presence  of  cobalt,  the  zinc  salt  is  colored  blue,  the  intensity  of  the 
coloration  depending  upon  the  amount  of  cobalt  present.  With  very  small  amounts 
the  color  is  exceedingly  faint  and  the  crystal  form  unchanged,  but  as  the  pro- 
portion of  cobalt  increases,  the  skeleton  crystals  of  the  zinc  s.alt  become  deeper 
and  deeper  blue,  simpler,  less  feathery,  and  gradually  assume  the  color  and 
appearance  of  the  normal  cobalt  mercuric  sulphocyanate.  As  in  the  case  of  the 
copper-zinc  compound,  these  blue  crystals  are  doubtless  cases  of  solid  solution, 
but  the  theory  of  isomorphous  mixture  is  more  tenable  in  this  case  than  in  that 
where  copper  is  present. 

Small  amounts  of  zinc  in  the  presence  of  much  cobalt  cannot  be  detected  by 
this  reagent. 

Cadmium  gives  long  prismatic  crystals  (Fig.  71),  which  are  more  soluble 
than  the  zinc  salt.  Even  a  small  amount  of  cadmium  destroys  the  feathery  and 
branched  character  of  the  skeletons  of  the  zinc-mercury  sulphocyanate,  owing 


and  Laboratory  Methods.  1453 

to  the  formation  of  mixed  crystals,  and  there  generally  result  crystallites  of  the 
shape  of  an  arrowhead.  Small  amounts  of  zinc  in  the  presence  of  much  cad- 
mium will  usually  escape  detection. 

The  presence  of  both  copper  and  cobalt  in  a  solution  containing  zinc  gives 
rise  to  the  formation  of  mixed  crystals  of  very  peculiar  color  and  form.  These 
peculiarities  are  accentuated  when  cadmium  is  also  present.  The  experienced 
worker  thus  will  have  little  difficulty  in  detecting  a  number  of  elements  in  one 
single  operation. 

Indium  forms  with  the  reagent  a  double  sulphocyanate,  crystallizing  in  forms 
resembling  those  of  the  corresponding  cadmium  double  salt.  The  reaction  is 
quite  slow  in  the  case  of  indium. 

When  iron  is  present  in  sufficient  amount  to  give  a  blood-red  color  to  the 
preparation  on  the  addition  of  the  reagent,  the  crystals  of  the  double  sulphocy- 
anate of  zinc  and  mercury,  separating  from  such  solutions,  are  colored  a  deep 
reddish  brown,  appear  jet  black  by  transmitted  light,  and  have  at  first  the  usual 
form  of  the  zinc  double  salt.  The  appearance  of  these  crystals  usually  changes 
rapidly,  and  in  a  few  seconds  bunches  and  masses  of  curving,  branching,  fili- 
form crystals  are  seen.  The  change  is  a  very  remarkable  one  and  takes  place 
rapidly. 

Lead,  unless  present  in  large  amount,  seems  to  have  little  or  no  effect  on  the 
zinc  reaction.  Under  some  conditions  it  seems  to  interfere,  however,  and  it  is, 
therefore,  always  best  to  first  remove  the  lead  by  means  of  dilute  sulphuric  acid. 
Add  the  acid,  draw  off  or  filter;  evaporate  the  clear  solution  to  dryness;  fume 
off  the  free  sulphuric  acid ;  dissolve  in  water  ;  add  ammonium  acetate,  and  test 
as  above. 

Silver  gives  with  the  reagent  a  white  amphorous  precipitate,  soon  crystalliz- 
ing in  the  form  of  small,  thin,  slender  prisms  with  square  or  oblique  ends, 
somewhat  resembling  those  of  the  cadmium-mercury  salt,  but  very  much  smaller 
than  the  latter.  In  the  presence  of  silver  the  test  for  zinc  is  sometimes  masked. 
In  such  an  event,  first  remove  the  silver  with  hydrochloric  acid  and  test,  after 
evaporation,  in  the  usual  manner. 

Exercises  for  Practice. 

Apply  the  reagent,  in  the  manner  indicated,  to  solutions  of  a  pure  Zn  salt  of 
different  degrees  of  concentration. 

To  a  Zn  solution  add  a  very  little  Cd  and  test.  Repeat  the  experiment, 
using  more  Cd. 

In  like  manner  try  mixtures  of  Zn  and  Cu  ;  Zn  and  Co  ;  Zn  and  Ni  ;  Zn 
and  Fe  ;  Zn  and  Mg  ;  Zn  and  Al  ;  Zn  and  Pb  ;  Zn  and  Ag. 

Then  try  more  complex  mixtures,  as  for  example  :  Zn,  Cd  ana  Cu  ;  Zn,  Cd 
and  Co  ;  Zn,  Cu  and  Co  ;  etc. 

In  each  case  prepare  several  slides  under  different  conditions  and  note  well 
the  changes  in  the  appearance  of  the  crystals  which  separate. 

See  also  remarks  and  suggestion  of  experiments  given  under  Cadmium, 
Copper  and  Cobalt. 


1454  Journal  of  Applied  Microscopy 

//.  Oxalic  Acid  added  to  solution  containing  Zinc  causes  the  separation  of  Zitic 
Oxalate. 

Z11SO4  +  H2C2O4  ==  ZnCoO^  .   2H2O  +  H2SO4. 

Method. — The  reagent  is  applied  to  the  test  drop,  as  in  previous  tests,  with 
oxalic  acid,  i.  e.,  employ  a  concentrated  solution  and  cause  it  to  flow  into  the 
test  drop. 

Small  double  spherulites,  pseudo-octahedra,  either  singly  or  united  in  twos, 
and  thin  rhombs  result.     (Fig.  69.) 

{\         ^  The  great   majority   of  the  crystals    separating  have 

^  ^^  ^^         their  angles   rounded.     It   is   tare  that   a   preparation  is 
O   ^P  '^ B,-^     obtained  yielding  clear-cut  crystals. 
^   ,,=po=^  -)  Remarks. — The   solution  to  be  tested  should  be  neu- 

^  vis^  /^A  (-i-^i  Qj-  Qi^]y  slightly  acid. 

"^  Crystals  of  zinc  oxalate,  when   examined  with  a  low 

NX)w--o.oVvyx«^  power,  often  bear  a  striking  resemblance  to  the  oxalates 
Fig.  69.  of  calcium   and   strontium  ;  for  this  reason   the  alkaline 

earths  should  be  first  removed. 

M  ignesium  interferes.  Under  certain  conditions  a  double  oxalate  of  zinc 
and  magnesium  separates  in  the  form  of  hexagonal  plates. 

Ammonium  salts  should  be  removed  before  adding  the  oxalic  acid. 

In  the  presence  of  cadmium  this  test  for  zinc  is  unreliable. 

Recrystallization  of  the  zinc  oxalate  from  a  solution  of  ammonium  hydroxide 
sometimes  yields  good  results,  and  will  aid  in  reaching  a  decision  as  to  what 
element  has  been  precipitated  by  the  oxalic  acid.  In  recrystallizing  proceed  as 
follows  after  adding  the  oxalic  acid  :  Carefully  separate  the  solution  from  the 
precipitate ;  add  to  the  latter  a  large  drop  of  ammonium  hydroxide ;  warm 
gently ;  cool  and  examine.  Zinc  oxalate  separates  from  such  solutions  in  the 
form  of  tufts  and  aggregates  of  very  fine  needles.  Occasionally  masses  of 
radiating,  curving  needles  are  seen.  In  most  preparations  the  crystals  separat- 
ing resemble  the  tufts  formed  by  calcium  sulphate.  These  crystals  are  not 
obtained  if  cadmium  or  magnesium  is  present. 

If  lead,  copper,  cobalt  or  nickel  should  be  present,  it  is  necessary  to  first 
effe'  t  a  separation  before  testing  for  zinc  with  oxalic  acid. 

Unless  present  in  very  small  amount,  iron  interferes. 

Oxalates  of  Group  I  also  yield  precipitates  consisting  of  normal  and  double 
oxalates,  but  these  are  of  little  value  as  tests  for  zinc. 

Exercises  for  Practice. 

See  suggestions  under  Cadmium. 

///.  Zinc  forms,  loith  P/imary  Sodium  Carbonate,  a  Double  Carbonate  of  Zinc 
and  Sodium  of  low  solubility. 

8  ZnSO^  +  22  HNaC03=  [3  (NajCOg)  .   8  (ZnCO.j)  •    8H2O]  -^  8  Na2S04 

+  11  CO2  +  11  H2O. 

Method. — Prepare  a  saturated  solution  of  the  reagent.     Place  a  large  drop 


and  Laboratory  Methods.  1455 

of  this  solution  next  to  the  drop  to  be  tested.  Tip  the  slide  a  very  little  and 
cause  the  reagent  to  flow  into  the  test  drop.  An  amorphous  precipitate  of  basic 
zinc  carbonate  is  generally  at  once  produced.  After  a  short  time,  if  the  reagent 
is  in  excess,  the  double  carbonate  will  appear  at  the  edges  of  the  test  drop 
nearest  the  reagent  as  small,  colorless,  triangular  and  tetrahedral  crystals.  (Fig. 
70.)  These  crystals  adhere  strongly  to  the  glass  and  are  very  characteristic  of 
zinc. 

Remarks. — It   is  essential   that   an   excess  of    the 
reagent  be  employed.     Failure  not  infrequently  results 
from  a  neglect  of  this  precaution.    This  is  particularly         V~y 
true  if  the   test  drop  is   acid.     Because  of  the  neces-  ^  A 

sity  of  adding  large  amounts  of  primary  sodium  car-  ^  ^  ^ 
bonate,  the  test  drop  must  be  of  greater  volume  than  >«  ^._^  \l 
is  usual  in   micro-chemical  testing  and   must  be  corre-  ^^ 

spondingly  dilute. 

The  formation  and  separation  of  the  double  salt  is 
rather  slow.  ^P' 

Other  carbonates,  as  for  example,  those  of  potas-  ■^^S"  ^^' 

slum  and  lithium,  can  be  substituted  for  primary  sodium  carbonate,  but  the  reac- 
tions are  not  so  satisfactory. 

Salts  of  ammonium  must  be  absent. 

It  is  unfortunate  that  this,  which  is  one  of  the  most  characteristic  as  well  as 
delicate  of  the  micro-chemical  tests  for  zine,  should  be  open  to  many  difficul- 
ties. The  chief  of  these  lies  in  the  fact  that  many  elements  are  precipitated  as 
carbonates,  and  that  these  often  bulky  precipitates  interfere  with  or  mask  the 
zinc  reaction.  Among  the  interfering  elements,  those  most  frequently  met  with 
are  doubtless  calcium,  strontium,  barium,  magnesium,  cadmium,  lead,  iron,  man- 
ganese, cobalt,  nickel.  Of  this  list,  calcium,  strontium,  barium  and  lead  will 
probably  have  been  removed  by  previous  treatment  with  sulphuric  acid.  For 
method  for  dealing  with  mixtures  containing  the  remaining  elements  of  the  list, 
see  Separation  of  the  Magnesium  Group. 

If  only  a  very  small  amount  of  cadmium  is  present,  it  is  precipitated  before 
the  zinc,  and  by  avoiding  the  addition  of  an  excess  of  the  reagent,  drawing  off 
the  clear  liquid  and  adding  to  the  decanted  liquid  a  fresh  portion  of  the  reagent 
in  sufficient  quantity,  the  zinc  can  be  precipitated  as  the  double  carbonate. 
When  considerable  cadmium  is  present  this  method  is  not  feasible.  In  such  an 
event  recourse  may  be  had  to  ammoniacal  solutions,  as  suggested  by  Behrens.* 
The  test  drop  is  made  strongly  ammoniacal  and  to  it  primary  sodium  carbonate 
is  added.  Cadmium  is  immediately  precipitated,  while  the  zinc  remains  in  solu- 
tion. The  clear  solution  is  separated  at  once  from  the  precipitate  and  allowed 
to  stand  for  a  short  time.  Zinc  separates  from  the  decanted  solution  as  the 
double  carbonate  in  the  forms  shown  in  Fig.  70.  Some  little  skill  and  experi- 
ence is  generally  necessary  in  order  to  obtain  good  results. 


*  Anleitung,  2  Auf.  p.  52. 


1456 


Journal  of  Applied  Microscopy 


Exercises  for  Practice. 

Try  precipitating  Zn  in  acid,  neutral  and  ammoniacal  solutions. 

Test  mixtures  of  Zn  and  Cd,  first  in  neutral,  then  in  ammoniacal  solutions. 

Experiment  with  Zn  in  the  presence  of  the  interfering  elements  noted  above. 


CADMIUM. 

Cadmium,  in  the  absence  of  zinc,  can  be  very  easily  and  satisfactorily 
detected  by  either  : 

I.  Ammonium  Mercuric  Sulphocyanate,  or 
II.  Oxalic  Acid. 

But  if  zinc  is  also  present,  great  care  must  be  exercised  to  avoid  being  led 
into  error,  for  these  two  elements  are  very  much  alike  in  their  chemical 
behavior. 

Several  other  reagents  have  been  suggested  for  the  detection  of  cadmium, 
but  it  can  be  said  of  all  of  them  that  the  results  are  not  satisfactory,  even  when 
working  with  pure  salts  of  cadmium,  and  that  they  fail  completely  when  dealing 
with  complex  mixtures. 

/.  Cadmium  forms,  ivith  Ammonimn  Mercuric  Sulphocyanate,  a  Double  Sulpho- 
cyanate of  Cathniutn  and  Mercury. 

CdSO^  +  [2  (NH^CNS)  •   Hg  (CNS)2]  =  [Cd  (CNS),  •  Hg(CNS)o]  + 

(NHjio,. 

Method. — Proceed  exactly  as  directed  under  Zinc,  Method  I,  avoiding  an 
excess  of  the  reagent.     Long,  highly  refractive  prisms  separate.     (Fig.  71.) 

/"^X  The  appearance   of  these    prisms   varies    with 

/     /       \\         \\  the  conditions  which   obtain  at  the   time   of  their 

^r  Jf^  \  \  n  \x  formation,  as,  for  example,  the  concentration,  depth 
of  the  test  drop,  amount  of  reagent  added,  acidity, 
etc.  These  variations  are,  however,  not  of  a  kind 
to  render  the  test  doubtful ;  long  prisms,  either 
singly  or  in  groups,  being  the  rule. 

Remarks. — The  remarks  made  under  zinc  are 
applicable  to  cadmium  in  every  case. 

The    double    sulphocyanate    of    cadmium    and 
>P''"°-°"^n>         mercury  is   more  soluble  than  that  of  zinc,  hence 
Fig.  71.  the  reaction  is  slower  and  more  concentrated  solu- 

tions should  be  employed. 

If  a  small  amount  of  zinc  is  also  present,  mixed  crystals  containing  zinc 
and  cadmium  first  separate  whose  crystal  form  can  be  described  as  non-feathery 
skeletons ;  soon  after  this  the  cadmium  double  salt  separates  in  its  usual  form. 
In  order  that  this  sequence  shall  be  brought  about,  it  is  best  to  employ  a  solu- 
tion somewhat  more  dilute  than  when  zinc  is  absent.  Much  zinc  usually  pre- 
vents the  formation  of  any  of  the  prismatic  crystals  of  the  cadmium  salt,  only 
mixed  crystals  resulting. 


and  Laboratory  Methods. 


1457 


Traces  of  copper  color  the  cadmium  crystals  a  faint  chocolate  brown  ;  this 
brown  color  intensifies  with  an  increase  in  the  amount  of  copper.  When  con- 
siderable copper  is  present,  the  copper  double  salt  first  separates,  since  it  is 
slightly  less  soluble  than  the  cadmium  compound  ;  then  mixed  crystals  form,  in 
which  the  copper  apparently  predominates  over  the  cadmium.  These  mixed 
crystals  are  of  a  deep  bluish-green  color.  By  this  time  most  of  the  copper  and 
but  little  of  the  cadmium  has  been  precipitated,  and  the  concentration  has  also 
reached  such  a  point  that  the  cadmium  double  salt  begins  to  separate  in  the 
crystal  forms  shown  in  Fig.  71.  These  are,  however,  still  mixed  crystals,  for 
they  are  colored  brown  by  the  small  amount  of  copper  yet  in  solution. 

It  is  improbable  that  these  brown  copper-cadmium-mercury  sulphocyanates 
are  isomorphous  mixtures. 

As  in  the  case  of  the  zinc  reaction,  iron  may  sometimes  color  the  cadmium 
salt  a  reddish  brown. 

Cobalt  colors  the  cadmium  salt  blue.  Much  cobalt  gives  an  intense  blue 
color  and  alters  the  crystal  form. 

Magnesium  and  aluminum  have  even  less  effect  than  in  the  case  of  zinc. 

Before  testing  for  cadmium  with  the  sulphocyanate  reagent,  it  is  best  to  first 
remove  any  lead  or  silver  which  may  be  present. 

See  also  remarks  under  Zinc,  Method  I. 

Exercises  for  Practice. 

Experiment  with  salts  of  cadmium  in  the  manner  suggested  under  "  Zinc," 
trying  all  the  exercises  mentioned,  but  having  cadmium  as  the  element  in  excess 
instead  of  zinc. 


//.    Oxalic  Acid  added  to  solutions  of  salts  of  Cadmium  pi-ecipitates  Cadmium 
Oxalate. 

CdSO^  +  H2C20^  =  C^CO^  '  3H2O  +  H2SO4. 

Method. — To  the  test  drop  add  a  solution  of  the  reagent  by  the  flowing  in 
method.  Clear,  colorless  monoclinic  prisms  and  tabular  crystals  separate,  either 
singly,  in  Xs,  or  in  clusters.  (Fig.  72.) 
The  tabular  crystals  have  the  appearance 
of  rhombs  and  rectangles. 

Frequently  very  concentrated  solutions 
yield   crystals  having  an  octahedral  aspect. 

Retnarks. —  The  solution  to  be  tested 
should  be  neutral  or  only  slightly  acid, 
and  rather  concentrated  with  respect  to 
cadmium. 

Dilute  solutions  fail  to  give  good  re- 
sults. 

The  typical  crystals  of  cadmium  oxa- 
late are  seen  only  when  working  with  al- 
most pure  salts  of  this  element.  Fig.  72. 


1458  Journal  of  Applied  Microscopy 

In  the  presence  of  zinc,  only  the  forms  of  zinc  oxalate  are  usually  obtained. 

Members  of  the  calcium  group  and  lead  are  first  removed  with  sulphuric  acid 
and  a  trace  of  alcohol.     Silver  with  hydrochloric  acid. 

In  the  presence  of  copper,  aluminum,  iron,  manganese,  chromium,  nickel 
and  cobalt,  the  reaction  with  oxalic  acid  is  not  reliable  and  in  most  cases 
worthless. 

Treated  with  ammonium  hydroxide  in  the  manner  described  under  Zinc, 
cadmium  oxalate  recrystallizes  in  the  form  of  rods  and  tables.  This  method  of 
procedure  is  often  of  value  in  arriving  at  a  decision  as  to  the  nature  of  a  pre- 
cipitate obtained  with  oxalic  acid.  Unfortunately,  zinc  prevents  the  formation 
of  these  rod-like  crystals. 

Exercises  for  Practice. 

Test  a  pure  salt  of  Zn  in  dilute  and  in  concentrated  solution.  Repeat  the 
experiments,  substituting  Cd  for  the  Zn. 

Make  a  preparation  of  ZnCoO^  •  ^H^O  ;  draw  off  the  supernatant  liquid  ; 
add  NH^OH  ;  warm  gently  and  study  the  preparation.  Prepare  slides  of  differ- 
ent degrees  of  concentration. 

Recrystallize  CdCgO^  •  '^Yi<fi  in  the  same  manner  as  the  Zn  salt. 

Test  mixtures  of  Zn  and  Cd. 

Recrystallize  the  mixed  oxalates  from  NH^OH. 

Make  mixtures  of  Zn  and  the  interfering  elements  listed  above.  Treat  the 
precipitated  oxalates  with  NH^OH.     Then  try  Cd  in  the  same  manner. 

Try  precipitating  Zn  with  HKCgO^,  K2C2O4,  (NH4)2C204,  etc.  Then 
try  Cd  in  like  manner.  E.  M.  Chamot. 

Cornell  University. 


Device  for  Leveling  the  Microscope. 

In    examining    objects    in    liquids    on    the    stage    of    a    microscope,    the 

want  of  true  level  annoys  by  keeping  up  cur- 
rents and  displacing,  often  disastrously,  the 
object  sought. 

By  this  simple  and  inexpensive  device  all 
this  trouble  could  be  overcome. 

One  need  not  even  add  the  expense  of  a 
small  "  spirit  "  level ;  for  a  glass  slip  on  which 
are  placed  a   few  drops   of  water  containing 

.r.  ThreaWd  hole"fo''r  leyeilng  screw.  particles    of    opaque    material    which    would 

b.  Microscope  pillar.  ,  .1  .  .  ,•  ■    -i  1 

render  any  tendency  to  current  motion  visible, 
=Y=  will  answer  the  purpose. 

I    :  k\  Place  this  trial  slip  on  the  stage,  and  level 

by  means  of  the  three  screws  until  no  currents 


A.  End  view  of  microscope  foot.  . 

B.  Milled  hefld  of  leveling  screw.  are   perceptible 


T.  O.  Reynolds. 


and  Laboratory  Methods.  1459 

Journal    of  The  Journal   has  been  called  the 

A          1  •     J     1\  yi  •  Clearing  House  for  Methods,   and    we 

Applied    Microscopy  hope  that  it  nils  a  place  among  scien- 

.     ,               ^"    n/i     1      1  tific  publications  such  that  the  term  may 

Laboratory   Methods.  be  truly  applicable,     a  clearing  house 


Edited  by  L.  B.  ELLIOTT.  ^^'^  methods,  just  as  a  clearing  house 

for    accounts,    is,  however,  dependent 


Issued  Monthly  from  the  Publication  Department  „»,„„     ^,,«.^;j„      ^^,,^^^„    f^^      ;+^     .^„;v, 

of  the  Bausch  &  Lomb  Optical  Co.,  upon   outside    sourccs   for     its    mam- 

ochester,    .    .  tcnance,  and  without  the  cooperation 


SUBSCRIPTIONS:  and  support  of  its  adherents  must  cer- 

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The    summer    months    have    closed 


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of  the  coming  year.  By  many  the  vacation  is  taken  as  an  opportunity  to 
do  original  work  in  some  summer  laboratory  ;  others  leave  their  own  laboratories 
for  the  purpose  of  securing  recreation  and  pleasure  ;  but  teachers,  wherever  they 
go,  seldom  forget  the  work  that  is  before  them,  and  are  constantly  alert  for 
methods  and  improvements  in  the  study  or  presentation  of  their  subject.  Visits 
to  strange  laboratories  and  contact  with  new  minds  give  new  and  valuable  sug- 
gestions for  work.  These  should  be  allowed  to  pass  the  clearing  house,  and 
their  helpfulness  made  as  general  as  possible. 

The  suggestions  you  have  received  from  some  other  worker  in  your  field, 
the  improvement  you  have  made  in  your  method  of  work  at  the  summer  or  field 
laboratory,  may  seem  of  little  importance  to  you,  but  may,  if  allowed  to  circulate, 
come  to  the  hands  of  some  who  need  just  what  you  have  to  give. 

If,  on  the  other  hand,  in  your  work  you  have  met  a  difficulty  which  you  have 
been  unable  to  solve,  through  the  clearing  house  you  may  expect  to  receive  an 
answer  to  your  question. 

No  doubt  there  are  few  of  our  readers  who  would  admit  that  they  had  spent 
the  entire  summer  without  having  learned  something  that  will  be  of  benefit  to 
them  during  the  coming  year.  Would  it  not  be  well  to  give  others  an  oppor- 
tunity to  profit  by  the  advancement  you  have  made  ? 

* 

The  article  on  Photo-micrography,  by  Dr.  D.  W.  Dennis,  which  appeared  in 
the  Department  of  Laboratory  Photography  last  month,  was  the  introduction  to 
a  series  of  articles  which  the  author  will  contribute  on  that  subject  during  the 
coming  year.  The  series  will  include  "  Apparatus,"  "  Illuminating  the  Object," 
"  Focusing  for  very  high  and  very  low  powers  with  long  bellows,"  "  The 
Negative  and  Positive,"  etc.,  and  the  author  will  endeavor  to  put  into  them  the 
most  valuable  things  now  known  on  the  subject. 


1460  Journal  of  Applied  Microscopy 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Wettstein    Dr.   R.  von.    Handbuch  der  Syste-  ^j^is  volume   deals  with    those   plants 

matischen    Botanik.     I :  v.  +    201.       Figs.  _  ^ 

762,  in    1 28  plates.    Franz  Deuticke,  Leip-.  which      are      usually     termed     Thallo- 

zig.  Germany,  iqoi.     7   marks.  phytes.       A  second    volume,  which  will 

be  ready  some  time  within  the  next  year,  will  treat  the  Bryophytes,  Pterido- 
phytes  and  Spermatophytes,  which  the  author  will  describe  under  the  term,  Cor- 
mophytes.  It  is  the  purpose  of  the  book  to  give  a  comprehensive  view  of  plant 
forms  with  particular  reference  to  development  and  phylogeny.  This  purpose  is 
accomplished  by  a  full  presentation  of  the  larger  divisions  and  by  giving  the  de- 
velopmental history  of  a  large  number  of  the  more  important  types. 

The  book  is  intended  for  those  who  would  know  systematic  botany  from  the 
phylogenetic  standpoint,  but  it  will  also  be  very  helpful  to  those  who  need  such 
a  taxonomic  background  for  morphological  and^  citological  work.  While  the 
author  is  indebted  toother  taxonomic  works,  and  especially  to  Engler  and  Prantl's 
Z)ie  Natiirlichen  Pfl anzetifamilien ,  the  work  is  by  no  means  a  compilation.  A 
chapter  on  the  history  of  taxonomy  gives  a  brief  summary  of  the  systems  of  Jus- 
sieu,  A.  P.  DeCandolle,  Endlicher,  Brogniart,  A.  Braun,  Eichler,  and  Engler. 

In  a  phylogenetic  classification  many  things  must  be  considered  and  it  is  not 
always  easy  to  decide  whether  a  plant  is  high  or  low  in  any  particular  respect. 
In  general,  the  lines  of  advance  are  the  same  as  those  given  in  Die  Ahxtiirlichen 
PjJanze7ifatnilie?i.  The  possibility  of  a  polyphyletic  origin  must  be  admitted  be- 
cause it  is  known  that  similar  life  conditions  tend  to  produce  similar  morphologi- 
cal structures.  The  fossil  record  shows  that  Angiosperms  are  more  recent  than 
Gymnosperms  and  Pteridophytes,  and  that  Pteridophytes  are  older  than  Gym- 
nosperms,  but  the  record  is  too  fragmentary  to  be  of  much  importance  in  deter- 
mining the  relative  positions  of  smaller  divisions.  Comparative  morphology 
must  be  the  principal  basis  for  classification.  The  evidence  of  geographical  dis- 
tribution, of  rudimentary  organs,  monstrous  forms,  juvenile  forms  and  anatomi- 
cal details  must  be  weighed,  and  it  must  be  remembered  the  ontogeny  of  a  form 
may  give  useful  hints  as  to  its  phylogeny. 

The  first  forty-five  pages  are  occupied  by  a  discussion  of  the  principles  of 
classification  ;  the  rest  of  the  book  is  devoted  to  plants  which  are  usually  desig- 
nated as  Thallophytes.  These  comprise  six  genetic  lines  (Stammen)  between 
which  it  is  not  possible  at  present  to  demonstrate  relationships,  although  such 
may  exist.  The  lines  are  Myxophyta^  Schizophyta,  Zygophyfa,  Euthallophyta^ 
Phceophyta  and  Rhodophyta.  The  term  "  Algai  "  is  usually  applied  to  the  inde- 
pendent members  of  these  groups,  and  "Fungi  "  to  the  parasitic  and  saprophytic 
forms.     Each  group  with  its  orders  and  families  is  clearly  characterized  and  the 


and  Laboratory  Methods.  1461 

life  histories  of  typical  forms  are  thoroughly  illustrated.  The  most  important 
genera  and  the  commonest  species  are  often  mentioned,  so  that  while  the  book 
does  not  pretend  to  be  a  manual  for  the  identification  of  genera  or  species,  it 
nevertheless  serves  this  purpose  in  many  cases.  The  large  number  of  excellent 
illustrations,  together  with  the  clear  style  in  which  the  book  is  written,  afford  the 
English  speaking  student  a  good  opportunity  for  improving  his  German  while 
increasing  his  knowledge  of  Algae  and  Fungi.  c.  j.  c. 

Bernard,  Ch.  Recherches  surles  spheres  attract-  For  the  past  five  or  six  years  many  in- 
ives  Chez  Lilium  candidum,  Helosis  guaya-      vestigators   have  denied   the  existence 

nensis,  etc.     Jour,  de  Botanique      14:    ii6-  ° 

124,  i77-[SS,  206-212,  pis.  4-5,  1900.  of    centrosomes  in   the   higher  plants, 

while  other  investigators,  working  with 
practically  the  same  material  and  employing  the  same  methods,  have  insisted 
that  the  centrosomes  are  present.  Prof.  Bernard  has  examined  Lilium  candidu7n, 
L.  Martagoii  and  Helosis  guayancnsis  and  has  convinced  himself  of  the  presence 
of  these  much  discussed  structures.  Material  was  fixed  in  alcohol  and  in  Flem- 
ming's  solution  and  was  stained  in  a  mixture  of  fuchsin  and  iodin  green  (1  per 
cent,  aqueous  solution  of  fuchsin,  '1  parts  ;  1  per  cent,  aqueous  solution  of  iodin 
green,  2  parts,  and  water  40  parts).  The  safranin-gentian-violet  orange  combi- 
nation did  not  give  as  good  results.  In  L.  cafididumthe  centrosomes  were  found 
tjuite  regularly  during  various  phases  in  the  germination  of  the  megaspore.  They 
resemble  the  structures  described  by  Guignard,  but  are  not  so  sharply  defined. 
The  centrosome  was  also  identified  in  the  gametophytes  of  Helosis.  In  Z.  Mar- 
tagon  centrosomes  were  found  in  the  female  gametophyte,  in  the  vegetative  cells 
of  the  ovule,  but  could  not  be  positively  identified  in  the  endosperm.  The  centro- 
some is  cytoplasmic  in  origin. 

Incidentally,  it  is  noted  that  there  are  sometimes  two  embryo  sacs  in  Z.  ca?idi- 
dum.  In  this  species  a  very  large  vacuole  develops  between  the  two  polar 
nuclei,  preventing  the  nuclei  from  fusing.  The  writer  suggests  that  this  may 
account  for  the  sterility  of  this  species.  It  is  also  noted  that  the  upper  polar 
nucleus  and  the  nuclei  of  the  egg  and  synergids  are  erythrophilous,  while  the  four 
nuclei  at  the  antipodal  end  of  the  sac  are  cyanophilous.  This  difference  in 
chromatophily  is  attributed  to  chemical  differences  due  to  sexuality,  the  nuclei  at 
the  antipodal  end  of  the  sac  having  lost  all  sexual  character.  c.  j.  c. 

Chodat,  R.,  and  Bernard,  C.  Sur  le  sac  embry-  Comparatively  little  is  known  of  the 
onnaire  de  I'Helosis  guayanensis.  Jour,  de  embryology  of  the  BalanophoraceJE, 
Botanique.     14:   72-79,  pis.  1-2,  1900.  ... 

but  it  is  certain  that  they  have  puzzling 
peculiarities.  Writers  agree  that  there  is  no  ovule  or  placenta  in  Balanophora^ 
but  that  the  megaspore  is  situated  in  a  tissue  at  the  base  of  a  prolongation  incor- 
rectly termed  a  "style."  Van  Tieghem  (1896)  found  that  in  B.  iudica  the  polar 
nuclei  do  not  fuse  and  that  fertilization  occurs  at  the  antipodal  end  of  the  sac  as 
often  as  at  the  upper  end.  According  to  Treub  (1898),  in  B.  elongata  the  mega- 
spore germinates  in  the  usual  manner.  The  polar  nuclei,  however,  do  not  fuse, 
but  each  divides  independently.  The  egg  apparatus  breaks  down  and  there  is 
no  fertilization,  but  an  embryo  develops  from  one  of  the  cells  of  the  endosperm. 
Lotsy  (1899)  investigated  B.  globosa  and  supported  Treub  in  every  particular, 
including  the  peculiar  origin  of  the  embryo. 


1462  Journal  of  Applied  Microscopy 

In  the  present  paper,  Chodat  and  Bernard  give  the  results  of  their  work  on 
Helosis  guayanensis.  The  archesporial  cell  becomes  the  megaspore  directly 
without  cutting  off  a  tapetal  cell  or  giving  rise  to  a  row  of  potential  megaspores. 
The  jacket  or  "  tapetum  "  surrounding  the  embryo-sac  is  sporogenous  tissue. 
The  two  daughter  nuclei  resulting  from  the  first  division  of  the  nucleus  of  the 
megaspore  are  quite  different  in  appearance,  the  one  at  the  upper  end  of  the  sac 
staining  much  more  deeply.  This  nucleus  gives  rise  to  the  egg,  two  synergids 
and  a  polar  nucleus  in  the  usual  manner.  The  other  nucleus  stains  faintly  and 
rarely  divides  at  all,  but  soon  degenerates,  so  that  no  antipodals  or  polar  nucleus 
are  formed.  According  to  Van  Tieghem  the  &gg  is  fertilized  in  Helosis  and 
Balanopho7-a.  The  present  writers  find  that  in  Helosis  the  ^gg  becomes  large, 
but  also  becomes  weak  and  feeble  in  appearance,  so  that,  while  they  were  not  able 
to  prove  or  disprove  the  occurrence  of  fertilization,  they  believe  that  the  feeble 
condition  of  the  egg,  together  with  the  position  of  the  embryo  in  the  endosperm, 
favor  Treub's  view  that  the  embryo  arises  apogamously  from  the  endosperm. 

C.J.  c. 


CYTOLOGY,  EMBRYOLOGY, 

AND  • 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole,  Cornell  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Hoffman.    Die  Rolle  des  Eisenbei  der  Blutbil-      ^he  efforts  of  the  author  were  directed 
dung.     Zugleich    ein  Beitrag    zur   Kentniss 

des  Wesens  der  Chlorose.    Virchow's  Arch.,       to    the    investigation    of    the    SO-called 
160:  235-306,  1900.  blood  forming  organs,  and  were  laid  in 

the  following  lines  :  Enumeration  of  the  blood  corpuscles  ;  determination  of  the 
hemaglobin ;  tracing  the  mjlnner  in  which  the  metal  is  taken  into  the  organism ; 
the  effect  of  different  preparations ;  the  effect  in  healthy  and  anaemic  animals, 
etc.  Ninety-eight  rabbits  in  all  were  used  in  the  investigation.  To  determine 
whether  the  entrance  of  the  metal  into  the  so-called  blood  forming  organs  could 
be  proved,  the  bone  marrow,  spleen,  and  the  mesenteric  lymph  glands  were 
examined  for  their  contained  iron.  The  liver  and  kidneys  were  usually  similarly 
tested.  The  bone  marrow  was  taken  out  as  entire  columns  of  ^-1  cm.,  depend 
ing  on  the  size  of  the  animal,  from  the  humerus,  radius,  femur,  and  tibia,  and 
put,  as  were  the  other  tissues,  into  70  percent,  alcohol  to  which  5  per  cent,  solu- 
tion of  ammonium  sulphide  was  added,  then  after  24  hours  into  absolute  alcohol 
plus  a  few  drops  of  sulphide.  In  all  animals  containing  iron  the  marrow  became 
after  one-half  or  more  hours,  of  a  gray  black,  then  of  a  distinct  green  color.  This 
was  especially  clear  to  the  eye  if  the  tissues  were  compared  with  some  from  an 
animal  not  containing  iron.  Since,  for  the  most  part,  parallel  investigations  were 
made  on  both  iron  and  non-iron  fed  animals,  it  was  easy  to  determine  macro- 


and  Laboratory  Methods.  1463 

scopically,  in  a  few  hours,  with  certainty  from  which  rabbit  the  piece  of  marrow 
was  taken.  Moreover,  the  marrow  of  the  non-iron  fed  animal  lost  much  of  its 
reddish  color,  passing  into  a  dirty  red,  yet  never  acquiring  the  looks  of  the  sul- 
phide of  iron  preparation.  The  spleen  always,  after  a  term  of  feeding  with  iron, 
was  the  quickest  to  color,  becoming  dark  green  in  a  few  minutes.  This  was 
sometimes  true  in  animals  which  had  received  no  iron,  but  the  difference  in  the 
intensity  of  the  reaction  was  a  ready  means  of  distinction  between  the  animals 
with  and  without  iron.  The  mesenteric  glands  also  showed  the  same  difference, 
taking  in  iron-fed  animals  a  clear  green  tone,  which  became  much  lessened,  or 
entirely  absent,  in  iron  free  cases.  The  pieces  of  tissue  were  hardened  24  hours 
in  absolute  alcohol,  embedded  in  paraffin,  sectioned  in  different  regions,  and 
fastened  with  albumenized  glycerin  on  the  slides  ;  paraffin  dissolved  out  with 
xylol,  and  the  sections  placed  for  an  hour  or  so  in  ammonium  sulphide,  washed 
rapidly  in  distilled  water,  and  mounted  in  glycerin.  In  every  iron-fed  case,  in 
the  bone  marrow  the  iron  is  readily  distinguished,  especially  in  thin  sections. 
The  iron  laden  transporting  cells,  diffusely  green,  have  two  to  five  black  green 
granules.  Usually  these  cells  are  most  abundant  in  the  red  marrow,  at  the  ends 
of  the  bone,  apparently  less  abundant  in  the  yellow  fatty  marrow,  though  abso- 
lutely more.  To  study  the  exact  position  of  these  iron  cells,  the  author  made 
preparations  after  Stieda's  method,  that  is,  Berlin  blue  with  alum  carmin.  The 
bone  marrow  of  the  animals  not  fed  with  iron  was  practically  iron  free.  The 
spleen  of  the  ordinary  plant  fed  rabbit  contained  a  fair  amount  of  iron,  contained 
exclusively  in  the  pulp.  The  amount  after  feeding  with  iron  rose  to  such  a 
degree  that  the  sections  became  stained  deep  green  in  a  second,  only  the  follicles 
appearing  as  light,  unstained  spots.  In  the  mesenteric  lymph  glands,  on  ordinary 
food,  solitary  green  leucocytes  can  be  found  ;  after  an  iron  diet  their  number 
increases  in  a  marked  degree.  The  liver  in  young  animals  gives  no  iron  reaction 
without  feeding  the  substance,  but  in  older  animals  a  small  amount  is  usually 
indicated.  On  feeding  iron  these  assume,  more  slowly  and  to  a  less  degree  than 
the  spleen,  the  characteristic  color  showing  the  presence  of  iron.  This  is  espec- 
ially true  in  the  portal  regions.  The  kidneys,  only  here  and  there,  even  with 
large  doses  of  iron,  show  single  green  epithelial  cells  in  the  convoluted  portions. 
On  the  contrary,  pieces  of  the  small  intestine  and  colon  washed  in  and  left  in 
ammonium  sulphide  for  a  short  time  become  green  to  deep  blackish  green.  The 
large  intestine  always  gives  the  strongest  reaction. 

The  enumeration  of  the  red  corpuscles  and  determination  of  the  hemaglobin 
were  also  made.  Cover-glass  preparations  were  stained  with  Ehrlich's  hsematoxy- 
lin  and  eosin  solution.  Sections  of  bone  marrow  were  hardened  in  alcohol  of 
irkcreasing  strength,  embedded  in  paraffin,  and  stained  with  eosin-haematoxylin 
and  alum  carmin.  The  spleen  and  mesenteric  glands  were  similarly  treated. 
For  investigation  of  the  special  kinds  of  cells  in  the  bone  marrow,  Neumann's 
process  was  discarded.  It  was  to  crush  a  piece  of  bone  and  receive  the  exuded 
marrow  pulp  into  a  capillary  tube,  and  bringing  very  small  drops  from  this  on 
to  the  cover-glass.  The  author  did  not  employ  this  method  because  it  seemed 
to  him  that  marrow  cells,  as  well  as  the  contents  of  the  larger  and  smaller  blood 
vessels,  were  obtained.     The  author  arrived  at  the  quantitative  relation  of  the 


1-lt^-i  Journal  of  Applied  Microscopy 

mature  non-nucleated  erythrocytes  to  the  nucleated  red  corpuscles  and  the 
remaining  marrow  cells  by  taking,  with  a  fine  pair  of  scissors,  as  nearly  as  possible 
equal  amounts  of  marrow  about  the  size  of  a  pin  head,  and  very  carefully  press- 
ing them  evenly  between  two  clean  cover-glasses.  With  the  very  cellular 
lymphoid  marrow  of  the  anaemic  animal  this  was  always  successful,  giving  a  thin 
smear,  a  little  thicker  in  places  where  librin  lay  about  heaps  of  cells,  which  did 
not  interfere  in  a  general  view  over  the  preparation.  The  air-dried  cover  smears 
were  fixed  in  an  alcohol  and  ether  mixture,  and  stained  with  Ehrlich's  eosin- 
haematoxylin  solution  or  the  triacid  stain.  In  the  same  manner  preparations 
were  made  of  spleen  and  in  a  few  cases  of  the  lymph  glands  also.  e.  j.  c. 

Danjeard,  P.  A.  Nuclear  division  in  Protozoa.  I"  this  paper  the  author  takes  excep- 
Le  Botaniste  7,  1900.  Extract  from  Royal  tion  to  the  usual  Statement  that  nuclear 
Mic.  Jour.  April,   looi.  ,...._,  .    .  .    ,  ,      ,. 

division  in  Protozoa  is  invariably  direct. 

Figures  and  descriptions  of  ordinary  division  as  shown  in  Avuvba  polypodia 
are  given  ;  that  of  Amoeba  crystalligera,  in  which  the  dividing  nucleus  is  drawn  to 
a  thread  at  the  division  plane  ;  in  Sappi/iia  pedata  in  which  the  nucleus  divides 
twice  without  cytoplasmic  division ;  finally  a  full  account  of  the  process  of 
division  in  Amceba  hyalhiia,  sp.  n,  in  which  no  karyokinesis  occurs.  In  this 
form  the  nucleus  contains  a  large  nucleolus  which  breaks  up  at  the  onset  of 
division  and  appears  to  give  rise  to  chromosomes.  Some  parts  of  the  nucleolus 
also  mingle  with  the  nucleoplasm  and  give  it  chromatic  properties.  This 
nucleoplasm  forms  a  spindle  in  which  very  fine  chromosomes  arrange  themselves 
in  an  equatorial  plate.  Later  they  separate  and  approach  the  poles  of  the  spindle. 
The  spindle  is  pulled  out  as  they  do  this,  which  process  is  continued  as  the 
chromosomes  migrate  to  the  poles  of  the  elongating  amoeba,  until  but  elongated 
threads  remain  to  represent  the  spindle.  The  author  holds  this  as  proof  that 
the  chromosomes  migrate  by  their  own  activity  here  as  elsewhere  ;  since  in  the 
present  case  no  spheres  exist  and  the  movement  of  the  chromosomes  continues 
after  the  poles  of  the  spindle  have  been  reached,  the  threads  of  the  latter  cannot 
be  active  agents.  As  the  new  cells  separate,  the  chromosomes  round  themselves 
off  and  form  the  nucleolus,  the  spindle  remains,  constituting  the  surrounding 
nucleoplasm.  This  is  clearly  a  karj'okinetic  process,  but  in  the  author's  opinion 
its  simplicity  shows  that  the  evident  process  is  merely  a  modification  of  the 
simpler  direct  division,  special  emphasis  being  laid  on  the  conditions  occurring 
in  Avui'ba  crystalligera.  a.  m.  c. 

Penard,  E.  Dr.  Experiments  in  Diiilugia,  ^^r.  Eugene  Penard  has  succeeded  in 
Rev.  Suisse  Zoo),  8,  1900.  Ext.  from  Jour.  separating  the  nucleus  intact  from  the 
Roy.  Mic.  See,  April,  igoi.  ,  .  ,  ,  ^ 

cytoplasm    in    several   cases,   three   of 

which  were  accomplished  without  any  other  material  injury  to  the  organism. 
Such  separate  nuclei  appear  healthy  for  9  to  '1\  hours  after  removal,  but  they  ulti- 
mately die  apparently  of  inanition.  The  non-nucleated  portions,  however,  lived 
and  moved  about  for  several  days  apparently  none  the  worse  for  the  operation. 
In  three  cases  the  specimens  were  killed  for  examination  and  consisted  to  all 
appearances  of  normal  protoplasm.  Non-nucleated  animals  were  not  seen  to 
take  food,  but  since  intact  forms  can  remain  without  food  for  weeks  uninjured 
there  seems  no  doubt  that  the  mutilated  specimens  could  digest  food.    a.  m.  c. 


and  Laboratory  .Methods.  1465 


CURRENT   ZOOLOGICAL   LITERATURE. 

Chakt.f.s  a.  KoFonx 

Books  and  separates  cf  papeis  on  zoSo^cal  snbfects  shoold  he  sent  for  review  to 
Chailes  A.  Kolrad.  Univeisty  of  Califcxnia,  Beikeief ,  Cafifomia. 

Ho'aard.  L.  0.    Mosqidtoes.    241  Rit,  irith  50      The    popular   interest   in  mosquitoes, 
fc-    r  :-r  reit.    New  York,  igoi.   McChue.  -  _^     ^^  ,^i_      j-  £  ^v   - 

r-     -;      Co     $1  -o  growing  out  of  the  discovery  ot  theur 

agenqf  in  causing  malaria  and  yellow 

fever,  makes  this  book  of  Dr.  Howard's  very  timely.     The  author  describes  the 

life  history,  the  feeding,  breeding,  and  liiring  habits,  and  the  transformations  of 

mosquitoes.      The  method  by  which  malaria,  yellow  fever,  and  filariasis   are 

transmitted  to  man,  and  the  species  concerned  in  this  process,  are  careful^ 

described.     American  mosquitoes  are  also  discussed,  and  a  key  to  all  of  the 

known  species  is  given.     Su^estions  are  giving  for  breeding  and  rearing  the 

lar\'2e,  and  directions  for  collecting  and  preserving  specimens  for  examination 

and  for  museum  piuposes  are  detailed.     Means  of  extermination  of  these  pests, 

and  the  precautions  necessary  to  prevent  the  spread  <rf  disease  by  them,  are 

given  in  the  light  of  recent  experiments.     This  book  should  find  its  way  into 

every   high-school  library,  and  will  be  of  value  to  physicians  and   travellers. 

Chapter  IX,  •'  How  to  Collect  and  Preserve  Mosquitoes."  :s  reprinted  herewith 

by  courtesy  of  the  publishers.  c  a.  k. 

•Adult  mosc  - :: :  es  are  very  fragile  creatures.  The  scales  upon  their  bodies  and 
legs  are  easily  rubbed  ott,  and  the  antennae,  and  especially  the  l^s,  break  with 
the  least  handling.  Even  in  their  ordinary  course  of  life  the  scales  rub  off,  and 
with  certain  species  an  adult  which  is  two  or  three  weeks  old  is  quite  different 
in  appearance  from  one  which  has  just  emerged  from  the  pupa.  Practically, 
they  cannot  be  handled  with  the  fingers,  or  their  value  as  cabinet  specimens  or  as 
specimens  for  study  is  lost.  With  some  forms  there  are  important  characters  in 
the  arrangement  of  the  scales  on  the  thorax.  With  others  the  scales  on  the  wing 
are  of  importance,  and  if  the  front  legs  are  accidentally  broken  off,  an  important 
character  to  which  I  have  referred  in  the  systematic  portion  of  this  book  as  exist- 
ing in  the  claws  of  the  fore  feet,  is  naturally  unavailable.  In  capturing  them, 
therefore,  they  must  not  be  handled,  and  I  have  found  the  most  satisfactory 
method  of  capture  to  consist  in  simpiy  placing  a  small,  open-mouthed  vial  over 
the  mosquito  while  at  rest.  On  the  wing,  it  cannot  be  caught,  even  with  a  deli- 
cate net.  without  rubbing  or  1^-breaking.  If  a  mosquito  lights  upon  your  hand, 
or  upon  a  twig,  or  a  leaf,  or  upon  a  wall  of  a  room,  it  is  quite  easy,  especially  if 
it  be  engaged  in  sucking  blood,  to  cover  it  adroitly  with  the  vial.  It  rises  almost 
instantly,  and  the  mouth  of  the  vial  is  plugged  with  a  plug  of  absorbent  cotton. 
A  drop  of  chloroform  on  the  cotton  will  stupefy  the  specimen  almost  immediately. 
and  another  drop  will  kill  it. 

The  specimen  may  be  kept  permanently  in  the  vial,  and  when  studied,  if  the 
study  goes  no  further  than  an  examination  of  the  coarser  characters.  In  an 
anempt  to  determine  the  species,  it  will  often  suffice  gently  to  slide  it  out  upon 
a  sheet  of  white  paper  and  examine  it  with  a  powerful  hand-lens.  With  the  one- 
quarter  inch  achromatic  triplet  lens,  made  by  different  firms,  I  have  found  it 
possible  to  distinguish  all  of  the  generic  and  specific  characters,  even  down  to  the 
teeth  of  the  tarsal  claws.     This,  however,  is  difficult  to  persons  not  accustomed 


1466  Journal  of  Applied  Microscopy 

to  the  use  of  high-power  hand  lenses,  and  in  such  instances  one  must  break  off 
a  tarsus  and  mount  it  upon  a  slide  in  glycerin  or  Canada  balsam  for  examination 
under  a  compound  microscope. 

It  is  not  advisable  to  mount  adult  mosquitoes  bodily  on  slides  in  any  medium 
whatever.  They  should  not  be  preserved  in  alcohol  or  formalin,  but  should  be 
kept  dry  in  vials.  Of  course  they  will  rattle  around  somewhat,  and  there  is 
danger  that  the  legs  and  the  antennee  will  be  lost ;  therefore,  if  they  are  moved 
from  the  vial  after  the  collecting  and  killing,  into  pill  boxes  with  cotton,  they  can. 
be  carried  safely,  or  can  be  sent  in  the  mails.  Several  of  the  pill  boxes  may  be 
placed  inside  a  tight  tin  or  wooden  box  and  mailed  with  perfect  security. 

A  collection  of  mosquitoes  should,  however,  not  be  kept  in  this  way,  provided 
that  it  is  intended  as  a  study  collection.  The  method  which  I  have  adopted,  and 
which  is  the  one  customarily  used  for  small  insects  that  are  not  too  small  for 
hand-lens  work,  is  the  triangular-tag  method.  Take  a  sheet  of  stiff  paper  or  very 
thin  cardboard,  and  cut  a  strip  say  tive-sixteenths  or  three-eighths  of  an  inch 
wide.  Then  from  this  strip,  by  slightly  oblique  cuts,  cut  a  series  of  triangles  that 
will  be  pointed  at  the  tip  and  a  little  less  than  an  eighth  of  an  inch  wide  at  the 
base.  Through  the  base  of  the  tag  may  be  run  an  insect  pin,  and  to  the  tip  the 
mosquito  should  be  glued,  white  or  yellow  shellac  being  the  best  medium  for  the 
gluing.  The  mosquito  should  be  glued  on  its  side,  just  behind  one  wing,  so 
that  its  back  is  away  from  the  pin.  This  enables  one  readily,  by  holding  the 
point  of  the  pin  in  one's  hand,  to  examine  with  a  lens,  all  legs,  antennae,  palpi,  one 
side,  and  the  back.  The  tag  should  be  pushed  up  on  the  pin  until  it  is  from 
two-thirds  to  three-quarters  of  the  length  of  the  pin  away  from  the  point.  To 
the  lower  part  of  the  pin  should  be  attached  a  small  label  giving  date,  exact 
locality,  and  name  of  the  collector,  and  below  this  may  be  pinned  another  small 
label  bearing  the  name  of  the  insect. 

Those  who  for  some  reason  do  not  like  the  paper  triangle  method  of  mount- 
ing, use  very  minute  pins,  made  by  Mueller  in  Vienna,  and  known  as  "minuten 
insekten  naedeln,"  which  are  sold  by  Queen  &  Co.,  Philadelphia,  and  other 
large  dealers  in  such  things.  These  pins  are  so  small  and  delicate  that  they  must 
be  thrust  through  the  thorax  of  the  mosquito  and  into  a  little  strip  of  cork,  the 
cork  strip  itself  being  pinned  upon  one  of  the  larger  and  longer  insect  pins. 

Some  collectors,  instead  of  using  the  chloroform  method  of  killing,  prefer  the 
cyanide  bottle.  The  cyanide  bottle  is  made  by  taking  a  wide-mouthed  flask, 
putting  a  small  lump  of  cyanide  of  potassium  at  the  bottom,  and  covering  it  with 
a  layer  of  liquid  plaster  of  Paris,  which,  when  allowed  to  set,  makes  a  complete 
layer  over  and  around  the  cyanide,  and  prevents  the  water  that  comes  from  the 
deliquescence  of  the  cyanide  from  injuring  specimens  that  are  placed  in  the  vial, 
but  which  at  the  same  time  is  sufficiently  porous  to  permit  the  escape  of  the 
deadly  cyanide  fumes.  Even  with  the  layer  of  plaster  of  Paris,  however,  the 
cyanide  bottle  will  sometimes  become  wet,  so  that  a  bit  of  blotting-paper  may 
with  advantage  be  inserted  to  cover  the  plaster  of  Paris,  and  to  absorb  the 
superfluous  moisture.  A  mosquito  captured  in  one  of  these  cyanide  fiasks  dies 
very  quickly,  and  is  in  good  condition  for  dry  mounting  or  for  transfer  to  pill 
boxes.  The  cyanide  bottle  is,  preferably,  stoppered  with  a  cork  stopper,  but 
rubber  stoppers  are  also  used. 

In  collecting  early  stages  of  mosquitoes,  it  is  only  necessary  to  hav^e  a  supply 
of  bottles,  a  little  coffee-strainer  with  a  handle,  and  a  large  reading  glass.  Other 
apparatus  is  cumbersome  and  unnecessary.  I  have  a  large  reading-glass  four 
inches  in  diameter,  with  a  strong  handle,  which  I  find  very  useful  in  examining 
the  surface  of  water-pools,  especially  for  Anopheles  larvae  The  dip  strainer 
used  is  an  ordinary  cheap  coffee-strainer,  which  has  been  mounted  upon  a  long 
handle,  so  that  one  can  reach  out  two  or  three  feet  from  the  shore  and  capture 
larvae  and  pupae.     Other  large  strainers  with  a  fine  mesh  are  sold  at  the  hard- 


and  Laboratory  Methods.  1467 

ware  stores,  and  may  be  purchased  cheaply.  In  bringing  larvae  and  pupae  in 
from  the  field,  too  much  jarring  about  in  a  bottle  may  result  in  their  death  by 
drowning.  It  is  desirable,  therefore,  to  put  moss  or  water-weed  in  the  bottle 
with  a  minimum  of  water,  provided  the  insects  are  transferred  to  an  aquarium  or 
a  still  jar  within  a  few  hours. 

Nuttall,  Cobbett,  and  Strangeways-Pigg,  who  have  done  a  great  deal  of  col- 
lecting of  mosquito  larvae  in  England,  as  shown  in  one  of  their  important  papers, 
entitled  "Studies  in  Relation  to  Malaria,"  published  in  Xho^  Journal  of  Hygiene, 
Vol.  1,  No.  1,  January,  1901,  used  as  their  collecting  apparatus  some  wide- 
mouthed  bottles  of  medium  size  with  cork  stoppers  ;  a  white  enamelled  dipper 
which,  when  required,  can  be  tied  with  a  piece  of  twine  to  a  long  bamboo  rod  ; 
a  small  pipette  with  a  rubber  bulb,  and  small  vials  containing  dilute  alcohol  for 
the  preservation  of  larvae  which  they  did  not  wish  to  keep  alive.  They  travelled 
over  England  on  their  collecting  trip  on  bicycles.  When  the  larvae  or  eggs  were 
captured  in  the  porcelain  dippers  they  were  removed  with  a  pipette  and  put  in 
bottles,  which  were  half  filled  with  water,  wrapped  in  cloths,  and  attached  to  the 
bicycle  frame.  They  found  that  they  could  be  transported  for  several  hours 
without  injury.  They  noted  also  that  the  large  larvae  did  not  withstand  the 
shaking  as  well  as  the  small  ones,  but  that  a  sufficient  number  could  always  be 
brought  back  for  studying  purposes.  On  expeditions  lasting  a  couple  of  days, 
they  took  precaution  to  remove  the  corks  occasionally  to  give  the  insects  fresh 
air.  White  dippers  were  used,  since  they  could  more  easily  detect  the  eggs  or 
larvae  on  the  white  background,  and  they  found  that  only  rarely  could  they 
detect  the  insects  by  direct  inspection  of  the  surface  of  the  water. 

Larvae  and  pupae,  when  it  is  desirable  to  preserve  them  in  these  stages,  and  it 
is  always  desirable  to  keep  a  small  set  of  each  species,  may  be  kept  in  vials  of 
alcohol  or  dilute  formalin  (5  to  10  per  cent.).  When  preserved  in  alcohol  they 
should  be  passed  through  different  strengths,  beginning  with  a  weak  mixture,  in 
order  that  they  may  not  shrivel ;  or,  what  is  still  better,  kill  the  larvae  or  pupae 
suddenly  in  a  cyanide  bottle,  then  bring  the  water  nearly  to  the  boiling  point  in 
a  little  porcelain  dish  over  an  alcohol  lamp,  and  drop  the  insects  in,  leave  them 
until  the  boiling  point  is  just  reached,  and  then  remove  them.  An  immersion  of 
only  a  few  moments  will  suffice.  Ordinarily  the  larvae  will  sink  at  once  to  the 
bottom  of  the  water,  and  very  soon  thereafter  rise  to  the  top.  This  rising  is  an 
indication  that  the  specimen  should  be  removed  at  once.  The  specimen  may 
then  be  preserved  in  ordinary  commercial  alcohol,  and  will  retain  perfectly  its 
color  and  shape.  This  method  is  used  successfully  with  the  larvae  of  many 
insects.  It  is  not  necessary  to  mount  either  larvae  or  pupae  whole  on  slides. 
One  of  these  preserved  specimens  can  be  put  in  a  cell  with  alcohol  or  glycerin 
and  studied  under  a  low  power  with  perfect  ease,  and  the  examination  of  minute 
details  of  its  anatomy,  external  and  internal,  may  readily  be  accomplished  by 
dissection,  and  the  parts  dissected  out  mounted  permanently  on  slides  in  any  of 
the  ordinary  media. 

In  rearing  different  species  of  mosquitoes  I  have  had  perfect  success  in  the 
use  of  large,  cylindrical  glass  jars,  known  as  battery  jars.  They  can  be  bought 
in  almost  any  city,  and  of  various  sizes.  The  size  which  I  find  most  convenient 
will  hold  about  a  gallon  of  water.  A  layer  of  sand  an  inch  or  two  deep  is  placed 
in  the  bottom  of  the  jar  and  a  quart  or  more  of  water  poured  over  it.  After  the 
sand  has  settled  and  the  water  has  cleared,  a  bit  of  almost  any  small  water-plant 
may  be  inserted  to  advantage,  provided  mosquitoes  of  the  genus  Culex  are  being 
reared.  If  the  experiment  is  with  Anopheles,  however,  some  fresh-water  alga  is 
introduced,  such  as  Spirogyra,  Mougeotia,  OEodogodium,  Cladophora,  or  Oscil- 
laria — almost  any  green  scum  from  stagnant  water,  in  fact.  Over  the  top  of  the 
jar  is  placed  a  piece  of  swiss,  or  other  fine,  translucent  cloth,  held  down  by  a 
large  rubber  band. 


1468  Journal  of  Applied  Microscopy 

The  eggs  of  Culex  may  be  had  with  ease  by  exposing  a  bucket  of  water  out 
of  doors  in  a  mosquito  locality  on  almost  any  summer  night.  If  the  egg  masses 
be  transferred  from  the  bucket  to  the  prepared  breeding-jar,  the  growth  of  the 
larvae  can  be  watched,  and  their  transformations  can  be  observed  with  perfect 
ease.  Occasional  specimens  can  be  taken  out  and  preserved,  to  illustrate 
variations  of  different  stages  of  growth.  Accurate  notes  can  be  kept  as  to 
temperature,  periods  of  transformation,  and  so  on.  A  series  of  dates,  provided 
several  jars  are  under  observation,  can  be  written  from  time  to  time  upon  a  slip 
of  paper,  which  may  be  pinned  to  the  edges  of  the  cloth  covering  of  each  jar. 

Where  the  eggs  of  Anopheles,  for  example,  have  not  been  found,  females 
collected  at  large  may  be  liberated  in  such  a  prepared  breeding-jar.  They  will 
rest  on  the  under  side  of  the  cloth  covering  during  the  day,  and  at  night  will  lay 
their  eggs  on  the  surface  of  the  water.  It  is  desirable  to  have  a  stick  in  the 
water,  or  a  leaf,  or  a  bit  of  cork  floating  on  the  surface.  I  have  had  no  difficulty 
in  obtaining  the  eggs  of  Anopheles  in  large  numbers  in  this  way,  and  the  eggs 
of  Culex  as  well,  but  although  as  many  as  fifty  females  of  Psorophora  have  been 
liberated  in  breeding-jars  prepared  in  this  way,  I  have  not  been  able  to  get  the 
eggs  of  this  genus,  which,  as  a  matter  of  fact,  are  yet  unknown.  It  is  possible 
that  Psorophora  does  not  deposit  its  eggs  upon  the  surface  of  water.  This,  how- 
ever, is  unlikely,  and  it  is  rather  to  be  supposed  that  the  females  used  in  my 
experiments  were  not  old  enough  for  oviposition,  and  died  from  the  confinement 
of  the  jar  before  the  egg-laying  period  arrived. 

When  one  wishes  to  study  closely  the  movements  and  intimate  habits  of  the 
early  stages  of  mosquitoes,  a  great  deal  may  be  observed  through  the  glass  sides 
of  the  jar,  by  using  a  coarse  lens  and  studying  those  near  the  side,  but  when  a 
closer  study  is  desired,  individual  larvae  or  pupae  may  be  lifted  out  with  a  strainer 
and  put  in  a  shallow  porcelain  vessel,  where  they  can  be  watched  with  ease  under 
a  dissecting  microscope.  Anopheles  larvae  may  be  studied  in  this  way  very 
easily,  and  no  nature  study  could  be  of  more  fascinating  interest  than  the  obser- 
vation of  these  creatures,  lying  as  they  do  with  the  body  practically  in  a  single 
plane,  so  that  they  may  be  easily  watched,  with  the  mouth  parts  in  constant 
action,  and  the  head  occasionally  turning  upside  down,  and  the  reverse,  with 
lightning-like  rapidity."  c.  a.  k. 


NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Sailer,  J.     Primary  Endothelioma  of  the  Left       At  the   orifice  of  the  left   superior   pul- 
Superior    Pulmonary   Vein.      Contributions  .        ,  ,  ^ 

from  the  William  Pepper  Laboratory  of  monary  vem  there  was  a  dense  mass  of 
Clinical  Medicine.  Philadelphia,  pp.  416-  fibrous  tissue,  almost  occluding  the 
444,  1900.  ,  ,  ,.  ,  .      , 

lumen,  and  extendmg  on  the  auricular 

wall  to  the  upper  end  of  the  left  inferior  pulmonary  vein.  The  wall  of  the 
superior  pulmonary  vein  was  nearly  uniformly  thickened  throughout  its  whole 
course  in  the  upper  lobe,  forming  a  round  cord  fifteen  mm.  in  diameter.  Upon 
section  it  was  seen  to  be  made  up  of  grayish  and  yellowish  tissue.  The  upper 
lobe  was  contracted,  pigmented  and  airless. 

There  was  found,  on  microscopical  examination  of  the  thickened  vein,  hyper- 
plasia of  the  connective  tissue   stroma  and  enlargement  of  the  lymphatic  spaces 


and  Laboratory  Methods.  1469 

and  of  the  vasa  vasorum.  The  pecuHar  feature  was  the  proUferation  of  the 
endotheUal  cells  in  these  spaces.  The  writer  regards  the  process  as  a  primary 
endothelioma,  although  he  admits  that  it  may  be  simply  the  result  of  chronic 
inflammation.     There  is  an  excellent  resume  of  the  literature  of   endothelioma. 

J.  H.  p. 

MacCallum,    W.    G.     On   the    Intravascular      MacCallum  Studied  a  malignant  tumor, 
Growth  of  Certain   Endotheliomata.     Con-  .    .      ^.        •     ^i      i    r.   .      .•   i         i,-   i    t,„ 

tributions  to  the  Science  of  Medicine,  dedi-       ongmatmg  m  the  left  testicle,  which  he 

cated  by  his  pupils  to  Dr.  W.  H.  Welch,      designates    lymphendothelioma    testis, 
imore,  pp.  497-509,  1900.  ^j^^  primary  tumor  formed  a  nodulated 

mass  about  6x5  cm.  in  size.  It  was  succulent,  myxomatous  in  places 
and  variegated  in  color  by  opaque  yellow  areas  of  necrotic  tissue.  Death 
occurred  four  months  after  the  removal  of  the  testicle.  The  growth 
had  extended  from  the  scrotum  in  a  remarkable  manner.  The  spermatic 
vein  was  packed  with  a  somewhat  cylindrical  tumor  mass  which  extended 
upward  through  the  left  renal  vein  into  the  vena  cava,  in  which  it  spread 
out  into  a  bunch  of  translucent  villus-like  processes  which  extended  throughout 
the  whole  length  of  the  vena  cava  and  projected  into  the  right  auricle.  These 
curious  formations  resembled  the  villi  of  hydatidiform  moles.  Similar  bundles 
were  also  found  in  the  pulmonary  arteries  and  in  the  pulmonary,  jugular  and 
subclavian  veins.  The  lungs,  liver,  intestine  and  brain  contained  tumor  nodules 
and  there  were  local  recurrences  in  the  scrotum  and  groin  which  formed  a  chain 
leading  up  to  a  large  tumor  mass  in  the  lumbar  region.  The  tumor  nodules  in 
the  lungs  and  liver  were  rounded  and  rather  sharply  outlined  ;  many  were  semi- 
translucent  and  appeared  to  be  made  up  of  small  cysts,  others  were  more  opaque 
and  consisted  of  very  soft,  succulent,  whitish  tissue. 

Histologically  the  neoplasm  consisted  of  a  framework  of  soft  myxomatous 
tissue  which  contained  cysts  and  tubules  of  various  sizes  and  shapes.  These 
spaces  were  lined  with  cells  which  varied  in  height  from  a  flat,  scale-like  form, 
exactly  resembling  the  endothelium  of  the  lymphatics,  through  all  gradations  to 
high  columnar  epithelium-like  cells.  In  places  the  cells  were  piled  up  two  or 
three  rows  deep  and  arranged  in  folds  and  papillary  masses  which  had  invaded 
the  surrounding  tissue  and  finally  had  broken  their  way  into  the  veins.  The 
intravascular  growths  were  covered  by  the  endothelium  of  the  veins,  just  as  an 
organizing  thrombus  would  be  covered,  and  continuing  in  their  development 
they  formed  papillary  masses  which  projected  along  the  lumen  of  the  vein  and 
occasionally  formed  secondary  attachments  to  the  walls.  Passing  through  the 
heart  into  the  pulmonary  arteries,  these  masses  became  attached  to  the  walls  of 
the  finest  arterioles,  and  breaking  through  these  gave  rise  to  the  secondary 
tumor  nodules  in  the  lung.  The  nodules  in  the  liver,  brain,  and  intestine  were 
probably  tertiary  in  origin. 

The  cyst-like  spaces  found  in  the  metastases  and  intravascular  masses,  as 
well  as  in  the  primary  tumor,  were  lined  by  cells  which  do  resemble  epithelial 
cells  and  have  been  regarded  as  epithelial  in  nature  by  the  few  other  investi- 
gators who  have  studied  this  type  of  tumor.  MacCallum,  however,  in  view  of 
the  facts — (a)  that  no  connection  with  the  well  defined  epithelium  of  the  seminal 


1470  Journal  of  Applied  Microscopy 

tubules  nor  with  any  other  epithelial  structure  can  be  traced  :  (b)  that  the 
morphology  of  such  cells,  as  shown  by  Volkmann,  Krompecher,  and  others,  is  of 
very  slight  importance  in  their  identification  with  epithelial  or  endothelial 
structures ;  (c)  that  direct  transitions  between  the  obviously  connective  tissue 
elements  of  the  stroma  and  these  cells  occur ;  (d)  that  intercellular  fibrils  can  be 
demonstrated  between  the  cells  of  such  masses  by  the  use  of  Van  Gieson's  stain  ; 
and  (e)  that  the  spaces  in  which  such  cell  masses  occasionally  lie  have  not,  like 
the  alveoli  in  carcinomata,  any  further  lining  endothelium — concludes  that  it  is 
justifiable  to  consider  these  cells  of  endothelial  rather  than  of  epithelial  nature. 
Hence  he  classes  the  tumor  as  an  endothelioma,  rather  than  a  carcinoma. 

J.  H.  p. 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Weinland,  E.     Zur  Magenverdauung  der   Hai-       This  paper  treats  in  considerable  detail 
fische.    Zeitschr.  f.  Biol.  41 :  35-68,  Taf.  I,      ^f  ^^le  digestive  processes  and  functions 
"^    ■  of  the  anterior  part  of  the  alimentary 

tract  in  the  selachians.  As  living  material  individuals  of  the  following  species 
were  used :  ScyUiiim  catidus  and  canicula ;  Torpedo  ocellata  and  fnarmorata  : 
and  J?aja  asterias^  clavata  and  glauca.  The  chemical  reactions  of  the  stomach 
contents  were  also  studied  in  dead  specimens  of  a  number  of  other  species. 
The  method  used  for  obtaining  the  secretion  of  the  gastric  glands  in  a  pure  con- 
dition from  the  living  animal  is  ingenious  and  seems  to  have  given  excellent 
results.  Briefly,  the  procedure  was  as  follows  :  one  end  of  a  glass  siphon  of 
from  10  to  15  mm.  diameter  was  thrust  down  through  the  mouth  well  into  the 
cavity  of  the  stomach,  and  allowed  to  remain  there  until  a  sufficient  amount  of 
the  fluid  contents  of  the  stomach  had  passed  out.  Meanwhile  artificial  respiration 
was  maintained  by  passing  a  current  of  water  over  the  gills.  The  oesophagus 
closed  tightly  over  the  siphon  so  that  there  was  no  risk  of  any  mixture  of  sea 
water  with  the  stomach  contents.  The  treatment  apparently  has  no  ill  effect  on 
the  animal,  as  the  author  states  that  he  has  in  some  cases  daily  emptied  the 
stomach  of  the  same  animal  by  this  method  for  considerable  periods  of  time 
(fourteen  days  and  over),  without  causing  any  injury.  This  method  should  be 
widely  applicable,  both  for  purposes  of  investigation  and  class  demonstration. 

The  principal  points  discussed  are  :  (1)  the  length  of  time  the  food  remains 
in  the  stomach,  and  (2)  the  chemical  reaction  of  the  stomach  contents.  It  was 
found  that  the  food  remains  in  the  stomach  for  a  considerable  time  ;  from  two  to 
three  days  in  the  majority  of  cases,  up  to  eighteen  days  in  one  instance  observed. 
The  food  is  generally  completely  disintegrated  in  the  stomach,  forming  a  fluid  or 
semi-fluid  mass.  The  animals  studied  were  able  to  live  for  several  months  at  a 
time  without  taking  food. 

It  appears  clearly  from  the  experiments  that  in  the  living   skate    (^Raja')  the 


and  Laboratory  Methods. 


1471 


reaction  of  the  stomach  may  be  either  acid  or  alkaline,  both  during  digestion 
and  when  empty.  The  reaction  is  influenced  by  the  nature  of  the  food.  For 
example,  when  the  animals  are  fed  crabs  the  reaction  of  the  stomach  contents  is 
alkaline,  while  with  fish  as  food  the  reaction  is  almost  invariably  acid.  In  the 
species  of  ScylUmn  and  Torpedo  studied  the  reaction  was  found  to  be  always  acid. 
That  the  reaction  of  the  stomach  contents  is  not  due  to  the  specific  reaction  of 
the  food  itself,  but  that  instead  there  are  both  acid  and  alkaline  gastric  secre- 
tions, is  proven  by  the  fact  that  the  alkaline  secretion  may  be  induced  by  the 
action  of  ergot  subcutaneously  injected.  This  drug  causes  certain  sphincter 
muscles  in  the  walls  of  the  blood  vessels  of  the  stomach  to  contract  strongly, 
and  at  the  same  time  the  secretion  becomes  alkaline.  After  a  time  recovery 
occurs  and  the  secretion  becomes  again  acid.  In  the  case  of  Scylliicm  and 
Torpedo  there  are  no  sphincter  muscles  in  the  walls  of  the  vessels,  and  injection 
of  ergot  gave  only  negative  results  ;  the  reaction  remained  acid.  Microscopical 
examination  of  the  walls  of  the  stomach  of  Raja  clavata,  in  which  the  reaction 
was  alkaline,  showed  the  sphincters  of  the  vessels  so  strongly  contracted  that 
there  was  only  a  very  minute  opening  in  the  center.  The  hindrance  of  the 
blood  flow  caused  by  this  contraction  of  the  walls  of  the  vessels  seems  to  be 
the  immediate  cause  of  the  pouring  out  of  the  alkaline  secretion.  r.  p. 


Oker=Bloni,  M.  Eine  Normal  Elektrode  fiir 
physiologische  Zwecke.  Arch.  f.  d.  ges. 
Physiol.     79:  534-536,1900. 


The  necessity  for  frequent  change  and 
renewal    of    its    contents    is    a    defect 
which   has  long  been  felt  in  the  ordi- 
nary physiological,  "  unpolarisable,"  zinc-sulphate  electrode.     These  electrodes 

dry  up  quickly,  and  if  it  becomes  necessary  to  use  them  in  contact  with  different 

fluids,  they  must  be   cleaned   and  refilled 

after  each  change  of  condition.     Oker-Blom 

has    devised  an   electrode  which   in    large 

measure  gets  rid  of  these  difficulties,  and 

is  constant  in  its  working. 

A  glass  tube,  A  (Fig.  1),  of   about   1.2 

cms.    diameter    and    (3    cms.   in    length,  is 

closed  at  one  end  and  has  fastened  to  the 

side  near  the  closed  end  a  smaller  tube,  B, 

of  the  form  shown  in  the  figure.     Over  the 

top  of  this  smaller  tube   is  fitted  a  small 

cap,  bearing  at    its    outer    end    a    camel's 

hair  brush.     The   upper   end  of  the  main 

tube  is  closed    by    a    rubber    tube    and   a 

spring  clamp.     Into  the  bottom  of  the  tube 

A  is  melted  a  platinum  wire,  through  which 

external    electrical    connection    is    made. 

This  platinum  wire  is  covered  with   about 

1  c.  c.  of  quicksilver,  over  which  is  placed 

some  calomel.     The  apparatus  is  then   filled  with  "  physiological   salt  solution  " 

(.7  per  cent.  NaCl).     This  can  be  done  most  easily  by  removing  the  cap  bear- 


Fig.  1. 


l-i72  Journal  of  Applied  Microscopy 

ing  the  brush  and  attaching  in  its  place  a  rubber  tube,  which  is  allowed  to  dip 
into  some  salt  solution.  Then  by  applying  suction  at  the  upper  end  of  the  main 
tube  A,  the  whole  electrode  may  be  filled.  When  desired  for  use,  the  cap 
bearing  the  brush  may  be  filled  with  the  salt  solution  and  slipped  over  the 
end  of  the  tube  B,  care  being  taken  not  to  allow  the  entrance  of  any  air  bubbles. 
When  not  in  use  the  end  of  the  tube  B  is  corked,  thus  preventing  any  evapora- 
tion of  the  solution.  The  author  says  :  "  Once  in  order,  the  electrodes  are 
always  ready  for  use  and  are  very  constant."  r.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Schultz.  Ueber  die  Lebensdauer  von  Bacillus  Schultz  has  had  an  opportunity  of 
pestis    iiominis  in   Reinkulturen.      Cent.  f.       .  .        .  ,  .  t       -n 

Bac.  u.  Par.  IT   27:  12  iqoi.  mvestigatmg  cultures  of  pest  bacillus, 

which  are  four  years  of  age.  He  finds 
that  these  cultures  are  still  filled  with  active  bacteria,  and  are  virulent.  The 
question  as  to  the  condition  assumed  by  the  bacteria  in  these  cultures  has  been 
carefully  studied,  with  the  following  conclusions  :  The  pest  bacillus  does  not 
produce  endogenous  spores.  Its  resisting  power,  lasting  for  four  years,  appeared 
to  be  due  to  a  condensation  of  the  protoplasm  of  the  bacteria  rods,  which  serve 
the  same  purpose  as  spores.  These  shriveled  bacilli  are  capable  of  resisting  ad- 
verse conditions.  H.  w.  c. 

Hinterberger.  Eine  Modifikation  des  Geis-  This  article  gives  a  convenient  and 
selfarbungsverfahrens        nach      Ermengen.  ,1  i-,-        •  r     1  1      1      r 

Centf.Bak.und  Par.  11,27:  597,  1901.    •         "seful  modification  of  the   method  of 

staining  flagella.    The  method   is  too 

detailed  to  be  given  here,  and  the  original   article  must  be   referred  to  by  those 

wishing  to  adopt  it.  h.  w.  c. 

Muller.  Uber  Tuberkelbacillen,  und  Sporen-  ^he  author  gives  an  improvement  of 
farbung  unter  Anwendung  von  Kalium  per-  the  method  of  Staining  tubercle  bacilli, 
karbonat  und  Wasserstoffsupero.xyd.    Cent.  1  •   1    1      ^,  •    ,      •     r  1 

f.  Bac.  u.  Par.  I  29:  701,  iqoi.  which  he  thinks  is  far  more  sure  to  de- 

tect the  presence  of  these  organisms 
than  the  one  commonly  in  use.  It  consists  in  the  use  of  calcium  percarbonate, 
or  hydrogen  peroxyde,  as  a  decolorizing  medium,  in  the  place  of  acid.  The 
method  of  use  is  simple.  The  material  containing  the  bacilli  is  fixed  upon  a 
cover-glass  in  the  ordinary  way,  and  stained  as  usual  in  carbol-fuchsin.  The  sur- 
plus stain  is  washed  ofi  with  60  to  70  per  cent,  alcohol,  and  then  with  water. 
Afterward,  the  preparation  is  placed  in  a  5  to  10  per  cent,  solution  of  calcium  per- 
carbonate. This  decolorizes  the  preparation,  a  quarter  of  an  hour  being  re- 
quired for  the  purpose.  After  this  a  counterstain  with  methyl  blue  follows.  In 
the  use  of  hydrogen  peroxyde,  essentially  the  same  method  is  followed,  hydrogen 
peroxyde  being  used  for  decolorizing   instead  of  calcium  percarbonate.     The 


and  Laboratory  Methods.  1^73 

hydrogen  peroxyde  acts  quickly,  only  a  few  moments  being  required  for  decolor- 
ization.  The  result  is  far  more  sure  than  by  decolorization  with  acid.  The  tuber- 
cle bacilli  are  never  decolorized,  and  will  be  found,  in  the  end,  fully  stained  with 
the  carbol-fuchsin,  whereas  the  decolorization  of  other  organisms  is  perfect.  The 
author  thinks  the  method  vastly  superior  to  the  methods  commonly  used  for  this 
purpose.  H.  w.  c. 

Dains.     A    Pseudo  Tetanus  Bacillus      Journ.       ^^le  author    studies    the  case  of   a  bov. 
Boston  Soc.  Med.  Science.     5:  506,  1901.  .        '  ' 

wounded  by  a  blank  cartridge,  m  re- 
gard to  whom  there  were  some  fears  of  tetanus.  For  the  purpose  of  study  the 
wound  was  examined  microscopically,  and  there  was  found  in  it  a  bacillus  hav- 
ing a  great  resemblance  to  tetanus.  The  patient,  however,  made  a  rapid  recov- 
ery and  never  showed  any  symptoms  of  the  disease.  This  led  to  a  special  study 
of  this  tetanus-like  bacillus,  which  is  given  in  the  article  referred  to.  The  resem- 
blance to  the  tetanus  bacillus  was  very  great,  the  organism  having  the  same  gen- 
eral appearance,  and  producing  spores  on  the  end  in  the  typical  manner.  It 
differed,  however,  from  the  tetanus  bacillus  chiefly  in  the  following  points  :  It  is 
decolorized  by  Gram's  method,  while  the  tetanus  bacillus  is  not.  The  flagella 
are  less  numerous  than  those  of  tetanus  bacilli.  It  is  not  pathogenic  for  guinea 
pigs,  while  the  tetanus  bacillus  is  markedly  pathogenic  for  these  animals.  Its 
growth  in  glucose  and  stab  culture  is  wholly  unlike  the  growth  of  the  tetanus 
bacillus,  and  it  does  not  liquefy  gelatin,  while  the  tetanus  bacillus  does.  The 
organism  is  quite  different,  evidently,  from  the  tetanus  organism  which  it  so 
closely  resembles.  h.  w.  c. 

Poynton  and  Paine.  The  Etiology  of  Rheumatic      These  authors  endeavor  to  confirm,  if 
Fever.     Lancet,  1900. 

possible,  the  claim  that  rheumatic  fever 

is  a  disease  due  to  micro-organisms.  By  proper  culture  methods  they  succeeded 
in  isolating  from  several  cases  of  rheumatic  fever  a  bacterium  in  the  form  of  a 
coccus,  with  a  diameter  of  .5  pi,  which  does  not  color  with  the  Gram  method,  and 
does  not  grow  in  ordinary  culture  media.  The  organism  does  grow  readily  in  a 
culture  medium  of  bouillon  and  milk,  with  the  addition  of  a  little  lactic  acid. 
This  organism  they  found  in  a  variety  of  exudates  in  the  bodies  studied,  in  that 
of  the  pericardium,  in  the  heart's  blood,  etc.  They  do  not  usually  find  it  in  the 
tissues  themselves.  Experiments  of  inoculating  animals  with  the  pericardial 
fluid  from  individuals  suffering  from  this  disease  resulted  in  the  development  in 
the  animals  of  an  infection  which  has  many  of  the  distinctive  characteristics  of 
rheumatic  fever,  and  which  the  authors  naturally  infer  is  the  same  disease.  Es- 
sentially the  same  results  were  obtained  by  inoculating  bacteria  cultures.  The 
coccus  grown  on  agar  tubes  was  inoculated  into  the  veins  of  rabbits,  and  this 
was  followed  by  manifest  disturbances  which  were  of  a  nature  to  indicate  to  the 
authors  that  they  were  dealing  with  an  infection  similar  to  rheumatic  fever,  and 
produced  by  the  coccus  in  question.  The  authors  think  their  organism  identi- 
cal with  that  previously  found  by  Achalme  and  others,  and  regard  their  observa- 
tions, therefore,  as  a  confirmation  of  the  view  that  this  disease  is  a  bacterial  dis- 
ease, produced  by  the  micro-organism  which  they  have  studied.  h.  w.  c. 


1474 


Journal  of  Applied  Microscopy 


NOTES   ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 


Viola,  C.  Ueber  optische  Erscheinung  am 
Quarz  und  am  Turmalin  von  Elba.  Zeit.  f. 
Kryst.  32:  551-560,  1899. 


Wulfung,  E.  A.  Ueber  die  Lichtbewegung  im 
Turmalin  Centralblatt.  f.  Min.  Geol.  u. 
Palaen.      Pp.  299-302,  1901. 


Viola  claims  that  the  indices  of  refrac- 
tion for  quartz  can  be  determined  from 
a  cut  plate  by  the  Abbe  refractometer 
with   exactitude    to    the  fifth    decimal 
place.     From    hoo    sections    cut    from 
the  same  crystal  of  quartz,  one  parallel,  the  other  perpendicular  to  the  optic  axis, 
he  obtained  for  the  ordinary  ray  with  sodium  light : 
00  (transmission  parallel  axis)  1.54426. 
CsD  (transmission  perpendicular  to  axis)  1.54442. 
That  is,  a  difference  of  0.0U(I16. 
From  which  he  concludes  that  the  wave  surface  consists  of  two  rotation 
ellipsoids,  and  that  the  Fresnel  theory  is  not  here  available. 

In  tourmaline,  using,  however,  the  prism  method,  but  using  two  different 
prisms  from  the  same  crystal,  he  obtains  for  the  ordinary  index  in  Elba  tour- 
malines with  sodium  light : 

Transmission 

Parallel 

axis. 

Yellow  crystals,  -         -         1.6494 

Colorless  crystals. 
Green  crystals, 

Wiilfing  objects  to  the  conclusions  of  Viola,  pointing  out  that  in  the  case  of 
quartz,  in  spite  of  great  care  in  observation  and  in  construction  of  the  apparatus, 
-L.,       it  is  not  possible  to  claim  absolute  freedom  from  error  to  the 
fourth  decimal.     In  case  of  tourmaline,  he   points  out  the 
well  known  variation  in  composition  in  different  portions  of 
same  crystal.     To  avoid  this  he  prepared  a  single  four-sided 
pyramid,  so  that  both  directions  of  transmission  were  through 
the  same  material,  as  in  the  figure,  the  dotted  lines  repre- 
senting the  portion  of  the  tourmaline  that  was  ground  away. 
With  such  pyramids  he  obtained  for  oj : 


1.6425 
1.6479 


Transmission 

Perpendicular 

axis 

1.6482 
1.6402 
1.6503 


Difference. 

0.0012 
0.0023 
0.0024 


Transmission 

Transmission 

Parallel  axis. 

Perpendicular  axis. 

Difference 

Elba  Colorless  I, 

1.6419 

1.6419 

0.0000 

Elba  Colorless  II, 

-     1.6418 

1.6418 

0.0000 

Elba  Colorless  III,    - 

1.6424 

1.6423 

0.0001 

Haddam  Green, 

-     1.6401 

1.6400 

0.0001 

That  is,  the  greatest  difference  was   yV  to  -,}-^  that  obtained   by   Viola,  and 
indicate  that,  at  least  for  tourmaline,  the  Fresnel  law  holds.  a.  j.  m. 


and  Laboratory  Methods.  1475 

Gareiss,   A.      Ueber    Pseudomorphosen  nach       Entirely    aside     from    synonyms,  some 
Cordierit  &  Tschermak's.      Min.   u.  petroe.       ^         ^  u  u  •  *. 

Mitth.  20:  1-39,  .900.  twenty    names    have    been    given    to 

pseudomorphs  after  cordierite  (iolite) 
which  are  simply  stages  and  phases  of  decomposition,  the  usual  final  product  of 
which  is  muscovite  or  biotite.  The  author  examines  most  of  these,  and  con- 
cludes that  the  decomposition  starts  from  a  network  of  clefts  in  the  cordierite, 
sometimes  arranged  irregularly,  at  other  times  parallel,  both  to  base  (OUl)  and 
the  cleavage  (Ul(»).  In  several  instances  clefts  were  also  observed  parallel  to 
the  prism  (110). 

These  alteration  clefts  in  some  instances  serve  only  as  central  canals  for  the 
transportation  of  material,  and  enclose  a  fine  grained  zone.  At  other  times  this 
zone  is  surrounded  by  another,  in  which  extinction  takes  place  in  perfect  unity 
with  the  cordierite,  but  differs  in  the  color  and  the  lowered  double  refraction, 
which  may  reach  isotropy.  Frequently  this  material  fills  the  entire  space 
between  the  canals,  and  the  entire  crystal  becomes  thus  "  intermediate  "  sub- 
stance composed  of  undeterminable  fibers  and  little  scales. 

A  third  type  of  cleft  shows  little  fibers  or  scales  perpendicular  to  the  central 
canal,  and  frequently  with  brilliant  interference  colors. 

The  final  products  of  the  decomposition  are  mica  and  chlorite,  and  some 
quartz.  The  MgO  of  the  iolite  gradually  diminishes,  and  water,  alkalies,  and  iron 
enter.  The  mica  is  usually  muscovite,  rarely  biotite,  and  in  one  case  paragonite. 
The  tendency  to  form  muscovite  is  shown  by  the  fact  that  this  exclusively  is 
formed  from  cordierite  in  granites  and  gneisses  (which  are  rich  in  potassium), 
whereas  in  cases  such  as  the  cordierite  in  quartz  lenses  in  mica  schist,  where 
little  potassium  is  obtainable,  the  pseudomorphs  consist  principally  of  chlorite. 

Upon  the  basis  of  alteration  product  the  author  proposes  the  following 
nomenclature,  which  is  practically  in  conformity  with  the  original  use  of  each 
name  : 

Finite. — Preponderating  final  product  mica,  and  without  lamellar  parting 
parallel  (001).  Here  are  included  the  occurrences  of  Schneeberg,  Auvergne, 
Silberberg,  Schonfeld,  and  Fichtelgebirge. 

Gigantolite. — Preponderating  final  product  mica,  and  with  lamellar  parting 
parallel  (001).  Here  are  included  the  pseudomorphs  from  Heidelberg  and 
Wasserhauseln. 

Prasiolite  (Praseolite,  wrong  orthog.). — Preponderating  final  product  chlorite, 
and  without  lamellar  parting  parallel  (001).  Here  are  included  the  occurrences 
from  Bamle,  Kragerde,  and  the  Alpine  pseudomorphs,  which  show  no  lamellar 
parting. 

ChlorophylUte. — The  preponderating  end  product  chlorite,  and  with  lamellar 
parting  parallel  (001).  Here  belong  the  occurrences  of  Haddam,  Unity,  and  the 
gigantolite  of  Tammela,  the  lamellar  fahlunite  of  the  Talkschiefer,  and  the 
Alpine  pinites  with  lamellar  parting. 

As  to  the  many  other  names  the  author  points  out  the  essential  identity  of 
several,  as  shown  by  descriptions.     These  may  be  summed  up  as  follows : 

Esmarkite  ==  ChlorophylUte. 

Raumite  =  Prasiolite. 

Weissite,  Triclasite  and  Huronite  ^  Fahlunite,  but  fahlunite  not  in  every 
instance  a  cordierite  pseudomorph. 

Iberite  =  Gigantolite. 

Bonsdorffite  =  Prasiolite. 

Micarrell  Frieskben  =  Kataspilite  Igelstrotn,  which  is  not  a  cordierite  pseudo- 
morph according  to  author.  a.  j.  m. 


1476  Journal  of  Applied  Microscopy 


MEDICAL  NOTES. 

Gram's  Method  for  Staining  Diphtheria  Bacilli. — Allow  the  fixed 
specimens  to  remain  for  20  to  SO  minutes  in  an  anilin-water  solution  of  gentian 
violet,  prepared  in  the  following  manner : 

To  100  c.  c.  of  distilled  water  add,  drop  by  drop,  anilin  oil  until  the  mixture 
is  opaque.  Shake  well  after  each  addition  of  anilin  oil.  Filter  through  moist- 
ened filter  paper  until  perfectl}'  clear.  To  100  c.  c.  of  the  filtrate  add  10  c.  c. 
of  absolute  alcohol  and  11  c.  c.  of  concentrated  alcoholic  solution  of  gentian 
violet. 

After  remaining  the  required  time  in  this  mixture  the  specimens  are  placed 
for  about  five  minutes  in  the  following  iodin  solution  : 

lodin, 1  gm. 

Potassium  iodide, 2  grns. 

Distilled  water, 300  c.  c. 

This  solution  should  be  allowed  to  act  until  the  specimens  are  black,  after 
which  they  are  thoroughly  washed  in  alcohol,  which  removes  the  black  color, 
causing  the  specimens  to  appear  pale  grey. 

Dry  and  mount  in  balsam,  or  contrast  stain  with  carmin  or  Bismark  brown. 

c.  w.  J. 

Piorhowska.     The  Staining  of  Diphtlieria  Or-      A  method  is  given  by  which  the  author 

ganisms.     Berliner  klin.  Wochens.,  Mar.  4,       ,     ,.  .,         ^^   ■,  ,       .^    a^^^    o«-     t-^ 

^  .  t.      believes    it    possible    to    demonstrate 

positively  the  existence  of  these  organ- 
isms by  staining.  Make  dry  cover-glass  preparations  from  a  culture  of  bacilli 
grown  on  either  glycerin-agar,  or  Loeffler's  blood  serum,  at  a  temperature  of 
37.7 °C.  for  15  to  20  hours.  Stain  for  20  to  30  seconds  with  methyl-blue.  Decol- 
orize in  3  per  cent,  solution  of  HCl-alcohol  for  5  seconds.  Counterstain  in  1  per 
cent,  aqueous  solution  of  eosin  for  5  seconds.  Polar  nuclei  stain  deeply,  and 
the  central  portion  takes  a  marked  red  color.  c.  w.  j. 

Boston,  L.  NapoleoB.     How  to  preserve  as  per-      Partially  fill  a  bottle  with  urine,  cork 
manent    specimens    casts    found    in   urine.         .    ,    ,  ,    „ii^„,    *.„    ^f„,,j    ;.,  „  ^^^1 

Rep.  N.  Y  Med.  Jour.,  Nov.  4,  1899.  lightly,   and   allow   to   stand   m  a  cool 

place  until  a  precipitate  collects  at  the 
bottom  of  the  liquid.  Decant  the  supernatant  urine,  add  an  equal  quantity  of 
distilled  water  to  the  precipitate,  and  allow  it  to  stand  until  it  collects  again  at 
the  bottom  of  the  liquid.  With  a  pipette  place  a  small  drop  of  the  thickest  of 
the  sediment  on  the  center  of  a  slide  and  examine  under  low  power.  If  casts 
are  present,  evaporate  the  mount  nearly  to  dryness  and  add  by  means  of  a  glass 
rod  to  the  center  of  the  drop  of  urine  a  drop  of  the  following  mixture  : 

Liquor  acidi  arseniosi  (U.  S.  P.),  one  fluid  ounce. 

Salicylic  acid,  half  a  grain. 

Glycerin,  two  fluid  drachms. 
Dissolve  by  heat  and  add  acacia   (whole  tears),  and  again  warm  until  the 
solution  is   saturated ;  after  subsidence,   decant  clear   supernatant   liquid,   and 
add  a  drop  of  40  per  cent,  formalin  if  desired. 

In  order  to  get  an  equal  distribution  of  casts  throughout  the  field,  it  is 
necessary  to  draw  a  fine  needle  from  the  outer  margin  of  the  urine  to  the  center 
of  the  medium  until  the  two  substances  show  no  tendency  to  separate.  A 
cover-glass  is  moistened  by  the  breath  and  then  allowed  to  fall  gently  on  the 
specimen.  Cool  the  slide  for  a  few  hours  in  order  to  harden  the  mount  com- 
pletely. Specimens  thus  prepared  may  be  kept  indefinitely  without  deteri- 
oration, c.  w.  J. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


Volume  IV. 


OCTOBER,  1901, 


Number  10 


The    Botanical    Laboratory   and   the   Botanical  Garden  of  the 
Tokyo  Imperial  University,  Japan. 

The  Botanical  Laboratory  of  the  Tokyo  Imperial  University  is  located  in  the 
Botanical  Garden,  about  three-quarters  of  a  mile  from  the  other  university  build- 
ings. It  was  removed  from  the  university  campus  to  its  present  site  three  years 
ago.  The  building  is  one  story  high,  and  consists  of  two  parts  which  are 
connected  by  a  covered  alley-way. 


Fig.  1. — Botanical  Laboratory  of  the  Tokyo  Imperial  University.     Front  view. 

The  front  part  of  the  double  building  contains  the  herbarium,  the  library, 
laboratories,  and  rooms  for  a  professor  and  three  assistants.  The  second  build- 
ing contains  the  museum  and  the  lecture  room.  Here  also  are  more  laboratories 
and  a  professor's  room.  In  addition,  here  are  found  the  dark  room,  the  room 
for  the  physiological  apparatus  and  chemical  balance,  the  store  room,  and  the 
room  for  the  incubators,  sterilizers,  etc.  Gas  and  water  are  conducted  to  all 
working  rooms. 

The  herbarium  is  well  represented  with  Japanese  flowering  plants  and  ferns, 

(1477) 


1478 


Journal  of  Applied  Microscopy 


including  tropical  plants  from  the  islands  of  Liu-Kiu  and  Formosa,  besides  some 
exotic  species.  The  lower  cryptogams,  though  constantly  added  to  the  collection, 
are  yet  far  from  complete. 

The  library  contains  the  leading  English,  German,  French,  and  Italian 
botanical  journals.  The  museum  contains  both  dried  and  alcoholic  specimens 
of  plants  for  morphological,  ecological,  and  pathological  purposes.  Some 
tropical  fruits  and  seeds  from  Java  and  Formosa  are  also  found  here. 

The  laboratory  is  quite  well  equipped  with  apparatus  and  literature  for 
research  work  in  plant  physiology.  Various  important  contributions  have  been 
made  here  along  this  line  during  the  last  few  years. 

A  good  microtome,  Zeiss'  microscopes  with  oil-immersion  objectives,  afford 
facility  in  the  study  of  cytology  and  embryology.  Some  good  work  has  also 
been  done  along  these  lines.  Among  them  Mr.  Hirase's  well  known  studies  on 
Ginkgo  should  especially  be  mentioned. 

The  specimens  and  literature  give  facility  for  the  study  of  systematic  botany 
also.     The  systematic  studies  of  tropical  and  subtropical  plants  from  the  islands 


^___^ 

|n|] 

B^ 

^^^^^^^H^WKBSESdtitefti^Bl 

Fig.  2. — Botanical  Laboratory  of  the  Tokyo  Imperial  University. 
End  view. 

of  Liu-Kiu  and  Formosa,  and  monographic  investigations  on  some  difficult 
phanerogamic  group,  e.  g.,  Bambusaceie,  are  the  present  features  along  this  line 
in  the  laboratory. 

The  apparatus  for  the  study  of  bacteriology  and  fermentation  is  also  well 
provided. 

The  following  lectures  and  laboratory  work  are  given  in  this  laboratory  for 
undergraduates : 

a.  Lectures. 

1.  General  botany  (morphology  and  physiology).     Three  hours   a  week 

throughout  the  year. 

2.  Systematic  botany.     Three  hours  a  week  throughout  the  year. 

3.  Advanced  plant  physiology.     One  hour  a  week  during  the  first  term. 

b.  Laboratory  work. 

L   Classification,  morphology,  histology,  and  embryology.     Twelve  hours 
weekly  throughout  the  year. 


and  Laboratory  Methods. 


1479 


2.  Morphology  and  histology.     Six  hours  weekly,  especially  for  geology 

students. 

3.  Plant  physiology.     Twelve  hours  weekly  throughout  the  year. 

4.  Research  work. 

There  are  now  six  graduates  and  about  fifteen  undergraduates  studying  in 
this  laboratory.  It  should  be  noticed  here  in  this  connection,  that  nearly  all  of 
the  studies  in  the  university  are  required,  and  students  who  specialize  in  botany 
are   required   to   study  zoology,  including   histology   and   embryology,  geology. 


Wall 


Uo-b. 


Gla.6S  IVi.viioiA' 


Coo.t  Janltc.. 

f?ccm   I         iTcom 


1 


1-^ 


Fig.  3. — Plan  of  the  building  of  the  Botanical  Laboratory  of  the  Tokyo  Imperial  University. 

paleontology,  mineralogy,  physiology,  physiological  chemistry  and  bacteriology, 
besides  the  above  mentioned  courses  in  botany.  Only  students  of  botany, 
zoology,  and  geology  come  to  this  laboratory  to  study.  Students  of  forestry  and 
agriculture  pursue  their  botanical  studies  at  the  botanical  laboratory  in  the 
agricultural  college  of  the  university. 

The  following  is  the  present  instructing  staff  of  the  botanical  laboratory : 
Prof.  J.  Matsumura,  professor  of  botany  and  the  director  of  the   botanical 
garden.     (Systematic  botany.) 

Prof.  M.  Miyoshi,  professor  of  botany.     (Plant  physiology.) 


1480 


Journal  of  Applied  Microscopy 


Mr.  K.  Fuji!,  assistant.     (In  charge  of  morphology  and  embryology.) 
Mr,  T.  Makino,  assistant.     (Systematic  botany.) 
Mr.  S.  Matsuda,  assistant.     (Systematic  botany.) 
Mr.  Y.  Yabe,  assistant,  and  secretary  of  the  botanical  garden. 
The  Botanical  Garden  is  the  great   source  of   living  materials  for  study.     It 
was  established  in  16(S1,  and  was  long  renowned  as  the  "  O  Yaku  Etr'  (garden 


Fig.  4 — Greenhouse  in  the  Botanical  Garden. 

of  medicinal  plants).  The  area  occupied  by  the  garden  is  more  than  five  acres. 
The  plants  are  placed  in  rows  according  to  the  natural  order,  each  with  labels 
having  Latin  and  common  names.  In  one  quarter  of  the  garden  medicinal 
plants  are  planted  in  groups.  In  the  greenhouse  the  tropical  plants  from  various 
parts  of  the  world  are  quite  well  represented. 


Fig.  5. — Cherry  trees  {Pni iius  pseudo-cerasii.':)  in  blossom  in  the  Botanical  Garden. 

A  row  of  large  cherry  trees  (^J''nni us pseii do-ccra sh s)  with  large  pink  blossoms 
is  a  very  beautiful  sight  at  the  flowering  time  in  April.  A  few  large  Ginkgo 
trees  {Ginkgo  bilobd)  and  several  hundred  tall  bamboos  would  be  a  new  sight  to 
the  Western  traveler. 

In  a  part  of  the  Botanical  Garden  we  have  a  genuine  Japanese  garden  with  a 
pond.  In  the  pond,  lotus  {Nclianbo  niicifcra),  water-lilies  {Nymphcea  odorata, 
etc.),  and  several  other  water  plants  are  growing.  It  is  also  a  good  collecting 
place  for  fresh  water  algae  and  planktons.     The  climbing  wistaria    {Wistaria 


and  Laboratory  Methods. 


1481 


c-hinc/isis),  from  which  hang  purple  branches  of  flowers  in  May,  and  plumb  trees 
{Priinus  miiftte),  renowned  for  their  sweet  fragrance  of  flowers,  are  found  in 
other  corners  of  the  garden. 

A  large  cycas  tree  {Cycas  rei'oluta'),  several  yuccas  {\'ucca  filamentosa,  V. 
gloriosa,  Y.  ahifolid),  Japanese  palms  {Trachycarpus  excelsa,  7.  fortunei'),  planted 
in  front  of  the  Botanical  Laboratory,  give  quite  a  tropical  aspect  to  that  corner. 


Fig.  6. — The   lotus    {NcliDiibo  micifera)  in  the  pond  of  the 
Botanical  Garden. 

Of  these  only  cycas  is  protected  from  the  cold  in  winter.     Japanese  banana  trees 
{Af/isa  bas/oo),  which  bear  no  fruit,  are  planted  in  groups  in  one  corner  of  the 
garden.     The  plant  dies  in  winter,  with  the  exception  of  the  lower  parts  of  the 
stem  and  the  root,  from  which,  new  shoots  come  out  in  the  spring. 
Cornell  University.  KlICHI    MiYAKE. 


The  Course  of   Study  in  Invertebrate  Zoology  in  the  Marine 
Biological  Laboratory  at  Wood's  Holl. 


Fig.  1. — Marine  Biological  Laboratory  at  Wood's  Holl. 


14S2  Journal  of  Applied  Microscopy 

The  laboratory  in  which  the  course  of  study  in  invertebrate  zoology  is  given 
at  Wood's  HoU  is  a  large  room,  twenty-eight  by  fifty-six  feet,  on  the  first  floor 
of  the  south  wing  of  the  main  building  of  the  Marine  Biological  Laboratory.  The 
room  receives  light  from  the  north,  west  and  south  sides  through  eighteen  large 
windows.  Directly  in  front  of  each  window  is  placed  a  laboratory  table  which 
is  arranged  to  accommodate  two  students.  On  the  middle  of  each  table  is  a  two 
shelved  rack  on  which  the  student  finds,  in  easy  reach,  a  set  of  the  reagents  and 
apparatus  he  will  need  for  the  work  of  the  course.  A  good  arrangement  of  the 
reagents  and  apparatus  on  the  shelves  is  that  given  below  in  the  table  : 


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One  finger  bowl,  "2  large  white  dishes,  1  paraffin  bottomed  tray,  2  pipettes, 
and  a  lamp. 

The  east  end  of  the  room  has  no  windows,  but  is  furnished  with  a  blackboard 
and  a  chart  rack.  In  addition  to  blackboard  sketches,  a  set  of  Leuckart  charts 
were  used  in  illustrating  the  lectures  of  the  course.  In  front  of  the  black- 
board, a  lecture  table  and  several  benches  are  placed.  The  laboratory  is  pro- 
vided with  running  water,  both  salt  and  fresh,  and  in  the  middle  of  the  room 
there  are  a  number  of  large  aquaria.  When  practical,  these  aquaria  were  sup- 
plied with  the  animals  being  studied,  thus  giving  the  student  the  opportunity 
of  seeing  the  living  animal  under  more  or  less  normal  conditions.  Considerable 
stress  was  laid  on  this  part  of  the  work  and  twice  each  week  excursions  were 
made  to  various  localities  for  the  purpose  of  seeing  the  animals   at   home.     On 


and  Laboratory  Methods. 


1483 


some  of  these  excursions  digging  and  shallow  water  collecting  was  the  object. 
In  others,  dredges  were  used  in  deeper  water.  On  one  afternoon  the  class 
accepted  the  invitation  from  Dr.  H.  M.  Smith  of  the  U.  S.  Fish  Commission,  to 
go  on  a  dredging  trip  on  the  Steamer  "  Fish  Hawk"  and  had  the  privilege  of 
seeing  how  dredging  is  done  by  means  of  the  most  efficient  and  up-to-date 
apparatus. 

The  course  of  study  covered  six  weeks,  beginning  July  3d,  and  ending 
August  13th.  It  consisted  in  both  lectures  and  laboratory  work.  The  lectures, 
which  were  given  daily,  from  9  to  10  a.  m.,  by  the  instructors,  had  for  their  subject 
matter  the  natural  history,  classification,  anatomy  and  development  of  the  ani- 
mals or  groups  of  animals  to  be  studied  in  the  laboratory  during  the  day.  The 
instructor  who  gave  the  morning  lecture  had  charge  of  the  laboratory  work 
which  followed.     In  this   he  was  assisted   by  two   other  instructors.     He  had 


Fig.  '1. — General  Laboratory. 


selected  the  forms  for  study  and  was  responsible  for  their  collection  and  prep- 
aration for  the  students.  He  had,  with  a  few  exceptions,  prepared  the  labora- 
tory outlines  which  were  used  by  the  students  in  their  studies  and  dissections. 
For  a  few  forms  Bumpus'  "  Invertebrate  Zoology  '"  was  used. 

In  addition  to  these  elementary  lectures  by  the  instructors  it  was  intended 
that  each  day's  work  should  close  with  a  lecture  by  an  investigator  of  the  group 
being  studied  on  some  subject  in  which  he  is  interested  relating  to  the  group. 
This  plan  was  given  up  for  two  reasons,  the  investigators  were  either  not  at 
Wood's  Holl  or  were  unwilling  to  lecture,  and  at  4  o'clock  the  greater  part  of 
the  students  wanted  to  go  bathing.  Those  lectures  which  had  been  promised, 
before  the  course  began,  were  given — the  three  by  Professor   C.   O.   Whitman 


1484 


Journal  of  Applied  Microscopy 


on  "  Metameric  Segmentation,"  at  the  originally  planned  hour,  4  p.  m.,  but  the 
three  lectures  by  Dr.  G.  N.  Calkins  on  the  "  Protozoa  '"  and  three  by  Prof.  G.  H. 
Parker  on  the  "  Sense  Organs  of  the  Crustacea  "  were  given  in  the  evening  at 
the  general  lecture  hall. 

The  laboratory  work  of  the  day  was  divided  into  two  periods  of  two  hours 
each,  the  first  from  10  a.  m.  to  12  m.,  the  second  from  2  to  4  p.  m.  During 
these  periods  it  was  the  duty  of  three  instructors  to  be  in  the  laboratory  for  the 
purpose  of  distributing  material  to  the  students  and  to  answer  such  questions  as 
they  might  be  asked  concerning  the  work  of  the  day.  Besides  these  regular 
working  hours  the  laboratory  was  open  at  all  times  to  the  students  and  many  of 
them  took  the  opportunity  to  carry  further  the  work  begun  in  the  course  or  to 
work  along  lines  in  which  they  were  especially  interested.  The  work  in  the  lab- 
oratory consisted  chiefly  in  gross  dissections  and  microscopical  studies  but  some 


Fig.  '.]. — Private  Laboratory. 

work  was  given  in  killing,  staining  and  mounting  small  animals  and  in  mounting 
and  interpreting  serial  sections.  Each  student  furnished  his  own  dissecting 
instruments,  drawing  materials  and  microscope.  In  many  cases  the  micro- 
scopes were  owned  by  the  students,  but  in  others  they  were  rented  for  the  course 
from  the  Bausch  ^r  Lomb  agency.  For  a  rental  of  five  dollars  a  student  was 
provided  with  a  B.B.  stand  furnished  with  an  Abbe  Condenser,  two  eyepieces 
and  two  objectives. 

Students  were  advised  to  make  accurate  drawings  of  what  they  saw,  and  in 
case  the  work  was  to  be  substituted  in  their  college  course  such  drawings  were 
required. 

The  table  which  follows  will  give  a  better  idea  of  the  actual  work  done  in 
the  course  and  the  manner  of  conducting  it,  than  could  be  gotten  from  a  lengthy 
description. 


and  Laboratory  Methods.  1485 


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1486  Journal  of  Applied  Microscopy 

Of  the  thirty-one  students  who  attended  the  course,  this  year  about  one-half 
were  teachers.  The  other  half  was  made  up  of  students  who  were  either  making 
credits  in  their  regular  college  work  or  who  were  preparing  for  a  course  in  med- 
icine. One  practicing  physician  took  the  course.  A  few  who  took  the  work 
had  come  to  Wood's  Holl  for  the  Nature  course  which  was  not  given  this  year. 

After  the  work  of  the  day  the  instructors  met  for  a  short  time  in  the 
director's  room  and  went  over  the  work  next  to  be  given.  Difficult  points  were 
demonstrated  by  the  instructor  who  would  be  in  charge  of  the  work  and  methods 
of  presenting  the  subject  were  discussed. 

Formerly,  at  the  time  each  group  was  being  taken  up,  an  investigator,  espec- 
ially interested  in  the  group  but  not  otherwise  interested  in  the  course,  has  been 
invited  to  lecture  on  his  specialty  before  the  class.  The  duty  of  the  instructors 
was  only  to  assist  in  the  laboratory  work,  while  the  director  in  charge  of  the 
course  was  responsible  for  the  selection  of  the  types  to  be  studied  and  for  the 
manner  in  which  each  group  was  presented  to  the  students.  While  this  method 
may  be  the  theoretically  ideal  way  of  introducing  students  to  the  study  of  zool- 
ogy it  is  lacking  in  practice  in  some  important  respects  ;  it  is  not  always  possi- 
ble to  get  investigators  of  certain  groups  of  animals  to  lecture  at  the  time  the 
lectures  are  needed,  and  often  the  work  goes  on  without  lectures.  Investigators, 
not  knowing  the  students  to  whom  they  are  lecturing,  nor  the  work  they  have 
done,  often  fail  to  present  their  subjects  in  the  best  manner  possible.  It  was  to 
ensure  a  coordination  between  lectures  and  laboratory  work,  and  lectures  adapted 
to  beginning  students,  that  the  director  this  year.  Dr.  G.  A.  Drew,  not  only  asked 
each  instructor  to  give  the  lectures  on  certain  groups  but  to  take  charge  of  the 
laboratory  work  on  the  same. 

The  success  of  the  plan  was  to  be  seen  in  the  active  interest  of  the  students 
in  their  work  which  continued  throughout  the  course.  Very  little  ciittvig  was 
done  and  on  the  last  day  twenty-eight  of  the  thirty-one  students  who  began  the 
course  reported  for  work.  Caswell  Grave. 

Johns  Hopkins  University. 

Botany  at  the  Biological  Laboratory  at  Wood's  Holl. 

BOTANICAL  STAFF. 

Bradley  Moure  Davls,  Instructor  in  Botany,  University  of  Chicago. 

Gkorge  T.  Moork,  Instructor  in  Botany,  Dartmouth  College. 

Rodney  H.  True,  Lecturer  at  Harvard  University. 

Charles  H.  Shaw,  Professor  of  Botany,  The  Temple  College. 

Anstruther  A.  Lawson,  Fellow  in  Botany,  University  of  Chicago. 

LiLiJAN  G.  MacRae,  Curator  and  Collector  in  Botany. 

Developments,  such  as  have  in  recent  years  taken  place  at  Wood's  Holl  in 
physiology  and  in  botany,  afiford  illustrations  of  the  all-embracing  love  of  knowl- 
edge so  characteristic  of  this  unique  station  by  the  sea,  the  Mecca  of  American 
biology.  More  than  ever,  during  the  past  season,  botany  has  made  itself  felt  as 
a  live  and  considerable  part  of  the  laboratory.  The  building  was  full  to  the 
last  table.  On  the  upper  floor  were  assembled  the  workers  in  cryptogamic  botany 
under  Dr.  Davis  and  Dr.  Moore.     These  were  divided  into  three  groups,  those 


and  Laboratory  Methods. 


1487 


devoting  the  whole  session  to  the  algre,  those  working  throughout  on  the  fungi, 
and  those  who  divided  the  time  between  the  two  classes. 

The  wetness  of  the  season  brought  on  an  abundant  supply  of  fungi;  twenty- 
six  species  of  myxomycetes  alone  were  found.  Expeditions  near  and  far  brought 
to  the  laboratory  tanks,  also,  the  usual  rich  assortment  of  algae. 

The  lower  floor  was  occupied  by  two  classes — in  physiology  and  in  ecology, 
respectively.  The  former  reached  a  circle  much  larger  than  the  class,  many 
zoologists  and  other  workers  attending  the  lectures.  The  ecology  people  spent 
much  of  their  time  in  the  field,  and  made  the  beginning  of  an  ecological  survey 
of  the  vicinity.  At  the  close  of  the  term  many  members  joined  in  a  botanizing 
party  to  the  White  Mountains. 

Year  by  year  the  work  in  botany  will  take  new   steps  forward,  and  seeking 
ever  to  turn  the  students  in  the  direction  of  research,  it  may  well  do   its   share 
in  upholding  the  Marine  Laboratory  ideal  of  productive  scholarship. 
Temple  College,  Philadelphia,  Pa.  C.  H.  Shaw. 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 
THE  ORTOL  DEVELOPER. 
It  may  be  of  some  interest  to  the  readers  of  the  Journal  who  are  interested 
in  laboratory  photography  to  be  put  in  touch  with  one  of  the  best  all  around 
developers. 


Fig.  1. — Viscera  of  Frog. 


1488 


Journal  of  Applied  Microscopy 


After  using  the  various  developers  put  on  the  market,  with  various  degrees  of 
success,  I  have  at  last  struck  upon  one  which  fulfills  all  of  my  requirements. 
This  salt  goes  by  the  name  of  Ortol,  and  is  purchased  in  sealed  tubes  with  an 
accompanying  cartridge  of  the  required  amount  of  soda.  The  contents  of  each 
are  dissolved  in  20  oz,  of  water  and  kept  in  separate  bottles. 

Various  grades  of  intensity  may  be  gotten  by  regulating  the  strength  of  the 
solution  and  by  the  use  of  more  or  less  soda  as  occasion  requires.  In  Fig.  1  the 
development  was  carried  slightly  too  far,  but  shows  great  contrast  and  clearness. 
In  the  second  illustration  we  have  a  transverse  section  of  the  human  sciatic  nerve 
developed  for  detail  rather  than  contrast. 


Fig.  2. — Transverse  section  of  human  sciatic  nerve. 


The  following  proportions  may  be  varied  somewhat  according  to  exposure, 
but  I  have  found  them  to  answer  for  all  purposes  by  using  more  or  less  bromide : 
For  negatives : 

Ortol,         -        -       -  -  2  parts  stock  solution, 

Soda,         .        -       .  .  2      "         " 

Water,       .        -        -  -  4      " 
For  lantern  slides  : 

Ortol,        -        -        -  -  2  parts  stock  solution. 

Soda,        -        -        -  -  1   part       "            " 

Water,  -  -  -  -  .3  parts. 
For  bromide  papers  : 

Ortol,        -        .        -  -  1   part  stock  solution, 

Soda,        ...  -  1      "         '< 

Water,      -        -        -  -  8  parts. 

I  do  not  believe  any  definite  rule  can  be  set  down  as  to  just  how  much 


and  Laboratory  Methods.  1489 

bromide  shall  be  used,  this  being  regulated  by  the  judgment  and  experience  of 
the  operator.  I  have  found  it  very  convenient  to  use  none  at  all  in  some  cases, 
and  again  in  others  to  stop  the  developer  back  so  that  an  image  will  not  appear 
in  less  than  six  or  eight  minutes. 

When  working  for  extreme  contrast,  as  in  reproducing  book  illustrations, 
mechanical  diagrams,  etc.,  a  plate  sensitized  specially  for  this  purpose  is  abso- 
lutely necessary.  Such  a  plate  is  manufactured  by  the  G.  Cramer  Dry  Plate  Co. 
of  St.  Louis,  and  may  be  obtained  of  any  dealer  in  photo  supplies.  They  are 
rather  slow  in  their  action  and  must  be  given  a  longer  exposure  than  ordinary. 

For  development,  the  solution  must  be  as  strong  as  possible  and  well  stopped 
back.  I  usually  make  up  the  developing  solution  in  200  c.  c.  quantities,  "  enough 
to  conveniently  fill  a  5x7  tray,"  after  the  following  formula  : 

Ortol,         -         -         -  75  c.  c.  stock    solution. 

Soda,  -         -         -       100  c.  c. 

l^romide,  10  per  cent,  sol.,  25  c.  c. 

After  exposure,  the  plates  should  be  brushed  off  and  immersed  in  the 
developer,  when  the  image  will  flash  up  rather  quickly,  and  stand  out  in  very 
strong  contrast  with  the  black  background  ;  but  do  not  remove  until  the  entire 
plate  is  perfectly  black,  even  when  held  up  to  the  red  light,  then  pass  it  into  acid 
fixing  bath.  The  black  background  will  be  found  to  be  perfectly  opaque,  even 
if  held  up  to  the  sun,  while  the  lines  will  stand  out  perfectly  transparent. 

Should  any  of  my  readers  start  using  Ortol,  I  feel  fully  satisfied  that  it  will 
be  some  time  before  they  change  again.  R.  P.  Woodford. 


Contributions  to  Our  Knowledge  of  Color  in  Photo- 
micrography.'^ 

One  of  the  most  perplexing  and  as  yet  unsolved  problems  of  Photo-micro- 
graphy is  that  of  color  values,  i.  e.,  how  to  reproduce  natural  colors  by 
means  of  the  sensitive  plate.  Of  the  plates  now  in  use,  the  orthochromatic  ap- 
proaches most  nearly  the  ideal  color  plate ;  yet  this  is  not  perfectly  satisfactory, 
as  it  does  not  give  sufficient  contrast. 

The  investigations  which  form  the  basis  of  this  article  were  undertaken  to 
determine  the  relative  merit  of  various  photographic  plates.  The  apparatus,  as 
illustrated  in  Fig.  1,  consists  of  a  direct  vision  spectroscope  so  mounted  in  the 
front  board  of  an  ordinary  camera  (with  lenses  removed)  that  the  spectrum, 
when  projected  on  the  plate,  will  come  in  the  center  horizontally  and  at  the  top 
of  the  plate.  The  back  of  the  camera  is  constructed  in  such  manner  as  to  allow 
of  its  being  moved  in  the  vertical  plane,  thus  making  it  possible  for  one  to  make 
four  exposures  on  the  same  plate,  and  by  so  doing  to  make  an  accurate  com- 
parison between  them. 

The  plates  examined  may  be  grouped  according  to  their  degree  of  perfection 
as  follows : 

Group  I — Characterized  by  a  very  high  degree  of  sensitiveness  a  little  above 

*  F.  L.  Richardson.     Journal  of  the  Boston  Society  of  Medical  Sciences,  S :  460-464. 


1490 


Journal  of  Applied  Microscopy 


line   D,  falling  oft"  abruptly  on   either  end,  and   only   slightly  sensitive  to   the 
greens  and  blues. 

Group   II — Characterized  by  two   distinct   maxima — one  a  little   above  the 


Fig.  1. 

Apparatus  for  making  spectrograplis.     A.  Spectroscope.     B.  Back  of  camera,  carrying  screen  and  plate- 
holder.     C.  Supports  upon  which  the  back  (B)  may  be  moved.     D.  Shutter. 
K.  Color  screen  in  color  screen  holder. 


D    line,    and    the    other    in    the    blue-green, 
sensitiveness  falls  very  considerably. 


Fig.  2. 

Kxplanation  to  Figure. — The  upper  curve  shows  the  visual  in- 
tensity of  the  spectrum  (from  Fraunhofer).  Curves  I-V  repre- 
sent the  photographic  intensity  of  the  sjjectrum  taken  on  plates 
from  group  of  corresponding  number.  C.koi'f  1 — Cranur 
isochromatic  (slow).  Gkoi'p  II — Standard  orthochromatic 
(slow) ;  Forbes  orthochromatic  (slow) ;  Carbutt  orthochro- 
matic (slow);  Otto  Perutz.  (Skoif  111  —  I.ovell  color-dif- 
ferentiating; American  spectrum  plate.  CIkoip  IV — Cadett 
!<;  Neal  spectrum  plate  (slow).  CiKoif  V  —  International 
"  F.rethro." 


Between   these  two   maxima  the 

Group  III — ^Characterized  by 
having  its  maximum  sensitiveness 
in  the  blue  (as  with  ordinary 
plates),  with  lesser  bands  of  sen- 
sitiveness extending  below  the  D 
line. 

Group  IV — Characterized  by 
bands  of  sensitiveness  extending 
below  line  D,  with  greatest  inten- 
sity in  the  yellow-green,  and  fall- 
ing off  at  the  violet  end  before 
H,. 

Group  V — This  group  most 
nearly  approaches  perfection.  It 
is  characterized  by  a  sensitive 
band  well  below  line  D,  and 
somewhat  below  the  red  end  of 
Groups  III  and  IV.  This  plate 
gives  an  almost  uniform  degree 
of  sensitiveness  with  a  maximum 
intensity  in  the  green. 

If  sensitiveness  to  the  spec- 


and  Laboratory  Methods. 


1491 


trum  were  the  only  feature  to  be  considered  in  the  selection  of  a  plate  for  photo- 
micrographic  work,  a  plate  from  Group  V  would  be  chosen,  but  the  general 
working  of  the  plate  as  well  as  the  keeping  qualities  are  factors  that  must  be 
considered.  For  practical  work  and  keeping  qualities  the  author  found  the 
Cadett  &  Neal  Special  Slow  Spectrum  Plate  of  Group  IV  most  satisfactory,  and 
used  it  in  the  preparation  of  the  spectrographs  illustrated  in  Fig.  3. 


Electric  arc.  -  - 
Picric  acid.  -  - 
Aurantia.  -  -  - 
Cresoidin.  -  -  - 
Congo  Red.  -  - 
Eosin.  -  -  -  - 
Carbol  Fuchsin. 
H<Eniatoxylin.  - 
INIethylen  Blue.  - 
Green  Glass.  -  - 
Methyl  Green.  - 
Quinine  Sulphate. 


Methylen  Blue. 


Methyl  Green. 
Picric  acid. 
Picric  acid. 
Methylen  Blue. 
Carbol  Fuchsin. 


Fig.  3. 

Explanation  to  Plate. — This  plate  is  a  reproduction  of  spectrographic  analyses  of  some  of  the  common 
stains.  The  red  end  of  the  spectrum  is  on  the  left.  The  principal  Fraunhofer's  lines  are  marked. 
The  name  of  the  stain  is  on  the  left,  while  on  the  right  is  the  name  of  the  proper  screen  to  use  to 
increase  the  photographic  intensity.  To  decrease  the  contrast  use  a  screen  of  the  same  color  as  the 
stain. 

The  perfect  photo-micrographic  plate  would  give  equal  photographic  in- 
tensity to  all  the  colors  of  the  visual  spectrum,  but  since  this  degree  of  excellence 
has  not  been  attained,  the  value  of  a  given  plate  may  be  enhanced  by  the  use  of 
color  screens,  or  ray  filters,  which  serve  to  increase  or  decrease  the  photographic 
intensity  of  a  color.  The  following  are  the  laws  upon  which  the  use  of  color 
screens  is  based : 

First.  To  increase  the  photographic  intensity  of  a  color,  a  screen  of 
complementary  color  should  be  used. 

Second.  To  decrease  the  photographic  intensity  of  a  color,  a  screen  of  the 
same  color  should  be  used. 

These  laws  hold  good  for  all  branches  of  photography,  whether  by  trans- 
mitted or  reflected  light,  and  are  not  dependent  upon  the  position  of  the  screen, 
i.  e,,  whether  the  screen  is  placed  between  the  source  of  illumination  and  the 
object,  between  the  object  and  the  objective  lens,  or  between  the  lens  and  the 
plate. 


1492  Journal  of  Applied  Microscopy 

In  determining  the  photographic  complement  of  a  color  it  is  more  accurate 
to  make  photographic  analyses  of  stains  and  color  screens  than  to  make  simple 
visual  analyses. 

The  same  plate  and  source  of  light  must  be  used  as  in  taking  the  photo- 
micrograph, because,  as  shown  above,  plates  of  different  makes  may  differ 
greatly  in  their  degree  of  sensitiveness  to  different  parts  of  the  spectrum,  and  it 
is  a  well  recognized  fact  that  diflferent  illuminating  agents  give  different  spectra. 
The  object  for  which  one  should  strive  in  the  analyses  of  stains  and  color  screens 
is  to  imitate  as  exactly  as  possible  the  conditions  that  will  exist  in  the  process  of 
photo-micrography.  The  spectrographs  illustrated  in  Fig.  3  were  made  with 
color  screens  in  preference  to  cells  of  fluid,  as  the  screens  are  much  more  con- 
venient than  cells,  and  are  quite  as  satisfactory.  The  color  screens  are  made 
by  soaking  a  cleared  lantern  slide  in  a  solution  of  the  desired  stain  until  the 
gelatin  is  saturated,  then  rinsing  and  removing  the  surface  liquid  with  a  cotton 
pad.  The  screen  is  then  dried  and  covered  with  a  cover-glass  as  in  mounting 
a  lantern  slide.  The  depth  of  color  in  these  screens  is  dependent  upon  the 
degree  of  concentration  of  the  staining  solution  rather  than  upon  the  length  of 
time  the  plate  is  soaked.  c.  w.  j. 


Kresylechtviolett. 

In  the  Centralblatt  fiir  Bakteriologie,  Vol.  27,  Homberger  describes  a  new 
method  of  staining  the  Gonococcus  by  which  it  may  be  distinguished  from  all 
other  micro-organisms.  The  stain  used  for  this  purpose  was  Kresylechtviolett,  a 
fluorescing  color  prepared  by  Leonhard.  In  dilutions  of  1-10,000  the  water 
solution  of  this  stain  colors  the  Gonococcus  a  reddish  violet  while  other  micro- 
organisms are  either  not  stained  at  all  or  take  a  faint  blue  tinge.  Homberger 
mentions  further  that  mast-cells,  amyloid,  mucin  and  colloid  are  stained  well  with 
this  dye  and  that  it  can  also  be  used  in  Gram's  method. 

It  was  found  on  working  with  the  stain  in  this  laboratory  that  the  water  solu- 
tions precipitate  on  standing  and  lose  their  staining  power.  The  alcoholic 
solutions  did  not  precipitate,  but  satisfactory  results  could  not  be  obtained  with 
them.  Since  the  reactions  obtained  with  the  stain  were  found  by  repeated  exper- 
iment to  be  very  characteristic,  and  as  it  promised  to  be  a  very  valuable  addition 
to  diagnostic  methods  the  present  work  was  undertaken  at  the  request  of  Dr. 
Warthin  to  find,  if  possible,  a  satisfactory  method  of  using  the  stain,  and  to  extend 
its  application  in  pathological  work. 

The  stain  used  was  prepared  by  Griibler.     It  is  soluble  in  water  and  alcohol. 
After  much  experimental  work  the  following  method  of  preparation  was  found  to 
be  satisfactory  in  all  respects,  and  to  meet  all  requirements,  and  is  the  one  used 
throughout  this  work  : 
Preparation  (Morse)  : 

Five  per  cent,  aqueous  solution  of    Phenol,  -         80  c.  c. 

Ninety-five  per  cent.  Ethyl  Alcohol,     -  -  -         20  c.  c. 

Stain,  - -  -  1  gm. 


and  Laboratory  Methods.  1493 

Mix  the  two  solutions,  then  add  the  stain,  stirring  thoroughly.  As  soon  as 
the  stain  is  all  dissolved,  filter.  This  solution  keeps  well,  gives  all  the  staining 
reactions,  and  may  be  diluted  to  any  degree  with  distilled  water  without  causing 
precipitation.  The  above  method  of  preparation  must,  however,  be  rigidly 
followed. 

Tissue  may  be  fixed  in  any  way,  but  formalin,  mercuric  chloride,  and  Zenker's 
give  the  best  results  in  the  order  named.  Parafiin  or  celloidin  embedding  may 
be  used.     Excellent  results  may  be  obtained  by  the  combination  plate-method. 

Staining  method  (Morse)  : 

1.  Stain  1-5  minutes. 

2.  Wash  thoroughly  in  distilled  water. 

3.  Blot  with  filter  paper. 

4.  Anilin-Xylol  (2:1). 

5.  Pure  Xylol. 

6.  Balsam. 

This  method  is  the  best  one  for  the  reactions  with  the  majority  of  patholog- 
ical substances.  The  variations  of  this  method  for  certain  specific  reactions 
will  be  mentioned  below.  Nearly  all  the  tissues  of  the  body  and  the  important 
pathological  conditions  have  been  worked  over  with  this  stain.  Its  most  impor- 
tant applications  are  as  follows  : 

For  use  as  a  simple  nuclear  stain  it  is  best  to  stain  only  two  minutes,  then 
wash  and  differentiate  in  alcohol.  A  contrast  stain  is  not  necessary,  as  the  deep 
violet  color  of  the  nuclei  is  easily  distinguished  from  the  pale  blue  tint  given  to 
other  structures.  Eosin  may,  however,  be  used  after  the  sections  have  been 
stained  and  differentiated  in  alcohol. 

Blood  smears  give  the  best  results  when  fixed  as  for  tri-acid  staining.  Heat 
fixation  gives  the  best  result.  When  stained  for  two  minutes  the  red  cells  are  a 
light  yellowish  green,  the  protoplasm  of  the  leucocytes  colorless,  and  the  nuclei 
rose-pink.  Blood-plates  and  basophile  granules  stain  like  the  nuclei.  The  mala- 
rial Plasmodium  stains  a  dull  pink,  not  as  deep  as  the  leucocyte  nucleus,  and  is 
easily  distinguished.  In  well  fixed  tissue  sections  blood  stains  similarly,  but  the 
basophile  granules  do  not  show. 

The  nuclei  of  young  connective  tissue  stain  a  light  purple,  while  the  intercel- 
lular substance  takes  a  dull  rose  pink;  in  old  connective  tissue  the  nuclei  stain  a 
deep  violet  or  purple,  while  the  intercellular  substance  does  not  stain  at  all  or 
takes  a  very  light  violet. 

Yellow  elastic  tissue  stains  a  sky  blue.  It  is  best  brought  out  by  longer 
staining. 

Voluntary  muscle  stains  a  pale  green  with  violet  nuclei,  the  protoplasm  of 
involuntary  muscle  a  light  purple  to  a  light  blue  with  nuclei  of  a  darker  shade. 

Nerve  cells  and  axis  cylinders  stain  purple,  the  nuclei  violet,  the  granules  of 
the  cell  body  are  very  distinctly  shown.  Neuroglia  stains  a  very  light  purple,  or 
not  at  all. 

Fibrin  stains  a  bluish  purple,  but  this  reaction  is  not  distinctive. 

Hyalin  in  corpora  fibrosa  does  not  stain  at  all  or  takes  faint  blue  tint. 

Colloid  takes  a  deep  indigo  blue,  which  is  very  characteristic. 


1494  Journal  of  Applied  Microscopy 

Calcification  takes  a  dirty,  cold  blue,  which  can  be  readily  distinguished  from 
colloid. 

Cartilage  takes  a  reddish  violet,  which  is  very  characteristic. 

Corpora  amylacea  also  stain  a  reddish  violet. 

The  most  valuable  reactions  with  this  stain  are  those  with  mucin,  amyloid, 
mast-cells,  and  the  so-called  cancer  parasite.  In  staining  for  these  the  sections 
should  be  dehydrated  in  alcohol  and  cleared  in  turpentine.  The  excess  of 
turpentine  must  be  thoroughly  blotted  off  before  mounting  in  balsam,  else  the 
stain  will  run  and  the  specimens  become  blotchy. 

The  reaction  with  amyloid  as  brought  out  by  the  method  given  above  is  very 
striking  and  characteristic,  and  is  particularly  valuable  in  teaching  work.  The 
amyloid  stains  a  ruby  red,  which  is  sharply  contrasted  with  the  clear  blue  nuclei 
and  faint  blue  protoplasm  of  the  living  tissues.  No  other  method  brings  out  so 
well  the  presence  and  location  of  small  quantities  of  amyloid.  The  stain  appears 
also  to  give  a  permanent  reaction  ;  in  the  ten  months  in  which  this  stain  has  been 
used  in  this  laboratory  no  fading  of  properly  prepared  specimens  has  been 
observed,  and  trial  specimens  exposed  daily  to  sunlight  have  retained  their  color 
for  months.  The  apparent  permanency  and  clearness  of  this  reaction,  as  well  as 
the  fact  that  the  sections  so  treated  can  be  preserved  in  balsam,  gives  this  stain 
a  high  place  over  the  anilin  dyes  usually  employed  for  amyloid  reactions. 

Mucin  stains  a  bright  rose  pink,  which  is  very  characteristic  and  brings  out 
the  presence  of  mucin  when  the  amounts  are  too  small  to  be  demonstrated  by 
other  methods.  It  also  distinguishes  mucin  from  pseudomucin,  the  latter  either 
not  staining  at  all  or  taking  a  very  light  blue  tint. 

The  method  is  also  a  most  excellent  one  for  the  staining  of  mast-cells.  The 
best  results  are  obtained  by  staining  only  ten  seconds,  washing  thoroughly  and 
differentiating  in  alcohol.  The  nucleus  stains  a  light  violet,  the  granules  a 
fuchsin  red. 

The  so-called  cancer  parasite  stains  a  rose  pink,  and  may  be  seen  as  a  point 
in  the  protoplasm  of  the  cell  or  as  a  slightly  larger  mass  surrounded  by  a  clear 
zone  containing  radiating  lines.  A  more  advanced  stage  is  seen  as  a  reticular 
mass  replacing  more  or  less  of  the  cell  protoplasm,  and,  in  some  instances,  con- 
tinuous with  the  mucin  surrounding  the  cell  nests.  The  resemblance  to  the 
reaction  with  mucin  may  be  noticed  in  passing. 

The  specific  reaction  with  the  Gonococcus  has  already  been  mentioned.  This 
stain  has,  however,  the  further  advantage  in  that  the  material  may  be  stained  in 
the  hanging  drop  and  the  life  of  the  cell  not  destroyed,  so  that  the  movements  of 
the  leucocyte  containing  the  micro-organisms  may  be  studied. 

From  these  investigations  it  seems  that  for  ease  of  manipulation,  wide  appli- 
cation and  specific  staining  reactions  Kresylechtviolett  holds  a  very  high  place 
among  differential  stains  for  diagnostic  purposes,  and  that  it  is  a  most  valuable 
addition  to  the  resources  of  the  teaching  laboratory. 
Pathological  Laboratory,  University  of  Michigan.  Ralph  L.  MorsE. 


and  Laboratory  Methods.  1495 

MICRO-CHEMICAL   ANALYSIS. 
XVII. 

MAGNESIUM  GROUP— SEPARATIONS. 

When  engaged  upon  the  examination  of  a  complex  mixture  of  unknown 
composition,  the  chemical  behavior  of  all  the  elements  and  salts  liable  to  be 
present  must  ever  be  borne  in  mind.  It  is  seldom  indeed  that  the  analyst  is 
called  upon  to  make  an  analysis  of  a  substance  or  mixture  of  absolutely  unknown 
composition.  The  chief  constituents  are  almost  always  known,  or  at  least  sus- 
pected ;  and  there  are  also  always  good  reasons  why  certain  other  substances 
cannot  be  present.  He  who  would  become  a  rapid  worker  must  learn  to  reason 
by  exclusion,  but  the  beginner  must  realize  that  there  is  a  vast  difference 
between  using  one's  judgment  and  common  sense,  and  hazarding  a  mere  guess. 
Thus  the  choice  of  methods  in  micro-chemical  work  will  depend  as  largely  upon 
what  substances  are  absent  as  upon  what  are  present. 

In  all  analytical  work  rapidity  is  to  be  striven  for,  but  such  rapidity  must 
never  be  gained  at  the  expense  of  accuracy. 

In  order  to  better  understand  the  chemistry  of  the  separation  methods  of 
this  group,  it  may  be  well  to  recall  the  most  important  of  the  chemical  properties 
of  the  elements  composing  it,  upon  which  our  procedures  will  depend. 

The  hydroxides  of  all  the  members  of  the  group  are  insoluble  in  pure  water. 

Glucinum  hydroxide  is  insoluble  in  excess  of  ammonium  hydroxide,  but  is 
soluble  in  ammonium  carbonate,  potassium  hydroxide,  and  sodium  hydroxide. 

Magnesium  hydroxide  is  soluble  in  the  presence  of  ammonium  salts, 
especially  ammonium  chloride,  but  is  insoluble  in  excess  of  the  fixed  alkalies. 

Zinc  hydroxide  behaves  like  that  of  glucinum  toward  fixed  alkalies,  but 
unlike  glucinum,  it  is  also  soluble  in  ammonium  hydroxide. 

Cadmium  hydroxide  is  insoluble  in  excess  of  sodium  or  potassium  hydroxides, 
but  is  soluble  in  ammonium  hydroxide. 

Magnesium  is  the  only  one  of  the  group  normally  yielding  a  crystalline 
precipitate  with  secondary  sodium  phosphate  in  ammoniacal  solution. 

Zinc  and  cadmium  are  readily  precipitated  by  oxalic  acid,  glucinum  with 
difficulty,  and  magnesium  only  when  much  acetic  acid  is  present  or  the  solution 
is  excessively  concentrated.  In  the  case  of  glucinum,  the  double  potassium 
oxalate  is  less  soluble  than  the  normal  oxalate. 

All  four  elements  are  precipitated  by  alkaline  carbonates. 

The  chlorides  and  oxides  of  zinc  and  cadmium  can  be  easily  volatilized. 


Zinc  and  Cad^nium  from  Magnesium^  etc.,  by  sublimation. 

A.  Place  a  small  portion  of  the  material  on  a  nickel  or  platinum  spatula,* 
moisten  with  nitric  acid,  evaporate  gently,  then  ignite  to  convert  into  oxides,  but 

*  This  Journal,  III,  p.  794,  Fig.  6. 


1496  Journal  of  Applied  Microscopy 

avoid  their  volatilization.  Convert  into  chlorides  by  evaporating  repeatedly 
with  hydrochloric  acid.  When  perfectly  dry,  the  flame  is  moved  nearer  and 
nearer  the  substance  and  a  glass  slide  bearing  on  its  upper  side  a  drop  of 
water**  is  held  directly  over  the  substance  being  tested.  If  a  sublimate  results 
it  may  consist  of  ZnCl2,  CdC^,  BiClg,  PbCU,  FeCl.^,  CuCU,  and  perhaps 
HgClg.  Any  arsenic,  antimony  and  probably  all  the  mercury  which  might  have 
been  present  will  have  been  driven  off  in  the  ignition  of  the  nitrates  to  oxides. 

Dissolve  the  sublimate  in  a  drop  of  water  and  test  with  ammonium  mercuric 
sulphocyanate.  In  the  event  of  the  sublimate  being  complex,  add  water  to 
remove  the  bismuth.  Decant.  Remove  the  lead  by  sulphuric  acid.  Add 
sodium  hydroxide,  cadmium  and  iron  are  precipitated  and  zinc  passes  into  solu- 
tion. Dissolve  the  precipitate  in  dilute  acid  and  test  for  cadmium.  To  the 
sodium  hydroxide  solution  add  ammonium  carbonate  and  examine  the  prepara- 
tion for  the  double  carbonate  of  zinc  and  sodium. 

When  much  copper  is  present  it  is  always  best  to  first  remove  it  by  placing  a 
drop  of  a  solution  of  the  substance  on  metallic  iron,  thus  causing  it  to  separate. 

B.  The  oxides  of  zinc  and  cadmium  can  be  sublimed  on  charcoal  before  the 
blowpipe.  Make  a  slight  depression  in  the  carbon,  place  in  it  a  moderate  quan- 
tity of  the  material,  moisten  with  water  and  heat  gently  till  dry,  then  strongly 
with  the  oxidizing  flame,  holding  the  coal  at  an  angle.  Zinc  oxide  forms  a  coat- 
ing on  the  charcoal,  yellow  while  hot,  pure  white  when  cold.  Cadmium  oxide 
yields  a  brown  variegated  coating.  The  film  on  the  charcoal  is  removed  by 
carefully  scraping  with  the  spatula,  transferred  to  a  slide,  heated  with  dilute  acid 
and  the  clear  liquid  drawn  off  from  the  residue  of  carbon.  The  solution  is  then 
tested  for  zinc  and  cadmium. 

In  addition  to  zinc  and  cadmium,  films  on  charcoal  are  given  by  antimony, 
tin,  bismuth,  lead,  and  rarely  by  arsenic  and  mercury. 


Removal  of  Phosphates. 

If  phosphates  are  present  in  the  substance  to  be  tested,  it  is  often  necessary 
to  first  remove  them  before  any  reliable  tests  can  be  obtained  for  magnesium 
and  glucinum.     One  of  the  simplest  methods  of  procedure  is  as  follows  : 

Add  strong  nitric  acid,  then  a  few  small  strips  of  thin,  pure  tin  foil  or  a  little 
powdered  tin.  Boil  until  all  the  tin  has  been  converted  into  the  oxide  and 
phosphate  ;  draw  off  or  filter,  and  repeat  the  process  until  no  test  for  phosphoric 
acid  is  obtained  with  ammonium  molybdate.  The  solution  is  now  evaporated  to 
dryness,  and  if  arsenates  are  also  present,  the  residue  is  ignited  to  drive  off  any 
arsenous  acid  which  may  have  resulted  from  any  reducing  action  of  the  metallic 
tin.  The  residue  is  extracted  with  water  acidified  with  nitric  acid  and  tested  as 
given  below. 

The  removal  of  phosphoric  acid  by  metallic  tin  is  simple,  expeditious  and 
satisfactory.  Only  a  few  cautions  are  necessary.  The  nitric  acid  must  be 
strong  acid,  the  size  of  the  drop,  and  the  amount  of  tin  added  must  correspond 


**  This  Journal,  III,  pp.  857-858. 


and  Laboratory  Methods.  1497 

to  the  amount  of  phosphoric  acid  present ;  that  is,  when  there  is  a  large  amount 
the  drop  must  be  large,  so  as  to  permit  of  sufficient  heating.  It  is  better  under 
such  conditions  to  employ  two  operations  to  remove  the  phosphates  rather  than 
attempt  it  in  a  single  one.  A  small  test  tube  will  generally  prove  more  satisfac- 
tory than  a  slide  or  watch  glass. 

GLUCINUM. 

A.  Ghicmnm  in  Simple  Substances. 

Add  ammonium  hydroxide  and  ammonium  carbonate  (see  page  1828)  in 
excess,  glucinum  hydroxide  is  dissolved  (also  U,  Zn,  Cd,  and  perhaps  Mg). 
Decant  or  filter,  and  to  the  solution  add  ammonium  chloride  in  moderate  amount, 
evaporate,  and  ignite  gently  until  all  ammonium  salts  are  removed.  The  residue 
is  dissolved  in  dilute  sulphuric  acid,  a  little  sodium  acetate  and  a  trace  of 
mercuric  chloride  added.  The  preparation  is  then  tested  for  glucinum  with 
potassium  oxalate.  Or,  treat  the  residue  with  uranyl  acetate  and  sodium  acetate. 
In  the  latter  case  Mg,  Zn,  Cd  must  be  absent. 

B.  Glucinum  from  Alagnesium. 

Add  to  the  solution  sodium  hydroxide,  warm,  evaporate  and  take  up  with 
water.  Magnesium  hydroxide  is  precipitated,  the  glucinum  passes  into  solution 
(also  Zn  and  Al). 

Wash  the  precipitate  and  treat  it  with  ammonium  chloride,  the  magnesium 
passes  into  solution  and  is  tested  with  sodium  phosphate  and  ammonium 
hydroxide. 

The  solution  containing  the  glucinum  is  evaporated,  extracted  with  hydro- 
chloric acid  and  tested  with  potassium  oxalate. 

C.  Glucinum  in  Complex  Mixtures. 

Remove  any  copper,  etc.,  by  iron  foil. 

Add  ammonium  hydroxide  and  hydrogen  peroxide.  Warm  for  a  time,  then 
evaporate.  Repeat  the  treatment.  Extract  the  residue  with  a  solution  of 
ammonium  carbonate.  Fe,  Mn,  Co,  Al  and  part  of  the  Mg  should  remain 
insoluble,  while  Gl,  Zn,  U,  Mg  will  be  dissolved.  The  solution  is  evaporated, 
ignited,  dissolved  in  dilute  acid  and  tested  with  potassium  oxalate  ;  or  if  much 
magnesium  is  present,  separate  with  sodium  hydroxide  as  in  B. 

MAGNESIUM. 

A.  Magncsiiwi  frot7i  Glucinum,  Zinc,  Cadmiiwi,  Alu?ninum. 

Precipitate  with  sodium  hydroxide,  warm,  evaporate,  and  extract  repeatedly 
with  water.  Mg  and  Cd  remain  behind.  Dissolve  the  residue  in  hydrochloric 
acid  and  divide  into  two  portions.  Test  one  part  for  magnesium  Avith  sodium 
phosphate,  and  the  other  for  cadmium  with  sulphocyanate  or  oxalic  acid. 

B.  Magnesium  ifi  Complex  Mixtures. 

Add  ammonium  chloride  and  ammonium  hydroxide  in  slight  excess,  then 
hydrogen  peroxide,  and  warm.     Evaporate  and  treat  again.     Extract  the  residue 


1498  Journal  of  Applied  Microscopy 

with  a  dilute  solution  of  ammonium  hydroxide;  Mg, Zn,  Cd,  Ni,  Cu  are  dissolved. 
To  the  solution  add  oxalic  acid  and  acetic  acid  and  from  the  precipitated 
oxalates  of  Zn,  Cd,  Ni,  and  Cu,  the  clear  solution  is  separated  by  decantation, 
filtration,  or  the  centrifuge.  This  solution  is  evaporated  with  sulphuric  acid  and 
heated  to  destroy  the  oxalic  acid.  The  residue  is  dissolved  in  acidulated  water, 
treated  with  sodium  hydroxide,  the  precipitate  carefully  washed,  dissolved  in 
hydrochloric  acid  and  tested  for  magnesium  with  sodium  phosphate. 

C.     Magnesium  from  Glucinitm. 

To  the  solution  to  be  tested  add  ammonium  chloride,  then  carefully,  a  little 
at  a  time,  ammonium  hydroxide  as  long  as  a  precipitate  results,  draw  off  at  once 
and  test  the  decanted  solution  with  sodium  phosphate  for  magnesium. 

ZINC. 

A.  Zinc  from  Gluciniim,  Magnesimn,  and  Cadmium. 

Ignite,  warm  gently  with  sodium  hydroxide  solution.  Zinc  and  glucinum  are 
dissolved,  magnesium  and  cadmium  remain  in  the  residue.  Separate  the  solu- 
tion, add  to  it  acetic  acid  and  precipitate  the  glucinum  with  potassium  oxalate 
(a  little  zinc  is  always  precipitated  with  the  glucinum).  Separate  the  clear  solu- 
tion, evaporate  and  destroy  the  oxalic  acid  by  means  of  sulphuric  acid  and  heat. 
Take  up  the  residue  with  water,  acidified  if  necessary,  add  ammonium  acetate, 
and  test  for  zinc  with  sulphocyanate ;  the  addition  of  a  trace  of  copper  will 
increase  the  delicacy  of  the  reaction.  Or,  instead  of  testing  with  sulphocyanate, 
to  the  sodium  hydroxide  solution  after  the  removal  of  the  magnesium  and  cad- 
mium, add  ammonium  carbonate,  zinc  will  separate  as  the  double  carbonate  of 
zinc  and  sodium. 

B.  Zinc fr inn  Cadmitcm. 

Precipitate  with  primary  sodium  carbonate  in  ammoniacal  solution,  cadmium 
separates  at  once,  draw  off  the  supernatant  solution  and  allow  to  stand  for  a 
short  time,  zinc  separates  as  the  double  carbonate.  Or,  acidify  the  solution  to 
be  tested,  and  to  it  add  powdered  metallic  magnesium.  Zinc  and  cadmium  are 
precipitated.  Wash  carefully  the  finely  divided  metallic  mass;  add  acetic  acid, 
heat  the  preparation  ;  zinc  passes  into  solution  with  but  very  little  cadmium, 
decant  and  test  for  zinc.  The  residual  metallic  cadmium  is  dissolved  in  hydro- 
chloric acid  and  tested. 

C.  Zinc  in  Complex  Afixtures. 

Treat  with  ammonium  hydroxide  and  hydrogen  peroxide.  Then  with 
ammonium  carbonate.  To  the  decanted  solution  add  oxalic  acid  and  acetic 
acid.  Separate  the  precipitated  oxalates,  ignite  and  extract  the  zinc  with  sodium 
hydroxide.  The  clear  solution  containing  the  zinc  is  heated  with  ammonium 
chloride  and  tested  for  zinc  with  primary  sodium  carbonate  or  acidified  and 
tested  with  sulphocyanate. 


and  Laboratory  Methods.  1499 

CADMIUM. 

A.      Cadf?ii urn  from  Magnesiian. 

The  bulk  of  the  magnesium  can  be  precipitated  as  magnesium  ammonium 
phosphate  by  adding  ammonium  hydroxide  and  sodium  phosphate,  the  latter 
very  carefully  and  only  a  very  little  at  a  time  so  as  to  avoid  precipitating  any 
cadmium.  The  clear  supernatant  solution  is  then  drawn  off  and  tested  for 
cadmium. 

B.  Cadmimn  from  Gluciniitn,  Zinc^  or  Aluminum. 

Precipitate  with  excess  of  sodium  or  potassium  hydroxide.  Glucinum,  zinc, 
and  aluminum  are  dissolved.  Cadmium  remains  insoluble.  Dissolve  the  pre- 
cipitate in  hydrochloric  acid  and  test  with  oxalic  acid  or  with  sulphocyanate. 

C.  Cadmium  in  Complex  Mixtures. 

Proceed  as  in  the  above  described  methods  in  which  ammonium  hydroxide 
and  ammonium  carbonate,  etc. ;  e.  g..  Zinc  C.  After  precipitation  and  ignition 
of  the  oxalates,  the  residue  is  extracted  with  sodium  hydroxide,  cadmium  remains 
insoluble.  This  residue  insoluble  in  sodium  hydroxide  is  washed,  dissolved  in 
hydrochloric  acid,  and  tested  for  cadmium. 

D.  Cadmium  frofn  Zinc. 

See  Zinc  B. 

E.  M.  Chamot. 

Cornell  University. 


A  Damp  Chamber  for  Use  on  the  Khnostat. 

Students  of  Plant  Physiology  have  often  found  need  for  a  damp  chamber  for 
use  on  the  klinostat  which  will  furnish  a  firm  support  for  seedlings  in  rapid 
rotation  and  will  retain  a  supply  of  moisture  for  several  hours.  The  original 
method  used  by  Knight  has  been  variously  modified  so  as  to  provide  moisture 
for  the  plants.  In  cases  where  the  desired  plane  of  rotation  is  horizontal  the 
experiment  may  be  successfully  performed  by  covering  with  a  bell-jar  a  rotating 
disk  whose  axis  projects  below  the  table  and  to  which  the  motive  power  is  applied. 
In  work  where  the  desired  plane  of  rotation  is  vertical,  some  experimenters  have 
arranged  a  reservoir  of  water  so  that  the  supply  of  moisture  falls  upon  the  rotat- 
ing seedlings  in  drops  ;  but  here  is  a  chance  that  the  thigmotropic  or  traumotropic 
stimulus  given  by  the  falling  drops  may  affect  the  results  of  the  experiment. 

The  apparatus  described  below  has  been  successfully  used  for  different 
kinds  of  work  in  the  Botanical  Laboratory  of  the  University  of  Michigan. 

A  narrow  board,  containing  about  eight  pegs  5  or  6  cm.  long,  is  fitted  into  the 
bottom  of  a  thin  glass  basin  of  about  22  cm.  diameter  and  G  cm.  depth.  The 
pegs  serve  for  the  attachment  of  the  small  bars  holding  the  seedlings.  The  glass 
dish  is  lined  with  moist  filter  paper  and  closed  with  a  circular  glass  plate  24  cm. 
in  diameter ;  the  inner  side  of  the  cover  is  completely  lined  with  a  piece  of 
moist  blotting  paper,  which  forms  an  almost  perfectly  tight  fitting  cover. 


1500 


Journal  of  Applied  Microscopy 


In  case  the  plate  of  the  klinostat  is  too  small  for  the  attachment  of  the  basin, 
it  must  be  fastened  to  a  circular  wooden  plate,  which  is  screwed  to  the  plate  of 
the  klinostat  and  also  serves  for  securing  the  cover  clamp.     The  cut  shows  the 

manner  of  securing  the  cover  and  the  dish. 
The  clamp  A  is  made  of  strong,  elastic  wood 
with  disks  of  rubber  under  the  extremities 
of  the  arms ;  one  end  of  it  slips  into  a  loop 
of  wire,  a  strip  of  brass  at  the  other  end 
hooks  to  the  wooden  plate  P.  The  dish  is 
held  from  slipping  sideways  by  four  pegs  in 
the  wooden  plate,  which  may  be  covered 
with  short  pieces  of  rubber  tubing.  The 
seedlings  for  use  should  be  attached  to  short 
wooden  bars,  which  are  fastened  to  the  pegs 
inside  of  the  dark  chamber  by  means  of 
two  rubber  bands.  The  strength  of  the  centrifugal  force,  in  terms  of  the  attrac- 
tion of  gravity,  may  be  calculated  for  the  pegs  from  the  following  formula  : 
4  TT  R  (in  meters)  4  tt^ 


gt2 

4.024 


X 


R 


^  4.024,  a  constant. 


=  no.   of  g   (gravity). 


R  =  radius  expressed  in  meters. 

t    =:  time  in  seconds  of  one  revolution  of  the  chamber. 

If  the  centrifugal  force  is  small,  i.  e.,  less  than  three  times  the  force  of 
gravity,  the  seedlings  may  be  attached  to  the  wooden  bars  in  the  ordinary  method 
by  the  use  of  rubber  bands  and  strips  of  blotting  paper  ;  but  if  the  centrifugal 
force  is  considerable,  I  have  found  it  better  to  pack  the  seedlings  in  short  pieces 
of  glass  tubing,  allowing  about  5  millimeters  of  the  root-tip  to  protrude,  then  to 
fasten  the  pieces  of  glass  tubing  to  the  wooden  bars.  The  pieces  of  tubing 
should  be  arranged  so  that  they  lie  in  a  tangential  plane. 

For  short  periods  of  time  the  supply  of  moisture  will  not  need  to  be  replenished 
if  the  chamber  is  well  saturated  before  beginning  ;  in  long  continued  experiments 
it  is  best  to  introduce  more  water  by  means  of  an  atomizer  every  six  or  eight 
hours. 

The  advantages  of  such  a  chamber  for  use  on  the  klinostat  are  that  in  the 
moist  air  the  roots  are  freer  to  respond  than  in  moist  sawdust,  where  the  higher 
rates  of  rotation  invariably  cause  the  sawdust  to  become  packed  at  the  circum- 
ference of  the  cylinder  ;  the  seedlings  can  be  easily  observed  during  the  progress 
of  the  experiment ;  it  retains  its  supply  of  moisture  well,^ — I  have  seen  corn  seed- 
lings grow  for  four  days  with  only  the  initial  supply  of  moisture  ;  it  can  be  rotated 
in  any  desired  plane  and  at  any  speed.  Howard  S.  Reed. 

University  of  Michigan. 


and  Laboratory  Methods.  1501 

Journal  of  ^^^  results  of  the  Denver  meeting 

of   the  American   Association  for  the 

Applied    Microscopy  Advancement  of  Science  undoubtedly 

gj^j  came  as  an  agreeable  surprise  to  any 

Laboratory   Methods.  ^,^^  may  have  had  fears  for  the  success 

^  of  a  meetmg  so  far  from  the  center  of 

Edited  by  L.  B.  ELLIOTT.  American  scientific  activity.     The  re- 

ports of  the  meeting  are  most  gratifying. 


Issued  Monthly  from  the  Publication  Department  The    rCSultS    accomplished    dunng    the 

of  the  Bausch  &  Lomb  Optical  Co.,  ^  i    ^u         i.  *.    i  £         iU 

Rochester,  N.  Y.  past  year,  and  the  steps  taken  for  the 

betterment  of  the  organization  in  com- 


SUBSCRIPTIONS.  ^^„  years,  are  of    such  mterest  as  to 

One  Dollar  per  Year.    To  Foreign  Countries,   $1.25        ,  "^  .■■  -.tu  ,  j       ,         ,    ■> 

per  Year,  in  Advance.  bear  repetition.     We  would  also  take 

"        "  "    "  this  opportunity  to  congratulate  those 

The  majority  of  our  subscribers  dislike  to   have  their  ,  j    .1  ■T_'i-i         £    i.i 

files  broken  in  case  they  fail  to  remit  at  the  expiration  of        wllO  aSSUmCd  the    responsibility  Ot    the 

their  paid  subscription.   We  therefore  assume  that  no      arrangement  of  the  rccent   meeting  at 

mterruption   m   the   series   is   desired,   unless  notice  to  *>  ,  .  o    _ 

discontinue  is  sent.  Dcnvcr  for  the  success  with  which  their 

SEPARATES  efforts  have  been  rewarded.    One  more 

One  hundred  separates  of  each  original  paper  accepted        Step     in    advaUCC    haS    been    taken,  and 

are  furnished  the  author,  gratis.  ^  closcr  and   more  efficient  organiza- 

Separates   are   bound   in   special   cover  with  title.     A  .  .         ,  ,  "^         . 

greater  number  can  be  had  at  cost  of  printing  the  extra        tlOn    haS   bCCn   attained.         1  hC    meeting 

copies  desired.  ^^^  ^  success   from  cvery   standpoint. 

The  attendance  approached  that  of  re- 
cent eastern  meetings,  the  registration  reaching  oil.  That  the  Association  is  rally- 
ing after  a  period  of  gradual  decrease  in  membership  is  assured  by  the  fact  that 
since  the  1900  meeting,  1500  new  members  have  been  added.  More  than  200 
papers  were  presented.  The  report  of  the  financial  condition  of  the  Association 
was  very  encouraging,  the  permanent  endowment  fund  having  been  increased 
during  the  year  by  over  one  thousand  dollars,  bringing  the  present  amount  of 
that  fund  to  something  over  eleven  thousand  dollars. 

Several  changes  of  policy  were  either  adopted  or  recommended  for  future 
consideration.  Of  these  the  most  important  is  that  of  a  change  in  the  time  of 
meeting,  to  conform  with  the  recent  action  taken  by  the  universities  to  establish 
a  Convocation  Week  during  the  winter.  It  was  recommended  that  the  Associa- 
tion, with  its  affiliated  societies,  meet  at  Washington  during  the  week  in  which 
New  Year's  day  of  1903  falls,  without,  however,  abandoning  the  summer  meet- 
ings. Thus  the  plan  for  winter  meetings  will  be  given  fair  trial,  and  its  perma- 
nent establishment  will  depend  largely  upon  the  success  of  the  first  winter 
meeting. 

An  amendment  for  the  mutual  advantage  of  the  Association  and  its  affiliated 
societies  provides  that  each  affiliated  society  be  entitled  to  elect  one  member 
(two  if  the  society  has  more  than  twenty-five  members)  as  its  representative  in 
the  council  of  the  Association ;  thus  forming  a  closer  organization  between  all 
the  societies  concerned. 

x\nother  step,  intended  to  add  strength  to  the  council,  is  the  provision  for 
the  election  each  year  of  three  councillors-at-large,  who  shall  serve  for  a  term  of 
three  years,  thus  giving  greater  permanency  and  efficiency  to  that  body.  It  is 
proposed  to  lengthen  the  term  of  ofiice  of  the  secretaries  of  sections  to  five  years, 
in  order  to  give  the  sections  the  benefit  of  experienced  service.  The  present 
term  gives  the  secretary  little  more  than  time  enough  to  become  familiar  with 
his  duties. 

Although  we  have  noted  only  a  very  small  part  of  the  proceedings  of  the 
Denver  meeting,  it  is  sufficient  to  show  that  the  American  Association  for  the 
Advancement  of  Science  is  growing  in  size  and  efficiency,  and  is  an  organization 
to  which  all  who  are  interested  in  scientific  work  can  well  afford  to  belong. 


1502  Journal  of  Applied  Microscopy 

CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chambeki_\ix. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  shotild  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Hill.  A.  W.     The  distribution  and  character  of       -j^j^i^  |^^.    y^^     ^111.    constitutes 

connecting  threads  in  the  tissues  of   /t»us  r    r  . 

..-.  .t vv.''v-f  "and  other  allied  species.  Phil.  Part  I  of  an  extensive  study  by  Gardi- 
Trans.  of  the  Roy.  Soc.  of  London.  Ser.  B..  ^er  and  Hill  of  "  The  Histolog\-  of  the 
194:  6^-125:  pis.  31-35.  iQOi.  °' 

Cell  Wall,  with  special  reference  to  the 

mode  of  connection  of  cells."  The  purpose  was  to  discover  the  extent  and 
distribution  of  the  connecting  threads  in  any  particular  plant.  The  embryo  and 
seedlings  of  Pinus  pirwj  and  the  mature  tissues  of  Pinus  syh-cstris  were  chosen 
as  particularly  favorable  material.  The  endosperm  was  also  studied  in  P.  pirua. 
The  endosperm  consists  chietiy  of  rather  large,  rounded  cells,  but  a  close 
examination  shows  that  in  many  cases  an  internal  division  has  occurred.  The 
threads  are  evenly  distributed  in  the  young  walls,  but  are  grouped  in  the  older 
walls.  Near  the  cot)ledons  the  cells  are  smaller,  the  threads  thicker,  and  there 
are  traces  of  ferment  action.  Ferments  from  the  cot}ledons  pass  into  the  endo- 
sperm through  the  threads,  and  by  the  same  route,  food  materials  pass  from  the 
endosperm  to  the  embr)-o. 

In  the  seedling  the  absorptive  side  of  the  cotyledon  is  more  abundantly  sup- 
plied with  threads  than  the  side  not  exposed  to  the  endosperm.  There  are  no 
threads  in  the  external  walls  of  the  epidermis,  and  but  ver)^  few  connecting  the 
guard  cells  with  their  neighbors.  All  parenchyma  cells  show  a  general  resem- 
blance in  the  character  of  their  threads,  the  threads  on  the  end  walls  being 
irregularly  scattered,  while  on  the  side  walls  they  are  grouped.  In  the  phloem, 
all  the  sieve  tube  threads  show  a  characteristic  median  dot.  The  albuminious 
cells  at  the  edge  of  the  phloem  of  the  leaf  have  their  threads  grouped  in  localized 
thickenings  on  the  walls,  and  serve  to  pass  materials  from  the  mesophyll  to  the 
phloem.  The  ver}-  numerous  threads  of  the  root  cap  form  a  connection  with  the 
free  surface  of  the  root  and  with  the  periblem. 

In  the  mature  tissue  of  P.  sylvestris  the  threads  in  the  cortical  tissue  are 
similar  to  those  of  the  seedling.  In  the  phloem  there  is  no  connection  between 
the  sieve  tubes  and  the  bast  parenchyma,  or  the  starch  medullary  ray  cells.  The 
sieve  tube  threads  on  the  radial  walls  have  a  median  dot.  The  torus  of  the 
bordered  pit  is  probably  traversed  by  threads  which  soon  disappear.  In  the  leaf. 
the  distribution  is  about  the  same  as  in  the  cotyledon.  The  endodermis.  with 
ver)-  numerous  threads,  is  in  close  connection  with  the  cortex  and  the  stele.  In 
the  pericycle,  living  cells  are  connected  by  threads,  but  there  is  no  connection 
between  the  pericycle  and  the  lignified  transfusion  tissue. 

In  general,  the  main  direction  of  threads  in  the  cortex  and  phloem  is  tangen- 
tial. The  transiton,"  nature  of  certain  threads  explains  the  absence  of  threads 
between  the  sieve  tubes  and  medullar)-  ray  cells.     Except  in  the  medullar)-  rays. 


and  Laboratory  Methods.  1503 

and  in  the  cork  cambium,  the  threads  are  chiefly  on  the  radial  walls.  This  sug- 
gests that  in  conifers  food  supplies  and  stimuli  are  conducted  mostly  in  a  tangen- 
tial and  vertical  direction.  c.  j.  c. 

Arnold!,  W.     Beitrage  zur  Morphologic  einiger       In    Gymnosperms,  aS    a    rule,  only   one 
Gymnospermen.     I.  Die  Entwickelung  des  ,  ^^    .  •  i        i  i 

Kndosperms    bei     Sequoia     sempervirens.       embryo    sac    attains    any   considerable 
Bull,  des  Natur.de  Moscou,  Pp.  1-13,  pis.      development.     Very  rarely  two  embryo 
'       ^'  sacs    develop    in    Taxiis,  and    in    one 

instance,  two  embryo  sacs  have  been  seen  in  Pinus  sylvestris.  In  Sequoia,  how- 
ever, several  embryo  sacs  begin  to  develop,  and  in  Gnetiim  it  is  the  rule  for 
several  sacs  to  develop  almost  to  maturity  before  one  of  them  secures  any  decided 
advantage.  Prof.  Arnoldi  has  taken  up  the  somewhat  incomplete  work  of  Shaw, 
and  has  made  a  careful  study  of  the  development  of  the  endosperm  of  Sequoia 
sempervirens.  Free  nuclear  division  takes  place  in  the  usual  manner  in  an  evenly 
distributed  peripheral  layer  of  protoplasm,  but  very  soon  there  is  a  denser 
accumulation  of  protoplasm  at  the  lower  end  of  the  sac.  When  the  formation  of 
walls  begins,  three  regions  of  the  endosperm  may  be  distinguished :  the  upper, 
the  lower,  and  the  middle.  The  upper,  and  particularly  the  lower,  develop 
faster  than  the  middle,  so  that  the  ends  of  the  sac  become  filled  with  a  solid 
tissue,  while  the  nuclei  are  still  almost  free  in  the  middle  portion.  Each  nucleus 
of  the  middle  portion  now  becomes  surrounded  by  a  wall  which  is  open  on  the 
inner  side ;  the  walls  grow  inward,  and  when  the  center  is  reached  walls  are 
formed  at  the  inner  ends  of  the  cells.  The  nucleus  now  begins  to  divide,  and 
each  of  these  cells  ("  alveoli  ")  becomes  divided  into  several  cells.  Archegonia 
are  formed  only  from  these  alveolar  cells  of  the  middle  region.  At  the  time  of 
fertilization,  the  upper  and  lower  portions  of  the  endosperm  consist  of  small- 
celled  tissue,  while  the  middle  portion  is  alveolar.  Sequoia  is  regarded  as  a 
connecting  link  between  Gnetum  and  the  Angiosperms  on  the  one  hand,  and 
between  Gymnosperms  and  the  Archegoniates  on  the  other.  c.  j.  c. 

Arnold!,  W.    Beitrage  zur  Morphologie    und  The  number  of  archegonia  in  Sequoia 

Entwickelungsgeschichte     einiger    Gymno-  .  ,  r    1  •       •      t 

spermen.     II.  Ueber   die   Corpuscula   und  IS  very  large,  SOme  of  the  writer  s  draw- 

Pollenschlauche  bei   Sequoia  sempervirens.  ings  showing  as  many  aS  sixty.      They 
Bull,  des  Natur.  de   Moscou,    Pp.  1-8,  pis.  ^.  •      1      i  r 

lo-ii   i8qq.  sometimes  occur  singly,  but  are  often 

grouped.  In  development  they  resem- 
ble the  archegonia  of  the  Cupressineae,  since  they  are  often  in  direct  contact 
with  each  other,  and  do  not  form  any  ventral  canal  cell.  There  are  no  proteid 
vacuoles.  The  neck  consists  of  two  cells,  in  this  respect  resembling  the  older 
Gymnosperms.  The  pollen  tube  grows  through  the  nucellus,  not  between  the 
nucellus  and  integument,  as  described  by  Shaw.  At  the  time  of  fertilization  the 
pollen  tube  contains  the  two  male  cells  of  equal  size,  and  two  small  nuclei,  one 
of  which  is  the  tube  nucleus  and  the  other  "  the  nucleus  of  the  cell  which  united 
the  generative  cell  with  the  microspore  wall."  The  general  structure  of  the 
pollen  tube  and  its  contents  agrees  with  the  Cupressineae.  The  morphological 
considerations,  together  with  the  geographical  distribution,  lead  to  the  conclusion 
that  Sequoia  is  nearly  related  to  the  ancient  type  from  which  the  modem  Arau- 
carias  and  Cupressineae  have  descended.  c.  j.  c. 


1504  Journal  of  Applied  Microscopy 


CYTOLOGY,   EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole,  Cornell  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Cade,  A.    Les  Elements  Secreteursdesglandes       ^j^jg   ^^^-^^^    contains    a    description  of 
gastriques    du   fond    chez   les  mammiferes.  _  ^ 

Arch.  d'Anatomie   Microscopique,  4 :  1-86,       the  histological  Structure  of  the  glands 

'9°'-  of  the   stomach  in  different   stages  of 

their  normal  activity,  the  changes  in  the  glands  during  hibernation,  the  effects 
produced  by  pilocarpin,  section  of  the  vagus  nerve,  isolation  of  a  part  of  the 
stomach,  and  the  formation  of  an  artificial  pylorus  in  the  region  of  the  fundus. 
The  cells  of  the  fundus  are  grouped  under  three  heads — cells  of  the  neck  of  the 
gland,  the  central  or  chief  cells,  and  the  parietal  cells.  The  material  studied 
was  obtained  from  the  dog,  cat,  rat,  mouse,  hedge-hog,  marmot,  and,  in  a  few 
cases,  from  man.  Methods  of  fixation  and  staining  are  given,  and  the  structure 
of  the  cells  described  in  detail.  The  central  or  chief  cells  frequently  contain 
two  or  more  nuclei,  and  divide  amitotically ;  careful  research  failed  to  discover  a 
single  case  of  karyokinesis.  Amitosis  is  apparently  the  chief  mode  of  division 
in  the  border  cells  also.  The  neck,  or  muciparous,  cells  exhibit  transitions  into 
the  central  cells,  and  the  author  inclines  to  the  view  that  the  one  type  may  be 
transformed  into  the  other,  although  it  is  admitted  that  this  is  not  proven.  A 
close  connection  exists  between  the  neck  cells  and  the  epithelial  cells  of  the  sur- 
face of  the  mucosa,  and  there  is  a  relationship  between  the  neck  cells  of  the 
fundus  glands  and  the  cells  of  the  body  of  the  pyloric  glands.  ("  II  y  a  entre 
les  cellules  des  grandes  pyloricques  et  les  cellules  principales  du  col  une  certaine 
parenti.")  There  is  no  evidence  of  a  transition  either  way  between  the  chief 
cells  and  the  border  cells  of  the  fundus ;  they  apparently  always  remain 
specifically  distinct  elements.  Contrary  to  the  opinion  held  by  Oppel  and  others 
that  the  cells  of  the  body  of  the  gland  in  the  lower  vertebrates  represent  only 
the  parietal  cells  of  the  mammals,  the  author  holds  that  they  correspond  to 
both  parietal  and  chief  cells  of  the  higher  forms.  The  theory  that  the  parietal 
cells  are  especially  concerned  in  the  production  of  hydrochloric  acid  is  considered 
devoid  of  adequate  foundation. 

Section  of  the  vagus  nerve  gives  rise  to  marked  structural  changes  in  the 
secreting  cells.  The  border  cells  become  less  granular.  The  cytoplasm  of  the 
chief  cells  stains  less  deeply,  and  the  cells  lose  the  differentiated  region  at  the 
base ;  the  nucleus  becomes  shrunken  and  wrinkled  in  outline.  Isolation  of  a 
part  of  the  stomach  gives  rise  to  changes  similar  to  those  produced  by  section 
of  the  vagus.  The  author  concludes  that  the  vagus  plays  an  important  role  in 
the  secretion  of  the  gastric  juice. 

The  experiment   of   forming   an    artificial    pylorus    in    the    fundus    of    the 


and  Laboratory  Methods.  1505 

stomach  gave  very  interesting  results.  The  stomach  of  a  cat  was  cut  com- 
pletely across  in  the  middle,  and  a  junction  was  made  between  the  fundus 
and  the  jejunum,  so  that  the  pyloric  region  was  entirely  excluded  from  the 
route  along  which  the  food  passed.  A  similar  operation  was  performed 
on  a  dog,  and  both  animals  were  killed  about  seven  months  afterward. 
A  careful  study  was  made  of  the  histological  changes  that  occurred  in 
the  region  of  the  fundus  near  the  line  of  juncture.  Near  the  junction  with  the 
intestine  the  fundus  glands  became  more  sinuous,  the  lumen  increased  in  size, 
and  the  interglandular  tissue  became  infiltrated  with  leucocytes.  The  character 
of  the  gland  cells  was  markedly  altered.  The  parietal  cells  disappeared  entirely  ; 
the  chief  cells  lost  their  characteristic  granulation,  their  basal  differentiation 
disappeared,  and  they  assumed  the  principal  features  of  the  cells  of  the  neck. 
Both  in  general  structure  and  in  the  character  of  the  cells  the  fundus  glands 
near  the  new  pylorus  became  strikingly  similar  to  the  ordinary  pyloric  glands. 
This  working  over  of  the  fundus  glands  into  glands  resembling  those  of  the 
pyloric  region  in  response  to  the  new  environing  conditions  imposed  is  a  feature 
of  considerable  theoretical  interest.  There  is  a  closing  section  on  deductions 
concerning  the  process  of  secretion  in  general.  s.  j.  h. 

Conklin,  E.  G.     Centrosome  and  Sphere  in  the       This    paper   is   a    summary  of    a    more 
Maturation,  Fertilization,  and   Cleavage  of  ^       i     i         i  i-      ^'  i  •   ^         -n 

Crepidula.     Anat.  Anz.  19:   280-287,1901.      extended  publication  which  will  soon 

appear  in  the  Journal  of  Morphology. 
In  fertilization  there  is  no  "  quadrille  "  of  the  centers.  The  o.^'g  sphere  and 
sperm  sphere,  however,  fuse  into  a  granular  mass.  Within  this  mass  the  centro- 
somes  of  the  first  cleavage  spindle  arise  apparently  independently  of  each  other. 
The  author  holds  that  "  there  is  good  evidence  that  the  cleavage  centrosomes 
are  not  derived  exclusively  either  from  a  sperm  centrosome  or  from  an  egg 
centrosome,  but  that  one  of  these  comes  from  the  egg  sphere,  the  other  from  the 
sperm  sphere."  s.  j.  h. 

Hoffmann,  R.  W.      Ueber  das  Orientiren  und      ^j^jig  ^j^g  consistency  of  yolk  depends 
Schneiden  mikroskopisch  kleiner,  undurch-  ■'  ■'  ^ 

sichtiger   und   dotterreicher  Objecte.     Zeit.       tO  a  certain  extent  upon  the  fixing  fluid 
wiss.  Mik.  4 :  443-448,  1901.  employed,  the  length  of  time  the  object 

remains  in  alcohol  and  the  duration  of  the  process  of  embedding  in  paraffin  were 
not  found  to  have  any  influence  on  the  ease  with  which  yolk  can  be  cut.  No 
reliable  method  of  treating  yolk  so  as  to  be  cut  easily  in  paraffin  was  hit  upon, 
so  resource  was  had  to  embedding  in  celloidin.  In  orienting  small  opaque  objects 
in  celloidin,  90  per  cent,  alcohol  is  a  serviceable  medium  in  which  to  operate,  as 
it  enables  one  to  see  easily  the  configuration  of  the  object,  and  only  hardens  the 
celloidin  very  slowly  ;  100  per  cent,  alcohol,  which  the  author  at  first  employed, 
dissolves  the  celloidin  too  much,  and  85  per  cent,  alcohol  hardens  it  too  rapidly. 
After  the  object  is  oriented  under  a  small  amount  of  the  alcohol  it  is  placed  in 
xylol  to  harden.  The  turbidity  that  appears  after  treatment  with  xylol  soon  dis- 
appears, and  the  mass  becomes  clear.  One  convenient  method  of  orienting  is  as 
follows :  A  number  of  objects  are  impregnated  with  thick  celloidin  solution  in  a 
shallow  glass  dish,  enough  celloidin  being  used  to  form,  after  it  is  hardened,  a 
mass  but  little  thicker  than  the  objects  embedded.  After  the  mass  is  hardened 
under  80  per  cent,  alcohol  it  is  cut  up  into  small,  square  pieces,  one  for  each 
object.  After  being  placed  for  some  time  in  90  per  cent,  alcohol,  the  objects 
are  stuck  to  a  block,  oriented  under  a  small  amount  of  90  per  cent,  alcohol,  and 
then  placed  in  xylol  until  hardened  and  cleared.  s.  j.  H. 


^^^6  Journal  of  Applied  Microscopy 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Siedlecki.    M.      Contribution    a    I'etude    des  Monocystis   ascidia   develops    within  a 

changements  cellulaires   provoques  par   les  cell  of  the  intestinal  epithelium  of  Cio7ia 

Gregarines.     Arch,   d' Anat.  Micros.  4 :  87-  ■,,•,■    r            1                        •■      • 

100.     Avec  9  figs.,  dans  le  texte.     1901.  intestinalis  from  the   sporozoite  issumg 

from  the  sporocyst  of  the  Gregarine  in 
the  intestine.  The  presence  of  the  parasite  in  the  cell  provokes  at  first  an 
hypertrophy  of  the  nucleus  and  the  cytoplasm.  The  chromatin  is  disorganized, 
and  the  nucleolus  increases  in  size.  These  changes  are  due  to  the  chemical 
action  of  the  parasite.  In  material  fixed  in  Flemming,  or  sublimate,  the  paras- 
itized cells  are  but  feebly  stained  by  safranin,  thionin,  the  various  haematoxylins, 
or  the  Biondi  mixture.  The  infected  and  greatly  hypertrophied  cell  is  pushed 
beneath  the  epithelial  layer  into  a  sac  of  connective  tissue  formed  by  the  base- 
ment membrane.  From  this  sac  the  Monocystis  escapes  by  passing  through  the 
epithelial  layer  between  the  cells  into  the  lumen  of  the  digestive  tract.  Here  it 
attaches  itself  by  means  of  its  small  amceboid  projection  upon  the  epithelial  cells, 
in  which,  however,  it  produces  no  pathological  condition.  Pterocephalus,  parasitic 
in  Scolopendra,  lies,  in  its  adult  stage,  between  the  cells  of  the  intestinal  epithelium 
attached  to  them  by  protoplasmic  prolongations  of  the  protomerite.  The 
changes  in  the  epithelium  due  to  its  presence  are  purely  mechanical.      c.  a.  k. 

Miall,  L.  C,  and  Hammond,  A.  R.  Structure  and  The  revival  of  interest  in  the  Diptera 
Life-history  of  the  Harlequin  Fly.  Pp.  vi,  which  has  resulted  from  the  discovery 
tq6.     With  127  figs.,  and  i  pi.     The  Clar-  ,  ^,  ,  .^  ,, 

endon  Press,  O.xford,  1900.  o*  the  agency  of  mosquitoes  and  house- 

flies  in  the  spreading  of  disease  makes 
the  publication  of  this  book  very  opportune.  Though  Chironomiis  is  not  itself 
an  obnoxious  form,  its  structure  and  life-history  are  in  many  points  so  similar  to 
those  of  the  Cidicidce^  that  this  work  may  well  serve  as  a  guide  to  students  of 
that  group.  The  anatomy  and  histology  of  both  the  larvae  and  the  adult  are 
treated  in  detail  with  abundant  original  illustrations,  and  comparisons  with  other 
Diptera  are  frequently  made.  The  habits,  parasites,  enemies,  and  the  life- 
history  are  fully  described. 

Chironomus  is  widely  distributed,  and  its  larvae  abound  in  every  body  of 
fresh  water,  and  also  in  some  places  in  salt  water  in  the  littoral  region.  Slow, 
muddy  streams  rich  in  decaying  organic  matter  constitute  the  best  collecting 
grounds,  and  a  long-handled  ladle  forms  a  good  collecting  instrument.  Larvae 
are  easily  reared  in  the  laboratory  in  shallow  aquaria  with  decaying  vegetation. 
High  temperatures  favor  their  rapid  transformation. 

The  study  of  the  living  larva  is  best  made  upon  half-grown  specimens 
entangled  in  a  nest  of  cotton-wool.  The  external  structure  is  best  seen  in  speci- 
mens killed  in  Flemming's  fluid,  and  the  detail  of  the  exoskeleton  in  specimens 
treated  for  three  days   in   a  ten   per  cent,  solution  of  caustic  potash,  and  then 


and  Laboratory  Methods.  1^^" 

mounted  in  glycerin  or  balsam.  For  histological  study  the  larvae  may  be  killed 
in  Flemming's  fluid ;  after  one  hour's  exposure  they  should  be  halved  and  again 
placed  in  the  fluid  for  another  hour,  and  then  thoroughly  washed  in  running 
water  in  a  washing  bottle  for  twenty-four  hours.  Perenyi's  fluid  for  six  hours 
also  fixes  the  tissues  very  well ;  after  three  hours  in  the  fluid  the  larvae  should 
be  halved.  Larvae  for  entire  mounts  should  be  prepared  as  follows :  Select 
transparent  specimens  and  keep  them  in  clear  water  for  a  day  or  two,  until  the 
alimentary  canal  is  emptied.  Then  place  the  larva  in  a  mixture  of  absolute 
alcohol  (9  parts)  and  ether  (1  part),  holding  it  in  the  desired  position  with  small 
sable  brushes  until  it  is  set  (three  to  ten  minutes).  After  several  hours'  exposure 
to  this  fluid  transfer  to  absolute  alcohol  so  as  to  remove  all  the  ether.  Pass 
through  oil  of  cloves  to  new  benzine  balsam.  Many  points  in  the  anatomy  may 
be  elucidated  by  fresh  specimens  teased  out  in  two  per  cent,  caustic  potash,  or 
by  those  killed  in  the  fluids  above  mentioned,  and  stained  in  carmin  or  haema- 
toxylin  before  teasing  out.  In  staining  in  toto  with  carmin  or  Delafield's  haema- 
toxylin  it  is  necessary  to  permit  the  stains  to  act  for  at  least  a  week.  Material 
prepared  by  the  methods  above  described  may  be  successfully  sectioned  in  either 
paraffin  or  celloidin.  Serial  sections  in  the  paraffin  are  facilitated  by  coating 
the  block  with  soft  paraffin. 

The  peculiar  nuclear  structures  of  the  salivary  glands  may  be  studied  in 
material  fixed  in  a  fluid  composed  of  equal  parts  of  one  per  cent,  osmic  and 
acetic  acids  allowed  to  act  for  several  minutes.  The  glands  are  then  stained  in 
acetized  methyl  green  followed  by  carmin,  and  are  mounted  in  glycerin.  The 
glands  are  easily  secured  from  decapitated  larvae,  passing  out  with  the  blood,  or 
being  readily  released  by  gentle  pressure. 

The  eggs  of  Chironotniis  are  very  favorable  objects  for  the  study  of  insect 
development ;  they  are  abundant,  quite  transparent,  and  the  larval  stage  is 
reached  in  the  brief  period  of  six  days.  The  authors  recommend  hot,  thirty 
per  cent,  alcohol  half  saturated  with  corrosive  sublimate  for  killing  the  egg-chain 
for  subsequent  sectioning  and  staining  by  the  Heidenhain  method.        c.  a.  k. 

Guiart,    J.     Les    Mollusques    Tectibranches.      This  is  a  resume  of  the  author's  mono- 
Causeries  Scientifiques  de  la  Soc.  Zool.  de      graph  of  these  mollusks,  and  is  devoted 

S  iJ, S;  7,;o''' "  ■"■    ''•"  ''  "''■     principally   to    the    Bum./,  and   the 

Aplysidce.  A  very  complete  outline  is 
given  of  the  gross  anatomy  of  several  types,  and  a  modified  classification  is 
proposed  for  the  group.  The  PleurobranchidcE  are  included  with  the  Nudi- 
branchs,  and  the  group,  as  a  whole,  are  derived  through  some  such  form  as 
Adeon  from  the  Prosobranchs  by  a  process  of  detorsion  as  shown  in  the  com- 
parative study  of  the  nervous  system.  The  author  recommends  the  subcutaneous 
injection  of  one  cubic  centimeter  of  five  or  ten  per  cent,  hydrochlorate  of 
cocaine  to  overcome  the  extreme  contractility  of  Aplysia.  Muscular  relaxation 
is  thus  secured  in  several  minutes.  Tissues  may  be  fixed  by  injection  of  sub- 
limate-acetic by  way  of  the  gill.  This  paper  is  one  of  an  excellent  series  of 
lectures  by  specialists  before  the  Zoological  Society  of  France.  c.  a.  k. 


1508  Journal  of  Applied  Microscopy 

NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

MIchaelis,  L.  Ueber  Fett-Farbstoffe.  Virch-  Much  that  relates  to  our  histological 
ow's  Archiv  fiir  path.  Anat.  164:  263-270,  staining  methods  is  purely  empirical. 
'^°''  Hence  Dr.  Michaelis'  attempt  to  deter- 

mine chemically  what  part  of  the  sudan  III  molecules  gives  the  fat-staining 
property,  and  from  this  knowledge  to  synthetically  prepare  a  more  satisfactory 
stain,  is  very  welcome.  But  why  do  we  require  a  new  stain  or  any  stain  for  fat, 
a  substance  so  easily  recognizable  both  morphologically  and  by  its  high  index 
of  refraction  ?  Unfortunately  other  substances,  as  the  zymogen  granules  of  the 
pancreas,  eosinophilic  granules,  etc.,  are  in  these  ways  indistinguishable  from  fat. 
Osmic  acid  is  not  specific  and  does  not  color  all  fat,  for  Altmann  has  shown  that 
osmic  acid  colors  only  the  oleic  acid  fats.  Sudan  III  has  not  proven  perfectly 
satisfactory. 

Sudan  III  is  azobenzol-azo  ft  naphthol.  Michaelis  first  assumes  that  the 
double  azo  group  lends  the  fat-coloring  property  to  the  molecule.  However,  he 
finds  that  a  stain  with  a  simpler  azo  group  (benzol-azo  /j  naphthol)  will  color  fat. 
Another  stain,  differing  from  last  only  in  being  a  instead  of  ft  naphthol, 
stains  tissues  diffusely.  This  latter  is  soluble  in  alkalies,  while  the  former  is 
soluble  neither  in  alkalies  nor  acids,  but  only  in  organic  solvents,  thus  resem- 
bling Sudan  III.  In  experimenting  he  finds  that  the  staining  reaction  of  the  ft 
compound  does  not  depend  on  the  orthoposition  of  the  azo  group  to  the  OH 
group,  but  in  the  lack  of  a  free  OH  group- — in  other  words  the  lack  of  a  salt- 
forming  group.  So  he  concludes  that  fat-stains  are  those  azo  bodies  which 
possess  no  salt- forming  group  —  indifferent  coloring  stuffs  in  opposition  to  acid 
and  basic. 

Knowing  this,  he  prepares  synthetically  azoorthotulolazo  ft  naphthol  (schar- 
lach  R  or  fettponceau),  which  gives  better  results  than  sudan  III,  coloring 
very  small  droplets  of  fat  bright  red.  This  stain  is  insoluble  in  water,  acids  and 
alkalies,  soluble  with  difficulty  in  alcohol,  and  easily  soluble  in  chloroform,  fatty 
oils  and  melted  paraffin. 

The  technique  for  its  employment  is — 

1.  Tissues  preserved  in  formol. 

2.  Freezing  microtome  sections. 

3.  Saturated  solution  of  scharlach  R    in  60-70%  alcohol  for  fifteen  to 

thirty  minutes. 

4.  Counterstain  with  haematoxylin. 

5.  Mount  in  glycerine  or  levulose  syrup.  H.  A.  Christian. 

Feldbausch,  F.  Ueber  das  Vorkommen  von  Goldmann,  Miiller,  Rieder,  and  Rein- 
eosinophilen  Leukocyten  in  Tumoren.    Vir-      bach  have  previously  called  attention 

chow's  Archiv.  f.  path.  Anat.  161 :  1-18,  inoo.       ^      ^,  r    1  u^„^    ^t 

^  ^  to  the   presence   of  large  numbers  of 

eosinophilic    leucocytes    in    certain    tumors. 


and  Laboratory  Methods.  1509 

Feldbausch  has  found  in  epidermoid  carcinomas  almost  constantly  a  marked 
increase  of  the  eosinophiles,  while  in  adeno-carcinomas  and  sarcomas  this  does 
not  occur.  The  eosinophiles  lie  chiefly  in  the  connective  tissue  surrounding  the 
masses  of  tumor  cells.  They  form  part  of  the  cellular  infiltration  which  owes 
its  origin  to  inflammatory  irritants  of  chemical,  bacterial,  or  mechanical  nature. 
They  are  present  in  greater  number  in  the  earlier  stages  of  the  development  of 
the  tumor,  and  also  in  the  beginning  of  inflammations,  than  later  when  degener- 
ation has  occurred. 

This  investigation  throws  little  if  any  light  on  the  origin  of  eosinophiles.  The 
author  does  not  believe  they  arise  in  loco.  Although  he  admits  that  Ehrlich's 
view  that  the  cells  develop  in  the  bone-marrow  may  be  correct,  he  thinks  they 
may  also  be  formed  in  the  blood.  The  eosinophilic  granules,  he  holds,  are  not 
formed  by  the  ingestion  of  broken-down  red  blood  corpuscles.  The  researches 
of  Arnold  have  shown  that  the  granules  belong  to  the  structural  elements  of  the 
cell,  and  hence  cannot  arise  through  phagocytosis.  Eosinophiles  are  often 
found  in  great  number  in  places  where  no  hemorrhage  has  occurred,  and  are 
often  not  found  where  destruction  of  red  blood  corpuscles  regularly  takes  place, 
as  in  the  liver  and  spleen,  or  where  hemorrhage  has  occurred.  j.  h.  p. 

Edmunds,  W.  The  Pathology  and  Diseases  of  Recent  researches  have  shown  that  the 
the  Thyroid  Gland.  Lancet,  1:  p.  131 7,  parathyroid  glands  are  of  great  im- 
^    ■  portance  to  the   organism.      Removal 

of  these  bodies  usually  causes  the  death  of  the  animal.  The  parathyroid  glands 
differ  in  structure  from  the  thyroid  gland  in  that  they  consist  wholly  of  cells  and 
contain  no  vesicles  and  no  colloid,  or  at  most  a  minute  droplet. 

It  is  not  easy  to  identify  the  parathyroid  glands  in  the  human  subject,  because 
some  of  the  minute  outlying  nodules  are  found  to  consist  of  ordinary  thyroid 
tissue,  and  to  be  therefore  accessory  glands.  The  anatomy  of  the  parathyroid 
gland  in  man  has  been  worked  out  by  Welsh  of  Edinburgh.  He  finds  that  there 
are  four  of  these  glands — one  anterior  and  inferior  to,  one  posterior  and  superior 
to,  each  thyroid  lobe. 

Although  no  symptoms  occur  as  a  consequence  of  the  removal  of  one  lobe 
of  the  thyroid  gland,  the  other  lobe,  as  pointed  out  by  Wagner,  hypertrophies. 
The  vesicles  enlarge  and  become  branched,  their  lining  membrane  becomes 
folded,  the  cubical  secreting  cells  become  columnar,  and  the  colloid  disappears 
and  is  replaced  by  a  mucous  secretion  which  takes  the  staining  reagents  badly. 

In  dogs,  if  one  parathyroid  gland  be  dissected  free  from  the  thyroid  lobe, 
taking  care  not  to  interfere  with  its  blood  supply,  and  the  entire  thyroid  gland 
with,  the  other  parathyroid  glands  be  excised,  so  that  only  one  parathyroid  be 
left  in  the  animal,  the  dog  will  live  and  no  obvious  effects  will  ensue.  The 
parathroid  that  is  left  in  these  experiments  shows  signs  of  more  active  growth 
than  the  normal,  but  it  does  not  develop  into  thyroid  tissue  proper.  No  vesicles 
form.  This  disposes  of  the  view  once  held  that  the  parathyroid  glands  are 
undeveloped  thyroid  tissue. 

Edmunds  excised  the  parathyroids  in  a  number  of  dogs,  leaving  the  thyroid 
intact.  Interesting  changes  occurred  in  the  thyroid.  The  colloid  diminished, 
or   completely   disappeared,  and  its   place  was  taken  by   a  watery  fluid ;    the 


1510  Journal  of  Applied  Microscopy 

vesicles,  instead  of  remaining  round  became  branched,  and  the  secreting  cells 
became  columnar,  or  multiplying,  filled  the  cavity  of  the  vesicles  with  round 
cells.  These  changes  are  identical  with  those  described  as  "  compensatory 
hypertrophy,"  but  the  thyroid  lobes  did  not  enlarge,  on  the  contrary,  sometimes 
they  seemed  to  become  smaller.  This  would  coincide  with  the  view  that  the 
parathyroids  manufacture  the  secretion,  and  the  thyroid  stores  it ;  when  the 
parathyroid  had  been  removed  there  would  be  no  secretion  for  the  thyroid  to 
store. 

In  a  number  of  dogs  important  histological  changes  followed  excision  of  a 
portion  of  the  superior  laryngeal  and  vaso-sympathetic  nerves  on  one  side,  and 
the  lateral  thyroid  lobe  on  the  other  side.  The  colloid  disappeared  from  the 
remainder  of  the  thyroid.  Usually  the  secreting  cells  multiplied  into  the  cavity 
of  the  vesicle.  In  one  case,  the  dog  having  survived  the  experiment,  the  lobe 
was  excised  49  days  later.  It  was  greatly  enlarged  and  weighed  35  grams, 
which  is  three  or  four  times  the  normal  weight.  The  proliferated  secreting  cells 
did  not  fill  the  cavities,  which  contained  instead  a  watery  secretion.  The  great 
size  of  the  lobe  was  due  to  a  growth  of  young  thyroid  tissue  between  the  vesicles. 
This  shows  the  possibility  of  defective  innervation  being  the  cause  of  serious 
symptoms  and  pathological  changes.  j.  h.  p. 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Dale,  H.  H.    Galvanotaxis  and  Chemotaxis  of      In     this    work    detailed    comparisons 
Ciliate   Infusoria.      Part    I.    Jour.    Physiol.  ,      .  ,  , 

26:  291-361,  1901.  were   made   between    the    chemotactic 

and  electrotactic  reactions  of  certain 
organisms,  for  the  purpose  of  determining  to  what  extent  the  electric  current 
stimulates  through  its  chemical  action.  The  experimental  work  was  done 
mainly  on  the  infusoria  parasitic  in  the  intestine  of  the  frog.  The  species  used 
were :  Balantidium  diiode7ii,  B.  elo?igatuvi,  B.  eiitozoon,  Nydothcnis  cordi/ormis, 
and  Opalina  ranarum.  On  account  of  the  high  osmotic  pressure  of  the  medium 
in  which  these  organisms  normally  live  it  was  necessary  to  examine  them  in  a 
solution  of  approximately  equal  concentration.  A  .()  per  cent,  solution  of  NaCl 
was  used  for  this  purpose,  the  organisms  being  shaken  directly  into  it  from  the 
intestine.  It  was  found  that  the  chemical  reaction  of  the  solution  in  which  the 
infusoria  were  placed  had  a  very  marked  influence  on  their  responses,  so  that  it 
was  necessary  to  conduct  parallel  experiments  with  solutions  carefully  made  acid, 
neutral  or  alkaline.  The  chemotaxis  was  tested  by  introducing  into  the  solu- 
tion containing  the  organisms,  either  on  a  slide  or  in  a  watch-glass,  capillary 
tubes  filled  with  the  test  solutions.  The  principal  solutions  employed  for  test- 
ing the  chemotaxis  were  an  organic  acid  (acetic  or  butyric),  a  mineral  acid 
(H.^SO^),  and  an  alkali  (NaOH  or  Na.^CO.,).  The  electrotactic  experiments 
were  performed  in  the  usual  way  with  a  stimulation  trough,  to  which  the  current 


and  Laboratory  Methods.  1511 

was  led  through  unpolarisable,  brush  electrodes.  The  current  was  obtained 
from  a  battery  of  small  bichromate  cells. 

The  reactions  of  Opalina  are  first  discussed.  It  was  found  that  this  form, 
when  in  an  alkalinated  or  neutralised  medium,  showed  an  "attraction"  to  (i.  e., 
formed  a  collection  in)  an  acid  test  solution  and  a  "repulsion"  from  an  alkaline, 
and,  in  response  to  the  electrical  stimulus,  collected  at  the  anode.  If  the  salt 
solution  containing  the  organisms  was  acidified  the  reactions  were  reversed,  col- 
lections being  formed  in  the  alkaline  test  solution  and  at  the  kathode  pole.  The 
reactions  of  Nyctotherus  showed  a  still  closer  dependence  on  the  chemical 
reaction  of  the  medium  than  did  those  of  Opalina.  In  a  strongly  alkalinated 
medium  Nyctotherus  collected  in  acids  and  at  the  anode  pole.  In  a  weakly 
alkalinated  medium  these  organisms  collected  in  weakly  acid  test  solutions  and 
were  "repulsed"  from  strong  acids  and  from  alkalis  of  all  strengths.  In  response 
to  the  electric  current  collections  were  formed  at  the  anode  except  in  very  strong 
currents,  when  there  was  a  transverse  orientation.  In  a  neutral  medium  there 
was  "repulsion"  from  both  acids  and  alkaline  test  solutions  and  the  electrotactic 
reaction  was  transverse  to  the  direction  of  the  current.  Passing  to  the  reactions 
in  acid  media,  it  was  found  that  when  in  a  weakly  acid  salt  solution,  the  organ- 
isms formed  collections  in  weakly  alkaline  test  solutions  and  at  the  kathode 
pole.  "Repulsion"  occurred  from  strong  alkalis  and  from  acids  of  any  strength. 
With  strong  electric  currents  there  occurred  again  the  transverse  orientation. 
In  a  strongly  acid  medium  collections  were  formed  in  alkaline  test  solutions  and 
at  the  kathode.  Under  all  conditions  except  when  in  strongly  alkalinated  media 
Balantidium  elongatum  collected  in  alkaline  test  solutions  and  at  the  kathode 
pole.  In  strongly  alkalinated  saline,  however,  this  form  collected  in  weakly 
acid  test  solutions  and  exhibited  a  diphasic  reaction  to  strong  solutions,  being 
first  "attracted"  to  the  acid  and  then  in  a  short  time  passing  over  to  the  alkali. 
In  this  strongly  alkaline  medium  there  was  also  a  diphasic  reaction  to  the  cur- 
rent. An  immediate  movement  to  the  anode  was  replaced — after  a  time  depend- 
ent on  the  strength  of  the  current — by  motion  towards  and  collection  at  the 
kathode.  Without  going  into  the  details  of  the  individual  cases  it  may  be  stated 
that  in  the  two  other  species  studied,  Balaiitidium  entozoon  and  B.  duodeui, 
essentially  similar  reactions  were  found.  In  all  cases  there  was  a  distinct  paral- 
lelism between  the  chemotaxis  and  the  electrotaxis. 

The  ciliary  action  in  these  responses  was  studied  principally  in  Opalina  and 
Nyctotherus.  Opalina  reacts  to  repellent  stimuli  by  a  response  like  the  "motor 
reflex"  of  the  free  living  infusoria  as  described  by  Jennings.  This  reaction 
brings  about  its  "repulsion"  from  alkalis.  Its  collections  in  acid  solutions  are, 
however,  the  result  of  a  different  sort  of  a  response.  When  the  anterior  end  of 
the  organism  comes  in  contact  with  a  weak  acid  the  ciliary  waves  change  their 
direction  in  such  a  way  as  to  directly  orient  the  body  along  the  lines  of  diffusion. 
The  organism  then  swims  toward  the  center  of  diffusion.  This  is  probably  the  first 
clear  case  recorded  in  the  literature  where  an  inf  usorian  becomes  directly  oriented 
along  the  path  of  diffusion  of  ions,  and  forms  collections  in  solutions  as  a  result 
of  such  a  response.  The  orientation  to  the  electric  current  is  brought  about 
by  a  rotation  as  in  the  ordinary  "motor  reflex"  until  the  anterior  end  is  towards 


1512  Journal  of  Applied  Microscopy 

the  anode  or  the  kathode  as  the  case  may  be.  There  is  no  ciliary  reversal  on 
the  kathode  half  of  the  body,  as  has  been  described  by  several  observers  in  the 
case  of  the  free  living  infusoria.  The  author  lays  stress  on  the  parallelism  in 
the  forms  he  has  studied  between  the  effect  of  chemical  and  electrical  stimuli  on 
the  ciliary  action. 

Experiments  were  made  with  media  of  different  electrical  conductivities.  It 
was  found  that  in  both  hyper-  and  hypotonic  solutions  Opalina  and  Nyctotherus 
show  a  tendency  to  pass  to  the  kathode  pole,  although  the  reaction  varies  some- 
what. Experiments  with  Paramecium  and  Colpidium  in  salt  solutions  showed 
that,  under  these  conditions,  both  of  these  ordinarily  kathodic  forms  went  to  the 
anode  when  the  current  was  passed.  It  is  maintained  that  probably  nearly  all 
the  ordinarily  described  electrotactic  reactions  are  conditioned  by  the  conduc- 
tivity of  the  solution  in  which  they  are  tested,  and  that  they  may  disappear  or 
be  replaced  by  very  different  responses  under  different  conditions. 

The  author  considers  the  general  phenomenon  of  electrotaxis  to  be  the  result 
of  two  factors ;  one,  a  rheotactic  reaction  to  the  current  of  fluid  produced  by  the 
kataphoric  action  of  the  electric  current,  and  the  other  a  chemotactic  reaction  to 
the  acid  continually  set  free  at  the  anodic,  and  the  alkali  at  the  kathodic  end  of 
the  body.  Whatever  may  be  one's  opinion  as  to  the  adequacy  of  this  theory,  the 
work  as  a  whole  is  an  extremely  important  and  well  developed  contribution  to 
the  discussion  of  the  phenomenon  of  electrotaxis.  r.  p. 

Jennings,  H.  S.      On   the  Significance  of   the      It  has   long  been   known   that   a  great 
Spiral    Swimming   of     Organisms.      Amer.  ,  .  , 

Nat.  35:  369-378,  1901.  i^^^Y    lower   organisms  (e.g.,   swarm- 

spores,  flagellate  and  ciliate  infusoria, 
rotifers,  and  others)  swim  in  a  spiral  path.  It  is  the  purpose  of  the  present 
paper  to  explain  the  biological  significance  of  this  form  of  progression.  It  is 
very  clearly  shown  by  Dr.  Jennings  that  the  purpose  and  result  of  this  movement 
in  a  spiral  is  to  keep  the  animal  on  a  straight  course.  Most  of  the  infusoria  are 
unsymmetrical,  and  as  they  start  to  move  forward  they  are  swerved  from  a 
straight  course  as  a  result  of  this  asymmetry.  This  swerving  is  always  towards 
the  same,  structurally  defined,  side  of  the  body.  If,  however,  as  is  in  fact  the 
case,  the  organism  rotates  on  its  long  axis  as  it  advances,  it  is  at  once  apparent 
that  any  tendency  to  swerve  to  one  side  from  the  straight  course  will  compensate 
itself,  thus  leaving  the  forward  component  of  the  motion  the  only  effective  one, 
and  making  the  path  a  spiral  with  a  straight  axis.  This  method  of  swimming 
is  closely  related  to  the  method  of  reaction  to  stimuli  of  these  organisms,  since 
the  side  of  the  body  towards  which  the  infusorian  turns  in  the  "  motor  reflex  " 
is  always  directed  away  from  the  axis  of  the  spiral.  Not  only  asymmetrical 
organisms  use  this  method  of  keeping  on  a  straight  course,  but  many  bilaterally 
symmetrical  rotifers  also  swim  in  a  spiral  path.  These  rotifers  have  a  marked 
tendency,  when  moving  freely  in  the  water,  to  swerve  towards  the  dorsal  side. 
This  tendency  is  the  one  compensated  for  by  the  spiral  swimming  in  this  case. 
Correlations  between  the  method  of  movement  and  the  form  of  the  body  in  other 
cases  are  discussed.  r.  p. 


and  Laboratory  Methods.  1513 

NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Clarke,  FW.,  and  Steiger,  George.    Experi-      ^he    article    treats    of    the    fractional 
ments  Relative  to  the  Constitution  of  Pecto- 

lite,  Pyrophyllite,  Calamine,  and  Analcite.      analysis  of  silicates  by  means  of  various 
Am.  Jour.  Sci.  iv,  8:  245,  1899.  reagents,    in    order   to   gain    evidence 

bearing  upon  their  chemical  structure. 

Pcctolite  proved  to  be  a  true  metasilicate  by  ignition  and  solution  in  sodium 
carbonate,  the  mineral  being  decomposed  and  losing  practically  \  of  its  SiOg, 
as  required  by  theory. 

PyropJiyllite  not  proved  to  be  a  metasilicate  by  same  test.  May  be  regarded 
as  having  formula,  SigOg^Al — OH, 

Calamijie  tests  as  a  rule  yielded-  negative  results,  but  supported  the  usual 
formula. 

Analcite  appeared  not  to  be  a  metasilicate,  but  may  be  a  mixture  of  ortho- 
and  tri-silicate,  represented  by  formula,  Al4Na4(Si04)2(Si3  0g)2  •  4H2O. 

Analcite    and    leucite    determined    by  authors   to   belong  to  garnet-sodalite 

group.  L.  IMcI.  L. 

Prior,  G.  T.  and  Spencer,  L.  J.     The   Identity      Binnite  possesses  the   same  degree  of 
of  Binnite  with  Tennantite,  and  the  Chemi-  i        /^        •   i 

cal  Composition  of  Fahlerz.    Min.  Mag.  12 :      symmetry    as    the    Cornish    tennantite, 
184,  1899.  being  simply  better  developed  and  hav- 

ing more  and  brighter  faces. 
Tetrahedrite  (Fahlerz)  appears  to  have  the  formula  3  CugS.  (Sb.As)2S.j   in 
the  case  of  the  simple  sulphantimonite  or  sulpharsenite  of  copper.     The  Fe  and 
Zn  appear  to  be  the  disturbing  elements,  producing  the  4  :  1  original  formula  of 
Rose. 

As  a  result  of  many  analyses  the  author  proposes  the  new  formula  3  R'gS  • 
R'"2S3+^[6R"S  .R'"2S,3],  in  which  R'=Cu,  Ag  ;  R"=Fe  Zn  ;  R"'==Sb,  As, 
Bi ;  and  .r=a  small  fraction,  often  ^  or  \,  but  rising  to  \  in  the  case  of  the 
highly  ferriferous  tetrahedrite  "  coppite." 

Many  tests  of  the  new  formula  are  given  by  reference  to  to  analyses. 

L.  McI.  L. 

Hartley,  E.  G.  G.     Communications   from  the  Fharmacosiderite. — As  a  result  of  care- 
Oxford    Mineralogical    Laboratory    on    the  ,   ,  ...  r^        •  ^  ^ 
Constitution  of  the  Natural  Arsenates  and  l^l  analysis,  m  green    Cornish  crystals, 
Phosphates.    Min.  Mag.  12:  152,  1899.  the  following  formula  was    proposed: 

2  Fe  ASO4  .  Fe[0(H,  K)]3  .  5  HgO. 

The  undoubted  presence  of  K  (new  to  pharmacosiderite)  is  of  interest. 

During  the  course  of  the  investigation,  the  apparent  permeability  of  the 
mineral  to  certain  liquids  was  shown.  A  green  transparent  crystal  immersed  in 
ammonia  turned  red,  and  became  green  again  in  hydrochloric  acid. 

L.  McI.  L. 


1514  Journal  of  Applied  Microscopy 

euwirth,  V.      Titanit   von   der   Hiittellehne       Asparagus    greei 
bei    Wermsdorf  un   Miihren.     Tschermak's       ^  ,,  ,  •       ,  •      • 

Min.  u.  petrog.  Mitth.  20:  178-1S0,  1901.         Crystallographic  description. 


Martin,  Fr.    Ueber  Scheinbar  spaltbaren  Quarz      Quartz  kernels  from    an  old   wall  are 
von  Karlsbad.    Tschermak's  Min.  u.  petrog.       .  ,  .         -^^     .  .  ^ 

Mitth.  20:  80-S2,  1900.  imperfect  crystals.     Under   action    of 

frost    certain    liquid    inclusions  which 

are   arranged  parallel   R,  go   P  and    sometimes    o  P,  have    produced    apparent 

cleavages.  a,  j.  m. 

Erben,  F.,  and  Ceipek,  L.    Analyse  des  Albits      Leading  to  formula  Ab^,  5 An,, 
von  Amelia.      Tschermak's  Min.  u.  petrog. 
Mitth.  20:  85,  1900.  A.  J.  M. 

Judd,  Hidden,  and  Pratt.    On  a  New  Mode  of      The  specimens   are  almost   similar   in 
Occurence    of    Ruby    in    North    Carolina.       ,  ,         ,  ,  r  1 

Min.  Mag.  12:  139,1899.  beauty  and   color  to   those   from    the 

Mogok  district  of  Burma,  and  certain 

garnet  (rhodolite)-bearing  basic  rocks  at  Cowee-Creek,  the  ruby  having  probably 

crystallized  out  from  the  basic  fluid  magma.     The  "  non-gem  "  corundum  occurs 

in  ordinary  crystalline  schists,  or  in  peridotites. 

The  rubies  frequently  contain  inclusions  of  various  kinds,  and  the  clearest 
crystals  almost  always  show  the  tabular  habit,  regarded  by  Lagoria  as  charac- 
teristic of  those  separating  from  an  igneous  magma. 

A  pseudomorphous  change  by  hydration  is  very  common,  as  in  the  case  of 
the  Burma  rubies,  and  it  is  hoped  that  more  investigation  will  bring  to  light  the 
similarity  in  rock  magma  producing  the  Burma  and  North  Carolina  rubies. 

L.  McI.  L. 

Pratt,  J.  H.    On  the  Crystallography  of  the      The  Cowee   Valley   crystals   have  two 
Rubies  from  Macon  Co.,  N.  C.     Min.  Mag.  ,  ,     ,. 

12:150,1899.  general  habits : 

(1)  Flat  tabular,  a  combination  of  base  and  unit  rhombohedron. 

(2)  Prism  a  (1120),  prominently  developed  with  base,  the  prism  being 
either  short  or  long.     Pyramidal  faces  ;/  (2243)  sometimes  show. 

Basal  planes  are  striated,  or  show  repeated  growth  of  unit  R  and  base. 
Similarity  in  development  is  noted  between  these  rubies  and  the  corundums 
from  Yogo  Gulch,  Mont.,  and  the  Burma  district.  l  Mci.  l. 


MEDICAL  NOTES. 

Blood  Examination. — The  following  method  for  the  preparation  of  speci- 
mens for  the  examination  of  blood  is  given  by  Dr.  W.  L.  Braddon,  of  the  Malay 
Peninsula :  The  mounts  may  be  made  either  between  two  square  cover-glasses, 
or  a  square  cover-glass  and  a  regular  size  slide.  The  covers  and  slides  are  first 
sterilized  by  a  method  recommended  by  Parker  and  Howard ;  viz.,  drop,  one  by 
one,  into  a  10  per  cent,  solution  of  chromic  acid,  contained  in  an  enamelled  iron 
dish,  and  boil  for  twenty  minutes.      They  are  then  poured,  altogether,  into  a 


and  Laboratory  Methods.  1515 

shallow  basin,  and  washed  with  ordinary  tap  water  until  no  trace  of  the  yellow 
color  of  chromic  acid  remains.  The  water  is  next  poured  off,  and  the  slips  are 
covered  with  rectified  spirit.  After  this  they  are  washed  in  absolute  alcohol, 
and  handled  with  clean  forceps. 

If  two  cover-glasses  are  used  for  the  mount,  they  are  accurately  superposed 
and  firmly  pressed  together.  An  edging  of  vaseline,  if  for  temporary  pur- 
poses, or  cement  if  for  permanent  purposes,  is  laid  over  all  the  edges, 
except  one,  and  a  very  small  portion  of  that  edge  which  is  opposite  the 
uncemented  one.  A  drop  of  blood  is  touched  with  the  free  edge  of  the  paired 
cover-glasses,  whereupon  the  blood  enters  between  the  glasses  in  an  exceedingly 
thin  film,  the  corpuscles  being  spread  out  with  beautiful  uniformity,  and  having 
suffered  a  minimum  amount  of  change  from  exposure  to  air  and  none  at  all  from 
handling  or  pressure.  When  the  blood  film  has  entered,  the  free  edges  may  be 
completely  closed,  and  the  examination  made. 

If  slide  and  cover-glass  are  used  the  latter  is  placed  on  the  slide  in  such  a 
position  that  one  of  its  edges  exactly  coincides  with  that  of  the  slide.  It  is  then 
firfnly  pressed,  and  sealed  with  vaseline  or  cement,  as  when  two  cover-glasses 
are  used,  and  the  subsequent  course  pursued  as  with  covers.  By  this  method 
a  number  of  mounts  may  be  made  and  stored  in  a  suitable  air-tight  bottle,  and 
thus  be  always  ready  for  use.  Fresh  blood  keeps  well  under  these  circum- 
stances.    No  special  skill   is   required  for  the   making  of  first-class  blood  film. 

This  method  has  been  carefully  tested,  and  it  was  found  necessary  to  put 
the  smallest  possible  amount  of  cement  between  the  covers  before  edging  them 
outside,  otherwise  the  cement  had  a  tendency  to  run  in. — Knowledge^  24:  183. 

c.  w.  J. 


Methods  of  Staining  the  Gonococcus. 
Schiitz  method  : 

1.  Stain  for  five  to  ten  minutes  in  sat.  sol.  methylen  blue  in  5  per  cent. 

carbolic  acid  water. 

2.  Differentiate  for  three  seconds  in  : 

Acetic  acid,         ...  1  part. 

Water,  dist.,  ....     4  parts. 

3.  Wash  in  distilled  water. 

4.  Counterstain  in  dilute  solution  of  safranin. 

Neisser's  method  : 

1.  Stain   in   cone.   ale.   sol.   of  eosin,  slightly  warmed,  for  two  or  three 

minutes. 

2.  Remove  excess  of  stain  with  filter  paper,  and  counterstain  with  cone. 

ale.  sol.  methylen  blue  for  fifteen  to  thirty  seconds. 

Chenzinski's  methylen  blue  and  eosin  : 

Methylen  blue,  sat.  aq.  sol.,  ...  2  parts. 

Eosin,  0.5  per  cent,  in  70  per  cent,  alcohol,     .     1  part. 
Distilled  water  or  glycerin,  ...  2  parts. 

With  this  solution  cocci  stain  blue,  pus  cells  pink. 


1516  Journal  of  Applied  Microscopy 


NEWS  AND  NOTES. 

The  University  of  Zurich  has  enlarged  its  anatomical  building.  A  dissecting 
room,  with  overhead  light,  to  accommodate  two  hundred  students,  has  been 
added,  and  on  the  floor  below  a  microscopical  room  of  the  same  size.  There  is 
also  a  demonstration  room  with  overhead  light,  a  laboratory  for  anthropology, 
and  a  laboratory  for  advanced  embryological  study,  together  with  rooms  for  the 
director.  The  old  part  of  the  building  will  be  rearranged  for  a  large  lecture 
room,  a  reading  and  study  room  for  the  students,  a  museum,  and  the  labora- 
tories for  assistants. — Science,  14  :  347. 


The  Bureau  of  Plant  Industry  of  the  U.  S.  Dept.  of  Agri.  has  recently  been 
reorganized,  and,  with  Beverly  T.  Galloway  as  chief,  now  embraces  the  following 
groups :  Vegetable  pathological  and  physiological  investigations,  Alferd  J.  Wood 
in  charge  ;  Botanical  investigations  and  experiments,  Frederick  V.  Coville  ;  Pomo- 
logical  investigations,  Gustavus  B.  Brachett ;  Grass  and  Forage  Plant  investiga- 
tions, F.  Lamson  Scribner ;  Experimental  Gardens  and  Grounds,  L.  C.  Corbett ; 
Congressional  Seed  Distribution,  Robert  J.  Whittleton  ;  Seed  and  Plant  introduc- 
tion, Ernst  A.  Bessey  ;  Tea  Culture  experiments,  Charles  U.  Shepard ;  and  the 
Arlington  Experimental  Farm,  L.  C.  Corbett. — Bot.  Gaz.  32 :  2. 


QUESTION   BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

13.  Is  there  a  simple,  satisfactory  method  of  determining  whether  or  not  a 
given  sample  of  milk  contains  bacteria,  that  can  be  performed  with  the  aid  of  a 
microscope  the  highest  power  of  which  is  a  one-fourth  inch  objective  ? 

I.  G.  B. 

14.  What  is  Tallquist's  method  of  blood  examination  and  estimation  of 
hemoglobin  ?  r.  c.  w. 

15.  Can  specimens  (animal)  preserved  in  alcohol  for  a  long  time  be  safel)"- 
transferred  to  formalin  ?  j.  d. 

16.  I  am  working  on  the  various  methods  of  fixing  or  producing  death  in 
the  animal  organism  and  cell.  My  work  ranges  from  micro-organisms  to  verte- 
brates, and  requires  specimens  for  dissection  killed  in  such  a  manner  as  not  to 
show  the  distortion  due  to  contraction  of  the  muscular  tissues,  such  as  occurs 
when  death  is  produced  by  chloroform,  ether,  or  other  anresthetic.  The  reagents 
I  employ  in  the  preparation  of  dissections  for  demonstrating  will  probably  react 
unfavorably  with  any  metallic  poisonous  compound  that  might  be  employed. 
The  substance  used  should  produce  death  as  immediately  as  possible,  in  order 
to  avoid  maceration  or  pathological  changes,  and  should  be  practically  tasteless, 
with  no  irritating  odor,  and  capable  of  being  used  in  minute  or  minimum 
quantities  with  delicate  water  animals,  etc.  If  you  can  suggest  such  a  sub- 
stance, or  anything  which  would  lead  to  similar  results,  it  would  be  greatly 
appreciated.  g.  w.  b. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 


Volume  IV. 


NOVEMBER,  1901, 


Number  1 1 


Studying  and  Photographing  the  Wild  Bird. 

The  problem  in  Bird  Photography*  is  how  to  see  and  not  be  seen.  If  a 
bird  is  actually  caught  and  kept  in  a  cage,  or  in  any  way  restrained,  its  behav- 
ior is  no  longer  perfectly  natural  and  free,  at  least  not  until  all  fear  has  been 
subdued,  and  it  is  no  longer  wild  but  tame.  What  is  most  needed  in  the  photog- 
raphy of  wild  birds  is  an  invisible  chain  to  hold  the  animals  to  some  fixed  spot 
which  can  be  approached  in 
disguise. 

Fortunately  for  the  student 
of  bird  habit  and  instinct,  all 
these  conditions  are  fulfilled  for 
a  most  important  and  interesting 
period,  that  of  life  at  the  nest. 
The  nest  is  the  given  fixed  point, 
and  parental  instinct  is  the  in- 
visible chain.  The  wild  bird, 
however,  is  bound  not  merely 
to  the  nest,  but  to  its  young. 
Wherever  the  young  go  the  old 
follow.  By  using  the  nearly 
fledged  young  as  a  lure,  some 
species  could,  I  believe,  be  led 
across  the  country  for  a  mile  or 
more.  I  have  taken  them  two 
hundred  feet  without  special 
effort. 

Hitherto    the    bird    photog- 

.    °  Fig.  I.— Nest-hole  of  Flicker  used  by  Bluebirds.     This  dead  stump 

rapher   has    had    to    rely     mamly  was  sawn  from  an  apple  tree  and  mounted  on  a  pivot  so  that  it 

could  be  easily  turned  at  any  angle  with  the  sun. 

*  The  following  paper  is  partly  taken  from  "  The  Home  Life  of  Wild  Birds :  A  New  Method 
of  the  Study  and  Photography  of  Birds,"  by  Francis  H.  Herrick,  with  141  original  illustrations 
from  nature  by  the  author,  and  published  by  Messrs.  G.  P.  Putnam's  Sons,  New  York  and 
London,  to  which  the  reader  is  referred  for  further  details.  It  also  contains  some  results  of  the 
author's  latest  experience  in  the  field. 

(1517) 


1518 


Journal  of  Applied  Microscopy 


upon  chance  in  getting  a  picture  of  the  nesting  scenes.  Most  land  birds  depend 
upon  concealment  for  protection  from  their  enemies  during  the  season  of  young. 
Their  nests  are  apt  to  be  shrouded  in  grass  or  foliage,  and,  if  easily  ap- 
proached, are  usually  inaccessible  to  the  camera.  If  the  nest  is  in  a  high  bush 
or  tree,  the  difificulties  of  the  position  and  light  are  usually  an  effectual  bar  to 
obtaining  good  pictures,  to  say  nothing  of  seeing  what  takes  place.  When  the 
nest  is  near  the  ground,  or  upon  it,  and  in  a  well  lighted  spot,  conditions  which 
are  rarely  fulfilled,  it  has  been  customary  to  set  up  the  camera,  and  attaching  a 
long  rubber  tube  or  thread  to  the  shutter,  to  retire  to  a  distance  and  wait  for  the 
birds  to  appear.  When  one  of  them  is  seen  to  go  to  the  nest,  the  plate  is  ex- 
posed by  pulling  the  thread  or 
pressing  the  pneumatic  bulb,  and, 
if  in  luck,  a  picture  may  thus  be 
obtained.  Many  plates,  however, 
are  sure  to  be  spoiled ;  little  can 
be  seen,  and  the  observer  has  no 
control  over  the  course  of  events. 
In  the  following  outline  a  method 
is  described  by  which  nesting  birds 
can,  in  most  cases,  be  successfully 
approached  and  studied  with  ease 
whatever  the  position  of  the  nest. 
The  usual  mode  of  procedure  is 
reversed,  and  instead  of  attempting 
to  carry  the  sensitive  plate  up  to 
the  bird,  the  camera  is  fixed  and 
the  bird  is  brought  directly  before 
it. 

It  is  a  comparatively  easy  mat- 
ter to  examine  and  photograph  the 

Fig.  2. — Tent  pitched  beside  Cedar-bird's  nest.     In  tliis  case  the  nCSt    the  eggS    Or  the  VOUno"  of  SUCh 

nesting  branch  was  sawn  from  a  neigliboring  apple  tree  and  '                         '                 _ 

mounted  upon  two  stakes  driven  into  the  ground,  on  a  hillside  SPCcicS  whOSC   dwellings   are  aCCCS- 

close  to  a  dwelling  house.  " 

sible  to  all ;  but,  to  portray  the  free 
behavior  of  the  adult  bird  in  the  shy  land  species  is  quite  another  question. 

The  method,  though  limited  in  its  application  from  the  necessities  of  the 
case,  is  based  on  the  solid  ground  of  animal  instinct,  and  may  confidently  be 
expected  to  have  a  wide  application. 

The  method  in  use  depends  mainly  upon  two  conditions :  (1)  The  control 
of  the  nesting  site,  and  (2)  the  concealment  of  the  observer. 

By  nesting  site  is  meant  the  nest  and  its  immediate  surroundings,  such  as  a 
twig,  branch,  hollow  trunk,  stem,  or  whatever  part  of  a  tree  the  nest  may  occupy, 
a  bush,  stub,  strip  of  sod,  or  tussock  of  sedge,  that  is — the  nest  with  its  imme- 
diate settings.  If  the  nest,  like  that  of  an  oriole,  is  fastened  to  the  leafy  branch 
of  a  tree,  the  nesting  bough  is  cut  off,  and  the  whole  is  then  carefully  lowered  to 
the  ground  and  set  up  in  a  good  light,  so  that  the  branch  with  the  nest  shall 
occupy  the  same  relative  positions  which  they  did  before.     The  nest,  however,  is 


ISJ^Ij 

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and  Laboratory  Methods. 


1519 


now  but  four  instead  of  forty  or  more  feet  from  the  ground.  The  nesting  bough 
is  carried  to  a  convenient  distance  from  the  tree,  and  firmly  fastened  to  two 
stakes,  driven  into  the  ground  and  placed  in  a  good  light.  If  the  nest  is  in  a 
tussock  in  a  shaded  swamp,  the  whole  is  cut  out  and  taken  to  the  nearest  well 
lighted  place  ;  if  in  the  woods,  it  is  carried  to  a  clearing  where  the  light  is  favor- 
able for  study.  Again,  when  a  nest  like  that  of  the  brown  thrush  occupies  the 
center  of  a  dense  thorn  bush  which  no  human  eye  can  penetrate  and  much 
less  that  of  the  camera,  its  main  supports  are  cut  off,  and  the  essential  parts  are 
removed  to  the  outside  of  the  clump  or  to  any  favorable  point  close  at  hand.  If 
the  nest  is  but  five  or  ten  feet  up,  the  main  stem  is  severed,  and  the  nesting 
branch  lowered  to  the  four-foot  mark,  a  convenient  working  height. 

This  sudden  displace- 
ment of  the  nesting  bough 
is  of  no  special  import- 
ance to  either  old  or 
young,  provided  certain 
precautions  are  taken. 
The  most  important  con- 
ditions for  success  ar^  as 
follows  :  the  change  of 
nesting  site  at  the  proper 
time,  or  when  parental 
instinct  is  approaching 
its  culmination  ;  the  pro- 
tection of  the  young  from 
excessive  heat  and  vio- 
lent storms,  and  the  pro- 
tection of  the  nesting 
bough  from  predacious 
enemies. 

When    the    nesting 
branch    is    vertical    and 
not  too  large,  it  can  be  easily  kept  fresh  for  days  by  placing  it  in  a  can  or  jug 
of  water,  which  should  be  set  in  the  ground. 

Young  birds  have  many  relentless  enemies,  among  the  worst  of  which  are 
cats,  jays,  squirrels,  and  small  boys.  On  page  15  of  "The  Home  Life  of  Wild 
Birds  "  this  subject  is  thus  referred  to  :  "I  feared  lest  prowling  cats  should  dis- 
cover the  young  whose  nest  and  branch  had  been  brought  down  from  the  tree 
top.  and  set  up  again  in  plain  sight  within  easy  reach  from  the  ground,  but  I 
was  happily  mistaken.  Predacious  animals  of  all  kinds  seem  to  avoid  such 
nests  as  if  they  were  new  devices  to  entrap  and  slay  them."  It  is  best  not  to 
stake  too  much  upon  this  assurance,  for  no  nest  of  young  birds  is  ever  safe, 
however  perfectly  concealed.  We  must  also  be  aware  that  cats  and  wild  depre- 
dators, like  the  birds  themselves,  soon  become  accustomed  to  new  objects  and 
surroundings.  The  nest  and  nesting  branch,  whether  moved  or  not,  should  be 
protected  whenever  possible  by  a  wire  net  of  ample  height,  secured  to  the  ground 


Fig.  3. — Female  Cedar-bird  astride  ucst,  sliielding  her  young,  wliich  ware 
then  six  day  old,  from  excessive  heat. 


1520 


Journal  of  Applied  Microscopy 


by  wire  staples.     It  is  impossible  to  overestimate  the  importance  of  this  screen, 
especially  in  a  country  overrun  with  cats. 

The  nest  might  be  taken  from  the  bough  or  from  the  sward,  but  this  would 
be  inadvisable,  chiefly  because  it  would  destroy  the  natural  site  or  the  exact  con- 
ditions selected  and  in  some  measure  determined  by  the  birds  themselves. 

For  an  observatory,  I  have  adopted  a  green  tent  which  effectually  conceals 
the  student,  together  with  his  camera  and  entire  outfit.  The  tent  is  pitched 
beside  the  nest,  and  when  in  operation  is  open  only  at  one  point,  marked  by  a 
small  square  window,  in  line  with  the  photographic  lens  and  nest. 

When  the  birds  approach  the  nest  in  its  new  position,  any  strange  objects, 
like  the  stakes  which  support  the  bough,  or  the  tent  which  is  pitched  beside  it, 

arouse  their  sense  of  fear  or 
suspicion  ;  they  may  keep  away 
for  a  time,  or  advance  with  cau- 
tion. If  very  shy,  like  many 
catbirds,  they  will  sometimes 
skirmish  about  the  tent  two 
hours  or  more  before  touching 
the  nest.  Their  fears,  however, 
are  usually  overcome  in  from 
twenty  minutes  to  an  hour,  and 
when  the  nest  has  once  been 
visited  in  its  new  site  the  vic- 
tory is  won.  I  have  known  a 
chipping  sparrow  and  red-eyed 
vireo  to  feed  their  young  in 
three  minutes  after  the  tent  was 
in  place. 

The  tent  which  I  have  used 
for  three  seasons  is  made  of 
stout  grass-green  denim,  and, 
with  the  frame,  weighs  only  six 
and  one-half  pounds.  It  can  be 
pitched  in  ten  minutes  almost 
anywhere,  and  may  be  compactly  rolled,  and  carried  for  miles  without  serious 
inconvenience.  One  may  spend  any  number  of  hours  in  it  by  day  or  night,  and 
with  a  fair  degree  of  comfort,  excepting  in  very  hot  or  sultry  weather,  when  ex- 
posed to  the  sun  on  all  sides.  It  is  also  a  welcome  shield  from  the  rain.  The 
green  color  of  the  material  renders  the  tent  an  inconspicuous  object  in  a  field  or 
open  pasture,  but  from  the  standpoint  of  the  bird  the  color  is  really  a  matter  of 
complete  indifference.  It  is  of  some  importance,  however,  when  we  consider  the 
attraction  which  a  tent  seems  to  possess  for  human  spectators,  whether  young  or 
old. 

The  front  of  the  tent  should  be  parallel  with  the  nesting  bough,  when  there 
is  one,  and  the  long  axis  of  the  latter  should  be  parallel  with  the  sun's  course. 
The  tent  is  so  placed  that  the  nest  is  in  direct  line,  not  with  the  middle  of  the 


Fig.  4. — Female  Robin  brooding  on  a  hot  July  day. 


and  Laboratory  Methods. 


1521 


tent,  but  with  the  window  to  one  side.  If  the  focal  length  of  the  lens  be  6)4 
inches,  the  nest  mounted  at  the  height  of  four  feet,  and  the  lens  be  28  inches 
from  the  rim  of  the  nest,  we  shall  get  a  picture  with  adequate  setting  on  a  4  x  5 
plate. 

When  the  nest  is  excavated  out  of  wood,  as  in  the  chickadees  and  wood- 
peckers, or  occupies  similar  cavities,  as  in  the  house  wrens  and  bluebirds,  the 
vertical  branch  or  stump  should  be  mounted  on  a  pivot,  so  that  it  can  be  readily 
turned  at  any  angle  with  the  sun.  Wherever  a  sky  background  is  not  available, 
it  is  of  great  advantage  to  use  a  large  screen  of  white  cloth,  which  should  be 
mounted  at  a  distance  of  five  or  six  feet  immediately  behind  the  nesting  bough. 
By  such  devices  one  can  obtain  serial  pictures  of  birds  performing  their  various 
acts  in  and  about  their  nests, 
in  front,  back,  or  profile 
views,  against  a  clear  white 
ground. 

After  birds  have  once 
adopted  the  changed  site, 
the  addition  of  the  white  or 
dark  screen  or  the  protect- 
ing wire  net  is  not  likely  to 
cause  the  least  annoyance. 
I  have  seen  a  Baltimore 
oriole  perch  on  the  top  of  a 
tall  screen  in  one  minute 
after  it  was  set  up,  and  the 
house  wren  come  to  her 
nest  almost  immediately 
after  the  screen  had  been 
torn  up  by  the  wind  and 
carried  with  a  crash  against 
a  neighboring  fence. 

Any  good  long-focus 
camera  with  reversible  back 
will  answer,  the  size  and 
weight  being  the  considerations  of  greatest  moment.  Most  naturalists  and 
sportsmen,  who  travel  long  distances  and  carry  their  own  traps,  find  a  camera 
which  takes  a  4  x  5  plate  the  most  convenient  and  economical.  I  have  used  this, 
but  for  work  with  the  tent  prefer  a  5  x  7  size,  because  it  gives  a  larger  and  better 
picture  of  the  object  sought.  For  work  outside  the  tent,  a  reflecting  camera  may 
be  used.     The  principal  requirement  in  either  form  is  a  long  bellows. 

In  photographing  a  moving  animal  at  the  close  range  of  from  twenty  to  thirty- 
six  inches,  the  difficulties  are  by  no  means  slight,  and  are  not  lessened  by  the 
use  of  long-focus  lenses.  A  lens  of  a  focal  length  of  ten  inches  or  more,  when 
used  so  close  to  the  object,  must  be  stopped  down  in  order  to  give  the  necessary 
depth,  and  bring  every  part  of  the  object  into  focus.  But  by  thus  cutting  off  the 
light  we  reduce  the   speed,  so  that  the   negative  with  an   exposure  of  1  /25  sec- 


..  S  (  iM  111  -!  h  ,1.  it  |iiiuii\  W  iiijilpn  Lli  iici-iiiiK(-l  li\  a  family  of 
Hiiusc  \\  icii--.  1  lie  leiiiale,  wIiilIi  hab  ju^t  fed  hei  hiood,  is  about 
to  re-enter  tlie  nest  tor  a  more  careful  inspection. 


1522 


Journal  of  Applied  Microscopy 


Fig.  6. — Nest-hole  of  Chickadees  appropriated  by   House  Wrens.     Front  view  of  circular 

entrance,  showing  the  female  approaching  it  with  niotli  miller.     No  screen  was 

used  here,  but  the  foliage  background  was  cut  out  of  the  picture. 

ond,"  the  maximum  time  usually  allowable,  is  too  weak  for  successful  printing 
even  after  the  intensifying  process  has  been  used. 

The    most  satisfactory  small  lens  with  which  I  have   worked  is  the  Zeiss 


Fif,.  7. — The  same  nest  turned  through  an   angle   of  cjo",  with   white   cloth   screen   at   back. 
.Stump  removed  from  tree,  mounted  on  pivot,  and  protected  by  a  fence  of  wire  netting. 


and  Laboratory  Methods. 


1523 


Anastigmat,  Ser.  11-a.  0  Xj-l  inch  focus,  speed  f/8,  when  used  with  a  4x  5  plate. 
Pictures  of  nearly  one-half  life  size  can  be  made  with  this  lens  without  stopping, 
in  full  sunlight,  with  an  exposure  of  1/25  second  of  the  iris  diaphragm  shutter, 
and  at  a  distance  of  eighteen  inches. 

Lenses  of  long  focus  are  not  available  for  work  at  very  close  range  unless  we 
are  able  to  allow  a  time  exposure  of  1/5  second  or  more,  but  at  distances  of 
eight  feet  and  upward  a  lens  of  9  or  1 0-inch  focus,  stopped  to  32,  with  a  speed 
of  f/6,  will  yield  satisfactory  results  with  an  exposure  of  1/5(1  second. 

When  a  clear,  perfect  image 
of  the  object  is  once  obtained, 
it  is  easy  to  make  pictures  of 
one-half  or  even  life  size  by 
the  well  known  process  of  en- 
largement. 

We.  thus  see  that  in  select- 
ing a  lens  for  photographing 
moving  objects  at  close  range, 
its  registered  speed  is  apt  to 
be  very  misleading.  We 
should  know  how  much  the 
lens  should  be  stopped  (or  how 
much  the  speed  must  be  re- 
duced) in  order  to  render  suf- 
ficient depth  or  detail. 

For  animal  photography 
the  most  rapid  plates  are  none 
too  fast,  and  any  of  the  best 
brands  can  be  recommended. 
Orthochromatic  plates  require 
careful  treatment,  but  in  skilled 
hands  offer  advantages  which 
should  not  be  neglected. 
When  used  out  of  doors  in 
full  sunlight  and  with  rapid 
exposure,   these  plates  do  not  seem  to  yield  their  best  results. 

We  have  thus  far  considered  the  wild  bird  during  the  period  of  young.  For 
photographing  inaccessible  nests,  and  for  approaching  birds  in  free  life  when  the 
sway  of  parental  instinct  is  over,  one  must  resort  to  other  methods.  For  fuller 
details  the  reader  is  referred  to  the  volume  from  which  the  preceding  paragraphs 
have  been  largely  drawn.  The  method  of  the  study  and  photography  of  birds 
which  is  here  illustrated  has  been  used,  as  the  case  of  each  required,  with  over 
forty  nests  of  the  common  land  birds  of  New  England,  and  its  value  has  been 
fully  demonstrated. 

Western  Reserve  University,  Cleveland,  Ohio.  FRANCIS    H.   HerriCK. 


Fig.  S. — Kingbirds  rending  a  troublesome  dragon-fly  preparatory 
to  serving  it  to  their  young.  The  female,  which  stands  at  the 
front,  was  brooding  when  the  prey  was  brought  in  by  the  male. 


1524  Journal  of  Applied  Microscopy 


A  Few  Remarks  on  the  Technic  of  Blood  Preparations. 

It  is  for  those  who  have  had  the  same  difficulty  as  myself  in  mastering  the 
technic  of  dried  and  heated  preparations  of  blood  for  clinical  examination  that 
these  remarks  are  intended.  While  I  will  not  say  that  I  have  not  sometimes 
succeeded  in  getting  beautiful  preparations  by  the  ordinary  method  of  drying  and 
heating  the  cover-glass  smears  and  staining  with  the  Biondi-Ehrlich  triacid  stain, 
I  may  say  that  to  make  a  perfect  slide  in  this  way  has  been  the  exception,  and 
I  have  often  had  to  try  over  and  over  again  before  accomplishing  creditable 
results.  This  I  will  not  say  is  the  fault  of  the  method,  but  I  imagine  from 
patient  work  that  all  are  not  able  to  acquire  the  requisite  skill  to  make  infallibly 
a  good  mounting.     My  own  results  have  been  far  from  uniform. 

The  method  which  I  am  now  using  is  in  no  wise  new,  but  it  is  the  application 
of  well  known  principles  that  I  would  call  attention  to.  With  a  little  care  and 
at  the  expense  of  less  time  than  the  usual  heat  method  employed,  I  have  been 
able  to  invariably  get  a  good  mounting.  Instead  of  the  cover-glass  preparation, 
the  method  of  spreading  the  blood  directly  on  the  slide,  as  pointed  out  by  Ewing 
in  his  new  work,  is  used.  This  consists  of  laying  the  slide  to  be  smeared  fiat  on 
the  table,  and  picking  up  the  drop  of  blood  from  the  finger  or  ear  on  the  end  of 
another  glass  slip  and  distributing  it  with  a  little  movement  along  the  edge  of 
the  end  of  the  slip  and  then  bringing  the  end  of  this  slide  in  contact  with  the  flat 
surface  of  the  other  at  an  angle  of  about  thirty  degrees  and  drawing  it  the 
length  of  the  slide  with  proper  pressure  to  produce  the  required  thickness  of 
film.  The  slide  is  then  hastily  placed  in  a  Naples  staining  jar  into  which  has 
previously  been  put  two  or  three  drops  of  one  per  cent,  osmic  acid  in 
one  per  cent,  chromic  acid  solution.  It  is  allowed  to  stay  in  this  vapor,  the 
cover  having  been  placed  on  the  jar,  for  from  forty  seconds  to  one  minute.  If 
allowed  to  remain  in  the  vapor  too  long,  it  will  not  take  the  stain.  The  object 
is  to  allow  it  to  remain  just  long  enough  that  when  removed  the  film  will  not 
wash  off  when  put  under  the  tap  of  water.  During  the  time  the  slide  remains 
in  the  jar  the  film  will  not  dry,  and  when  removed  it  should  be  dried  carefully 
over  the  lamp,  and  may  be  held  as  long  as  the  hand  will  bear  the  heat.  Without 
any  washing  now,  the  film  is  flooded  with  an  aqueous  solution  of  eosin  (  quite 
strong  )  and  allowed  to  remain  thus  for  from  three  to  ten  minutes.  The  time 
will  depend  on  the  strength  of  the  eosin  solution  and  the  fixation.  It  is  then 
washed  under  the  tap  for  a  considerable  length  of  time,  flooded  with  distilled 
water  and  stained  with  a  full  strength  solution  of  Mayer's  haemalum  for  about 
ten  or  fifteen  minutes.  It  is  then  washed  off  in  the  hydrant  and  the  tap  water 
allowed  to  run  over  it  as  long  as  desired.  W^ith  this  method  all  cells  are 
characteristically  stained,  and  everything  is  distinct  and  in  good  contrast. 
Haematoxylin  may  of  course  be  used  for  the  nuclear  stain  instead  of  haemalum, 
but  it  appears  that  the  latter  makes  by  far  the  most  beautiful  stain. 

As  when  one  has  once  learned  to  distinguish  the  various  elements  of  the 
blood  the  oil-immersion  lens  is  no  longer  necessary,  there  is  no  advantage  in 
using  balsam  as  a  rriounting  medium,  the  slide  may  be  allowed  to  drain  and 
be  covered  with  a  cover-glass  and  examined  at  once. 

While  the  above  method  does  not  fill  all  the  requirements  of  the  triacid  stain 
in  pathological  specimens,  perhaps,  it  makes  the  differential   count   easier   and 
shows  the  different  elements  of  the  normal  histology  of  the  blood  perfectly. 
Chicago,  111.  B.  L.  Rawlins. 


and  Laboratory  Methods. 


1525 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 

PHOTOMICROGRAPHY. 

II.     An  Apparatus  Adapted  to  All  Kinds  of  Work. 

The  apparatus  with  which  my  work  in  photomicrography  is  at  present  clone 
is  in  one  of  the  private  offices  of  Dr.  C.  S.  Bond  of  Richmond,  Ind.  ;  he  has  not 
only  by  his  material  help  made  it  possible  for  me  to  have  such  an  apparatus 
with  which  to  work,  but  he  has  also  worked  with  me  from  the  first ;  everything 
that  has  been  done  with  this  apparatus  has  been  our  joint  work. 

The  essential  parts  of  the  apparatus  are  shown  in  Fig.  1.  It  rests  on  an 
unshakable  stone  tioor,  and  consists  of  two  tables  supported  on  adjustable  metal 


Fig.  I. — Photomicrograpliic  apparatus. 

legs.  Their  combined  length  is  ten  and  a  half  feet.  One,  four  feet  long,  carries 
the  arc  light  and  illuminating  accessories;  the  other  carries  the  microscope  and 
camera.  The  microscope  stand  is  the  1899  Zeiss  model,  expressly  made  for 
photomicrography.  It  is  fitted  with  apochromatic  objectives  of  from  70  mm.  to 
2  mm.  and  compensating  and  projecting  eyepieces.  The  fine  adjustment  screw 
is  controlled  by  a  brass  rod,  which  lies  on  the  bench  under  the  camera  and  has 
a  pulley  and  cord  attachment  (ct)  with  the  milled  head  of  the  micrometer  screw. 
The  microscope  is  so  supported  by  an  adjustable  brass  pillar  (/;)  that  this  pulley 
cannot  in  the  least  affect  it. 

The  camera  is  carried  on  two  nickeled  steel  tubes  (r)  which  rest  on  adjustable 
metal  supports.  The  board  Q/)  on  which  the  microscope  rests  is  bound  also  by 
clamps  to  these  same  tubes.  Four  strong,  adjustable  brass  pillars  (e)  hold  the 
board  firmly  at  one  distance  from  the  table.  These  arrangements  may  be 
summed  in  the  statement  that  the  microscope  and  its  supports  are  immovable. 


1526 


Journal  of  Applied  Microscopy 


The  movable  stage  is  also  controlled  from  the  ground  glass  six  feet  away  by  brass 
rods  with  milled  heads  and  cord  and  pulley  attachment  (/),  and  the  stage  is  sup- 
ported against  the  strain  of  these  by  an  adjustable  brass  pillar  (g-).  The  stage 
can  thus  easily  and  quickly  be  searched  over  a  space  three-eighths  of  an  inch 
square.     The  course  adjustment  of  the  microscope  is  similarly  controlled. 

Some  may  think  that  these  arrangements  are  mere  conveniences ;  they  are, 
however,  indispensable,  for  the  reason  that  without  them  photomicrography 
ranging  in  powers  from  5  to  5000  diameters  consumes  so  much  time  that  the 
game  is  not  worth  the  ammunition. 


^ 

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A 

Fk;.  2. — Photomicrograph  of  a  starfish,  fixed  and  decalcified  in  picro-sulpliuric  acid,  and,  after  washing,  stained  in 
acid  carmine.  A  35  mm.  apocliromatic  gave  the  necessary  resohition  and  depth  of  focus,  and  a  camera  extension 
of  four  feet  gave  a  magnification  of  forty  diameters  determined  by  measuring  the  object  and  the  image. 

The  arrangement  for  controlling,  from  the  ground  glass,  the  coarse  adjust- 
ment— necessary  in  low  power  work  ;  that  for  controlling  the  stage — so  conven- 
ient as  to  be  necessary  in  all  classes  of  work;  the  adjustable  pillars  under  the 
microscope  bench  ;  the  adjustable  pillar  under  the  microscope  to  offset  the  pull 
of  the  cord  on  the  fine  adjustment  screw ;  the  adjustable  pillar  under  the  stage, 
and  such  a  scale  on  both  the  camera  table  and  the  optical  bench  that  all  parts 
of  the  apparatus  can  quickly  be  brought  into  any  desired  relationship,  are  addi- 
tions which  we  have  made  to  the  apparatus  since  setting  it  up. 

When  work  of  all  powers  is  to  be  done  on  the  same  instrument,  two  features 


and  Laboratory  Methods. 


1527 


Fig.  3. — Malignant  (Edema,  2  mm.  oil  immersion,  apochiomatic 
objective,  and  a  No.  4  projection  eyepiece  with  camera  e.\ten- 
sion  of  66  inches.     The  magnification  is  3000  diameters. 


of  our  microscope  stand  are  necessary,  namely,  the  large  tube,  two  inches  in 
diameter  into  which  the  objectives  screw  without  collars,  and  the  improved  fine 
adjustment  which  lowers  or  raises  the  objective  only  .04  of  a  millimeter  for  an 
entire  round.  As  it  can  easily  be  turned  less  than  a  degree,  the  distance  from 
the  object  to  the  objective  can  easily  be  varied  .0001  mm. 

The  camera  is  large  enough 
to  carry  a  six  and  one-half  by 
eight  and  one-half  plate  and 
can  be  extended  six  feet. 

The  optical  bench  carries 
the  arc  light  and  all  the  illumi- 
nating accessories  somewhat 
as  the  camera  is  carried  ;  all 
these  are  adjustable  up  and 
down,  to  and  from  the  light, 
and  from  side  to  side. 

The  necessary  accessories 
are  a  pair  of  condensers  (//), 
a  cooling  cell  (/),  two  ray 
filters  (y),  a  field  diaphragm 
(/'),  and  a  double  convex  lens 
not  shown  in  the  cut,  as  the 
instrument  was  arranged  for  low  power  work  at  the  time  the  photograph  was 
made  ;  these  are  necessary,  in  the  sense  that  one  pays  more  in  time  and  failures 
for  not  having  them  than  they  cost. 

These  tables,  benches,  condensers,  and  cells  should  all  be  carefully  levelled ; 
this  is  done  by  means  of  a  spirit  level  and  adjustable  feet  and  clamps,  one  or  the 
other  of  which  they  all  have.  Our  cooling  cell  is  three  and  a  half  inches  long 
and  four  and  a  half  inches  in  diameter  ;  we  keep  it  filled  with  water  and  have 
never  had  either  a  slide  or  an  objective  perceptibly  warm,  though  we  have  kept 
them  exposed  for  hours  together.  The  tradition  that  calls  for  alum  in  the  cell  is 
not  valuable.  In  a  future  article  on  "  Illuminating  the  Object,"  the  use  of  the 
other  accessories  will  be  explained.  It  follows  from  what  I  have  said,  that  a 
laboratory  costing  some  thousands  of  dollars  is  necessary  for  the  best  results  in 
photomicrography.  Experience  convinces  me  that  it  is  equally  necessary  for  an 
expert  microscopist  and  photographer  to  be  in  charge  of  it ;  he  then  could  do 
all  work  of  this  sort  in  conjunction  with  all  departments  of  a  university,  or  pos- 
sibly of  more  than  one  university.  A  joint  laboratory  used  by  a  dozen  different 
men,  all  mainly  interested  in  something  else,  will  yield  in  the  future  results  simi- 
lar to  what  it  has  in  the  past.  The  discouragement  one  hears  on  every  hand  is 
not  well  founded  ;  it  is  traceable  to  the  notion  that  photomicrography  is  a  simple 
art  that  any  one  can  practice.  If  courses  of  instruction  were  given  in  our  leading 
universities  in  connection  with  such  laboratories,  it  would  soon  come  to  pass  that 
we  should  be  as  well  off  in  photomicrographic  manipulators  as  we  are  now  in 
microscopists. 

Recently,  at  the  seaside  laboratory  of  the  Misses  Foot  and  Strobell,  I  saw 
some  excellent  work  done  with  very  simple  apparatus;  their  work  follows  entirely 
new  lines  ;  in  a  future  article  on  "  Focusing  the  Instrument,"  I  shall  describe 
their  arrangement. 
Earlham  College.  D.  W.  DenniS. 


1528  Journal  of  Applied  Microscopy 


Staining  Bacteria  in  the  Root-tubercles  of  Leguminous  Plants. 

In  a  paper  presently  to  be  published  in  the  Proceedings  of  the  California 
Academy  of  Sciences,  Third  Series,  Botany,  I  have  discussed  some  of  the  rela- 
tions of  the  bacteria  which  cause  the  formation  of  the  root-tubercles  of  legumi- 
nous plants  to  the  cells  in  which  they  occur.  One  or  two  matters  of  technique 
developed  in  the  course  of  the  study,  and  reported  in  the  paper  above  referred 
to,  may  be  of  some  interest  to  the  readers  of  the  Journal. 

When  I  once  casually  made  some  hand-sections  of  the  root-tubercles  of  Bur 
Clover  {Medicago  denticulata  IV/lld.),  the  plant  which  I  have  especially  studied, 
these  objects  seemed  to  me  unusually  favorable  for  treatment  with  paraffin  and 
the  microtome.  I  tried  fixing  them  in  a  concentrated  35  per  cent,  alcoholic 
solution  of  corrosive  sublimate,  and  found  that  they  were  easily  penetrated  by 
paraffin  from  xylol  solution,  embedded,  and  sectioned.  They  stain  readily  by 
the  usual  anilin  stains.  I  was  particularly  interested  in  demonstrating  the  man- 
ner of  infection  of  the  cells  of  the  root,  and  of  the  tubercle  subsequently  formed, 
and  in  order  to  produce  the  best  conditions  for  cytological  study,  I  fixed  a  fresh 
lot  of  young  and  growing  tubercles  in  dilute  FJemming's  chrom-osmic-acetic 
mixture.  This  visibly  browns  the  tubercles  of  any  considerable  size,  blackens  the 
oldest,  and  darkens  all.  Of  this  change  in  color  I  took  no  active  notice  until 
just  before  staining  the  sections  on  the  slide.  Then  I  bleached  by  immersing 
the  slide  for  half  an  hour  in  a  solution  of  one  part  Marchand's  hydrogen  per- 
oxide in  twenty  parts  80  per  cent,  alcohol. 

Since  the  tubercles  are  composed  almost  exclusively  of  soft  tissues,  paraffin 
melting  at  54°C.  is  hard  enough  for  embedding  and  sectioning,  provided  of 
course  that  the  room  temperature  is  suitable.  I  cut  very  thin  sections.  1  f.i  in 
some  cases,  with  perfect  success. 

The  staining  method  was  fundamentally  that  described  by  Hof^.  For  mak- 
ing up  the  stains  I  used  the  proportions  given  in  Humphrey's  translation  of 
Zimmermann's  Botanical  Microtechnique,  anilin  safranin,  anilin  gentian-violet, 
orange  G.  The  sections  were  attached  to  the  slide  by  albumen  fixative.  Hof's 
directions  for  staining,  followed  without  modification,  give  excellent  preparations, 
showing  the  degeneration  of  the  nucleus  and  cytoplasm  as  the  bacteria  multiply 
in  the  cells.  This  method,  however,  does  not  show  the  infection  threads  by 
means  of  which  the  root-hairs,  root,  and  new  tubercle  cells  are  infected,  for  it 
does  not  clearly  differentiate  the  bacteria  from  the  cytoplasm.  This  can  be 
readily  done  by  treating  the  slide,  washed  with  water  as  it  comes  from  the  anilin- 
gentian-violet  solution,  with  Gramm's  iodine  solution,  for  a  half  hour  or  longer, 
before  staining  with  orange  (i. 

This  method  consists  simply  in  applying  to  bacteria  in  tissues  the  well  known 
bacteriological  method  used  in  differentiating  cover-glass  preparations  stained  by 
Ehrlich's  anilin-gentian-violet.  By  this  means  the  infection  threads  running 
from  older   infected  cells  toward  and  into  the   daughter  cells  of  the  tubercle 

^  Hof,  A.  C.  Histologische  Studien  an  Vegetationspunkten.  Botan.  Centralb.,  Bd.  76,  No. 
3.  1898. 


and  Laboratory  Methods.  1529 

meristem  can  be  shown  in  all  growing  tubercles.  In  sufficiently  young  and 
recently  infected  roots,  the  course  of  the  infection  threads  from  the  root-hairs  to 
the  pericycle  can  be  clearly  demonstrated. 

The  success  in  embedding,  sectioning,  and  staining  root-tubercles  which 
follows  the  application  of  the  methods  just  described,  makes  it  difficult  to  under- 
stand the  difficulties  which  prompted  Miss  Dawson  ^  to  declare  "  the  tubercle 
tissues  very  difficult  objects  to  stain  upon  the  slide,"  and  that  "ordinarily  thin 
hand-sections  serve  better  for  the  examination  of  the  filaments  within  the  cells." 
Miss  Dawson  used  with  success  the  following  method,  but  it  lacks  some  of  the 
advantages  possessed  by  the  one  I  have  used.  She  placed  "  sections  hardened  in 
alcohol  (best  without  previous  treatment  with  chromic  or  osmic  acid)  "  "  for  about 
two  hours  in  alcoholic  potash  (one  part  5  per  cent,  potash  to  three  parts  absolute 
alcohol)  and  then  passed  into  Eau  de  Javelle  for  ten  minutes.  From  this  solution 
they  are  transferred  to  the  dye,  which  is  prepared  by  mixing  an  alcoholic  solution 
of  anilin  blue  with  orseillin,  drop  by  drop,  until  a  violet  solution  is  obtained. 
This  mixture  is  acidulated  with  a  few  drops  of  glacial  acetic  acid.  The  sections 
remain  in  the  stain  for  two  hours,  and  are  then  transferred  directly  to  dilute 
glycerine,  and  finally  mounted  in  glycerine."  This  method  of  Miss  Dawson's  is 
merely  an  improvement  in  definiteness  of  statement  of  the  one  described  by 
Strasburger  in  his  "  Praktikum."  One  of  the  stains  which  she  used,  Orseillin, 
is  not  obtainable  under  that  name  in  this  country,  and  I  do  not  know  whether 
she  means  Orcein  or  Orseille,  two  stains  made  by  Griibler  and  carried  in  stock 
here. 

However,  it  is  not  my  intention  to  criticise  Miss  Dawson's  method  or  her 
description  of  it,  but  rather  merely  to  describe  my  own,  which  anyone  sufficiently 
interested  to  try  it  will  find  practicable. 
Leland  Stanford  Jr.  University.  "  George   J.  Peirce. 


MICRO-CHEMICAL  ANALYSIS. 
XVIII. 

In  order  to  be  consistent  with  a  former  statement  we  should  properly  con- 
sider in  this  article  the  analytical  reactions  of  the  element  mercury  ;  this  element 
falling  in  the  same  group  in  the  periodic  system  as  the  elements  last  considered. 
Unfortunately  there  is  still  a  missing  element  between  cadmium  and  mercury, 
thus  causing  a  serious  break  in  the  series.  The  change  in  chemical  behavior 
which  we  find,  in  passing  from  cadmium  to  mercury,  is  such  that  so  far  as  our 
micro-chemical  tests  are  concerned,  there  are  practically  no  analogous  reactions 
existing  between  mercury  and  the  other  members  of  the  group. 

On  the  other  hand,  so  many  of  the  properties  of  aluminum,  the  horizontal 
analogue  of  magnesium,  are  closely  related  to  those  of  the  group  last  discussed, 
that  it  has  been  thought  best  to  take  up  aluminum  at  this  point.     Moreover,  we 


2  Dawson,  Maria.     Nitrogin  and  the  Nodules  of  Leguminous  Plants.     Philos.  Trans.  Royal 
Soc,  London,  1899. 


1530  Journal  of  Applied  Microscopy 

are  now  reaching  a  part  of  the  periodic  system  containing  so  many  rare  elements 
that  a  strict  adherence  to  the  order  of  the  periodic  system  is  no  longer  practi- 
cable if  the  plan  outlined  in  VII  of  this  series  of  papers  is  followed — namely,  to 
merely  discuss  the  tests  employed  for  the  detection  of  the  elements  most  fre- 
quently met  with  in  ordinary  analytical  work. 

The  remaining  articles  of  the  series  will,  therefore,  be  devoted  to  the  common 
metals — mercury,  lead,  silver,  arsenic,  antimony,  bismuth,  tin,  copper,  cobalt, 
nickel,  iron,  manganese,  chromium.  Then  will  follow  the  tests  for  the  common 
acids,  and  finally  tests  for  several  of  the  less  common  acid  forming  elements. 

ALUMINUM. 

Reference  has  already  been  made  a  number  of  times  to  this  element  in  pre- 
vious articles,  as  seriously  interfering  with  many  tests  ;  thus,  it  frequently  happens 
that  an  indication  of  its  presence  will  be  obtained  while  engaged  in  testing  for 
other  elements. 

The  separation  of  aluminum  from  most  of  the  other  elements  has  been  hinted 
at  in  the  last  article  (XVII).  Like  glucinum  and  zinc,  its  hydroxide  is  precipi- 
tated by  alkalies  and  is  soluble  in  excess  of  sodium  or  potassium  hydroxides, 
an  aluminate  of  the  general  formula  A1(0M)3  being  formed.  In  this  connection 
it  should  be  borne  in  mind  that  aluminum  phosphate  may  often  separate  in  the 
course  of  micro-chemical  analyses  when  the  material  containing  phosphates  is 
made  alkaline,  or  when  sodium  phosphate  is  being  used  as  a  reagent.  Aluminum 
phosphate  (AIPO4  •  4H0O)  is  soluble  in  potassium  and  sodium  hydroxides,  diffi- 
cultly soluble  is  ammonium  hydroxide,  and  insoluble  in  these  hydroxides  in  the 
presence  of  ammonium  salts.  Unlike  the  hydroxide,  aluminum  phosphate  is 
insoluble  in  acetic  acid. 

The  following  reagents  have  been  suggested  for  the  micro-chemical  detection 
of  aluminum  : 

I.  Cesium  Sulphate. 
II.  Ammonium  Fluoride. 

III.  Primary  Potassium  Sulphate. 

IV.  Staining  Aluminum  Hydroxide  with  Dyes. 

/.  Cesium  Sulphate  added  to  solutions  containing  Aluminum  SuIpJiate  leads  to 
the  formation  of  Cesium  Almn. 

Al2(S04)3  +  CS2SO4  =  [AL^CSOJ;,  .  Cs.^S04  •  24H,0]. 

Method. — To  a  drop  of  the  solution  to  be  tested,  add  a  drop  of  ammonium 
hydroxide.  Draw  off  or  filter  off  the  supernatant  solution.  Wash  the  precipitate 
once  with  water.  Then  add  a  single  drop  of  water  and  a  trace  of  dilute  sul- 
phuric acid,  only  just  enough  to  dissolve  the  aluminum  hydroxide.  Warm 
gently ;  cool,  and  to  the  drop  add  a  fragment  of  the  reagent.  After  a  few 
seconds,  beautiful  large  crystals  of  cesium  alum  separate  (Fig.  To).  The  crystals 
are  regular  octahedra,  and  the  usual  combinations  of  octahedron  and  cube,  etc. 

Remarks. — Cesium  chloride  can  be  employed  as  reagent,  providing  that  the 
solution  to  be  tested  contains  a  little  free  sulphuric  acid.     The  chloride  is,  how- 


and  Laboratory  Methods.  15:>1 

ever,  not  as  satisfactory  as  the  sulphate,  particularly 
in  the  hands  of  beginners,  for  cesium  chloride  crystal- 
lizes in  the  isometric  system,  thus  sometimes  leading 
to  confusion.  Cesium  sulphate,  on  the  contrary, 
crystallizes  in  the  orthorhombic  system.  An  examina- 
tion of  a  preparation  with  the  latter  salt,  between 
crossed  nicols,  will  therefore  permit  of  an  easy  differ- 
entiation between  crystals  of  cesium  sulphate  and 
those  of  cesium  alum. 

Cesium  sulphate  is  not  found  in  the  list  of  reagents 
heretofore  given.  It  is  made  from  the  chloride  as 
follows :  Place  a  drop  of  sulphuric  acid  at  the  corner 
of  a  slide  or  on  platinum  foil.  Add  a  small  crystal  of  cesium  chloride,  and 
evaporate  to  dryness.  If  no  fumes  of  sulphur  trioxide  escape,  add  another  drop 
of  acid  and  heat  again.  It  is  evident  that  by  this  method  of  treatment,  in  the 
majority  of  cases,  it  is  primary  cesium  sulphate  that  is  formed,  and  not  the 
normal  sulphate  as  indicated  in  the  reaction  given  above. 

Test  drops  containing  cesium  alum  have  a  great  tendency  to  remain  in  a 
state  of  supersaturation.  Often  a  single  large  crystal  only  will  appear.  In  such 
an  event,  crushing  the  crystal  and  drawing  its  fragments  through  the  drop  will 
almost  invariably  yield  a  large  crop  of  well  formed  crystals. 

Testing  for  aluminum  with  cesium  sulphate  leaves  little  to  be  desired  as  to 
accuracy  and  elegance,  but  requires  a  little  practice  to  learn  just  the  proper  con- 
centration. Too  dilute  a  test  drop  requires  very  long  waiting.  Spontaneous 
evaporation  leads  almost  invariably  to  supersaturation.  Evaporation  over  the 
"micro"  flame  is  very  unsatisfactory.  On  the  other  hand,  the  addition  of  the 
reagent  to  too  concentrated  a  test  drop  gives  rise  to  the  immediate  formation  of 
dendritic  masses  and  skeleton  crystals.  It  is  true  that  the  experienced  worker 
^'ill  usually  at  once  recognize  these  dendrites  as  due  to  the  presence  of  aluminum, 
but  in  view  of  the  fact  that  beautiful  and  far  more  characteristic  crystals  can  be 
obtained,  the  worker  should  not  be  satisfied  with  an  unsightly  preparation. 

It  is  because  of  the  difficulties  just  mentioned  that  the  method  of  first 
precipitating  the  aluminum  as  hydroxide  has  been  suggested.  By  this  method 
the  operator  always  knows  the  concentration  of  the  test  drop  and  the  probable 
amount  of  free  sulphuric  acid.  Moreover,  all  other  free  acids  have  been  removed 
as  well  as  many  objectionable  salts,  a  matter  of  not  a  little  importance. 

In  the  presence  of  magnesium  sulphate  there  is  formed  a  double  sulphate  of 
magnesium  and  cesium,  hence  in  dealing  with  such  cases  it  is  necessary  to  add 
a  sufficient  amount  of  cesium  sulphate  to  permit  of  the  formation  of  both  the 
cesium  magnesium  sulphate  and  the  cesium  alum.  It  is  very  seldom  that  the 
cesium  magnesium  double  sulphate  separates;  when  it  does  its  crystals  are  to  be 
referred  to  the  monoclinic  system. 

It  is  of  course  obvious  that  in  the  case  of  simple  substances  it  is  merely 
necessary  to  acidify  with  sulphuric  acid  and  add  the  reagent.  Excellent  results 
can  be  thus  obtained.  But  this  method  of  procedure  requires  (1)  just  the  proper 
concentration,  (2)  the  absence  of  much  free  sulphuric  acid,  (3)  the  absence  of 
free  acids  other  than  sulphuric. 


1532  Journal  of  Applied  Microscopy 

Cesium  alum  is  one  of  a  group  of  double  sulphates  known  as  "  alums,"  hav- 
ing the  general  formula  M^(SO^)^  •  7V^2^04  •  24H2O,  where  -Jlf-  can  be  Al,  Cr, 
Mn,  Fe,  In,  Cxa,  Tl ;  and  -A-  Na,  K,  Rb,  Cs,  NH^,  Ag,  or  Tl.  All  alums  are 
isomorphous,  and  are  to  be  referred  to  the  isometric  system.  Theoretically, 
therefore,  one  would  be  led  to  expect  that  the  presence  of  elements  capable  of 
taking  the  place  of  aluminum  in  alums  would  be  liable  to  interfere  with  the  test 
for  aluminum.  But  in  addition  to  their  property  of  being  able  to  replace 
aluminum  in  these  double  sulphates,  we  must  consider  the  crystallizing  power  of 
the  compounds  formed.  It  is  herein  that  lies  the  explanation  of  the  value  of 
cesium  sulphate  over  and  above  that  of  any  other  of  the  sulphates  we  might  be 
inclined  to  select.  Of  the  above  listed  alum  forming  elements,  aluminum  is  the 
only  one  which  unites  with  cesium  or  rubidium  sulphates  to  form  easily  crystal- 
lizable  alums.  The  other  elements  unite  with  these  two  sulphates  only  with 
difficulty,  and  the  alums  formed  can  be  regarded,  from  a  micro-chemical  stand- 
point, as  practically  uncrystallizable.  Sodium,  potassium,  and  ammonium  sul- 
phates readily  unite  to  form  more  or  less  crystallizable  alums  with  the  other  alum 
forming  elements  as  well  as  with  aluminum. 

Exercises  for  Practice. 

To  a  test  drop  consisting  of  a  solution  of  aluminum  sulphate  add  a  fragment 
of  the  reagent. 

Precipitate  another  drop  with  ammonium  hydroxide,  draw  off,  wash  the 
precipitate,  dissolve  in  the  least  possible  amount  of  sulphuric  acid,  and  test. 

Try  rubidium  sulphate  as  reagent ;  then  potassium  sulphate  ;  sodium  sulphate  ; 
ammonium  sulphate.     Try  cesium  chloride. 

Test  for  Al  in  the  presence  of  free  hydrochloric  acid ;  free  nitric  acid. 

Test  preparations  containing  Al  and  Fe ;  Al  and  Cr ;  Al  and  Mn  ;  Al,  Fe, 
Cr  ;  Al  and  Mg;  Al  and  Gl;  Al  in  the  presence  of  phosphates. 

Prepare  slides  of  chrome  alum,  iron  alum,  etc.,  then  mixtures  of  these  various 
alums ;  note  isomorphism. 

//.  Ammouiian  Fluoride  in  excess  leads  to  the  separatiofi  of  a  Double  Fluoride 
of  Aluminufn  and  Ammonium. 

Al2(SOj3  4-  12  NH4F  =  2(A1F3.  3NH4F)  +  3(NH4)2S04. 

Method. — Place  on  a  celluloid  slip  a  drop  of  a  moderately  dilute  neutral  solu- 
tion of  the  substance   to  be  tested,  and  to  it  add  several  small 
^  ^  S         fragments  of  ammonium  fluoride.     Very  minute  crystals   im- 
'  ^   ^  c>  mediately   separate.     The   preparation   is   set  aside  for  a  few 

^  ^  m    ^         seconds,  and  is  then  examined  near  the   circumference   of  the 
I  I  I  I  I  drop.     Small  but  clear  cut  octahedral  crystals  of  the  double 

\p\V.»O.Ol»x\»y». 

Fie  74  fluoride  of  ammonium  and  aluminum  will  be  seen  (Fig.  74). 

/Remarks. — The  solution  must  contain  no  appreciable  amount 
of  free  mineral  acid.  The  best  results  seem  to  be  obtained  when  the  test  drop 
is  neutral. 

Unless  the  reagent  is  present  in  excess,  a  compound  of  different  composition, 


and  Laboratory  Methods.  l-')3o 

containing  a  higher  percentage  of  aluminum,  separates  in  the  form  of    tiny  rods. 

Double  fluorides  of  aluminum  and  sodium,  potassium,  rubidium  and  cesium, 
having  the  general  formula  AlFg  •  3RF,  are  also  known.  Or  we  can  indicate  the 
composition  by  the  formula  R.^AlFg,  calling  the  compounds  fiuoaluminates,  a 
term  preferred  by  some  chemists. 

With  lithium  fluoride  the  double  fluoride  formed  is  less  soluble  than  in  the 
case  of  the  alkali  metals ;  its  crystallizing  power  is  also  considerably  less. 

Crystalline  double  fluorides  of  aluminum  with  copper,  nickel,  and  zinc  have 
been  described,  but  these  are  too  soluble  to  appear  under  the  conditions  which 
usually  obtain  in  an  analysis. 

In  testing  for  aluminum  with  ammonium  fluoride,  salts  of  lithium,  sodium, 
and  iron  must  be  absent. 

The  presence  of  silicon  and  analogous  elements  will  generally  seriously  com- 
plicate matters,  and  may  ruin  the  test,  owing  to  the  formation  of  fluosilicates. 
(See  ammonium  fluosilicate  tests,  under  Sodium  and  Barium.)  Aluminum  fluo- 
silicate  is  gelatinous,  and  does  not  crystallize. 

Testing  for  aluminum  with  ammonium  fluoride  generally  yields  results  a 
trifle  quicker  than  Method  I,  but  the  delicacy  of  the  reaction  is  but  very  little 
greater.  Moreover,  Method  II  is  subject  to  many  complications  and  interfer- 
ences, and  there  is  always  danger,  in  spite  of  great  care,  of  damaging  objectives 
by  the  corrosive  vapors  arising  from  the  test  drop.  For  these  reasons,  testing 
with  ammonium  fluoride  will  never  be  considered  as  being  as  satisfactory  as  the 
cesium  method.  One  of  the  chief  reasons  for  inserting  the  test  in  this  series  is 
the  fact  that  crystals  of  ammonium  fluoaluminate  may  occasionally  appear  when 
this  reagent  is  being  employed  for  other  purposes,  and  the  presence  of  aluminum 
is  not  yet  suspected. 

///.    Wif/i  Primary  Potassium  Sulphate,  HKSO4. 

This  salt,  added  to  sulphate  solutions  of  aluminum,  leads  to  the  formation 
and  separation  of  beautiful,  large  crystals  of  potassium  alum.  This  reaction  is 
an  elegant  and  satisfactory  one,  but  is  not  nearly  so  good  as  that  with  cesium 
sulphate,  for  the  reasons  which  have  already  been  stated  above,  still  with  due 
observance  of  the  precautions,  etc.,  there  given,  testing  for  aluminum  with 
primary  potassium  sulphate,  in  the  absence  of  the  cesium  salt,  can  be  depended 
upon  to  give  neat  and  satisfactory  tests. 

IV.   Staining  the  Precipitated  Hydroxide. 

Owing  to  the  fact  that  aluminum  hydroxide  has  the  property  of  uniting  with 
various  pigments  to  form  colored  compounds,  it  is  possible  to  detect  this  element 
by  staining  methods. 

Of  the  various  dyes  proposed,  Congo  red  and  cochineal  (Carmine)  have 
been  most  favorably  received,  the  former  being  the  better. 

An  aqueous  solution  of  the  dye,  added  to  freshly  precipitated  aluminum 
hydroxide,  stains  the  latter  a  more  or  less  deep  red. 

The  reaction  is  subject  to  many  errors,  is  of  very  limited  application,  and  is 
unsatisfactory  in  the  routine  work  of  chemical  analysis. 

Cornell  University,  E.  M.  ChamOT, 


1534  Journal  of  Applied  Microscopy 

Journal  of  ^^^  twenty-fourth  annual  meeting 

of  the  American  Microscopical  Society 
Applied    Microscopy  was  held  in  Denver,  Colorado,  on  Xhurs- 

day,   Friday  and   Saturday,  August  29 

lahoratorv   Methods  ^°'^^-     While  in  attendance  the  meeting 
LaOOraiOry   memous.           ^    ^^^  ^j^^   smallest  but   one  which   the 


Edited  by  L.  B.  ELLIOTT.  organization    has    held  ;    Prof.    H.    B. 

Ward,  the   secretary  of  the  society,  in- 


issued  Monthly  from  the  Publication  Department       forms  US  that  the  papers  presented  were 

of  the  Bausch  &  Lomb  Optical  Co.,  i.     •    r      •  •  i.  Ti.      i. 

Rochester,  N.  Y.  "ot   mfcnor  in   number  or   quality  to 

those  of  any  previous  meeting. 


SUBSCRIPTIONS  :  'Phg  Thursday  evening  meeting  took 

One  Dollar  per  Year.    To  Foreign  Countries,   $1.25       .,  „  r .„  „r ,; i  ,     .1  „  /^„i„„„  J„ 

per  Year,  in  Advance.  the  torm  oi  a  rcccption  by  the  Lolorado 

,     Microscopical  Society,  on  whose  invita- 

The  majority  of  our  subscribers  dislike  to  have  their  ,.  ,1         .  •  -.f  'lo' 

files  broken  in  case  they  fail  to  remit  at  the  expiration  of  tlOn    the  AmCnCaU    MlCrOSCOpiCal   SOCl- 

their  paid  subscription.     We   therefore  assume   that  no  g|-y  j^g^    Jj^    DenVCr.        AftCT     addrCSS    of 
interruption   in   the   series   is   desired,   unless   notice  to  .*  . 

discontinue  is  sent. wclcome  by  Dr.  A.  M.  Holmcs,  presi- 

SEPARATES  ^^"^  ^^  ^^^  Colorado   Society,   and  a 

One  hundred  separates  of  each  original  paper  accepted      respOUSC  by  Dr.  A.  M.  Bleilc,  the  retiring 

are  furnished  the  author,  gratis.  president  of  the  American  Microscopi- 

Separates  are   bound   in   special  cover  with  title.     A       *  .  .  .  .  ,  _? 

greater  number  can  be  had  at  cost  of  printing  the  extra      Cal  SoClCty,  the  inCOming  president.  Dr. 

copies  desired.  ^   pj   Eigcumann,  gave  the  annual  ad- 

dress on  the  solution  of  the  eel  problem. 
The  society  enjoyed  several  fine  musical  numbers  furnished  by  friends  of  the 
Colorado  Microscopical  Society,  and  at  the  close  of  the  program  a  very  pleasant 
informal  reception  with  refreshments  was  tendered  by  the  latter  organization. 

On  Friday  the  general  sessions  of  the  society  were  occupied  by  the  reading 
of  papers,  a  noteworthy  feature  of  which  was  an  address  by  Ex-President  Dr.  W. 
C.  Krauss  of  BufTalo,  on  "  The  Debt  of  American  Microscopy  to  Spencer  and 
ToUes."  The  committee  appointed  at  the  New  York  meeting  announced  the 
completion  of  the  Spencer-Tolles  Fund  to  the  limit  of  $1200,  as  set  a  year  ago, 
and  other  members  spoke  in  a  congratulatory  tone  on  the  completion  of  the 
fund.  The  report  of  the  committee  which  provided  that  a  specific  sum  should 
be  set  aside  yearly  from  the  interest  of  this  fund  for  the  encouragement  of 
microscopical  research  was  adopted,  and  the  conditions  of  the  grant  ordered 
printed  in  the  annual  volume. 

Resolutions  of  regret  at  the  death  of  Ex-President  E.  W.  Claypole  were  read 
and  ordered  spread  upon  the  minutes  of  the  society. 

The  following  officers  were  elected  for  the  year  1901-2  : 

President,  Charles  E.  Bessey,  University  of  Nebraska,  Lincoln,  Nebr. 

First  Vice-President,  E.  A.  Birge,  University  of  Wisconsin,  Madison,  Wis. 

Second  Vice-President,  John  Aspinwall,  New  York  City. 

Elective  members  of  the  Executive  Committee  :  Dr.  A.  M.  Holmes,  Denver, 
Colorado ;  Dr.  V.  A.  Latham,  Chicago,  III;  Mr.  G.  C.  Whipple,  New  York  City. 

Secretary,  Henry  B.  Ward,  University  of  Nebraska,  Lincoln,  Nebr. 

Treasurer,  J.  C.  Smith,  New  Orleans,  Louisiana. 

Custodian,  Magnus  Pfiaum,  Pittsburg,  Pa. 

The  session  of  Saturday  morning  was  held  while  en  route  to  Colorado 
Springs  on  an  excursion  tendered  the  society,  in  the  course  of  which  a  banquet 
at  the  Antlers  Hotel,  a  visit  to  Colorado  College,  a  carriage  ride  through  the 
Garden  of  the  Gods,  and  Manitou  were  enjoyed.  The  society  is  indebted  to  the 
Commercial  Clubs  of  Denver  and  Colorado  Springs  and  to  the  Colorado 
Microscopical  Society  for  the  many  hospitalities  extended  to  it. 

The  next  meeting  will  probably  be  held  in  Pittsburg.  We  would  urge  the 
desirability  of  every  one  interested  in  microscopical  work  becoming  a  member 
of  this  society. 


and  Laboratory  Methods.  1535 

CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Dangeard,  P.  A.  La  reproduction  sexuelle  des  The  most  interesting  portion  of  this 
Champignons.    Etude  critique  Le  Botaniste,  •       ,i     .        i  ■    i      j      i  -^i      .1 

7:  89-130  igoo.  paper    is    that    which    deals    with   the 

question  of    sexuaHty    in   Sph(zrotheca. 

Several  botanists  believe  that  in  Sphmrotheca  there  is  a  fusion  of  the  nucleus  of 

the  antheridium  with  that  of  the  oosphere,  and  that  this  fusion  is  followed  by  a 

fusion  of  the  two  nuclei  of  the  ascogonium  cell.     Prof.  Dangeard  claims  that  no 

nucleus  passes  from  the  antheridium  into  the  oosphere,  but  that  the  antheridium 

cell  with  its  nucleus  soon  disorganizes.     He    points  out  that  in    the  stage  of 

development   in  which  the    ascogonium   contains   two   nuclei,  the   antheridium 

should  not  have  any  nucleus,  if   the  theory    of  a    fusion    of    o.^^  nucleus  and 

antheridium  nucleus  is  correct.    According  to  his  observations  the  antheridium, 

at  this  stage,  still  retains  its  nucleus.     The  author  attempts,  on  other  grounds, 

to  disprove  a  repeated  nuclear  fusion  in  Spharotheca.  c.  j.  c. 

Stephani,  F.  Species  Hepaticarum.  Bull,  de  This  portion  of  the  writer's  work  on 
r  Herbier  Boissier,  pp.  27i;-3t;3.     Dec.  i8qq       tt         ,•  ^   •  r   n  ^ 

and  Apr.  1 900.  <  ^  ^^^  y^      Hepaticae  contains  a  very  full  account 

of    the     genus     Metzgeria,    sixty-four 

species  being  described.     Of  these,  two  species  are  cosmopolitan,  one  belongs  to 

northern   forests,  nine  are  native  in  tropical   and   sub-tropical   Africa,  eight  in 

tropical   Asia   and  Oceanica,  twenty-nine   in  tropical  America,  and  fifteen   in 

antarctic  regions.     Another  paper  (Extrait  des  Memoires  de  1'  Herbier  Boissier, 

pp.  1-46,  1900)   contains  a  full  account  of  Fossombronia,  with   descriptions   of 

forty  species.     Several  other  genera  are  described  in  this  paper.     The  writer 

believes  that  Fossombronia  is  the  connecting  link  between  the  thallose  and  leafy 

liverworts.       This  paper,   which    completes   Vol.   I,  Acrogyneae  der    "  Species 

Hepaticarum,"  has  an  index  of  thirteen  pages.  c.  j.  c. 

Butters,  F.  K.  A  Preliminary  List  of  Minne-  Material  from  which  this  list  is  made 
sota  Xylariaceae.      Minn.  Bot.  Studies,  Sec-       ,        ,  1    ^-        r       rr. 

ond  Ser.  3 :  563-567,  1901.  has  been  accumulating  for  fifteen  years. 

Specimens  of  all  the  forms  listed  have 
been  deposited  in  the  herbarium  of  the  University  of  Minnesota.  The  list  con- 
tains nineteen  species  distributed  among  five  genera,  as  follows  :  Niimmidaria, 
3;  Ustilina,  1;  ffypoxylofi,  12;  Daldinia,  2;  Xylaria,  1.  The  list  is  accom- 
panied by  notes.  c.  j.  c. 

Hirn,  Karl  E.  Monographie  und  Iconographie  This  is  the  most  important  work  on  the 
der  Oedogoniaceen.     Acta  Societatis  scien-  ,     ,  j       ^  ^c     t-u^ 

tiarum  Pennies,  27:  1-394,  pis.  1-64,  1900.      morphology    and      taxonomy    of    the 

Oedogoniaceae  which  has  yet  appeared. 

The  first  forty-seven  pages  are  devoted  to  structure  and  development.     Special 


1536  Journal  of  Applied  Microscopy 

attention  is  given  to  tlie  development  of  the  ring.  The  analytical  key  of  twenty- 
two  pages  is  in  Latin,  supplemented  by  notes  in  (ierman.  Descriptions  are 
given  of  244  species,  of  which  about  4(i  species,  with  o5  varieties,  are  new.  The 
illustrations  form  a  valuable  feature  of  the  work,  '2:>!l  of  the  244  species  being 
figured.     About  two-thirds  of  the  illustrations  are  original.  c.  j.  c. 

Buller,  AH.  R.    Contributions  to  our  Knowl-      Besides  malic  acid  and  its  salts,  many 

edge  of   the  Physiology  of  the  Spermatozoa  _  _  '  -' 

of  Ferns.    Ann.  of  Botany,  14 :    543-582,      Organic  and  inorganic  salts  in  the  cell 
'9°°-  sap  have  a  positive  chemotactic  stimu- 

lus for  the  spermatozoa  of  ferns,  but  malic  acid  exerts  a  stronger  influence  than 
any  other  substance  tested.  Sugar,  alcohols,  asparagin,  and  urea  do  not  attract. 
The  cell  sap  attracts  spermatozoids,  but  this  does  not  prove  that  the  sap  contains 
malic  acid  compounds,  because  the  attraction  takes  place  in  their  absence. 
Withdrawal  of  water  brings  the  spermatozoids  to  rest,  but  they  may  recover 
upon  the  reabsorption  of  water.  The  swarm  period  for  spermatozoa  of  Gymuo- 
gramme  Martens i  is  about  two  hours,  much  longer  than  was  previously  supposed. 
The  starch  in  the  vesicles  of  spermatozoa  disappears  during  the  swarm  period. 

c.  J.  c. 

Golden,  Katherine  E.     Aspergillus  oryzje  (Ahl-      Aspergillus  oryzcB  \s  a  mold  of  consid- 
burg)  Cohn.  Proc.  Indiana  Acad,  of  Science.  ,  ,  .•      i    •    ,  .    ^  •     ■ 

pp.  1-15,  12  figs.  i8q8.  erable   practical  mterest  because  it  is 

claimed  that  under  certain  conditions 
it  can  be  converted  into  a  yeast  and  that  it  can  give  rise  to  alcoholic  fermenta- 
tion. In  Japan  it  is  used  in  the  manufacture  of  sake',  and  Takamine,  a  Japanese 
chemist,  introduced  the  mold  into  the  United  States  hoping  to  do  away  with  the 
malting  of  grain  in  breweries.  He  took  out  a  patent  and  introduced  it  into  a 
brewery,  but  while  fermentation  took  place,  the  mold  has  not  superseded  yeast. 
The  present  paper  traces  the  life  history  in  some  detail.  Good  figures  are 
given  of  the  conidia  and  mycelium,  but  an  ascosporic  stage  could  not  be  found. 
Pure  cultures  made  from  material  obtained  from  Takamine  and  a  series  of  experi- 
ments have  led  the  writer  to  conclude  that  this  mold  is  never,  under  any  cir- 
cumstances, converted  into  a  yeast  and  that  it  does  not  have  the  power  of 
inducing  alcoholic  fermentation.  It  has  been  admitted  by  previous  investigators 
that  their  cultures  were  not  quite  pure.  c.  j.  c. 

Du  Sablon,  Leclerc.    Recherches  sur  les  fleurs      Violets,    and   especially   Viola   odorata, 
cleistogames.      Revue  Generale  de   Botan-       ,  .       •      1        1    •  ^  n 

ique.    12:  305-318,  figs.  11,1900.  ^ave    typical    cleistogamous     flowers. 

The  normal  flower,  which  appears 
early  in  the  spring,  has  a  handsome  corolla,  but  it  seldom  produces  good  seed. 
The  inconspicuous  cleistogamous  flowers  which  come  later,  usually  after  the  nor- 
mal flowers  have  disappeared,  produce  an  abundance  of  good  seed.  The  sta- 
mens are  larger  in  the  normal  iiowers  than  in  the  cleistogamous,  but  the  size  of 
the  pollen  grains  is  about  the  same  in  both.  The  structure  of  the  anther  wall  is 
quite  different,  the  normal  anther  having  the  usual  endothecium  with  lignified 
thickenings,  while  in  the  cleistogamous  flowers  the  endothecial  layer  retains  its 
nucleus  and  cytoplasm.  After  the  pollen  is  mature  there  is  a  resting  period  of 
various  duration.  Pollen  tubes  are  then  put  out  which  penetrate  the  wall  of  the 
anther  at  its  upper  part  where  there  is  a  region  of  small  cells  rich  in  protoplasm, 
a  tissue  comparable  to  the  conductive  tissue  of  the  style.  Oxalis  acctosella, 
Linaria  spuria  and  Leersia  oryzoides  were  also  studied.  In  typical  cleistogamous 
flowers  the  pollen  germinates  within  the  pollen  sac  and  the  structure  of  the  an- 
ther wall  is  modified  to  meet  the  new  mode  of  pollination.  In  Li/iaria  and 
Leersia,  where  the  pollen  was  not  observed  to  germinate  within  the  pollen  sac, 
the  anther  wall  has  the  same  structure  as  in  the  normal  flower.  c.  j.  c. 


and  Laboratory  Methods.  1537 


CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypoi.e,  Cornell  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Cloetta,  M.    Kann  das  Medicamentose  Eisen      The    white    mouse   was   used    for   the 
nur  im  Duodenum  resorbirt  werden  ?    Arch.       .  ^.        .  ,^,       ^  ^    •  1 

f.Exp.  Path.  u.  Pharm.  64:  363-367.  1900.      investigation.     The  form  of  iron  used 

was  a  specially  prepared  iron  nuclein. 

The  animals  were  fed  for  two  weeks  on  a  food  poor  in  iron  ;  then  the  alimentary 

canal  is  to  be  considered  free  from  iron.     For  several  days  succeeding,  the  food 

was  mixed  with  iron  nuclein.     The  animals  were  killed  with  ether,  the  intestine 

hardened  at  once  in  absolute  alcohol,  and   subsequently  treated  by  Quincke's 

method  (see  Arch.  f.  Exp.  Path,  and  Pharm.  37:  183). 

After  staining  with   ammonium  sulphide  it  was  found   necessary  to  let  the 

tissue  lie  in  glycerin  before  examination,  since  the  granules  became  more  distinct. 

An  aqueous  solution  of  safranin,  which  is  not  changed  by  alkalies,  was  used  for 

a  double  stain.     The  dark  green  granules  stood   out  markedly  from  the  yellow 

red  protoplasm.     In  applying  the  Berlin  blue  reaction  it  is  necessary  to  put  the 

section  first  into  weak   alcohol  containing  hydrogen  dioxide,  for  24  hours.     The 

intestine  hardened  in  absolute   alcohol  was  cut   into  1  cm.  pieces.     These  were 

embedded  in  paraffin,  and   the  whole  canal  thus  cut   serially.     All   preparations 

proved  that  the  iron  reaction  extended  far  beyond  the  limits  of  the  duodenum. 

E.  J.  c. 

Wilson,  J.  T.  -^  A    New   System   of   Obtaining  r^^^  method  was  suggested  while  using 

Directing  Marks  in  Microscopical    Sections  °°                                   ° 

for  the  Purpose  of  Reconstruction  by  Wax-  the  Born-Peter  process,  and  is  a  modifi- 

plate  Modeling.     Zeit.f.wiss.  Micros,  u.  f.  ^^^^^^    ^f    ^^is   in    the    following   way: 

Mikros.  lechn.  17:   169-177,  1900.  _                              °          ■' 

Instead  of  depending  entirely  upon  the 
filling  of  ruled  lines  in  a  glass  plate  with  pigment,  darkly  colored,  perfectly 
straight  organic  filaments  are  placed  in  the  paraffin  together  with  the  tissue  to 
be  embedded.  The  materials  used  were  some  of  the  long,  slender  root  bundles 
of  the  human  cauda  equina  ;  the  intraspinal  roots  of  the  fifth  sacral  and  coccygeal 
nerves  are  very  long  and  fine,  but  if  more  delicate  strands  are  required  they  may 
easily  be  separated  from  other  nerve  roots.  The  absence  of  branching  and 
uniform  caliber,  together  with  their  delicacy,  make  these  very  favorable  for  the 
purpose.  Portions  of  such  bundles  of  10-12  centimeters  in  length  are  suspended 
by  a  thread,  and  carry  on  a  thread  at  the  other  end  weight  enough  to  keep  the 
strand  perfectly  straight  after  immersion  in  fluid,  but  not  enough  to  stretch  it. 
These  pieces  are  now  hung  in  a  vessel  of  1  per  cent,  osmic  acid  to  blacken  the 
myelin  of  the  nerve  fibers.  Then  they  are  carried  in  a  like  manner  through  the 
alcohols  and  xylol,  and  infiltrated  with  paraffin  in  a  test  tube.     These  strands 


1538  Journal  of  Applied  Microscopy 

are  lifted  carefully  out  and  allowed  to  harden,  and  can  be  kept  till  required  for 
use. 

For  embedding,  a  glass  base  plate  and  the  usual  Naples  L-shaped  embed- 
ding bars  are  required.  The  glass  plate  may  be  constructed  in  the  laboratory 
from  plane-surfaced  glass.  It  is  convenient  to  have  it  of  such  a  shape  as  to 
replace  easily  the  stage  of  a  dissecting  microscope,  but  it  should  not  be  more 
than  2-0  mm.  thick.  The  surfaces  should  be  plane,  and  it  is  advantageous  to 
have  a  central,  rectangular  outline  on  the  upper  surface  with  the  sides  measur- 
ing 2  cm.  each.  This  outline  should  be  blackened.  On  the  under  side  of  this 
area  a  series  of  deep  lines  should  be  engraved  and  blackened ;  they  must  be 
accurately  parallel  to  two  of  the  sides  of  the  quadilateral  figure  on  the  upper  side 
and  to  each  other,  and  placed  at  intervals  of  1-2  mm.  The  embedding  bars 
should  be  exactly  rectangular  throughout,  and  have  their  arms  2  cm.  in  length. 

Before  proceeding  to  embed  the  object,  the  glass  plate  must  be  so  placed 
that  it  can  be  heated  from  below,  and  it  and  the  bars  are  slightly  rubbed  with 
glycerin  to  facilitate  the  removal  of  the  paraftin  block.  Two  or  more  of  the 
blackened  strands  of  nerve  tissue  are  laid  very  carefully  on  the  glass  coincident 
with  two  of  the  parallel  blackened  lines.  The  base  plate  is  now  heated  to  fix 
the  paraffin-covered  strands  in  place,  and  to  arrange  them  perfectly  coincident 
with  the  engraved  lines.  One  of  the  embedding  bars  is  then  so  placed  that  the 
cross-arm  will  limit  the  basal  plane  of  the  future  paraffin  block,  corresponding 
to  or  parallel  to  the  plane  of  sectioning.  Both  bars  slightly  cover  the  ends  of 
the  nerve  strands.  A  moderate  weight  of  a  kilo,  in  the  shape  of  an  iron  bar,  is 
laid  on  the  upper  surface  of  the  embedding  bars,  and  the  plate  is  again  heated 
till  the  paraffin  is  melted,  and  either  allowed  to  cool  or  the  process  of  embedding 
completed.  The  weighting  of  the  bars  allows  complete  flattening  of  the  ends  of 
the  strands  of  tissue,  which  are  held  in  place  by  the  bases  of  the  bars,  and  hence 
their  very  slight  thickness  does  not  interfere  with  the  angle  of  the  surface  of  the 
bars.  If  there  is  objection  to  this  process,  the  filaments  may  be  held  down  by 
two  pieces  of  lead  placed  inside  the  bars.  If  the  plate  has  been  allowed  to  cool 
it  must  be  again  warmed  to  the  melting  point  of  paraffin  ;  after  filling  the 
chamber  with  melted  paraffin  the  object  must  be  carefully  oriented,  with  refer- 
ence to  the  lines  and  strands,  under  a  dissecting  microscope,  if  necessary,  but 
the  plate  must  be  kept  warm  till  everything  is  completed.  Rapid  cooling  in  iced 
water  follows,  with  care  to  prevent  cupping  of  the  block  by  the  addition  of  drops 
of  melted  paraffin,  and  manipulation  with  a  hot  needle. 

The  advantages  of  this  method  are  :  1.  No  special  apparatus  is  required  be- 
yond what  is  found  in  any  laboratory,  even  the  glass  plate  may  be  prepared  by  an 
ordinary  engraving  diamond  if  necessary.  2.  None  of  the  operations  require 
any  special  dexterity,  and  all  may  be  accomplished  by  anyone  with  certainty. 
3.  Very  little  expenditure  of  time  is  required  beyond  that  of  ordinary  embedding 
after  the  stock  of  prepared  nerve  is  laid  in.  4.  The  actual  directing  marks  in 
each  section  are  brought  as  close  as  desired  to  the  object.  5.  Whenever  the 
embedding  has  taken  place  the  importance  of  the  directing  plane  disappears,  the 
only  plane  of  importance  being  the  future  base  of  the  object  block.  6.  The 
necessity  for  scratching  the  paraffin  block  by  a  "  Ritzer  "  or  for  the  alternative 


and  Laboratory  Methods.  1539 

Born-Peter  ridges.  7.  There  is  no  necessity  for  filling  up  the  scratches,  or  for 
coating  the  ridges  with  amorphous  color,  nor  for  the  addition  of  color,  lacquer, 
or  any  foreign  substance.  8.  Each  section  bears  its  directing  marks  in  the  shape 
of  circumscribed  black  spots.  9.  The  detecting  strands  cause  no  inconvenience 
at  any  time  in  the  processes,  and  their  axes  are  for  all  practical  purposes  as 
accurately  perpendicular  to  the  plane  of  section  as  are  the  colored  ridges  of  the 
Born-Peter  block.  10.  It  is  possible,  but  not  yet  fully  tested,  to  apply  the  pro- 
cess to  celloidin.  a.  m.  c. 

Stepanow,  E.  M.    Eine  neue  Einbettungsmeth-      The  author  uses  a  solution  of  celloidin 
od  in   Celloidin.      Zeit.  f.  wiss.   Mikros.   u.       .         ,  ....         ,  111 

f.  Mikros.  Techn.  17:   185-191,  1900.  1"   clo^e  Oil  with   ether  and  absolute 

alcohol  in  the  following  proportions : 
Celloidin  (shavings  very  fine  and  well  dried),  1.5  gr.;  clove  oil,  5.0  c.  c;  ether, 
20.0  (of  0.720  sp.  gr.)  ;  alcohol  absolute,  added  by  drops,  1.0  c.c.  One  c.  c.  of  this 
mixture  contains  more  than  6  per  cent,  celloidin,  corresponding  to  the  weakest 
used  solution.  By  the  addition  of  ether  and  alcohol  much  thinner  liquid  can  be 
obtained,  and  by  concentration  thicker  up  to  35  per  cent.  The  process  with  the 
"  normal  "  solution  (6  per  cent.)  is  as  follows  :  Tissue  well  hardened  in  alcohol, 
dehydrated,  and  freed  from  superfluous  alcohol  by  touching  it  lightly  with  filter 
paper,  is  put  in  a  glass-stoppered  bottle  containing  4-5  c.  c.  of  clove-oil-ether- 
celloidin.  According  to  the  size  of  the  pieces,  it  is  kept  here  from  one  to  six 
hours  or  more,  then  the  bottle  is  uncorked  and  put  under  an  inverted  glass, 
leaving  the  solution  to  evaporate  for  four  to  six  or  more  hours.  This  thickened 
mixture  is  poured  into  a  small,  freely  hanging  filter  of  fine  silk  paper ;  the  mass 
is  then  either  left  open  or  loosely  closed  to  reduce  it  to  embedding  consistency. 
The  process  may  be  hastened  by  keeping  the  filter  in  a  warm  place.  The  clear- 
ness and  dryness  of  the  substances  are  the  best  assurances  for  a  good  embedding 
matrix.  This  thickening  takes  place  in  from  four  to  six  hours  ;  the  object  is 
then  cut  out  from  the  surrounding  mass.  Further  preparations  may  follow  one 
of  several  lines :  1.  If  the  sections  are  to  be  cut  in  alcohol  the  material  is 
mounted  on  a  cork  which  has  been  well  coated  with  celloidin  and  then  put 
for  twenty-four  hours  in  70-85  per  cent,  alcohol.  Treatment  for  two  to  three 
hours  in  chloroform,  is  equally  sure  and  much  quicker.  2.  The  object,  fastened 
on  a  piece  of  wood,  is  made  firm,  by  means  of  a  needle,  to  the  cork  of  a  bottle 
containing  chloroform,  for  two  to  six  hours,  then  the  sections  are  cut  with  a  dry 
knife  and  transferred  with  oil  to  a  slide.  Sections  10,  7.5,  and  5 /<  can  be  cut 
this  way,  and  the  block  is  always  transparent.  3.  The  best  method  is  to  put  the 
freshly  embedded  object  into  benzol,  and  there  it  hardens.  Such  an  object  may 
be  put  directly  into  anethol  (later  into  anethol-paraffin)  ;  into  a  solution  of 
paraffin  in  benzol,  and  then  into  liquid  paraffin  ;  into  cedar  oil  for  dry  sections, 
or  into  85  per  cent,  alcohol  for  wet  sections. 

The  chief  advantages  of  the  method  are:  1.  The  manipulations  are  as  sim- 
ple as  in  the  ordinary  methods,  and  fewer.  2.  The  imbibition  is  more  quickly 
completed  (twenty-four  hours  or  fewer).  3.  The  embedding  is  so  thorough  that 
sections  can  be  cut  3  //  in  thickness.  4.  The  control  of  the  embedding  processes 
is  easily  indicated  by  transparency.     5.  After  these  preliminaries  the  tissues  may 


1540  Journal  of  Applied  Microscopy 

be  finished  by  many  various  methods.  6.  The  possibility  of  embedding  by  anilin 
oil  without  higher  grades  of  alcohol  than  70-80  per  cent.  7.  The  short  time 
needed  in  each  embedding  solution.  a.  m.  c. 

Eisen,  G.     The  Spermatogenesis  of  Batrach-      Earlier    investigations    were  made   on 

oseps.     Jour.  Morph.  Vol.   17,  1900.  ^     •    1    1       j         j  •      -1-1  •       >  1 

•^  material  hardened  m  Flemming  s  and 

Hermann's  fluids  ;  later  Heidenhain's  sublimate-acetic  mixture  with  and  without 
formol  was  tested.  Others  also,  as  Hermann's  and  Flemming's  fluids  mixed  with 
sublimate  or  palladium  chloride,  vanadium  chloride,  uranium  chloride  and  osmium 
chloride.  Except  the  latter  the  author  discarded  them  all.  He  believes  to  have 
proved  that  every  mixture  containing  platinum  chloride  or  osmic  acid  completely 
destroyed  the  outer  cells.  As  the  testis  of  Batrachoseps  is  very  small  and  pos- 
sesses but  few  cell  layers,  all  such  fixatives  must  be  rejected.  Platinum  chlor- 
ide is  more  injurious  than  osmic  acid,  since  it  destroys  the  chromatin,  while  the 
latter  injures  the  fine  structure  of  the  cytoplasm.  Osmium  chloride  is  a  very  val- 
uable fixative,  especially  in  1/2  to  1/10  per  cent,  solutions,  although  it  also  pos- 
sesses the  property  of  blackening  the  tissues  to  a  less  degree  than  osmic  acid. 
Three  to  twelve  hours  are  necessary  for  proper  fixation,  no  shrinking  and  no 
blackening  occurs,  and  the  outer  layers  of  cells  are  in  good  condition  as  well  as 
the  inner  ones.  An  hour's  washing  in  water  follows  the  treatment  with  alcohol 
and  bergamot  oil  and  xylol,  again  into  bergamot  oil  and  embedded  in  paraffin. 
Sections  4  to  6  //  thick  are  cut  that  every  cell  may  be  sectioned.  This  the  author 
holds  to  be  an  essential  for  good  staining. 

Benda's  iron  haematoxylin  combined  with  congo-red  was  largely  used.  The 
sections  left  for  24  hours  in  the  following  solution  :  ferric  sulphate  according  to 
the  German  pharmacopeia  diluted  with  six  times  its  volume  of  water,  then  in 
concentrated  haematoxylin  solution  containing  10  per  cent,  alcohol  for  48  to  72 
hours.  The  best  results  came  from  the  longer  action  of  the  stain.  The 
differentiation  is  effected  by  10  per  cent,  acetic  acid  containing  a  very 
small  quantity  of  liquor  ferri,  in  10  to  20  minutes,  washed  as  rapidly  as  pos- 
sible, cleared  in  bergamot  oil  and  mounted  in  xylol  balsam.  A  triple  stain  with 
congo-red,  thionin  and  ruthenium  red  can  be  used  also.  The  sections  remain  a 
few  seconds  in  a  weak  aqueous  solution  of  congo-red,  then  about  10  minutes  in 
thionin  in  water,  and  finally  differentiated  by  a  very  weak  aqueous  solution  of 
ruthenium  red.  a.  m.  c. 

Ra'dl,  Em.    Arthropod  Vision.    Zeitschr.  wiss.      The  author  considers  that  not  enough 
Zool.  67:   557-598,  I   pi-,   1900  (review  in      importance  has  been  laid  on  the  study 

Journ.  Roy.  Micr.  Soc.  pt.  i,  pi.  ^6,  igoi).  r    ^l  i.  c  4.1, 

■'  •'  K     .  F    J  '    ^    /  q£   l-]^g    nerve   centers    of  the    eye    as 

well  as  of  its  dioptric  apparatus.  He  believes  the  key  to  the  problem  of 
arthropod  vision  to  lie  in  the  central  rather  than  peripheral  organs.  He  has 
exhaustively  studied  the  eye  and  optic  tract  in  Squilla  mantis.  After  a  brief  de- 
scription of  the  external  appearances  of  the  eye,  the  phenomenon  of  "  double 
eyes  "  in  arthropods  is  fully  considered.  The  eye  itself  is  described  briefly. 
Each  ommatidium  gives  off  seven  nerve  fibrils,  which  unite  in  a  bundle ;  as 
these  pass  across  the  space  between  the  basal  membrane  of  the  eye  and  the  first 
ganglion,  those  from  neighboring  ommatidia  unite  to  form  larger  bundles.     In 


and  Laboratory  Methods.  1541 

the  first  ganglion  these  bundles  break  up  into  constitutional  bundles.  This 
ganglion,  like  the  eye,  is  made  up  of  two  halves  connected  by  a  thick  bunch  of 
vertical  nerve  fibers.  The  ganglia  are  complex.  An  especially  important 
element  is  the  granular  layer,  containing  darkly  staining  bodies,  "  nerve  nodes  " 
(Nervennoden),  corresponding  to  the  ommatidia  in  number.  These  consist  of 
neuroglia  fibrils,  which  come  from  several  ommatidia ;  the  fibrils  do  not  end,  but 
pass  on  to  other  ganglia.  As  the  fibers  leave  the  first  ganglion  to  pass  on  to  the 
•second,  they  cross  so  that  the  right  becomes  left  and  vice  versa.  The  import- 
ance of  this  crossing  lies,  according  to  the  author,  in  the  varying  lengths  of  the 
fibers,  which  have  a  physiological  significance,  explained  as  follows  :  Suppose 
the  eye  to  be  stimulated  in  such  a  way  that  a  certain  set  of  retinulae  receive  an 
equal  impulse.  These  impulses  pass  down  the  fibrils  to  the  second  ganglion, 
but  owing  to  variation  in  the  length  of  these  fibers  will  arrive  at  different  times. 
If  it  is  supposed  that  a  stimulus  affects  one  ommatidium  only,  a  successive 
series  of  changes  in  the  nerve  centers  would  follow,  since  each  ommatidium  has 
seven  nerve  fibrils  and  each  has  a  different  length  from  the  others.  This  theory 
of  arthropod  vision  was  reached  by  the  author  by  a  process  of  induction,  but  he 
believes  he  is  supported  by  the  theories  of  other  authors  who  have  based  their 
conclusions  on  theoretical  grounds.  a.  m.  c. 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Seeliger,  0.  Tierleben  der  Tiefsee.  49  pp.,  i  This  brief  treatise  on  the  abyssal  life 
Taf.,  Verlag  von  W.  Engelmann,  Leipzig,  ^f  ^.^g  ^^^^^  ^^^^^^  ^^g  subject  in  SUC- 
1901.     Preis  Mk.  2. 

cinct  fashion  in  the  light  of  the  latest 

investigations  in  this  field  of  zoological  exploration.     A  short  historical  sketch 

is  followed   by  an   explanation  of  the  factors   of  the  environment,  such  as  the 

chemical  condition,  pressure,  temperature,  and  light.     The  problems  that  center 

about  the  coloration,  phosphorescence,  and  vision  of  deep-sea  animals  are  also 

discussed.  c.  a.  k. 

Nutting,  C.  C.  The  Hydroids  of  the  Wood's  Students  at  marine  laboratories  will 
Hon  Region.    Bull.  U.  S.  Fish  Commission      welcome  Professor  Nutting's  paper  on 

for   I099,  pp.  325-306.       I9OI.  o         r     r 

these  favorite  forms  of  seaside  study. 
The  descriptions  are  brief,  but  illustrations,  mainly  original,  are  abundant,  and 
very  full  keys  are  provided  for  both  hydroid  and  medusa  stages.  In  all,  112 
different  forms  are  described  from  Wood's  HoU  and  Newport  indicating  the 
richness  of  the  hydroid  fauna  in  that  region.  In  providing  for  the  preparation 
and  publication  of  a  series  of  papers,  of  which  this  is  one,  upon  the  local  fauna 
at  the  Wood's  Holl  Station,  the  United  States  Fish  Commission  is  rendering 
valuable  aid  to  all  American  students  of  marine  life.  c.  a.  k. 


1542 


Journal  of  Applied  Microscopy 


Petersen,  C.  G.  J.    An  Otter-Seine  for  the  Ex-      The  problem   of  capturing  the  larger 
ploration  of  the  Deeper  Seas.    Rep.  Danish  ,  ^.        •    u    !_•..      ^        c  ^\ 

Biol.  Sta.,  8:  24  pp.,  4to,  10  figs.,  1899.  and  more  active  inhabitants  of  the  sea 

bottom  has  been  attacked  in  the  past 
by  large  beam  trawls.  These  are  limited  in  size,  the  largest  being  10  to  15  feet 
beam,  and  are  difficult  to  manceuver,  even  with  large  vessels.  The  beamless 
trawling  gear  used  of  late  by  North  Sea  trawlers  has  been  adapted  by  Dr. 
Petersen  to  biological  work.     With  a  32-foot  steam  launch  he  operated,  in  depths 


Sailing  Vessel  with  Otter  Drag-Seine. 

of  1  to  300  fathoms,  a  trawling  gear  of  this  pattern  whose  spread  at  ordinary 
speed  was  12  to  16  feet.  With  a  more  powerful  craft  and  larger  trawl  a  much 
greater  speed  can  be  secured.  Two  boards,  29  x  32  inches,  with  iron  runners, 
are  attached  to  the  ends  of  the  wings  of  the  bag,  which  is  provided  with  a  light, 
collapsible  funnel.  The  mouth  of  the  bag  is  kept  in  shape  by  suitable  weights 
and  Norwegian  glass  floats.  From  the  boards  pass  bridles,  eight  fathoms  long,  to 
the  vertex  of  the  crow-foot,  where  a  shackle,  float,  and  lead  keep  the  bridles 
from  twisting.  From  the  vertex  a  single  line  passes  aboard  ship.  A  proper  ad- 
justment of  speed  is  necessary  to  secure  the  most  successful  operation  of  the 
trawl.  The  figure  gives  some  idea  of  the  trawl  in  action.  Full  directions  for 
knitting  the  bag  are  given  in  the  original  article.  The  catches  of  this  apparatus 
are  said  to  be  phenomenal.  c.  a.  k. 

Bock,  M.  de.     Observations  Anatomiques  et      ^^^e  author  seeks  to  clear  up  some  con- 
Histologiques  sur  les  Oligochetes  Speciale- 

ment  sur  leur  Systeme   Musculaire.    Rev.      troverted  points  concerning  the  muscu- 
Suisse  de  Zool.  9:  1-41,  pl.  i,  2,  1901.  \^x^^x^  of   the   OUgochceta,  employing  as 

objects  of  study  a  number  of  different  terricolous  and  limicolous  species.  The 
study  of  sections  was  supplemented  by  the  examination  of  material  prepared 
by  maceration  for  several  months  in  a  ^  to  1  per  cent,  solution  of  bichromate 
of  potash  and  then  disassociated,  after  several  weeks,  in  glycerin.  The  silver 
nitrate  method  of  Dekhuyzen  was  employed  to  demonstrate  the  membrane  of  the 
so-called  sarcolemma.  The  muscle  columns  (colonnes  musculaires  of  Cerfon- 
taine)  which  constitute  the  musculature  of  the  body  wall  of  the  Oligochceta  are 
composed  of  bundles,  each  containing  a  small  number  of  fibers,  and  are  enclosed 


and  Laboratory  Methods.  1543 

in  a  delicate  membrane.  The  fibers  in  turn  are  made  up  of  muscle  elements 
which  cannot  be  further  divided.  The  muscle  elements  arise  in  myogenous 
cells,  each  cell  producing  several  of  the  elements,  though  neither  the  fiber  nor 
the  muscle  column  represents  a  single  cell.  In  the  LumbricidiT;  the  muscle  col- 
umns unite  in  well  defined  compartments,  most  pronounced  in  the  longitudinal 
series,  each  with  distinct  connective  tissue  membrane  in  which  nuclei  but  no 
cell  boundaries  are  found.  This  connective  tissue  serves  to  reduce  the  pres- 
sure and  friction  in  muscular  movement,  and  in  limicolous  species  it  forms  a 
compact  layer  beneath  the  peritoneum.  The  nuclei  of  the  muscular  tissue  are 
distinguished  from  those  of  the  connective  tissue  by  their  larger  size,  and  in 
limicolous  forms  are  often  pedunculate  and  grouped  along  the  lateral  line.  No 
nerve  runs  along  this  line,  though  a  fine  canal,  probably  a  lymphatic  vessel,  lies 
among  the  nuclei  cells.  c.  a.  k. 

Sabin  Dr.  Florence  R.    An  Atlas  of  the  Me-      ^^is  atlas  was   prepared  for  the  study 
duUa  and  Mid-brain.     A   Laboratory   Man-  . 

ual.      Pp.  123;  52  figs.;    8  pi.,  1901.    The      of  the  human  brain,  and  it  will  prove 
Friedenwald  Co.,  Baltimore.    ^1.75.  to  be  a  valuable   aid  in  the  laboratory 

for  the  study  of  the  brain  of  lower  types.  The  abundant  drawings  of  typical 
sections,  and  above  all  the  elegant  colored  plates  of  the  medulla  and  mid-brain 
regions,  with  their  several  parts  shown  in  relief,  will  serve  to  elucidate  these 
difficult  and  complicated  parts  of  the  brain.  It  is  stated  that  reproductions  in 
wax  from  the  studio  of  Zeigler,  in  Freiburg,  will  be  available  witliin  the  year. 
The  material  studied  was  preserved  in  Mliller's  fluid  and  stained  by  the  Wright- 
Pal  method.  Sections  of  70  jx  thickness  were  made  in  a  horizontal  plane  and 
every  other  one  used  as  the  basis  for  reconstruction  in  wax  by  the  Born  method. 
Wax  plates  two  millimeters  in  thickness  were  used,  thus  giving  a  magnification 
of  14.5.  The  wax  was  composed  of  19  parts  ordinary  beeswax  and  1  part  resin. 
To  facilitate  the  counting  of  the  sections  in  the  model,  every  fifth  plate  was  made 
black  by  an  admixture  of  lampblack.  Melted  wax  of  a  weight  sufficient  to  cast 
a  plate  of  the  desired  size  is  poured  through  a  strainer  into  a  tarred  receptacle, 
and  then  emptied  upon  a  pan  of  hot  water,  bubbles  being  removed  by  a  strong 
gas  flame.  When  firm,  the  plate  is  removed  to  a  level  surface  to  harden.  The 
drawings  from  the  sections  were  made  by  the  aid  of  a  projection  apparatus 
and  an  electric  lamp,  the  image  being  received  upon  a  rigid  but  movable  screen, 
and  care  being  taken  to  preserve  a  uniform  magnification  and  orientation  of  the 
sections.  Drawings  are  then  transferred  to  the  wax  plates  by  carbon  paper,  and 
finished  in  oil  paints.  The  sections  thus  outlined  are  cut  from  the  plates, 
which  are  slightly  warmed,  and  placed  upon  a  sheet  of  glass,  a  thin,  narrow- 
bladed  knife  being  used  for  the  cutting.  The  sections,  and  the  shells  also,  are 
then  piled  up  in  proper  relation  and  their  edges  fused,  thus  giving  a  model  of  the 
external  form  of  the  organ  and  a  mould  for  a  plaster  cast  of  the  same.  The 
different  structures  of  the  organs  were  then  modeled  separately,  and  the  whole 
so  united  as  to  display  the  true  spatial  relations  of  the  various  nuclei  and  fiber 
tracts.  The  result,  even  as  shown  in  the  figures,  will  serve  to  elucidate  and 
simplify  greatly  the  study  and  the  demonstration  of  the  structure  of  these  im- 
portant but  very  complex  organs.  c.  a.  k. 


1544  Journal  of  Applied  Microscopy 

NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Hirschmann,  A.  Pathologisch-anatomische  Stu-  This  paper  is  based  upon  a  histological 
dien  iiber  acute  u.chronische  laryngitis  nicht-  ,         ,  r  ,  ,  .   , 

specifischen  Ursprungs  nebst  Bemerkungen  Study   of    twenty-four   larynges     which 

iiber  Vorkommen  von  Plasma-  und  Mastzel-  were  the  seat  either  of  acute  or  chronic 

len.     Virchow's  Archiv  fiir  path.  Anat.  164 :  .    ^  .  ^  r         i  i 

541-569,  1901.  inflammation.      Cases    of    tuberculous 

or  syphilitic  laryngitis  were  excluded. 
Formalin  was  the  fixing  agent  employed.  It  was  found  that  mast-cells  are  as 
well  preserved  by  formalin  as  by  alcohol.  Earlier  writers  have  claimed  that 
alcohol  yields  the  best  results  in  the  study  of  mast-cells.  The  tissues  were 
embedded  in  parafBn.  Sections  were  stained  with  haematoxylin  and  eosin, 
orcein,  thionin,  and  polychrome  methylen  blue. 

Plasma  cells  were  not  found  either  in  the  normal  or  inflamed  larynx. 
Laryngitis  is  usually  due  to  an  irritant  which  is  too  weak  to  cause  an  extensive 
destruction  of  cells.  There  is  generally  a  marked  emigration  of  leucocytes  and 
proliferation  of  cells.  The  author  found  mast-cells  in  every  case  of  laryngitis 
examined.  This  agrees  with  the  view  that  these  cells  are  found  especially  in 
those  organs  which  are  the  seat  of  a  mild,  chronic  inflammation.  He  holds  that 
mast-cells  are  due  to  the  long  continued  action  of  a  mild  irritant,  while  plasma 
cells  are  due  to  the  long  continued  action  of  a  strong  irritant. 

Hirschmann  claims  that  mast-cells  are  derived  from  leucocytes.  Large  mono- 
nuclear leucocytes  wander  from  the  blood  vessels  into  an  inflammatory  area  and 
are  there  converted  into  mast-cells  by  ingesting  the  products  of  inflammation.  It 
is  these  products  of  inflammation  which  give  the  cell  its  characteristic  color. 
The  different  forms  of  mast-cells  which  have  been  described  are  simply  different 
stages  in  the  development  of  the  cell.  For  the  demonstration  of  mast-cells 
either  thionin  or  aqueous  methylen  blue  gives  as  good  results  as  polychrome 
methylen  blue.  j.  h.  p. 

Melnikow=Raswedenkow.     Studien    uber  den      The  black  jaundice  of  Tyrol  is  herein 
Jb^chinococcus  alveolans  sive  multilocularis.  •'  ^ 

Ziegler's  Beitriige  zur  path.  Anat.,  Supple-      pretty  clearly  established  as  a  separate 
mentheft4:  1-295.     i9°'-  type  of  echinococcus  disease,  endemic 

in  Tyrol.  The  multilocular  type  is  found  also  in  various  parts  of  Germany  and 
Russia  in  such  degree,  as  to  play  some  part  in  the  differential  diagnosis  of  liver 
affections,  such  as  cancer  and  cirrhosis.  In  all,  2;>5  cases  are  reported.  Mel- 
nikow-Raswedenkow  presents  the  protocols  of  101  cases,  besides  8  cases  in 
animals,  and  seeks  to  establish  the  parasitology,  general  pathology,  and  patho- 
logical anatomy  of  the  affection  which  he  prefers  to  call  alveolar  echinococcus 
disease. 

As  early  as  185G,  Virchow  had  made  clear  the  parasitic  nature  of  what  was 
before  confused  with  colloid,  or  even  with  colloid  cancer.  It  is  interesting  to 
find  that  at  least  the  alveolar  type  of  echinococcus  disease   is  hardly  surpassed 


and  Laboratory  Methods.  1545 

in  malignancy  by  eitlier  cancer  or  tuberculosis.  This  is  the  more  surprising  in 
that  the  cestodes  are  as  a  rule,  though  dangerous,  still  far  from  malignant.  But 
for  many  years  the  differences  between  the  many  small  chambers  of  the  alveolar 
type  and  the  great  hydatids  were  set  down  as  the  effects  of  individual  variation, 
and  the  Taenia  echinococcus  v.  Siebold  was  held  responsible  for  both  species  of 
reaction. 

It  is  of  course  somewhat  out  of  fashion  in  these  days  to  work  upon  parasites 
without  recourse  to  experiment.  Fresh  material  was,  however,  not  accessible  to 
Malnikow-Raswedenkow.  And  by  histological  study  alone,  several  capital  points 
have  been  brought  out.  No  intermediate  host  appears  to  be  required  (a  char- 
acter resembling  the  trematode  rather  than  the  usual  cestode  type  of  attack). 
The  embryo,  doubtless  of  intestinal  origin,  makes  its  way  by  the  blood  stream 
to  its  favorite  site  in  some  small  vein  just  beneath  Glisson's  capsule.  Here  a 
multilocular  chitinous  structure  is  formed,  wholly  analagous  with  the  mature 
segment  (proglottis)  of  the  tape-worm.  The  chitinous  walls  are  lined  not  only 
within,  as  in  the  great  single  hydatids,  but  also  externally  with  a  layer  of 
granular  protoplasm  in  which  are  produced  not  only  scoleces,  as  in  the  hydatid, 
but  also  young  parasite  forms,  without  capsule,  and  ovoid  embryos,  with  capsule. 

By  release  from  the  outer  wall  of  the  cyst,  metastasis  in  this  form  is  rendered 
much  easier  than  in  the  unilocular  type.  The  discharged  embryos,  in  case  they 
do  not  forthwith  succumb  to  phagocytosis  within  the  tissue  spaces,  gain  entrance 
to  some  blood  vessel,  or  perhaps  a  bronchiole,  and  there  form  more  chitinous 
cysts.  As  a  consequence  of  their  more  intimate  contact  with  the  body  fluids, 
the  new  cysts  lose  in  virulence  and  usually  remain  sterile.  It  is  probable,  more- 
over, that  feeding  experiments  may  fail  for  similar  reasons  if  the  material  is 
metastatic. 

The  affection  works  by  no  means  simply  through  pressure  or  mere  mechanical 
destruction,  but  toxically  as  well.  Proliferation,  phagocytosis,  and  local  tissue- 
necrosis  occur,  and  in  places  true  granulomata  are  formed,  characterized  by  the 
presence  of  lymphoid  cells,  epithelioid  cells,  and  giant  cells,  with  caseous 
degeneration. 

The  technique  employed  is  in  brief  as  follows  : 

1.  Fix  in  4  per  cent,  formaldehyde,  24  hours. 

2.  Harden  in  alcohols  of  increasing  strength,  cut  from  celloidin. 

3.  Place  from  water  into  Weigert's  elastic  tissue  stain,  3(J  minutes. 

4.  Wash,  decolorize  in   90    per    cent,   alcohol  2   minutes,  dip  in   weak 

lithium  carbonate  solution,  and  wash. 

5.  Stain  with  alum-haematoxylin  and  either  eosin  or  Van  Gieson's  mixture. 
The  histological  appearances   are  adequately   shown    in  colored  plates,  of 

which  a  good  example  is  Taf.  iii,  Fig.  25,  showing  penetration  of  the  elastica  by 
young  forms  of  the  parasite  in  the  act  of  invading  an  hepatic  vessel. 

E.  E.  Southard. 


1546  Journal  of  Applied  Microscopy 

GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Dewitz,  J.    Verhinderung  der  Verpuppung  bei      In  a  brief  but  interesting  paper,  Dewitz 
Insektenlarven.       Arch.    f.    Entwickelung-  .  .        r       i  ^  r 

smech.  11:  690-699,  1901.  gives    an    account    of    the    results    of 

experiments  on  the]_efl[ect  of  a  limited 
amount  of  air  on  the  time  of  pupation  of  the  larvae  of  flies  and  other  insects. 
The  method  of  experimentation  was  to  place  active  larvae  in  small  medicine  vials 
which  were  filled  to  different  heights  with  sand.  These  vials  were  corked  and 
sealed  with  wax,  and  the  number  of  cubic  centimeters  of  contained  air  recorded. 
After  some  days  they  were  opened  and  the  results  noted.  In  case  of  the  larvae 
of  Luciliaccesar,  which  normally  pupates  in  two  days,  it  was  found  that  after  a 
stay  of  five  days  in  the  corked  vials  only  three  larvae  out  of  ninety-five  had 
pupated ;  eighteen  were  dead,  and  the  remaining  seventy-four  were  alive  but  had 
not  pupated.  Left  with  free  access  to  air  these  all  transformed  in  two  days. 
Musca  larvae  were  not  influenced  in  their  time  of  pupation  by  the  amount  of  air, 
those  in  the  closed  tubes  transforming  as  soon  as  the  controls.  The  author 
correlates  this  difference  in  behavior  with  the  fact  that  Lticilia  larvae  do  not 
pupate  under  natural  conditions  later  in  the  year  than  the  end  of  October,  while 
Musca  larvae  pupate  up  to  the  end  of  November,  and  indoors  throughout  the 
winter.  The  caterpillars  of  Pieris  brassicce  were  prevented  from  pupating  by 
limiting  the  supply  of  air.  The  transformation  of  the  larvae  of  the  ichneumonid 
Microgaster  glomeratus  was  prevented  by  placing  them  in  a  very  moist  atmos- 
phere. R.  P. 

Bickel,  A.   Beitrage  zur  Gehirnphysiologie  der       In   continuation  of   his  earlier  work  on 
Schildkrote.  Arch.  f.  Anat.  u.  Physiol.  Phys-       ,,  1       •    1  r    .,  •      1  1       r 

iol.  Abth.,  1901,  Pp.  52-80.  the  physiology   of  the  spinal   cord  of 

the  turtle,  the  author  presents  this  con- 
tribution on  the  functions  of  the  brain  of  the  same  animal.  The  results  were 
gained  from  operation  experiments,  in  which  different  parts  of  the  brain  were 
isolated  or  extirpated,  and  from  stimulating  the  surface  of  the  brain  by  electrical 
or  chemical  means.  The  wounds  from  the  operations  were  covered  with  gela- 
tine mixed  with  tannin,  the  latter  preventing  the  gelatine  from  dissolving  in  the 
water.  Most  of  the  work  was  done  on  Emys  curopcEa,  although  in  a  few  experi- 
ments the  terrestrial  form,  Testudo  graeca,  was  used.  The  operations  consisted 
of  complete  extirpation  by  transection  of  each  of  the  five  principal  divisions  of 
the  brain  (forebrain,  'tweenbrain,  midbrain,  cerebellum,  and  medulla),  and  of 
transverse  cuts  extending  to  the  middle  line  at  the  posterior  boundaries  of  each 
of  these  divisions. 

Loss  of  the  forebrain  causes  a  decrease  in  the  frequency  with  which  spon- 
taneous movements  are  executed,  although  there  is  no  difference  in  the  char- 
acter of  the  movements  themselves  under  these  circumstances.  An  animal  in 
which  the  'tweenbrain  has  been  extirpated,  shows  a  tendency  to  hold  the  legs  in 


and  Laboratory  Methods.  1547 

abnormal,  cramped  positions  for  long  periods  of  time.  Movements  are  normal 
in  character,  but  spontaneous  movements  are  again  less  frequent  than  in  the 
normal  animal.  The  primary  function  of  both  the  forebrain  and  the  'tweenbrain 
is  to  stimulate  the  animal  to  spontaneous  movement,  /.  e.,  furnishes  motor 
impulses.  The  olfactory  lobes  alone  have  this  power  to  some  extent.  The 
forebrain  is  lacking  in  any  appreciable  regulatory  effect  on  the  movements,  but 
the  'tweenbrain  has,  in  a  small  degree,  such  an  effect.  Removal  of  the  mid- 
brain causes  a  pronounced  increase  in  the  activity  of  the  animal.  All  directive 
influence  over  the  movement  is  lost,  the  animal  proceeding  in  a  straight  line 
until  it  is  stopped  by  some  obstacle.  The  co-ordination  between  the  different 
extremities  is  preserved,  but  the  movements  of  the  individual  appendages  are 
wild  and  exaggerated.  The  chief  function  of  the  midbrain,  in  its  relation  to 
the  movement  of  the  animal,  is  evidently  an  inhibitory  and  regulatory  one. 
Removal  of  the  cerebellum  has  no  observable  effect  on  the  animal.  Turtles  in 
which  the  nervous  system  has  been  transected  at  the  point  of  junction  of  the 
medulla  with  the  cord  show  only  very  slight  spontaneous  movements  of  single 
appendages.  There  is  no  spontaneous  locomotion.  The  reflex  irritability  of 
the  posterior  part  of  the  body  is  greatly  increased,  and  various  forced  move- 
ments appear.  Locomotion  in  a  straight  line  forward  can  only  be  induced  by 
very  strong  stimulation  at  the  posterior  end  of  the  body.  In  this  movement 
the  different  appendages  are  fairly  well  co-ordinated.  The  most  important 
function  of  the  medulla  is  the  inhibition  of  spinal  reflexes. 

Electrical  or  chemical  stimulation  of  the  surface  of  the  cerebral  hemispheres 
causes  no  muscular  movement,  or  tonic  cramps,  or  convulsions,  such  as  result 
from  similar  stimulations  of  the  mammalian  brain.  r.  p. 

Holmes,  S.  J.    Phototaxis  in  the  Amphipoda.      ^he    author    investigated    the    photo- 
Amer.  Jour.  Physiol.    5:  211-234,  1901.  °  '^ 

tactic  response  in  about  twenty  species 

of  aquatic  and  terrestrial  amphipods.  The  aquatic  Gammaridea  were  found  to 
be  uniformly  negatively  phototactic.  This  reaction  may  be  modified  and 
obscured  by  the  thigmotactic  reaction,  but  positive  phototaxis  does  not  appear 
under  any  conditions.  The  terrestrial  forms  most  studied  were  Talorchestia 
longtcornis,  Onhestia  agilis,  and  Orchestia  palustris.  All  three  species  are 
positively  phototactic  under  ordinary  conditions,  the  intensity  and  precision  of 
the  reaction  in  each  case  being  correlated  with  the  general  habits  of  the  organ- 
ism. The  positive  reaction  is  less  decided  in  those  species  which  are  habitually 
exposed  to  the  most  light.  Talorchestia  longicor?iis  always  reacts  positively 
both  in  weak  and  in  strong  light.  Nevertheless  this  animal  generally  comes  to 
rest  in  shaded  areas,  presumably  because  it  is  less  stimulated  in  the  shade. 
The  normal  positive  reaction  of  Orchestia  agilis  is  temporarily  changed  to 
negative  by  keeping  the  animals  for  a  time  in  the  dark.  When  returned  to  the 
light  they  again  react  positively.  A  rather  remarkable  fact  was  brought  out  by 
the  experiments  on  this  form,  it  being  found  that  if  specimens  that  are  exhibiting 
a  well  marked  positive  reaction  in  strong  light,  are  suddenly  brought  into  weak 
light,  their  reaction  becomes  immediately  strongly  negative.  This  reversal  is 
independent  of  changes  of  temperature.     The  phototaxis  of  Orchestia  palustris 


1548  Journal  of  Applied  Microscopy 

is  positive  in  sense,  thougli  much  less  pronounced  than  the  reaction  of  the  other 
two  terrestrial  species  studied. 

The  positive  phototaxis  of  Talorchestia  longicornis  and  Ordiestia  agilis  is 
changed  to  negative  if  these  animals  are  placed  in  water,  the  permanence  of  the 
change  apparently  depending  to  some  extent  on  the  degree  of  salinity  of  the 
water.  In  sea  water  the  change  persists  until  death,  while  in  fresh  water 
Orchestias  become  again  positive  some  time  before  they  die.  Experiments  in 
which  one  eye  was  blackened  over  with  asphalt  varnish,  or  extirpated,  were 
performed  on  several  species  of  amphipods  and  insects.  These  operations 
caused  the  animals  to  perform  circus  movements,  which  differed  in  direction 
according  as  the  specimen  was  positively  or  negatively  phototactic.  Positively 
phototactic  forms  turn  continually  in  this  movement  towards  the  side  bearing 
the  blackened  eye,  while  negative  forms  turn  in  the  opposite  direction.  Hemi- 
section  of  the  brain  caused  a  complete  loss  of  the  power  of  orientation  to  light 
in  all  cases  where  the  experiment  was  tried,  although  the  animals  are  still 
afifected  by  light,  as  is  shown  by  their  general  behavior. 

The  closing  section  of  the  paper  is  devoted  to  a  discussion  of  the  relations 
of  phototaxis  and  photopathy.  The  author  sharply  criticises  the  position 
recently  taken  by  Holt  and  Lee  {Amer.  Jour.  FhysioL  iv.  p.  479.  Review  in 
this  Journal,  p.  1264)  that  there  is  no  proper  basis  for  the  separation  of  reactions 
to  intensity  of  light  from  reactions  to  direction  of  ray.  Dr.  Holmes  maintains 
that  there  are  different  forms  of  behavior  towards  light,  which  may  be  conven- 
iently designated  by  the  terms  "phototaxis  "  and  "photopathy." 

Throughout  the  paper  there  appear  numerous  interesting  references  to  the 
general  habits  and  behavior  of  the  organisms  discussed.  r.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

The  Reception  of  Prof.  Koch's   New  Views   concerning   Bovine   and 

Human  Tuberculosis. 

The  paper  of  Prof.  Koch,  delivered  at  the  Tuberculosis  Congress  in  London, 
was  a  veritable  bombshell  in  the  camp  of  the  bacteriologists.  This  paper  has 
been  widely  read  and  much  discussed.  The  address  of  Prof.  Koch  can  be 
ioMw^m.\.\\&  British  Medical  Journal,  July  27,  TOOL  The  reputation  of  Prof. 
Koch  as  the  discoverer  of  the  tuberculosis  bacillus  lends,  of  course,  to  his  con- 
clusions a  weight  greater  than  would  be  given  those  of  any  other  bacteriologist. 
The  general  conclusions  of  this  remarkable  address  are  already  well  known. 
They  are  essentially  two  :  1.  Bovi?ic  tuberculosis  and  human  tuberculosis  are 
produced  by  quite  different  bacteria.  This  he  concludes  from  the  fact  that  the 
inoculation  of  cattle  with  human  tuberculosis  does  not  produce  the  typical  bovine 
disease.     '1.  Human  tuberculosis  is  to  be  attributed  to  injection  Jrom  other  human 


and  Laboratory  Methods.  1549 

beijigs  and  very  rarely  fro7n  cattle.  This  belief  he  bases  upon  his  first  conclusion, 
and  also  upon  the  fact  that  in  mankind  primary  tuberculosis  in  the  intestinal 
tract  is  quite  rare,  while,  if  the  disease  were  commonly  due  to  the  consumption 
of  flesh  or  milk,  primary  intestinal  tuberculosis  should  be  frequent. 

It  was  inevitable  that  these  bold  conclusions  should  be  received  by  the 
members  of  the  congress  with  consternation  and  disapproval.  Many  of  the 
members  of  the  congress  had  appeared  especially  prepared  to  discuss  the 
dangers  to  mankind  of  the  distribution  of  tuberculosis  by  milk  or  flesh  of  cattle, 
and  the  sweeping  conclusions  of  Prof.  Koch  inevitably  destroyed,  in  a  large 
degree,  the  significance  of  many  of  the  papers  read  before  the  congress.  The 
members  of  the  congress  did  not  accept  the  conclusions  of  Prof.  Koch,  and 
nearly  all  of  the  remarks  which  referred  to  the  paper  took  a  position  quite 
opposite  to  that  occupied  by  the  discoverer  of  the  tubercle  bacillus.  The 
opinion  was  expressed  that  Prof.  Koch  had  done  the  cause  of  public  health  a 
great  injury  by  advancing  unproved  conclusions  which  would  tend  to  decrease 
the  care  given  to  the  methods  of  preventing  the  use  of  tuberculous  material  as 
food,  and  thus  making  the  work  of  sanitary  boards  more  difficult.  Indeed,  a 
resolution  was  passed  in  the  State  and  Municipal  Section  to  the  effect  that  the 
conclusions  of  Prof.  Koch  were  not  demonstrated,  and  that  the  same  amount  of 
care  should  be  exercised  in  preventing  the  use  of  tuberculous  material  as  before 
the  publication  of  the  address  of  Prof.  Koch. 

Since  the  closing  of  the  congress  bacteriologists  of  repute  have  expressed  in 
public  opinions  as  to  the  conclusions  taken  by  Prof.  Koch.  These  are  too 
numerous  to  be  mentioned  in  this  place,  but  the  attitude  taken  by  some  of  the 
more  prominent  bacteriologists  may  be  properly  mentioned. 

It  must  be  noticed  at  the  outset  that  the  first  conclusion  is  not  new  with 
Prof.  Koch,  for  Theobold  Smith  of  Harvard  University  had  already  some  years 
ago  demonstrated  conclusively  that  the  human  bacillus  is  only  slightly,  if  at  all, 
pathogenic  for  cattle.  This  conclusion  was,  therefore,  well  known,  and  the  only 
novelty  in  Prof.  Koch's  address  is  in  the  claim  that  bovine  tuberculosis  is  not  a 
source  of  human  tuberculosis.  In  regard  to  Prof.  Koch's  claims,  wide 
divergence  of  opinion  may  be  found  among  bacteriologists  who  have  commented 
on  the  matter.  Prof.  Virchow  (  Ber.  K/m.  Woc/i.,  p.  8i8,  JQOi)  expresses  himself 
as  of  the  opinion  that  there  is  a  difference  between  the  bovine  and  human 
bacillus,  though  not  so  great  a  one  as  Prof.  Koch  is  inclined  to  think.  He 
believes  that  many  of  the  tubercles  which  have  been  described  as  due  to  tuber- 
culosis are  not  properly  described,  and  that  histological  study  of  the  tubercles 
alone  can  be  depended  upon  to  determine  the  presence  of  this  disease,  and  not 
the  simple  presence  of  a  tubercle  which  stains  properly.  He  insists  that  the 
second  conclusion  of  Prof.  Koch  is  not  justified,  and  that  there  are  cases  on 
record  which  show  that  the  disease  may  pass  from  cattle  to  men,  although  the 
danger  is  slight.  He  thinks  that  more  attention  must  be  paid  to  the  tiumber  of 
bacteria  inoculated  than  has  been  paid  hitherto.  Prof.  Klebs  {Milchztg.,  p.  501, 
1901  )  very  violently  attacks  Koch's  position,  claiming  that  both  of  Koch's  con- 
clusions are  erroneous  ;  that  the  bacillus  is  the  same  in  cattle  and  men,  and  the 
milk  and  flesh  of  tuberculosis  animals  are  a  prominent  source  of  danger  to  man. 


1550  Journal  of  Applied  Microscopy 

Prof.  Heuppe  (^Ber.  Klin.  Woch.,  Aug.  2)  is  also  positive  in  his  opposition  to 
Prof.  Koch's  views,  insisting  that  the  evidence  in  our  possession  is  quite 
sufficient  to  demonstrate  that  the  disease  may  pass  from  animals  to  men,  and  insist- 
ing that  the  differences  between  the  bacilli  in  the  two  animals  are  far  less  than  the 
differences  between  the  avian  and  bovine  bacillus,  which  experiment  has  shown 
to  be  only  cultural  conditions  of  the  same  organism.  Among  others  who  hold 
a  similar  position  may  be  mentioned  McFadyen,  Ravenel,  Nocard,  Brouardel, 
Bang,  Boullanger.  Without  giving  further  references  of  this  sort,  it  may  be 
stated  that  the  majority  of  bacteriologists  who  have  expressed  any  opinion  at 
the  present  time  hold  a  view  somewhat  as  follows  :  The  bacillus  from  man  is 
very  slightly,  if  at  all,  pathogenic  for  cattle.  This,  however,  does  not  indicate 
that  they  are  different  species  of  bacteria,  but  simply  that  they  are  different  cul- 
tural varieties  of  the  same  organism  due  to  growth  in  different  environment. 
The  second  conclusion  of  Prof.  Koch,  that  human  tuberculosis  is  not  derived 
from  cattle,  is  quite  generally  discredited.  It  is  insisted  that  Prof.  Koch  drew 
this  conclusion  without  sufficient  evidence ;  that  primary  intestinal  tuberculosis 
is  common  among  children ;  and  that  there  are  sufficient  instances  of  direct 
transference  from  cattle  to  man  to  show  that  such  a  source  of  the  disease  is 
possible.  There  is  thus  a  general  tendency  to  discredit  the  second  position  of 
Prof.  Koch. 

On  the  other  hand,  some  have  expressed  themselves  as  agreeing  in  general 
with  Prof.  Koch's  views.  Prof.  Baumgarten  ( Bcr.  Klin.  IVoch.,  Sept.  2 )  is 
inclined  to  accept  the  position  of  Koch.  He  had  in  1893  found  it  impossible  to 
produce  the  bovine  disease  with  human  bacilli.  He  instances  a  long  series  of 
attempts  made  to  inoculate  a  certain  patient  suffering  from  cancer  with  tubercu- 
losis by  the  use  of  a  culture  from  cattle.  These  all  proved  futile  because,  as  he 
believes,  the  bovine  bacillus  was  used  rather  than  the  human  bacillus.  He,  how- 
ever, is  inclined  to  regard  the  organisms  as  of  the  same  species,  though  different 
cultural  varieties,  but  he  believes  that  the  danger  of  transference  of  the  disease 
from  cattle  to  man  is  very  small.  Heubner  is  inclined  to  side  with  Prof.  Koch, 
thinking  with  him  that  the  danger  to  man  from  bovine  tuberculosis  is  slight 
although  perhaps  it  is  too  early  to  make  generalizations. 

Dr.  Ostertag  {Zeif.  f.  Fl.  u.  Milch  ffyg.,  XI,  353)  has  given  one  of  the 
most  complete  discussions  of  the  present  aspect  of  the  question.  While  very 
careful  to  make  no  positive  statements,  he  points  out  an  unfortunate  result  that 
Prof.  Koch's  lecture  has  had  in  tending  to  allay  the  care  taken  by  farmers  in 
regard  to  the  treatment  of  tuberculous  animals.  He  emphasizes  the  fact  that 
we  have  as  yet  no  proof,  indeed,  no  good  reason,  for  believing  that  Prof.  Koch's 
position  is  a  correct  one,  and  until  this  question  can  be  positively  settled  we 
should  proceed  exactly  as  we  have  done  in  the  last  few  years,  upon  the  assump- 
tion that  the  disease  can  be  transmitted  from  cattle  to  men,  and  that  bovine 
tuberculosis  is  therefore  a  serious  danger  for  mankind. 

All  who  have  discussed  the  question  recognize  that  the  conclusions  which 
Prof.  Koch  advanced  can  only  be  settled  by  further  experiment  and  discussion. 
Already  a  number  of  persons  have  offered  themselves  for  experiment  and  have 
expressed  their  willingness  to  be  inoculated  with  bovine  bacilli  in  order  to  dem- 


and  Laboratory  Methods.  1551 

onstrate,  if  possible,  the  truth  or  falsity  of  Prof.  Koch's  position.  A  committee 
has  been  appointed  recently  in  England,  consisting  of  the  most  prominent 
experts  among  English  scientists,  to  investigate  the  questions  concerned.  The 
great  importance  of  these  conclusions  rests  upon  the  fact  that  the  belief  in  the 
possibility  of  transference  of  the  disease  from  animals  to  man  has  been  the 
basis  of  widely  adopted  public  laws  and  sanitary  rules,  connected  with  the  care 
of  cattle  and  the  distribution  of  milk  in  all  civilized  communities,  and  if 
Prof.  Koch's  views  should  be  accepted  as  correct  it  would  result  in  almost  a 
revolution  in  conducting  sanitary  inspection.  The  extreme  importance  of  the 
subject  makes  it  certain  that  in  the  next  few  years  many  contributions  will  be  • 
given  on  the  question,  and  we  may  in  a  short  time  expect  a  satisfactory 
demonstration  or  refutation  of  the  two  positions  advanced  by  Prof.  Koch. 

H.  w.  c. 

Nikolsky.  Charbon  chez  des  animaux  nourris  r^^^  question  as  tO  the  distribution  of 
avec  leur  ailments  habituel,  meles  de  spores  ^ 

charbonneuses.  Ann.  d.  1.  Inst.  Past.  14:  794,      the  anthrax  baciUus  has,  ever  Since  the 

'900-  days  of  Pasteur,  been  subject  to  a  con- 

siderable degree  of  uncertainty.  The  author  endeavors  to  determine  whether 
the  anthrax  spores,  mixed  with  ordinary  food,  are  capable  of  giving  rise  to  the 
disease.  His  conclusion  is  positive.  The  spores,  mixed  with  the  ordinary  food, 
were  not  only  able  to  resist  the  action  of  the  ordinary  intestinal  bacteria,  but 
made  their  way  through  the  intestinal  walls,  and  in  a  short  time  produced  typi- 
cal cases  of  anthrax.  This,  of  course,  is  a  factor  which  explains  in  a  measure 
the  appearance  of  anthrax  in  old  pastures  where  the  bodies  of  animals  that  have 
suffered  from  this  disease  have  been  buried.  h.  w.  c. 

Smith.  The  Nodule  Organism  of  the  Legum-  The  author  has  made  a  more  careful 
inosae.     Proc.    Linn.    Sec.    of    New   South         .     1         r   ^1  •  ^.v    .  j 

Wales.    P.  653, 1899.  st^dy  °^  ^^^  organism  that   produces 

the  tubercle  in  legumes  than  has  hith- 
erto been  made.  Previous  observers  have  dwelt  almost  wholly  upon  the  action 
of  the  nodule  in  producing  the  tubercles,  without  making  a  sufficiently  careful 
study  of  the  organism  itself.  Smith  studies  and  gives  a  thorough  description  of 
the  tubercle  organism.  His  conclusions  are,  essentially,  as  follows  :  1.  The 
nodule  organism  is  a  yeast,  possessing  a  vacuole,  and  not  a  bacterium.  2.  It 
multiplies  by  budding,  and  this,  together  with  a  persistent  mucilaginous  capsule, 
indicates  its  relations  to  yeasts,  although  the  organism  has  a  variety 
of  forms.  3.  Vigorous  motor  forms  are  found  and  the  motile 
organ  in  each  consists  of  a  single  terminal  or  tufted  flagellum.  4.  The 
organism  grows  best  in  a  slightly  acid  glucose  medium.  5.  It  does  not  fix  nitro- 
gen in  an  artificial  medium,  at  least,  so  far  as  the  author's  experiments  show.  6. 
It  is  always  accompanied  in  the  nodule  by  other  bacteria,  but  whether  they  have 
anything  to  do  with  the  formation  of  the  nodule,  the  author  is  not  sure. 

H.  W.    C. 

The  Exclusion  and  Elimination  of  Pathogenic  An  editorial  article  discusses  the 
Bacteria  from  Sewage.  Brit.  Med.  Jour.  hygienic  value  of  the  modern  accepted 
p.  902,  1901.  method  of  the  treatment  of  sewage  as 

adopted  chiefly  in  England.     The  bacterial  treatment  of  sewage  produces  a  very 


1552  Journal  of  Applied  Microscopy 

great  chemical  purification  of  the  material,  but  the  results  of  such  careful  exper- 
iments have  seemed  to  indicate  that  ordinarily  this  treatment  does  not  materially 
reduce  the  bacteria,  and  it  is  very  questionable  whether  it  lessens  the  danger  of 
the  sewage  material  distributing  diseases.  The  author  of  the  present  article 
points  out  that  no  satisfactory  means  has  yet  been  employed  for  destroying  the 
bacteria  in  sewage  sufficiently  to  reduce  in  any  great  measure  its  pathogenic 
nature.  It  should  be  noted  that  the  results  obtained  in  the  bacterial  purification 
of  sewage  are  not  always  in  harmony,  and  certainly  in  many  of  the  sewage  plants, 
particularly  in  this  country,  there  is  a  very  remarkable  reduction  in  bacteria, 
which  surely  renders  the  sewage  far  less  liable  to  distribute  disease.  In  the  filter 
beds  used  in  the  vicinity  of  London,  however,  such  a  reduction  is  not  very  great, 
and  the  author  is  of  the  opinion  that  entirely  new  methods  must  be  adopted  in 
the  treatment  of  sewage  before  we  can  be  satisfied  that  the  problem  has  been 
mastered.     His  general  conclusions  are  five,  as  follows  : 

1.  The  lines  of  defense  which  protect  us  from  invasion  by  sewage  borne 
disease  germs  are  defective  and  uncertain.  Consequently,  it  is  ever  necessary 
to  strengthen  these  deficiencies  by  all  means  in  our  power. 

2.  The  presence  of  disease  germs  in  sewage  and  the  possibility  of  their  sur- 
viving the  various  processes  for  sewage  purification  cannot  be  ignored. 

3.  We  are  ignorant  with  regard  to  the  fate  of  these  germs  before,  during,  and 
after  the  processes  of  purification,  and  can  only  say  that,  so  far  as  the  evidence 
hitherto  acquired  shows  anything,  it  tends  to  prove  that  the  disease  germs  are  not 
necessarily  destroyed  by  the  purification  processes. 

4.  It  is  imperative  that  investigations  (similar  to  those  described  above)  should 
be  continued,  developed,  and  applied  to  the  various  systems  of  purification  by 
precipitation,  by  "  bacteria  beds,"  etc.,  and  by  land  filtration  and  irrigation. 

5.  Any  effluent  which  has  been  so  far  purified  that  it  is  free  from  putrescible 
matter  and  incapable  of  giving  rise  to  offensive  nuisance  must  still  be  regarded 
as  capable  of  giving  rise  to  disease  until  it  has  been  shown  that  the  disease  germs 
have  been  eliminated  from  it.  h.  w.  c. 

Weissenfeld.     Uer  Befund  des  Bakterium  coli  -pj^g   ^^^j^qj.   investigates    the   question 
in   Wasser   und  das  Thierexpenment    sind  °  ^ 

keine  brauchbaren    Hilfsmittel  fur  die  hy-  as  tO  whether  the  presence  of  the  com- 

gienische  Beurteilung  des  Wassers.     Zeit.f.  j^^^   B.   coll  in  water  is  an   indication 
Hyg.  34:   70,   1900. 

of  the  unhealthfulness  of  the  water.  It 
has  generally  been  assumed  that  the  presence  of  this  organism  in  quantity  is  an 
indication  of  sewage  contamination,  and  consequently  an  indication  that  the 
water  in  question  is  unwholesome.  The  author  uses  modifications  of  Pariettii's 
fluid,  and  having  isolated  his  organisms  injects  them  into  guinea  pigs.  The 
conclusions  that  he  reaches  in  regard  the  B.  coli  from  the  waters  which  he 
studied  were,  that  the  organisms  isolated  from  the  best  waters  were  commonly 
pathogenic  for  guinea  pigs,  whereas  those  isolated  from  the  suspicious  waters, 
and  waters  of  a  clearly  undesirable  character,  were  less  pathogenic,  or  indifferent 
in  their  action  upon  guinea  pigs.  He  therefore  is  of  the  conclusion  that  the  dis- 
covery of  B.  coli  in  water  is  not  necessarily  an  indication  of  sewage  contamina- 
tion. H.  w,  c. 


and  Laboratory  Methods.  1553 

Whipple,  Geo.  C.     Changes  that  Take  Place  in      The  author  has    undertaken  a  study  of 
the    Bacterial    Contents  of  Waters    during       ^,  j.^.  ,  i_-   i    ^i  i 

Transportation.    Tech.  Quart.  14  :  21,  1900       the  conditions  under  which  the  number 

of  bacteria  in  samples  of  water  for 
analysis  increase  or  decrease  during  transportation  from  the  point  of  collection 
to  the  laboratory,  a  subject  of  considerable  interest  to  those  engaged  in  bacteri- 
ological analysis  of  drinking  waters.  He  reaches  two  conclusions  :  1.  After  a 
sample  of  water  is  collected,  either  in  large  or  small  bottles,  there  is,  first,  a 
slight  reduction  in  the  number  of  bacteria,  due  to  the  change  in  environment. 
The  reduction  is  greater  when  small  volumes  of  water  are  collected.  Subse- 
quently, there  is  an  increase  in  the  number  of  bacteria,  which  is  greater  in  a 
small  bottle  than  in  a  large  one,  and  is  more  rapid  when  the  bottle  is  but  par- 
tially filled.  With  bottles  of  the  same  size  the  growth  is  more  rapid  in  small  vol- 
umes of  water  than  in  large  volumes.  2.  An  agitation  of  the  water  exercises 
a  slight  retarding  influence  upon  the  multiplication  of  bacteria,  but  the  shaking 
to  which  the  water  samples  are  liable  during  transportation  is  so  slight,  that  it  is 
of  practically  no  importance  in  affecting  the  number  of  bacteria  in  the  water. 

H.  w.  c. 


NOTES  ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  AKred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Vernadsky    W.      Zur    Theorie    der    Silicate.      ^  theoretical  discussion  limited  to  the 
Zeit.  f.  Kryst,  34:  37-66,  1901. 

simpler  and  better  known  compounds. 

All  historical  and  bibliographic  data  are  omitted,  though  included  in  a  previous 

article  in  a  Russian  journal. 

Natural  silicates  are  usually  isomorphic  mixtures,  that  is,  are  analagous  to 
solid  solutions,  some  predominating  substance  (the  "  solvent ")  containing  dis- 
solved in  it  other  substances,  necessarily  crystallizing  i?i  the  same  one  of  the  thirty- 
two  classes,  but  possibly  of  very  different  type  of  formula. 

If  the  "solvent"  contain  no  R2O3  we  may  call  the  silicate  a  simple 
(einfache)  silicate. 

If  the  "solvent"  contain  R2O3  the  silicate  may  be  called  an  alumosilicate, 
ferrisilicate,  borosilicate,  etc.  Only  the  alumosilicates  are  directly  discussed,  the 
others  may  be  considered  by  analogy. 

There  is  a  sharp  line  between  simple  and  alumosilicates  : 

{a)  There  is  no  known  reaction  by  which  the  metals  of  RO  can  be  replaced 
by  Al,  or  conversely. 

if)  There  is  no  known  reaction  by  which  the  alumosilicates  can  be  directly 
changed  into  silica  hydrate,  (opal)  or  conversely. 

(<:)  The  alteration  products  of  simple  silicates  often  include  opal  and  quartz  ; 
the  alumosilicates  can  only  with  very  uncommon  proportions  yield  opal  (and 
aluminum  hydrate),  but  usually  yield  only  clay  and  minerals  of  the  chlorite  group. 


1554  Journal  of  Applied  Microscopy 

{d)  Simple  and  alumosilicates,  under  action  of  heat,  either  unite  to  a  com- 
plicated substance  or  there  is  an  interchange  of  metals  of  the  RO  group. 

{e)  With  complicated  alumosilicates  containing  RO  there  are  many  known 
reactions  which  produce  aluminates  of  RO. 

The  simple  silicates  are  salts  of  known  acids,  but,  while  a  few  alumosilicates, 
such  as  leucite  KgAlgSi^Oj  2,  could  be  considered  double  salts  of  known  acids, 
most  are  classed  as  salts  of  complicated  hypothetical  acids,  and  for  some  no 
satisfactory  acid  has  been  found. 

Alumina  may  be  regarded  either  as  an  anhydrous  acid  or  as  a  weak  base. 
In  the  latter  case  the  salts  show  many  characters  of  so-called  complex  acids,  and 
as  indicated  by  the  following  experimental  data,  it  is  much  more  probable  the 
alumosilicates  are  anhydrides,  hydrates,  and  salts  of  complex  alumosilicic  acids  : 

{a)  Aluminates  form  under  the  same  conditions  as  alumosilicates. 

(b)  By  splitting  up  of  alumosilicates  at  high  temperature  aluminates  are  formed. 

(/)  The  action  of  water  or  carbonate  solution  often  results  in  destruction  of 
alumosilicates,  and  formation  of  aluminates  or  alumina  hydrates. 

The  complex  structure  of  the  alumosilica  nucleus  is  shown  by  the  properties 
of  the  compounds  ;  for  instance  : 

{a)  Compounds  of  only  AI2O3  and  SiOg  correspond  in  properties  to  acid 
anhydrides;  e.  g.,  heated  with  carbonates  they  swell  and  evolve  CO 2  rapidly, 
and  form  an  alumosilicate.  Similar  results  are  obtained  by  heating  with  sul- 
phates, haloids,  etc. 

(b)  Kaolin  and  other  clays  act  like  acids,  destroying  haloid  salts  at  compara- 
tively low  temperatures. 

In  nature  and  the  laboratory  many  substitution  reactions  occur  in  which  the 
alumosilica  kernel  is  not  destroyed.     The  general  scheme  is : 

Mx-j-MjAls^M^x+MAls,  in  which  x  is  the  acid  anhydride,  M  and  M^ 
different  metals,  and  Als  the  alumosilica  kernel. 

Clay  (alumosilicic  acid)  and  minerals  of  sillimanite  group  (alumosilica  anhy- 
dride) form  by  destruction  of  alumosilicates  under  the  same  conditions  as  hydrates 
and  anhydrides,  by  the  destruction  of  their  salts.  This  is  best  seen  by  a  com- 
parison with  silicates. 

(1)  By   heating  opal  we  obtain  silica  ;  by   heating  clay  we  obtain 

minerals  of  sillimanite  group. 

(2)  By  destruction  of  simple  silicates  under  the  action  of  water  and 

CO 2  in   nature   we  obtain  opal;    by   similar   destruction   of 
alumosilicates  we  obtain  clay. 

(3)  At  high  temperatures  in  fusions  rich  in  alumina,  corundum  or 

sillimanite    separates,    just    as    from    fusions    rich    in   silica, 
tridymite  or  quartz  separates. 

THE  SIMPLE  SILICATES. 

These  are  salts  of  known  acids,  and  their  derivatives,  the  two  great  groups, 

RO.  .OR 

being  the  orthosilicatcs,  with  a  structure  formula  /Si\  ^"*^    the    7neta- 

RO/        \0R 
RO. 
silicates,  with  a  structure   formula  of  \Si=0. 

RO/ 


and  Laboratory  Methods.  1555 

It  is  given  as  a  general  rule  that  with  strong  mineral  acids  orthosilicates 
gelatinize,  metasilicates  yield  pulverulent  silica,  and  slimy  silica  indicates  the 
presence  of  both. 

The  Orthosilicates. — Sa/fs. — -The  only  great  family  of  normal  salts  is  the 
chrysolite  group.     Sepiolite  is  probably  an  acid  salt. 

Derivatives. — The  general  formula  is  m  R^Si04  •  nA,  in  which  A  is  an  added 
group  of  atoms.  That  these  derivatives  may  fairly  be  said  to  consist  of  an 
orthosilicate  nucleus  and  added  groups,  is  evidenced  by : 

{(.i)  The  groups  A  may  be  added  by  influence  of  water  or  of  heat. 

{U)  Heat  will  split  the  derivative  into  nucleus  and  A  if  latter  is  volatile. 

0  One  derivative  passes  into  another  by  simple  exchange  of  elements  in  A, 
without  affecting  the  nucleus. 

{d')  If  A  involves  a  metasilicate  the  jelly  of  Si02  becomes  slimy. 

These  derivatives  cannot  be  regarded  as  ordinary  double  compounds  of  A 
and  orthosilicate,  for  the  properties  of  A  are  very  variable;  e.g.,  humite, 
SMgjSiO^  •  MgF2  does  not  simply  split,  but  on  heating  yields  SiF^.  Serpen- 
tine yields  its  water  only  at  red  heat. 

Only  a  few  proportions  between  m  and  71  are  possible.  For  instance,  the 
orthosilicate  Mg2Si04  and  A=MgF2,  if  w=l  then  n^\  or  2  only. 

/^\      /^\ 
Mg<       >Si<       >MgorMg2Si04. 

\o/     \o/ 

O.        /0-MgF. 
Mg<      >Si<  orMg2Si04.MgF2. 

\0/        \0— MgF 
F— Mg— O.  .0— MgF. 

yS'i/  or  MgoSi04  .  2MgF<,. 

F— Mg— 0/       ^O— MgF. 
Isomeres  may  be  expected,  for  the  unsymmetrical  character  of  the  structure 
formula  of  Mg2Si04  .  MgFjbecomes  symmetrical  if  doubled. 

There  are  known  the  following  five  series  of  derivatives  of  orthosilicates : 
1        M     Q'n       A  J  A=metasilicate.     Serpentine  group. 

1.  n  Mg2biU4  .  A  I  A=MgF2Mg(OH)2.     Chondrodite  group. 

2.  ?i  Ni2Si04  .  A.      A=H2  0.     Garnierite  group. 

3.  ;;  Cu2Si04  .A.      A^H20.     Chrysocolla  group. 

4.  H  Zn2  Sio4  .  A.      A==H20.     Calamine  group. 

5.  fi  Mn2Si04  .  A.     A=MnS,  MnCl2,  etc.     Helvite  group. 

The  Metasilicates.— iVdV/t/rrr/  Sa/fs. — The  very  stable  group  of  pyroxenes 
and  amphiboles  not  only  are  neutral  salts  but  form  derivatives,  for  it  is  an 
excellent  solvent  for  different  alumosilicates  and  ferrisilicates,  the  best  known 
being  R"Al2Si06  and  R'2Al2Si40i2  (or  R'2Fe2Si40i  2)- 

And  Salts. — The  metasilicates  differ  from  orthosilicates  in  the  formation  of 
chain-like  compounds.  For  instance,  the  structure  formula  of  talc  H2Mg3Si40j  2 
may  be  written 

HO— Si— O— Mg— O— Si— O— Mg— O— Si— O— Mg— O— Si— OH 

II  II  II  11 

0  0  0  0 

whereas  rensselaerite,  with  two  more  atoms  of  water  than  talc,  is  intermediate 
between  talc  and  serpentine,  and  with  still  two  more  atoms  would  pass  into  the 
orthosilicates. 

(Continued  in  December.) 


1556 


Journal  of  Applied  Microscopy 


Publications  Received  for  Journal  Library. 


The  Neocene  Lake  Beds  of   Western   Montana 

and   Descriptions  of  Some  New  Vertebrates 

from  the  Loup  Forlt.    Earl  Douglass,  Univer- 
sity of  Montana. 
Proceedings  of  the  Indiana  Academy  of  Science, 

Vols.  1898  and  1899. 
Annual  Report  of  the  Board  of  Regents  of  the 

Smithsonian  Institution,  1898. 
Library    Expedients     in    Microscopy.     R.    H. 

Ward,  M.  D.     Reprint  from  Transactions  of 

the  American  Microscopical  Society.  Twenty- 
second    annual   meeting,    Aug.    17,   18,  and 

19,  1899. 
Peach   Leaf   Curl,   its    Nature   and   Treatment* 

Newton    B.    Pierce.     Bull.    No.    20,    U.    S- 

Dept.  of  Agri. 
Key  to  Land  Mammals  of  Northeastern  North 

America.      Bull,    of    the    New    York    State 

Museum,  Vol.  VIII,  No.  38. 
Agriculture  Year  Book,  University  of  Tennessee 

Record.     Vol.  IV,  No.  i. 
Report  of  Committee  on  the  Protection  of  North 

American  Birds.     Witmer  Stone. 
Concerning  the  New  Formation  of  Elastic  Fibers, 

Especially   in   the    Stroma   of    Carcinomata. 

Herbert  W.  Williams,  M.  D. 
Report  of  Laboratory  of  Pathology  of  the  Uni= 

versity  of  Buffalo.     No.  i. 
Course   in   Biology   in   the   Horace  Mann  High 

School.     Teachers  College  Record,  Vol.  II, 

No.  I. 
Columbia  University  Quarterly.     Vol.  II,  No.  3. 
A    Fresh    Water    Sponge    from  Sable   Island. 

A.  H.  Mackay,  LL.  D.,  Halifax. 
Transactions  of  the  Texas  Academy  of  Science. 

Vol.  III. 
A  Catalogue  of  Publications  of  the  University  of 

Chicago  Press. 
Sixteenth  Report  of  the  State  Board  of  Health 

of  the  State  of  New  Hampshire. 
Proceedings  of  the  American  Association  for  the 

Advancement  of  Science.     Forty-ninth  meet- 
ing- 
Bulletin   of   the    Illinois   State    Laboratory   of 

Natural  History,  Vol.  V. 

Art.  XI.  Notes  on  species  of  North  Amer- 
ican Oligochasta.  IV.  On  a  new  Lum- 
briculid  genus  from  Florida,  with  addi- 
tional notes  on  the  nephridial  and  circula- 
tory systems  of  Mesoporodrilus  asym- 
metricus  Smith. 

Art.  XII.  The  Hirudinea  of  Illinois. 
High  School  Department,  University  of  the  State 

of  New  York.     Bull.  2,  Feb.  1901. 
Wakker's    Hyacinth   Germ.     Erwin    F.    Smith. 

Bull.  No.  26,  U.  S.  Dept.  of  Agri.,  Division 

of  Veg.  Phys.  and  Path. 
Report  of  the  Connecticut  Agri.  Expt.  Sta.  for 

year  ending   Oct.  31,    1900.     Part  II,  Food 

Products. 
Thirty-first  Annual  Report  of  the  Entomological 

Society  of  Ontario. 
Maryland  Agricultural  Experiment  Station. 

Bulls.  Nos.  68,  69,  70,  71,  72,  73,  74,  75,  77. 


Tennessee  Agricultural  Experiment  Station. 

Bulls.  Nos.  I  (Vol.  XIV),  4  (Vol.  XIII). 
Second  Preliminary  Report  of  the  Behring  Sea 
Fur    Seal    Investigations.     David  Starr  Jor- 
dan, 1897. 

New  York  Agricultural  Experiment  Station. 

Bulls.  Nos.  177,  178,  179,  180,  181,  182,  183, 
184,  185,  1S6,  187,  188,  189,  190,  191. 
Fourth  Annual  Report  of  the  Forest  Preserve 
Board,  1900. 

New  Jersey  Agricultural  Experiment  Station. 

Bulls.  Nos.  148,  149. 
Florida  Lichens.     P.  H.   Rolfs.     Transactions 
of   the    Academy   of   Science  of  St.  Louis, 
Vol.  XI,  No.  2. 

Pineapple    Fertilizers.     P.   H.  Rolfs.     Reprint 

from    Proc.    12th    annual    meeting     Florida 

State  Horticultural  Society,  1899. 
The  Wilson  Bulletin,  Nos.  34,  35,  36. 
Vanderbilt  University  Quarterly,  Vol.  I,  No.  i. 
Bericht   uber    die   Pestepidemie   in    Kobe   und 

(Jsaka  von  Nov.  1899  bis  Januar  1900.     Prof. 

Dr.  S.  Kitasato. 
Bulletino  del  Laboratorio  ed  orto  Botanico.  Vol. 

terzo,  Fasc.  III-IV.     Fl.  Tassi. 
Annual  Report  of  the  Smithsonian  Institution, 

1897.     The  U.  S.  National  Museum,  II. 
Publications  of  the  University  of  Pennsylvania. 

Proceedings    of   "  University    Day."      Bull. 

No.  9,  new  series. 
Commercial    Fertilizers.     J.  H.  Stewart  and  B. 

H.  Hite.     West   Virginia   University   Agri. 

Expt.  Sta.    Bull.  72. 

Spraying.  L.  C.  Corbett.  West  Virginia 
University  Agri.  Expt.  Sta.     Bull.  70. 

University  of  the  State  of  New  York.  Bulls.  15, 
16,  35.  52. 

Mosses  with  a  Hand  Lens.     A.  J.  Grout. 

The  Use  of  the  Rontgen  Ray  by  the  Medical 
Department  of  the  U.  S.  Army  in  the  War 
with  Spain.  This  is  a  volume  of  98  pages, 
prepared  by  W.  C.  Borden,  under  the  direc- 
tion of  Surgeon  General  Geo.  M.  Sternberg, 
U.  S.  Army.  The  first  30  pages  are  descrip- 
tive of  apparatus,  following  which  is  a  series 
of  specific  cases  in  which  the  Rontgen  Ray 
was  used  in  the  study  of  wounds,  fractures, 
and  other  effects  produced  by  missiles.  These 
cases  are  illustrated  by  38  full-page  helio- 
type  plates  which  represent  accurately  the 
positions  of  Mauser  bullets  or  other  missiles 
in  eveiy  part  of  the  body,  and  the  effects 
they  produce.  The  great  distinctness  with 
which  these  plates  reveal  the  location  of 
bullets  deeply  embedded  in  the  chest,  pelvis, 
and  even  within  the  skull,  is  ample  proof 
of  the  extraordinary  character  of  the  radio- 
graphs from  which  they  were  taken. 

The  concluding  chapter  is  devoted  to 
radiographic  technic,  embracing  the  manipu- 
lation of  machines,  photographic  plates,  and 
methods  in  development  of  negatives,  and 
in  printing. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods. 

VOLUME  IV.  DECEMBER,  1901.  Number  12 


General  Methods  for  the  Study  of  the  Nervous 

System. 

Biologists  were  accused,  not  so  very  long  ago,  with  a  tendency  to  neglect  the 
nervous  system  in  their  laboratory  courses.  Whatever  may  have  been  the  reasons 
for  the  existence  of  such  a  condition,  it  is  certain  that  there  now  are  means 
whereby  adequate  corrective  measures  can  be  applied.  The  literature  of 
neurological  methods  has  grown  into  bulk  of  indeed  formidable  proportions.  In 
fact,  the  beginner  in  the  field  of  neurology  is  very  apt  to  find  himself  bewildered 
by  the  opportunities  for  choice  between  the  many  and  widely  divergent  schemes 
recommended  by  their  enthusiastic  advocates. 

This  paper  has  been  written  for  the  purpose  of  describing  those  methods 
which  may  properly  claim  a  place  in  the  general  laboratory  study  of  the  verte- 
brate nervous  system.  Since  this  field  necessarily  involves  certain  technical 
difficulties,  the  outlines  have  been  given  some  degree  of  detail  wherever  it  has 
seemed  desirable  to  do  so.  The  attempt  has  not  been  made  to  anticipate  the 
special  needs  of  the  investigator,  but,  rather,  to  serve  the  purposes  of  those  who 
desire  to  demonstrate,  for  the  class  room,  the  microscopical  structure  of  the 
nervous  system  of  some  vertebrate. 

1.      Staining  with  Tro?i  Hiematoxylin  and  Orange-G. 

While  originally  intended  for  an  altogether  different  purpose,  iron  haematoxylin 
may  now  be  claimed  as  a  neurological  reagent  of  distinct  value.  It  may  be 
applied  advantageously  wherever  it  is  desired  to  obtain  a  comprehensive  picture 
of  structural  elements,  such,  e.  g.,  as  would  appear  in  a  longitudinal  section  of  the 
entire  brain  of  the  frog;  in  transverse  sections  through  typical  regions  of  a  small 
mammalian  brain;  or  in  sections  at  different  levels  of  the  spinal  cord.  This 
stain  exhibits  the  structural  elements  without  confusing  the  eye  with  too  much 
detail,  outlining  groups  of  nerve-cells,  and  bringing  tracts  of  nerve-fibres  into 
sharp  relief.  Its  value,  therefore,  lies  in  the  basis  which  it  gives  for  a  conception 
of  the  relations  between  important  parts  of  the  nervous  system. 

For  this  general  purpose,  the  whole  brain  of  a  rat  or  mouse,  or  segments   of 

(1557) 


1558  Journal  of  Applied  Microscopy 

the  spinal  cord  of  a  cat  or  dog,  should  be  hardened  for  several  days  in  an  excess 
of  a  solution  of  formaldehyde  : 

Commercial  formaldehyde     -         -         -         -      10  vols. 
Water      -         - 100  vols. 

After  the  nervous  tissue  has  been  well  hardened,  take  pieces  of  suitable  size 
and  character,  wash  them  in  water  to  extract  the  formaldehyde,  dehydrate,  and 
finally  imbed  them  in  celloidin  in  the  usual  manner.  The  sections  should  be  cut 
as  thick  as  '20  to  30//,  since  a  considerable  thickness  is  necessary  to  define  general 
relations. 

The  sections  are  to  be  brought  from  distilled  water  into  the  mordant  of  iron 
alum: 

Ammonia-ferric  alum         -         .         -         .         4  grams. 
Distilled  water  -  .         .         .  IQQ  c.  c. 

The  duration  of  the  mordanting  is  not  of  especial  consequence,  but    may    be 
some  two  to  four  hours  in  length.     The    free  mordant  must  now  be  rinsed  away 
with  distilled  water,  a  moment  only.     Then  bring  the  sections  into  the  stain  : 
Haematoxylin  crystals         -         -         -         -       0.5  gram. 
Distilled  water         .         .         .         .         .     lOO.O  c.  c. 
Staining  will  require  at  least  four  hours  for  entirely  satisfactory  definition. 

The  uncombined  stain  is  to  be  washed  away  with  water.  Clean  tap-water  is 
best,  since  this  appears  to  fix  the  lake  more  firmly. 

The  stain  now  requires  dififerentiation.  Dilute  some  of  the  iron  alum  solution 
used  as  a  mordant  with  an  equal  volume  of  distilled  water,  and  pour  this  over 
the  sections.  The  full  strength,  four  per  cent.,  may  be  used  later  if  necessary. 
Only  a  few  sections  should  be  decolorized  at  a  time,  and  these  should  be  moved 
about  in  the  fluid  to  secure  even  results.  Observe  the  process  of  decolorizing 
with  the  microscope  from  time  to  time,  and  when  the  desired  effect  has  been 
obtained,  transfer  the  sections  to  tap-water.  Nerve-cells  and  nerve-fibres  should 
appear  blue  on  a  light  field. 

Thorough  washing  of  the  sections  after  staining  is  necessary  to  prevent 
subsequent  fading.  The  slight  alkalinity  of  ordinary  tap-water  appears  to  be  a 
factor  aiding  in  the  preservation  of  the  stain. 

Light  counter-staining  with  orange-G  is  almost  always  desirable.  After 
washing,  bring  the  sections  for  one  to  two  minutes  into  a  nearly  saturated  aqueous 
solution  of  orange-G  (Griibler's).  Dehydrate  rapidly  with  ninety-five  per  cent, 
alcohol,  clear,  and  mount  in  balsam. 

Iron  haematoxylin  may  also  be  employed  in  the  study  of  the  minute  structure 
of  the  nerve-cell.  Small  pieces  of  the  brain  or  cord  should  be  fixed  with  the 
fluid  of  Flemming,  or  with  the  chrome-oxalic  mixture  given  below  (2).  Imbed 
in  paraffin,  and  cut  the  sections  quite  thin.  For  such  cytological  study,  it  has 
been  found  preferable  to  omit  the  counter-staining  with  orange-G. 

2.      The  Staining  Method  of  Nissl. 

The  staining  method  of  Nissl  is  invaluable  for  demonstrating  the  organiza- 
tion of  the  nerve-cell  in  general,  as  well  as  for  the  comparative  study  of  different 
types  of  nerve-cells.     The  cervical  cord  of  the  rabbit  is  a  particularly    favorable 


and  Laboratory  Methods.  1559 

object  for  the  former  purpose  ;  while  for  the  latter,  material  should  be  chosen 
from  sensory  ganglia,  from  the  spinal  cord,  and  from  the  several  regions  of  the 
brain. 

For  the  fixation  of  the  tissues,  the  writer  has  found  nothing  so  admirable  as 
the  chrome-oxalic  mixture  of  Graf  : 

Oxalic  acid,  S  per  cent.  aq.  sol.         -         -  200  c.  c. 

Alcohol,  05  per  cent.    -----     150  c.  c. 
Chromic  acid,  1  percent,  aq.  sol.       -         -         150  c.  c. 
Mix  in  the  order  as  named. 
This  fluid  has  been  given  a  thorough  trial  with  nervous  tissue  from  many    verte- 
brates, and  it  has  proven  uniformly  satisfactory.     Quite  small  pieces  of  perfectly 
fresh  tissue  should  be  fixed  for  some  six  hours.     It  is  preferable  to  wash  out  the 
fixing  agent  with  seventy  per  cent,  alcohol,  changed  repeatedly. 

Thin  sections  should  be  made  by  the  paraffin  method.  It  is  quite  desirable 
that  the  sections  be  mounted  in  serial  order,  so  that  the  organization  of  any 
given  nerve-cell  can  be  traced  completely.  Mayer's  albumen  is  the  safest  thing 
to  use  for  sticking  the  sections  to  the  slide,  since  some  of  the  subsequent  steps 
make  strong  demands  on  the  tenacity  of  the  medium.  If  the  sections  are  at  all 
wrinkled,  a  film  of  water  should  be  interposed  in  the  usual  manner. 

Dissolve  the  paraffin  from  the  sections  with  xylol,  and  carry  the  slide  through 
descending  grades  of  alcohol  to  distilled  water.  The  sections  are  now  ready  for 
staining. 

The  stain  of  Nissl  has  the  following  composition  : 

Methylen-blue  (Grlibler's  BX)         -         -       3.75  grams. 
Olive-oil  soap      -----  1.75       " 

Distilled  water         -         -         -         .         1000.00  c.  c. 
Use  any  pure  soap  from  one  of  the  southern  countries  of  Europe  where  olive-oil 
is  employed  in  soap-making.     The  soap  should  be  cut  into  thin  shavings ;  gentle 
heat  may  be  employed  for  hastening  its  solution.     The  stain  should   be    filtered 
just  before  using. 

Rest  the  slide  to  be  stained  on  a  watch-glass  so  that  its  surface  will  be  level. 
Heat  five  c.  c.  of  the  Nissl  stain  in  a  test-tube  until  it  almost  boils.  Pour  the  hot 
stain  over  the  slide,  and  allow  it  to  act  for  five  minutes.  Have  at  hand  a  quantity 
of  bibulous  paper.  Flood  the  slide  w'ith  distilled  water  for  the  briefest  practi- 
cable time,  just  enough  to  rinse  away  the  free  stain.  Absorb  as  much  of  the 
water  clinging  to  the  slide  as  possible  with  the  porous  paper,  and  immediately 
pour  on  the  decolorizer  : 

Alcohol,  95  per  cent.       -         -         -         -         -     90  c.  c. 
Aniline  oil  -         -         -         -         -         -  10    c.  c. 

The  process  of  decolorizing  is  the  only  part  of  this  method  involving  especial 
difficulty,  as  it  lasts  only  20  to  30  seconds  and  must  be  stopped  at  just  the  right 
point.  Hold  the  slide  between  the  thumb  and  finger,  and  rock  it  back  and  forth 
so  as  to  move  the  decolorizer  over  the  entire  field.  This  reagent  should  be 
applied  liberally.  Watch  the  process  closely.  Just  as  soon  as  the  sections 
begin  to  look  nearly  colorless  and  take  on  a  delicate  rose  tint,  stop  the  decolor- 
izing by  flooding  the  slide  with  oil  of  cajeput. 


1560  Journal  of  Applied  Microscopy 

The  oil  of  cajeput  will  clear  the  sections  within  a  few  seconds  in  perfectly 
dry  air ;  but  under  the  usual  conditions  of  the  laboratory,  it  is  desirable  to 
hasten  the  action  by  warming  the  slide  gently.  Mount  in  a  thick  solution  of 
colophonium  dissolved  in  xylol. 

A  word  concerning  colophonium  may  not  be  out  of  place  here.  Colophonium 
is  the  ordinary  •'  rosin  ''  of  commerce.  It  is  best  to  secure  the  privilege  from 
the  dealer  of  selecting  those  lumps  which  have  the  least  possible  color.  When 
held  up  to  the  light,  a  suitable  lump  should  not  show  crystals  on  the  inside. 
The  powdered  rosin  of  dealers  should  be  avoided,  since  its  character  cannot  be 
determined. 

When  the  several  steps  of  the  method  as  outlined  are  followed  faithfully,  the 
staining  wnll  be  so  precise  that  the  nerve-cells  will  appear  bright  blue  on  a  per- 
fectly colorless  field.  Moreover,  the  tigroid  bodies  should  not  appear  blurred  in 
any  part  of  the  ner\-e-cell.  As  to  the  permanence  of  the  stain,  slides  stained  by 
me  several  years  ago  have  shown  no  tendency  to  fade. 

3.     Methylcn-Blue  and  Eryth rosin. 

Staining  with  methylen-blue  alone,  as  just  noted,  simply  outlines  the  bodies 
of  nerve-cells.  As  an  accessory  demonstration,  it  is  often  quite  desirable  that 
the  remaining  structures  present  be  given  a  light  stain.  This  may  be  accom- 
plished through  the  use  of  the  following  double-staining  method. 

The  tissues  should  be  treated,  and  the  sections  should  be  cut  and  affixed  to  the 
slide  as  in  the  piocedure  given  for  the  method  of  Xissl.  But,  instead  of  staining 
with  methylen-blue  at  once,  this  is  preceded  by  the  erjthrosin  mixture  of  Held  : 

Erythrosin  (^Griibler)  .         .         .         .  l  gram. 

Distilled  water        -----  150  c.  c. 

Glacial  acetic  acid     -----  2  drops. 

The  acetic  acid  tends  to  precipitate  the  erj'throsin,  and  so  the  stain  should  be 
made  shortly  before  using.  Warm  the  mixture,  pour  it  over  the  slide,  and  allow 
it  to  act  for  some  ten  seconds,  only.  Rinse  with  distilled  water  for  about  thirty 
seconds  ;  the  red  color  should  be  made  distinctly  lighter. 

Now  dilute  some  of  the  regular  stain  of  Xissl  with  an  equal  volume  of  a  five 
per  cent,  aqueous  solution  of  acetone.  Heat  the  mixture  rather  strongly,  flood 
the  slide  with  it,  and  allow  staining  to  proceed  for  five  minutes.  Then  difier- 
entiate  the  stain  with  : 

Distilled  water         -----         100.0  c.  c. 

Common  alum     ------  .1  gram. 

The  solution  of  alum  extracts  the  blue  color  gradually,  so  that  the  effect  is  readily 
controlled.  From  one  to  two  minutes  will  usually  sufiice  to  bring  the  red  of  the 
erythrosin  distinctly  into  view.  Follow  with  a  brief  rinsing  in  water.  Dehydrate 
rapidly  with  absolute  alcohol,  clear  in  xylol,  and  mount  in  xylol-colophonium. 

A  successful  preparation  should  have  the  nerve-cells  outlined  in  bright  blue 
on  a  pale  red  field.     Too  deep  a  stain  with  erythrosin  must  be  avoided. 

4.      Chro7)U'Silver  Impregnation. 

The  principle  of  this  remarkable  method  involves  the  formation  of  a  deposit 
of  chrome-silver  in  the  elements  of  the  nervous  system.     This  is  accomplished 


and  Laboratory  Methods.  15(51 

by  hardening  in  potassium  bichromate,  followed  by  impregnation  with  silver 
nitrate.  The  production  of  the  metallic  deposit  is  not  in  the  nature  of  a  staining 
process  ;  it  is  the  outcome,  rather,  of  an  interaction  between  the  potassium 
bichromate  and  the  silver  nitrate  employed,  in  conjunction  with  the  substances 
present  in  the  nervous  elements  themselves.  Since  the  chemistry  of  nervous 
tissue  is  continually  changing  during  life,  it  follows  that  the  results  of  this  method 
will  be  far  from  uniform.  In  one  animal,  certain  elements  may  be  brought  out 
with  all  the  crisp  sharpness  of  a  silhouette,  while  in  another  individual  the 
corresponding  structures  may  not  appear  at  all.  The  reactions  of  the  tissues  in 
the  two  instances  have  differed  sufficiently  to  thus  affect  the  result.  Hence  the 
apparent  qapriciousness  of  the  method  is  due  to  the  presence  of  physiological 
factors  often  quite  unknown.  The  character  of  some  of  these  physiological 
conditions  has  been  discussed  by  me  elsewhere.* 

Freshness  of  tissue  and  active  metabolism  are  factors  indispensable  to  suc- 
cess. A  young  animal  will  yield  better  results  than  an  old  one  ;  and  one  direct 
from  the  field  is  preferable  to  one  which  has  been  kept  for  a  time  in  confine- 
ment. In  any  case,  the  parts  of  the  nervous  system  to  be  studied  should  be 
placed  in  the  hardening  mixture  as  soon  as  possible  after  the  death  of  the 
animal. 

a.  The  Rapid  Method  of  Golgi. — Of  the  many  schemes  for  securing  the  forma- 
tion of  the  chrome-silver  deposit,  the  one  which  has  given  me  the  largest  returns 
is  the  so-called  "  rapid  "  method  of  Golgi.  For  this  purpose,  the  pieces  should 
be  taken  small,  not  over  two  or  three  millimeters  in  thickness  for  a  slice  of  the 
brain.     Place  these  fresh  slices  directly  in  the  hardening  mixture : 

Potassium  bichromate,  8.5  per  cent.  sol.  -       4  vols. 

Osmic  acid,  1  per  cent.  sol.         .         .         .  1  vol. 

This  reagent,  while  somewhat  expensive,  must  be  used  liberally.  There  should 
always  be  a  large  volume  in  proportion  to  the  mass  of  the  nervous  matter  present, 
and  the  solution  should  be  renewed  before  it  shows  the  least  signs  of  becoming 
turbid.     Hardening  must  take  place  in  the  dark. 

The  duration  of  the  hardening  is  the  all-important  factor  which  we  have 
under  control.  If  the  tissues  be  hardened  for  too  short  a  time,  the  impregna- 
tion will  be  diffuse  ;  while  an  overhardened  specimen  may  permit  of  no  impregna- 
tion at  all.  The  proper  length  of  hardening  must  be  determined  by  experiment 
for  each  animal ;  it  will  most  often  be  found  to  lie  between  the  limits  of  two  to 
three  days  where  supporting  elements  are  to  be  shown,  three  to  five  days  for 
neurones,  and  five  to  seven  days  for  nerve-fibres. 

Impregnation  is  secured  by  bringing  the  hardened  slices  into  a  solution  of 
silver  nitrate  : 

Silver  nitrate  crystals         .         .         .         .     0.75  gram. 

Distilled  water       -----      100.00  c.  c. 

A  solution  which  has  been  used  once  will    answer    for   washing   the    specimens 

until  the  copious  precipitate  ceases  to  form.     Then  pour  on  a  liberal  quantity  of 

the  fresh  solution,  and  change  it  frequently.     It  should  not  be    allowed    to   take 

*  Houser,  1901.  The  Neurones  and  Supporting  Elements  of  the  Brain  of  a  Selachian:  Jour- 
nal of  Comparative  Neurology,  Vol.  XI. 


150-2  Journal  of  Applied  Microscopy 

on  a  yellowish  tinge.  Keep  the  specimens  in  the  dark.  The  duration  of  the 
silver-bath  is  not  of  importance ;  it  may  be  one  to  three  days  in  length,  or  even 
longer  if  the  solution  be  renewed. 

When  the  steps  which  follow  can  be  carried  through  without  a  pause,  replace 
the  silver  solution  with  ninety-five  per  cent,  alcohol.  Change  the  alcohol 
repeatedly  during  the  course  of  half  an  hour,  in  order  that  all  free  silver  nitrate 
may  be  washed  away.  Follow  with  absolute  alcohol,  also  renewed,  thirty  minutes  ; 
alcohol  and  ether,  fifteen  minutes  ;  thin  celloidin,  thirty  minutes  ;  and  thick  cel- 
loidin,  five  minutes.  Then  mount  the  specimen  on  a  wooden  block,  and  harden 
the  celloidin  with  chloroform.  The  imbedding  has  been  too  hastily  done,  of 
course,  to  permit  true  infiltration  with  celloidin,  but  this  cannot  be  secured  with- 
out destroying  the  impregnation. 

Clearing  of  the  entire  mass,  as  imbedded,  is  advantageous.  For  this  purpose, 
place  the  block  in  a  mixture  of  oil  bergamot,  oil  cedar-wood,  and  melted  car- 
bolic acid  crystals,  equal  parts.  This  mixture  clears  rapidly,  it  may  be  used 
over  and  over  again,  and  it  has  the  additional  advantage  of  allowing  the  prepara- 
tions to  be  kept  in  it  for  a  little  while  without  impairing  the  impregnation. 

Sections  should  be  cut  as  thick  as  75;^.  In  cutting,  keep  the  knife  flooded 
with  the  clearing  mixture.  Place  the  sections  on  slides,  as  desired.  The  sur- 
plus oil  may  be  removed  neatly  by  pressing  on  the  sections  with  tissue  paper 
backed  with  a  blotter.  Mount  in  either  colophonium,  dammar,  or  balsam,  with- 
out a  cover-slip.  The  mounting  medium  should  cover  the  section  with  a  thin 
and  even  coating.  Hasten  the  drying  of  the  preparation  with  gentle  heat  for  a 
few  hours. 

Successful  preparations  should  have  the  neurones,  to  their  finest  ramifications, 
a  perfectly  opaque  black  on  a  nearly  transparent  field.  If  the  field  becomes 
clouded  with  fine  specks  later  on,  the  silver  nitrate  was  not  all  removed 
in  the  washing. 

Under  certain  conditions,  pure  formaldehyde  may  be  substituted  for  the 
osmic  acid  in  the  hardening  mixture  given  above,  a  good  proportion  being  three 
parts  of  commercial  formaldehyde  to  one  hundred  parts  of  the  bichromate  solu- 
tion. The  formaldehyde  tends  to  reduce  the  potassium  bichromate  quite  rapidly, 
and  so  the  two  should  be  mixed  just  before  using.  The  application  of  formal- 
dehyde in  this  way  often  yields  beautiful  results  with  the  mammalian  nervous- 
system,  but  I  have  not  been  so  successful  with  it  in  the  field  of  the  lower  verte- 
brates. 

b.  Previous  Hardening  with  Formaldehyde. — It  is  often  quite  desirable  to  be 
able  to  harden  a  brain  some  time  in  advance  of  its  final  use  for  impregnation,  as 
in  planning  for  classwork,  or  in  the  transportation  of  specimens  from  the  field  to 
the  laboratory.  This  may  be  accomplished  through  preliminary  hardening  with 
formaldehyde. 

The  whole  brain  is  hardened  in  ten  per  cent,  formaldehyde,  and  should  be 
kept  in  this  strength  of  solution  until  it  is  wanted.  Slices  cut  from  the  desired 
regions  are  washed  with  water  for  half  an  hour  to  extract  the  formaldehyde,  and 
are  then  to  be  saturated  with  potassium  bichromate. 

Place  the  pieces  in  a  three  and  five-tenths  per   cent,    solution    of   potassium 


and  Laboratory  Methods.  15(>3 

bichromate  in  an  oven  kept  at  60°C.  for  twelve  hours.  Change  the  fluid  fre- 
quently. Continue  the  saturation  with  potassium  bichromate  in  the  dark  box  for 
five  to  seven  days  ;  the  proper  duration  must  be  determined  by  tests  made  from 
time  to  time.     Then  transfer  to  the  solution  of  silver  nitrate. 

From  this  point,  the  procedure  is  the  same  as  for  the  "  rapid  "  method  of 
Golgi  previously  described.  This  plan  is  especially  successful  for  the  forebrain 
and  spinal  cord.  In  any  case  where  there  is  good  impregnation,  the  beauty  of 
the  preparation  is  enhanced  by  the  perfect  clearness  of  the  field. 

It  may  be  worth  noting  here  that  this  same  method  is  particularly  desirable 
for  demonstrating  the  bile-capillaries  of  the  mammalian  liver.  A  shortening  of 
the  bichromate-bath  is,  of  course,  necessary  in  this  case. 

5.      JVeigerfs  Myelin  Stain. 

The  study  of  the  central  nervous  system  cannot  be  complete  without 
preparations  to  illustrate  the  course  of  medullated  nerve-fibres.  The  many 
schemes  extant  for  staining  the  myelin  sheath  depend  upon  the  formation  in  the 
nervous  elements  of  either  a  chrome-  or  copper-lake  of  hiEmatoxylin.  This  lake 
combines  so  firmly  with  myelin  that  the  decolorizing  process  does  not  remove  it. 
Doubtless  the  chrome-lake  yields  the  more  delicate  results,  and  hence  some  plan 
for  securing  it  is  to  be  recommended  for  purposes  of  •special  investigation.  But 
for  general  classwork,  the  writer  has  found  a  copper  method  preferable  because 
of  its  entire  certainty  under  all  conditions. 

Harden  the  brain  and  spinal  cord  of  the  animal  chosen  for  study  in  a  bichro- 
mate solution.  This  will,  of  course,  require  several  months  if  the  well-known 
fluid  of  Miiller,  or  a  simple  solution  of  a  bichromate  salt  be  used.  The  time  may 
be  shortened  to  a  more  convenient  length  by  preliminary  hardening  with  formal- 
dehyde ;  or,  formaldehyde  may  be  added  directly  to  the  bichromate  solution.  In 
any  case,  the  hardening  should  take  place  in  the  dark. 

When  the  hardening  has  been  completed,  cut  slices  from  the  regions  desired, 
and,  without  washing  in  water,  transfer  them  to  graded  alcohols  for  dehydration, 
still  in  the  dark. 

After  the  pieces  have  been  dehydrated,  imbed  them  in  celloidin,  allowing 
plenty  of  time  for  thorough  penetration.  Harden  the  celloidin  in  eighty  per 
cent,  alcohol. 

The  celloidin  blocks  are  now  to  be  mordanted  for  two  days  in  a  half-saturated 
solution  of  neutral  acetate  of  copper.  At  the  end  of  this  time,  place  the  blocks 
back  in  eighty  per  cent,  alcohol  for  a  day,  and  then  cut  sections  at  once. 

Sections  should  have  a  thickness  of  25//.  Rinse  the  sections  rapidly  with 
distilled  water,  and  bring  them  directly  into  the  stain  : 

.   \  Distilled  water         -         -         -       93  c.  c.     )  q      , 

(  Lithium  carbonate,  sat.  aq.  sol.     -     7  c.  c.      ^ 
^  <  Absolute  alcohol         -         -         -     10  c.  c.     )  ..        , 
( Haematoxylin  crystals     -         -  1  gram.  ) 

Mix  A  and  B  just  before  using. 
The  duration  of   the  staining  will  depend  upon  the  region  involved.     For  sections 
of  the  spinal  cord,  two  hours  will  be  sufficient ;  but  for  sections   of   the   brain, 
twenty-four  hours  will  be  required. 


1-564  Journal  of  Applied  Microscopy 

Rinse  away  the  excess  stain  for  a  few  minutes  with  distilled  water.  Then 
transfer  the  sections  to  the  decolorizer  : 

Distilled  water         -----      1000.0  c.  c. 

Potassium  ferricyanide         -         -         -  12.5  grams. 

Borax     -------  10.0     " 

The  decolorizing  must  be  watched  carefully  so  that  it  may  be  stopped  at  just  the 
right  moment.  The  gray  matter  should  be  bleached  to  a  yellow  tint,  while  the 
medullated  nerve-fibres  should  remain  a  bright  blue-black.  When  the  action  has 
gone  far  enough,  transfer  the  sections  to  clean  tap-water,  running,  if  possible,  in 
which  they  must  be  washed  for  some  time.  If  the  washing  be  thorough,  the 
stain  will  not  fade. 

Dehydrate,  clear,  and  mount  the  sections  in  the  usual  manner. 

6.      Staifihig  Isolated  Nerve-Cells. 

Some  plan  for  making  preparations  of  entire  nerve-cells  is  altogether  desirable 
for  class  demonstrations.  For  this  purpose,  the  writer  is  about  to  risk  his  repu- 
tation as  a  modern  microscopist  by  advocating  the  use  of  a  stain  which  the  older 
workers  are  supposed  to  have  forgotten,  and  of  which  the  younger  ones  have 
never  even  heard.  This  stain  is  Beale's  Carmine.  No  other  method  known  to 
the  writer  is  capable  of  yielding  preparations  of  nerve-cells  rivaling  in  clearness 
and  beauty  the  results  of  the  procedure  here  indicated. 

Secure  the  cooperation  of  a  butcher,  and  have  him  furnish  the  lumbar  cord 
of  the  ox  in  a  perfectly  fresh  condition.     Slit  the  cord  lengthwise  so  as  to  expose 
the  gray  matter.     Scoop  out  quite  small  pieces  from  the  anterior  cornua.     Place 
these  fragments  directly  in  the  following  stain  : 

Carmine         --..-.  3  grams. 

Ammonia  -         -         -         -         -         -       10  c.  c. 

Glycerine      -         -         -         -  '       -         -         300  c.  c. 

Water        -         -         -         -         -         -         -     300  c.  c. 

Alcohol  -         -         -         -         -         -  75  c.  c. 

(Dissolve  the  carmine  in  the  ammonia  with  the  aid  of  gentle  heat ;  allow  the  solu- 
tion to  stand  in  an  open  dish  for  an  hour ;  then  add  to  the  other  ingredients. 
Filter.) 

The  pieces  of  nervous  matter  are  to  lie  in  this  stain  for  several  weeks.  Then 
transfer  them  to  dilute  glycerine  acidulated  with  a  trace  of  acetic  acid.  After  a 
few  days,  replace  this  medium  with  pure  glycerine. 

Place  a  bit  of  the  stained  tissue  on  a  slide,  in  a  drop  of  glycerine.  Press  a 
cover-slip  straight  downward  on  the  mass  so  as  to  spread  the  nervous  matter  in 
an  even  film.  Avoid  rotating  the  cover-slip.  Some  of  the  preparations  will 
necessarily  show  so  few  cells  as  to  be  practically  valueless,  but  others  will  be 
crowded  with  brilliantly  stained  nerve-cells  in  all  their  completeness.  Aside 
from  the  value  of  such  a  preparation  for  purposes  of  demonstration,  it  is  worth 
noting  that  the  fibrillar  elements  of  the  neurone  are  made  particularly  evident. 

7.     Peripheral  Neri'e-Fibres. 

Place  short  lengths  of  the  sciatic  nerve  of  the  frog  in  five-tenths  per  cent, 
osmic  acid   for   about  twenty-four  hours.     Keep  in  the  dark.     The  osmic   acid 


and  Laboratory  Methods.  1565 

blackens  the  myelin  sheath  of  the  nerve-fibre.     When  the  degree    of   blackening 
desired  has  been  obtained,  wash  the  nerve  thoroughly  with  distilled  water. 

A  piece  of  the  nerve  may  now  be  transferred  to  a  drop  of  glycerine  on  a  slide, 
teased  apart,  and  used  at  once  for  a  demonstration.  Other  pieces  should  be 
imbedded  in  paraffin  for  sectioning.  It  is  convenient  to  arrange  the  pieces  so 
that,  in  the  same  block,  some  will  be  cut  lengthwise  and  others  transversely. 
The  sections  should  be  counter-stained  with  a  saturated  aqueous  solution  of 
fuchsin-S  for  twenty-four  hours.  Gilbert  L.  Houser. 

Laboratory  of  Animal  Morphology,  The  State  University  of  Iowa. 


A  Short  Method  for  the  Widal  Test. 

One  of  the  inconveniences  of  this  test  is  the  length  of  time  required  to  get  a 
broth  culture  in  which  the  typhoid  bacilli  are  sufficiently  numerous  and  active  to 
permit  of  use  for  the  test.  If  the  bacilli  are  too  few  in  number  the  "  clumps  " 
are  not  only  small,  but  are  also  slow  in  forming;  and  if  they  are  not  active 
agglutination  does  not  take  place  readily.  Eighteen  hours  is  the  time  usually 
specified  as  being  necessary  to  get  a  broth  culture  in  proper  condition  for  use, 
but  this  length  of  time  makes  it  impossible  to  apply  the  test  during  the  day  on 
which  the  culture  was  started,  so  the  tube  must  be  inoculated  late  in  the  after- 
noon of  the  day  before  it  is  to  be  used  in  order  that  it  may  be  ready  for  the  test 
at  a  seasonable  hour  the  next  morning.  If  the  culture  is  not  then  used  rather 
promptly  at  the  specified  time,  agglutination  is  likely  to  be  found  to  have  taken 
place  spontaneously  in  the  tube.  The  culture  is  thus  rendered  useless  and  a 
delay  of  eighteen  hours  more  is  required  to  get  another  one  ready.  The  writer 
has  found  that  the  time  can  easily  be  shortened  to  six  or  eight  hours  and  that 
cultures  started  in  the  morning  can  be  used  in  the  afternoon  of  the  same  day. 
The  essential  point  of  the  method  is  to  start  the  culture  in  wann  broth.  The 
details  are  so  simple  as  scarcely  to  require  explanation.  By  means  of  the 
platinum  loop  a  small  mass  of  the  growth  is  removed  from  the  stock  culture  on 
agar  and  is  well  mixed  with  eight  or  ten  cubic  centimeters  of  broth  by  rubbing 
the  mass  against  the  inside  of  the  tube  below  the  surface  of  the  liquid.  The 
tube  is  then  gently  tapped  for  a  minute  or  so  in  order  to  insure  the  thorough 
and  even  dissemination  of  the  bacilli  throughout  the  broth,  and  is  heated  until 
it  feels  comfortably  warm  to  the  hand  by  holding  well  above  the  gas  flame.  A 
tumbler  is  then  heated  still  warmer  by  holding  it  inverted  over  the  flame  and  the 
culture  placed  in  it  resting  on  a  wad  of  cotton  and  the  whole  put  into  the  incu- 
bator for  six  or  more  hours.  It  is  sometimes  well  to  remove  the  tube  once  from 
the  incubator  and  tap  it  gently  for  a  few  seconds  in  order  to  mix  the  denser 
growth  in  the  bottom  with  the  upper  portion  of  the  broth.  The  opacity  and 
opalescence  of  the  liquid  will  show  when  sufficient  growth  has  taken  place.  On 
several  occasions  cultures  only  six  hours  old  have  been  used  with  perfect  satis- 
faction, but,  of  course,  the  bacilli  are  neither  so  active  nor  so  numerous  as  in 
cultures  which  have  stood  in  the  incubator  for  two  or  three  hours  longer. 
University  of  Rochester.  CharLE.S   Wright   Dodge. 


1566  Journal  of  Applied  Microscopy 


Method  for  Rearing  Amoeba. 

It  is  frequently  very  difficult  to  secure  a  large  number  of  Amoebaj  satisfactory 
for  class  work  when  wanted.  The  usual  method  of  visiting  the  pools  of  stagnant 
water  is  very  uncertain  and  laboratory  cultures  made  from  dacaying  Algae  are 
equally  unsatisfactory.  For  some  time  the  writer  has  been  experimenting  with 
protozoan  cultures  and  has  been  successful  in  securing  Amoebae  in  the  following 
simple  manner  :  A  nutrient  medium  is  made  by  boiling  a  lot  of  dead  leaves. 
As  soon  as  cool,  both  liquid  and  leaves  are  placed  in  an  ordinary  battery  jar 
and  a  lot  of  unboiled  leaves  and  enough  water  to  stand  about  one  inch  above 
the  leaves  added.  In  two  or  three  days  a  scum  will  form  and  in  from  five  to 
ten  days,  depending  upon  the  temperature  of  the  room,  Amcebae  will  be  found  in 
the  scum  in  large  numbers.  They  will  be  small,  but  very  satisfactory  for  class 
work.  The  writer  has  tried  this  method  a  great  many  times,  and  in  different 
localities  and  at  different  seasons  of  the  year,  and  always  been  successful. 
De  Pauw  University,  Greencastle,  Indiana.  Mel   T.    Cook. 


The  Arrangement  of  Cilia  on  Paramecium. 

The  cilia  of  many  of  the  Infusoria  are  so  fine  and  so  closely  set,  notably  on 
Paramecium,  that  it  is  very  difficult  for  the  student  to  determine  their  exact 
arrangement.  Careful  focusing  will  sometimes  reveal  their  order  on  favorable 
specimens,  but  the  use  of  Loeffler's  alkaline  methylen  blue  as  a  stain  has  been 
found  to  be  a  very  reliable  method  of  demonstrating  their  arrangement  on 
almost  every  specimen.  A  drop  of  the  stain  is  mixed  with  the  drop  of  water  in 
which  the  animalcules  are  swimming  on  the  slide  and  the  cover  glass  placed  in 
position.  Intra  vitam  staining  takes  place  in  many  of  the  individuals,  but  they 
soon  die  and  the  cuticle  separates  more  or  less  completely  from  the  cytoplasm 
and  forms  a  halo  around  the  deeply  stained  body.  The  perforations  in  the  cu- 
ticle are  thus  brought  very  distinctly  into  view  and  the  plan  of  arrangement  of 
the  cilia  revealed.  The  stain  is  prepared  as  follows  :  Add  8(1  c.c.  of  a  concen- 
trated alcoholic  (05  per  cent.)  solution  of  methylen  blue  to  100  c.c.  of  a  .0001 
solution  of  caustic  potash.  The  caustic  potash  solution  may  be  prepared  by 
adding  1  c.c.  of  a  one  per  cent,  solution  of  potash  to  100  c.c.  of  distilled  water. 
University  of  Rochester.  CharlES   Wright   Dt)J)(;E. 


Labeling  Mounted  Specimens. — The  following  method  is  given  as  useful 
where  specimens  mounted  on  cover-glasses  or  slides  are  to  be  passed  through 
various  solutions  :  Mix  with  equal  parts  of  ^^g  albumen  and  glycerin,  sufficient 
lampblack  to  make  a  good  black  fluid.  This  may  be  used  with  a  steel  pen  for 
writing  on  the  glass,  after  which  the  glass  should  be  held  over  a  flame  until  the 
characters  are  dry.  The  markings  are  not  removed  when  the  specimens  are 
passed  through  solutions. — Knmvledge,  24  :  191. 


and  Laboratory  Methods.  1567 

Immersion  Oil  in  Collapsible  Tubes. 

The  immersion  oil  bottle  shares  with  the  Canada  balsam  bottle  the  distinc- 
tion of  being  the  stickiest  and  dirtiest  piece  of  apparatus  on  the  work  table. 
A  portion  of  the  first  drop  of  oil  to  be  removed  is  almost  invariably  left  on  the 
mouth  of  the  bottle  and  thereafter  the  stopper  never  fits  tightly ;  the  oil  runs  over 
the  neck  of  the  bottle  and  smears  the  fingers  in  spite  of  almost  all  precautions ; 
in  the  course  of  time  the  oil  thickens,  turns  yellow,  becomes  turbid  as  the  result 
of  exposure  to  the  light  and  air,  and  its  refractive  index  is  thus  changed.  A 
container  which  excludes  both  light  and  air,  holds  the  oil  in  such  a  manner  as 
to  reduce  the  inconvenience  of  handling  to  a  minimum,  delivers  the  exact  amount 
needed  each  time,  and  is  made  of  a  material  which  causes  no  change  in  the 
optical  properties  of  the  oil,  is  the  ordinary  collapsible  tube  which  has  long  been 
used  for  holding  moist  water  color  and  oil  paints.  During  the  past  few  years 
certain  dealers  in  microscope  supplies  have  used  this  form  of  tube  as  a  container 
for  Canada  balsam  with  most  satisfactory  results.  After  several  annoying  ex- 
periences with  bottles,  the  writer  determined  to  have  immersion  oil  put  up  in 
tubes  although  warned  that  the  metal  of  which  the  tubes  are  made  might  have 
some  deleterious  effect  upon  the  optical  properties  of  the  oil.  During  the  past 
year  the  tubes  have  been  used  daily  in  the  routine  bacteriological  examinations 
attending  health  department  work  and  have  proved  to  be  so  free  from  the  faults 
which  characterize  the  bottles  that  on  no  account  will  the  latter  be  again  em- 
ployed. Immersion  oil  which  has  been  stored  in  these  tubes  for  more  than  a 
year  is  now  being  used  and  no  signs  of  deterioration  have  been  detected.  The 
oil  is  as  clear,  colorless  and  thin  as  when  first  made,  and  there  is  no  indication  of 
change  in  its  refractive  index.  Charles  Wright  Dodge. 

University  of  Rochester. 


The  minute  adopted  by  the  council  of  Columbia  University  on  the  resigna- 
tion of  President  Low  gave  a  few  very  interesting  facts  concerning  the  develop- 
ment of  that  institution  in  the  twelve  years  during  which  Mr.  Low  has  been  at 
the  head  of  its  administration.  In  the  academic  year  1889-90  the  institution 
consisted  of  four  faculties,  in  charge  respectively  of  the  schools  of  arts,  laws, 
mines,  and  political  science.  These  faculties  numbered  122  officers  of  instruc- 
tion, and  the  schools  were  attended  by  1134  students.  The  library  of  the 
college  contained  91,000  volumes,  and  the  wealth  of  the  corporation  was 
estimated  at  $10,500,000.  The  faculties,  schools,  library,  and  entire  equipment 
were  crowded  into  narrow  and  noisy  quarters,  bordering  upon  the  tracks  of  the 
New  York  Central  Railway. 

To-day  the  university  consists  of  nine  faculties,  in  charge  respectively  of 
Columbia  College,  Barnard  College,  Teachers' College,  and  the  university  schools 
of  law,  medicine,  applied  science,  pure  science,  philosophy,  and  political  science. 
The  faculties  now  number  385  officers  of  instruction,  and  the  colleges  and  schools 
are  now  attended  by  4500  students.  The  library  of  the  university  now  contains 
311,000  volumes,  and  the  wealth  of  the  corporation  is  now  estimated  at 
$18,000,000,  of  which  $1,500,000,  in  round  numbers,  were  given  by  Mr.  Low 
himself.  And  finally,  the  university  is  now  located  upon  a  site,  and  possesses 
a  physical  equipment,  unsurpassed  in  beauty,  comfort,  and  completeness,  by 
those  of  any  institution  of  learning  in  the  world. — Science,  14 :  35G. 


1508 


Journal  of  Applied  Microscopy 


LABORATORY  PHOTOGRAPHY. 

Devoted  to  methods  and  apparatus  for  converting  an  object  into  an  illustration. 

FURTHER  NOTES  ON  THE  USE  OF  THE  TELEPHOTO  LENS. 

In  Volume  IV,  No.  4,  of  the  Journal  of  Applied  Microscopy,  I  gave  an 
illustration  of  the  use  of  the  telephoto  lens  in  taking  a  nest  from  the  top  of  a 
dead  pine  tree.  During  the  past  summer  I  made  further  efforts  to  test  the  value 
of  a  long  focus  lens,  and  give  some  of  the  results. 

My  telephoto  lens  is  fitted  to  a  wide  angle  Zeiss  Anastigmat,  series  IV. 
The  illustrations  presented  in  this  paper  are  from  experiments  on  mountain 
ranges  and  peaks,  to  be  used  geologically  and  physiographically. 

The  Mission  range  is  in  western  Montana,  extending  north  and  south  for  a 
distance  of  about  a  hundred  miles ;  its  western  slopes  are  abrupt  and  craggy. 
Many  peaks  in  the  range  are  unexplored,  and  few  are  named.  The  difficulties 
one  encounters  in  making  a  study  of  the  range  are  many,  and  a  complete  study 
has  not  yet  been  made. 


Fic;.  1. — View  of  a  Portion  of  the  Mission  Range  of  Mountains  in  Western  Montana. 


Fig.  1  is  a  portion  of  the  southern  end  of  the  range,  taken  from  a  high  hill 
to  the  southwest  of  the  range  proper. 

The  mountain  indicated  by  A  is  McDonald  peak,  distance  from  the  place 
where  the  picture  was  taken  about  fifteen  miles.  The  height  is  about  ten  thousand 
feet,  or  about  seven  thousand  above  the  village  of  St.  Ignatius  on  the  plain. 
The  negative  was  made  early  in  June,  with  an  orthochromatic  plate  and  ray  filter. 
The  atmosphere  was  clear,  and  the  mountains  are  shown  with  much  more  than 
ordinary  distinctness. 

The  mountain  indicated  by  B  is  Sin-yale-a-min  mountain,  distance  about  ten 
to  twelve  miles  on  an  air  line,  with  an  elevation  of  9"20(l  feet.  This  mountain  is 
nearer  than  McDonald,  owing  to  the  position  of  the  photographer  with  regard  to 


and  Laboratory  Methods. 


15G9 


the  range.  The  direction  toward  Sin-yale-a-min  is  almost  due  east,  toward 
McDonald  northeast.  The  photographs  were  taken  at  about  eleven  o'clock  in 
the  morning,  and  hence  the  sun  is  in  the  rear,  or  behind  the  operator.  A  slight 
wind  was  blowing,  requiring  considerable  time  to  secure  the  negatives.  When 
the  wind  blew  so  as  to  shake  the  instrument  the  cap  was  placed  over  the  lens, 
and  was  removed  when  the  instrument  again  became  quiet. 

Fig.  '2  shows  distinctly  many  features  not  made  out  in  Fig.  1,  and  not 
discernible  to  the  naked  eye.  McDonald  peak  is  the  double  peak  in  the  left  of 
the  illustration,  the  peak  to  the  right  being  much  lower,  nearer,  and  not  con- 
nected with  McDonald.  The  ridge  extending  to  the  left  immediately  in  front  of 
McDonald  is  a  separate  peak,  and  between  it  and  McDonald  is  a  deep  canyon 
through  which  fiows  a  good  sized  stream. 

McDonald  peak  is  plainly  seen  to  be  double.  The  distant  peak,  the  right 
hand  snow-capped  peak,  is  about  a  thousand  feet  higher  than  the  nearer  one. 


Fig.  2. — A  Portion  of  the  Range  shown  in  Fig.  1,  indicated  by  A. 

So  far  as  known  it  has  not  been  ascended  from  the  side  seen,  and  is  apparently 
inaccessible,  although  experience  may  prove  it  to  be  reached  from  this  side. 

The  strata  are  clearly  shown.  In  a  photograph  printed  lighter  the  details  of 
the  mountain  side  are  shown  more  clearly,  but  the  snow  summit  is  less  distinct. 
I  do  not  know  that  the  two  peaks  shown  in  the  foreground  and  to  the  right  of 
McDonald  have  ever  been  ascended,  and  I  have  heard  of  no  names  for  them. 
In  this  connection  I  may  say  that  land  snails  are  living  on  the  shoulder  of 
McDonald  shown  in  the  left.  The  plate  used  in  making  Fig.  2  was  an  ortho- 
chromatic,  with  ray  filter.     The  magnification  is  about  eight  diameters. 

The  camera  was  next  turned  toward  Sin-yale-a-min  mountain,  shown  in  Fig. 
3.  The  conditions  were  precisely  the  same,  and  the  position  unchanged. 
By  an  examination  of  Fig.  4,  Sin-yaie-a-min  mountain,  the  snow-capped  peak 
to  the  left,  is  seen  to  be  double.  The  nearer  peak  is  the  lower,  by  some  few 
hundred  feet.  At  a  point  half  way  up  the  peak  along  the  ridge,  snails  similar  to 
to  those  mentioned  on  McDonald  were  found. 

By  printing  for  detail  in  any  one  place  in  the  negative  various  features  may 
be  brought  out  with  much  distinctness. 


l''>"0  Journal  of  Applied  Microscopy 

By  following  up  the  creek  along  the  canyon  to  the  right,  as  shown  in  Fig.  2, 
the  position  from  which  Fig.  4  was  taken  is  found.  The  peak  is  the  same 
as  shown  in  P^ig.  3,  the  direction  of  the  peak  being  east  of  north.  It  was 
several  days  after  taking  the  negatives  from  which  the  first  three  figures  were 
taken.  The  peak  is  distant  from  the  camera  from  five  to  seven  miles.  To 
reach  the  summit  from  the  position  given  in  the  illustration  would  require  five 
or  six  hours  of  stiff  climbing.  The  wooded  peak  in  the  foreground,  immediately 
behind  the  small  tree  in  the  center,  is  a  mile  or  more  nearer  than  the  main  peak, 
and  is  the  end  of  the  slope  shown  in  Fig.  3.  The  day  was  bright  and  almost 
cloudless,  following  a  light  rain.  The  sun  was  in  the  rear.  The  plate  used  was 
an  orthochromatic,  with  ray  filter  as  usual. 

In  Fig.  5  the  telephoto  was  turned  on  the  wooded  ridge  to  the  right  of  the 
middle  of  Fig.  4,  with  the  peak  on  the  left.  By  one  of  those  peculiar  acci- 
dents which  often  occur  while  exchanging  plates  under  a  blanket  in  the  field,  the 


Fig.  3. — A  Portion  of  the  Range  shown  in  Fig.  1,  indicated  by  B. 

plate  was  reversed  in  the  camera,  but  the  result  is  sufficient  to  show  all  that  was 
desired.  The  ridge  to  the  right  should  be  to  the  left.  By  a  close  examination 
of  Fig.  5,  and  comparison  with  Fig.  4,  many  details  will  appear  in  the  former 
not  seen  in  the  latter. 

The  advantages  of  such  photographs  will  be  at  once  apparent  to  the  reader. 
It  is  possible  to  make  out  details  of  structure  in  mountain  ranges  without  neces- 
sarily making  laborious  climbs,  and  it  is  possible  to  make  out  details  where  ascent 
is  impossible. 

The  difficulties  in  taking  such  photographs  are  not  few.  In  mountainous 
regions,  where  weight  is  a  great  consideration,  it  becomes  necessary  to  carry  a 
small  and  light  tripod.  This  is  not  sufficient  for  work  with  the  telephoto,  where 
there  must  be  great  strength  and  rigidity  in  the  tripod  to  prevent  motion.  The 
days  when  wind  is  not  blowing  from  some  direction  are  not  numerous  while  the 
traveller  is  among  mountain  peaks.  Long  exposures  are  necessary,  and  a  very 
small  vibration  makes  the  result  a  failure.  Another  difficulty  is  that  of  getting  a 
satisfactory  focus.  The  light  coming  through  the  lens  from  so  small  a  portion 
of  the  horizon  is  not  great.     The   landscape  on   the  ground  glass  is  indistinct, 


and  Laboratory  Methods. 


1571 


and  it  becomes  a  difficult  task  sometimes  to  determine  when  the  instrument  is 
in  focus.  Often  the  developed  plate  shows  the  unexpected,  and  failure  is  the 
result. 

A  long  focus  lens  will,  of  course,  produce  results  similar  to  those  here  given, 


Fig.  4. — Sin-yale-a-min  Mountain,  Mission  Range,  Montana. 

but  on  a  smaller  scale.  The  advantage  of  a  telephoto  attachment  is  that  the 
magnification  may  be  made  such  as  will  bring  out  the  features  desired.  With  a 
telephoto  attachment,  with  magnification  up  to  eight  diameters,  an  ordinary  lens 
is  increased  in  value  many  fold.     With  a  long  focus  or  short  focus  lens  but  one 


Fig.  5. — View  of  Portion  of  Fig.  4,  magnified  eight  diameters.     Plate  accidentally  reversed. 

size  of  photograph  may  be  taken,  that  of  the  long  or  short  focus.  With  tele- 
photo attachment  the  focus  may  be  lengthened,  and  the  size  of  the  image  cor- 
respondingly increased,  at  pleasure,  resulting  in  a  very  great  increase  in  the 
usefulness  of  the  lens.  Morton  J.  Elrod. 

University  of  Montana. 


157-  Journal  of  Applied  Microscopy 

11      c  The  present  number  closes  the  fourth 

d     U  volume,  and  fourth  year  of  our  publica- 

Applied    Microscopy  ^^°"-     beginning  with  a  few  closely  set 

*^  '^                                         '^  -^  pages    and    hmited    corps    of   regular 

y     ,               ^"    A/i     u    ^  assistants,  the  scope  and  volume  of  the 

Laboratory   iVietnOUS.  Journal  has  been  gradually  increased 

_  ,.^   ,  .     '    D   CT  T  Tz-k-r-r  to  the  present  time,  the   demand    for 
Edited  by  L.  B.  ELLIOTT.  ^   ,                      •    .         r 
'     more,    and    more    varied     information 


Issued  Monthly  from  the  Publication  Department  resulting  in  the  addition  of  the  VarioUS 

of  the  Bausch  &  Lomb  Optical  Co.,  departments    which     have    succeeded 

Rochester,  N.  Y.  '^ 

each    other.     Although    the   Journal 


SUBSCRIPTIONS:  has  received  from   the   beginning  the 

One  Dollar  per  Year,  Strictly  in  Advance.     To  Foreign      loyal  SUpport  of    those    mOSt    interested 
Countries,  $  1.25  per  Year,  in  Advance.  .       ,,  ,  ,       ,         .         .^  , 

in  the  advancement  of  scientific  work, 

NOTICE    Subscribers  will  be  notified  on  expiration      the    CXpCnSe    of    introducing    it    tO    the 

of  subscription  and,  unless  remittance  for  i  ,■  j      r    i  •  ^i.  i  i- 

ensuing  year  is  received  prompdy,  the  Journal  will  be      publlC,  and  Ot    keeping  Up  the    publlCa- 

discontinued.  ^jon  until  its  merits  became  sufficiently 

SEPARATES.  w^H  known   to  give   it   the    favorable 

One  hundred  separates  of  each  original  paper  accepted      pOSition  which  it  haS  nOW  attained,   haS 

"llrrit^'are%:ot°d"'irsS^^^^^  A     bccn  SO  great  that,  had   its  publishers 

greater  number  can  be  had  at  cost  of  printing  the  extra      bcCn   leSS    in  CamCSt    in  their    dcsirC    tO 
copies  desired.  i  i  •    ,     ■  »  • 

=    establish  in   America  a  representative 

journal  for  the  sciences  in  which  the 

microscope  is   used,  they   might  many  times  have  been   discouraged,  and   the 

thanks  of  the  users  of  the  microscope  are  certainly  due  them  for  their  efforts  in 

this  direction. 

From  this  time  forward,  however,  the  Journal  will  have  to  stand  or  fall  on 
its  own  intrinsic  value  to  the  public.  It  will  therefore  be  necessary  to  cancel  all 
subscriptions  which  are  not  renewed  upon  expiration.  The  subscription  price 
will  not,  however,  be  advanced  for  the  year,  and  we  trust  that  sufficient  new  sub- 
scribers will  be  added  to  our  list  during  that  time  to  make  it  unnecessary  for  us 
to  do  so  later. 

A  number  of  interesting  series  of  articles  will  be  published  during  the  com- 
ing year,  among  which  may  be  mentioned  the  conclusion  of  Professor  Dennis's 
series  on  photomicrography,  illustrated  with  some  of  the  best  examples  of  his 
work.  A  series  which,  when  concluded,  will  be  a  monograph  on  medical  micro- 
technique for  the  use  of  practicing  physicians,  will  be  begun  in  the  January  num- 
ber, and  we  hope  to  add  a  number  of  features  of  special  interest  to  physicians. 
We  have  also  received  a  valuable  addition  to  our  review  department  in  the 
cooperation  of  M.  Girauld,  of  the  Laboratorie  de  Kacteriologie  de  la  Ville  de 
Paris,  who  will  contribute  each  month  reviews  of  current  French  literature.  It 
is  our  desire  to  improve  the  Journal  just  as  rapidly  as  circumstances  will  war- 
rant, and  to  this  end  we  welcome  suggestions,  and  respectfully  solicit  your 
contributions  for  publication,  in  order  that  the  Journal  may  become,  as  intended, 
a  repository  of  American  microscopical  literature,  accessible  not  only  to  the 
specialist  and  the  richly  endowed  library,  but  to  all  who  may  desire  it. 

Our  observation  shows  that  a  large  number  of  the  methods  published  during 
the  year  have  been  put  into  practical  use  in  many  institutions,  and  our  contribu- 
tors have  the  satisfaction  of  knowing  that  every  article  contributed,  whether  the 
result  of  original  investigation  or  an  improved  reworking  of  an  old  process, 
assists  many  of  our  readers  who  are  so  situated  as  not  to  be  able  to  pursue  these 
researches  for  themselves. 

The  Journal  has  also  met  with  a  kindly  reception  in  foreign  countries,  as  is 
shown  by  the  large  number  of  citations  and  reprinted  articles  in  such  leading 
publications  as  Zeitschrifi  fiir  IVissenschaftUihe  AfikroskoJ>ic,  Journal  of  the  Royal 


and  Laboratory  Methods.  1573 

Microscopical  Society,  London,  and  many  others.  It  now  numbers  its  sub- 
scribers in  England,  Canada,  Japan,  Mexico,  Germany,  Cuba,  New  Zealand, 
Brazil,  Cape  Colony,  France,  Portugal,  Hawaii,  Belgium,  New  South  Wales, 
Bermuda,  Sweden,  Queensland,  Victoria,  Costa  Rica,  Alaska,  St.  Lucia,  Hayti, 
San  Marino,  Equador,  Jamaica,  India,  Argentine  Republic,  Venezuela,  Formosa, 
Chili,  Greece,  Austria,  Servia,  Russia,  Italy,  and  Natal,  in  the  order  given. 


CURRENT  BOTANICAL  LITERATURE. 

Charles  J.  Chamberlain. 

Books  for  review  and  separates  of  papers  on  botanical  subjects  should  be  sent  to 

Charles  J.  Chamberlain,  University  of  Chicago, 

Chicago,  111. 

REVIEWS. 

Arnoldi,  W.    Beitrage  zur  Morphologie  einiger      ^^^  previous  papers  of  this  series  have 
Gymnospermen.     V. Weitere  Untersuchung  '■  '■    ^ 

en  der  Embryogenie  in  der  Familie  der  already  been  reviewed  in  the  JOURNAL. 
Sequoiaceen.  Bull,  des  Nat.  de  Moscow.  '^^6  present  paper  deals  with  Scaiwia 
Pp.  1-28,   pis.  7-8,  190 1.  f  1    f  y 

and  other  members  of  the  Sequoiaceae, 

viz.  :  Taxodium,  Cryptomeria,  Cuiininghamia^  Arthrofaxis,  G/yptosirobiis,  and 
Sciadopitys.  As  might  be  expected  in  a  paper  dealing  with  so  many  and  such 
inaccessible  genera,  the  series  are  often  incomplete,  but  the  results  are  never- 
theless interesting  and  important.  In  Cunninghamia  sinensis  there  are  numerous 
archesporial  cells,  and  several  embryo-sacs  attain  a  considerable  degree  of 
development.  In  Sequoia  gigantea  the  endosperm  develops  uniformly,  thus  differ- 
ing decidedly  from  S.  sempervi?'e?is,  in  which  the  development  at  the  middle  of 
the  endosperm  differs  from  that  at  both  ends.  The  archegonia  occur  singly  or 
in  groups,  but  are  not  so  numerous  as  in  S.  sempei'virens.  There  are  two  neck 
cells  and  no  ventral  canal  cell.  In  Taxodiiim,  Cryptomeria  and  Cunninghamia 
the  archegonia  are  grouped  as  in  Cupressinese  and  have  a  common  jacket,  but 
sometimes  there  is  a  layer  of  endosperm  between  the  archegonia.  In  Sciadopitys 
the  neck  is  very  peculiar,  consisting  of  from  four  to  eight  vertically  elongated 
cells.  Proteid  vacuoles  are  present  in  the  archegonium  and  they  probably  arise 
from  the  jacket  cells.  These  vacuoles  are  not  found  in  any  other  members  of 
the  Sequoiaceae.  No  ventral  canal  cell  was  identified,  but  it  may  yet  be  found. 
In  Cryptomeria  the  upper  end  of  the  egg  becomes  mucilaginous  and  sometimes 
separates  from  the  rest  of  the  o^g^,  but  no  ventral  canal  is  formed. 

In  Sequoia  sempervirens  at  the  time  of  fertilization  the  pollen  tube  contains 
two  male  cells  and  two  free  nuclei,  one,  the  nucleus  of  the  pollen  tube,  and  the 
other  the  nucleus  of  a  disorganized  cell  which  Belajeff  called  the  sterile  cell  of 
the  generative  complex.  No  vegetative  cell  of  the  male  prothallium  is  formed. 
The  body  cell  contains  starch.  In  S.  gigantea  the  pollen  tube  presses  between 
the  endosperm  and  the  nucellus.  The  pollen  tubes  of  Taxodium  and  Cryptomeria 
behave  as  in  the  Cupressineae.  The  upper  part  of  the  egg  becomes  mucilaginous 
and  presses  upon  the  neck  cells  from  beneath,  while  an  outgrowth  from  the  pollen 
tube  presses  from  above  and  forces  its  way  into  the  egg.     In  Seqtwiasejupervirens 


1574  Journal  of  Applied  Microscopy 

the  round  male  cell  becomes  elongated,  one  figure  showing  it  spirally  wound, 
but  this  may  not  be  the  normal  form.  In  Taxodium,  however,  the  spiral  form  is 
the  usual  one,  and  this  is  probably  the  case  in  the  Cupressineae  also.  The  form 
is  probably  due  to  the  narrow  entrance,  the  male  cell  having  a  greater  diameter 
than  the  neck  of  the  archegonium.  The  behavior  of  the  chromatin  during 
fertilization  is  not  described. 

In  Sequoia  sempervi reus  the.  sex  nuclei  fuse  at  the  middle  of  the  archegonium, 
then  sink  to  the  bottom  and  divide.  Two  cells  are  organized  about  the  nuclei 
and  the  lower  nucleus  divides  again,  thus  giving  rise  to  a  row  of  three  cells,  the 
lowest  of  which  becomes  the  embryo  and  the  middle  the  suspensor.  The  upper 
soon  disorganizes  and  at  this  stage  the  embryo  appears  to  consist  of  two  cells. 
The  first  division  of  the  embryo  is  longitudinal.  In  Cryptomeria  and  Taxodium 
the  fertilized  egg  nucleus  passes  to  the  base  of  the  archegonium,  where  two  or 
three  divisions  occur.  Cells  are  formed  about  the  lower  nuclei,  but  the  upper 
ones  remain  free.  Two  or  three  tiers  are  organized,  the  lower  one  or  two  tiers 
forming  the  embryo,  and  the  tier  next  above,  the  suspensor.  This  agrees  with 
Strasburger's  diccouwt  oi /iinipenis,  except  that  the  free  cells  were  not  described. 
Cunninghamia  agrees  with  Taxodium,  Cryptomeria  and  the  Cupressineae.  In 
Sciadflpifys,  the  series  was  very  incomplete,  but  enough  was  obtained  to  show 
that  the  embryology  is  very  peculiar.  The  earliest  stage  found  shows  four  free 
nuclei  at  the  base  of  the  archegonium,  as  in  all  the  Abietineae.  A  later  stage 
shows  a  "  rosette,"  suspensors,  and  a  loose  tissue  of  embryonic  cells.  The  lowest 
of  these  cells  form  the  embryo,  those  next  above  develop  into  a  second  set  of 
suspensors,  still  leaving  some  of  the  embryonic  cells  between  the  two  suspensor 
systems.  The  figures  bear  some  resemblance  to  Strasburger's  figures  oiAraucaria, 
but  in  Strasburger's  account  the  second  set  of  suspensors,  as  described  by 
Arnoldi,  form  a  cap  which  is  cast  off  while  the  part  between  the  two  suspensor 
systems — or  between  the  suspensor  and  cap — develops  into  the  embryo. 

Arnoldi  believes  that  the  two  species  of  Sequoia  should  constitute  a  family, 
the  Sequoiaceae ;  that  Taxodium,  Cryptomeria  and,  perhaps,  Cunninghamia 
should  be  included  in  the  Cupressineae ;  and  that  Sciadopitys  is  best  regarded  as 
constituting  a  special  family,  the  Sciadopiteae.  c.  j.  c. 

Livingston,   B.  E.     On  the  Nature    of    the  The  effect  of  external  osmotic  pressure 

Stimulus  which  Causes  the  Change  of  Form  upon    green    algae   has    recently    been 
in   Polymorphic    Green    Aleae.       Bot.  Gaz.  j     ^i  i  •      ^     /•        •  .•      .-        i 

30:  289-317,     pis.  17-18,  1901.  i^ade  the  subject  of  an  mvestigation  by 

B.  E.  Livingston.  A  polymorphic  form 
of  Stigeoclonium  was  chosen.  Knop's  solution  was  used  as  a  nutrient  medium. 
In  a  weak  solution  (up  to  0.1  per  cent.)  swarm  spores  are  produced  in 
great  numbers  which  germinate  to  form  branching  filaments.  In  a 
solution  whose  concentration  is  0.5  per  cent,  to  '1  per  cent,  no  swarm 
spores  are  produced,  but  the  cells  grow  and  multiply  by  divisions  in  all  directions, 
the  daughter  cells  separating  almost  as  soon  as  formed.  This  process  results  in 
a  mass  of  rounded  cells  like  those  of  Palmclla.  If  filaments  are  placed  in  the 
stronger  solution  their  cells  change  from  the  cylindrical  to  the  spherical  form 
and  break  apart.     In  solutions  of  intermediate  strengths,  swarm    spores    may  be 


and  Laboratory  Methods.  1575 

produced,  but  fail  to  germinate  as  filaments,  growing  directly  into  the  round 
celled  form.  By  varying  the  porportions  of  the  four  salts  contained  in  the 
nutrient  fluid  and  yet  keeping  its  osmotic  pressure  the  same,  an  attempt  was  made 
to  determine  whether  these  changes  in  the  form  of  the  alga  were  due  to  a  physi- 
cal or  chemical  cause.  Many  cultures  were  made  in  this  way  with  a  uniform 
result ;  the  form  of  the  plant  is  always  determined  by  the  osmotic  pressure  of 
the  surrounding  medium  and  is  never  influenced  by  the  varying  proportions  of 
the  constituent  compounds.  Some  considerations  follow  concerning  the  probable 
way  in  which  variations  in  the  osmotic  pressure  of  the  medium  may  affect  the 
organism.  The  author  believes  that  the  effect  is  produced  by  a  change  in  the 
water  content  in  the  cell.  Whether  or  not  the  rounding  up  of  the  cells  in  the 
strong  solution  has  any  connection  with  the  process  of  plasmolysis  is  a  question 
which  he  discusses  at  some  length.  The  results  of  the  different  experiments  are 
shown  by  full  tables  and  by  two  excellent  plates  of  half-tone  reproductions  of 
photomicrographs.  This  work  opens  up  an  entirely  new  field  in  plant  physiology, 
and  it  is  to  be  hoped  that  further  research  on  the  influence  of  osmotic  pressure 
in  nutrient  media  may  be  forthcoming.  J.  B.  Overton. 

Chicago. 

Guignard,  L.  La  double  ficondation  dans  le  The  details  of  fertilization  in  Naias 
Naias  Major.     Jour  de  Bot.  15:  i-g,  igoi.  ,  ^      ,.„  ^.    ,,       , 

^     ^  major  do  not   diner    essentially    from 

those  in  other  forms.     The   chromosomes    are  very  long  and  the   gametophyte 

number  is  six,  being  the  smallest  number  yet  observed  in  any  plant.     The   male 

nuclei  are  elongated,  but  do  not  assume  the  vermiform  appearance  so  conspicuous 

in  those  of  the  Compositae.     One  synergid  disintegrates  soon  after   the  entrance 

of  the  pollen  tube,    the    other,    in    general,    remains    intact    until    after    several 

divisions  of  the  embryo.     The  fertilized  egg  does  not  rest  for  a  time,  as  is  usual 

in    other   forms,    but   divides    immediately  after  fusion  is  effected.     One    figure 

shows  the  fertilized  endosperm  nucleus  in  the  spirem  stage,  and  the  embryo  two 

celled.     In  all  previously  described  cases  of  double  fertilization,  the    endosperm 

nucleus  invariably  divides  first.     Many  times  two  embryos  were    observed  lying 

side  by  side,   the  unfertilized  endosperm    nucleus  lying  between   them.     One  of 

the    synergids   had    evidently    functioned   as    an    egg,    and  endosperm  was  not 

produced.     Two  of  the  antipodals  soon  show  signs  of  disintegration.     The  upper 

one  continues  to  enlarge  long  after  fertilization.     The  article  is  illustrated    with 

fourteen  text  figures.  W.  J.  G.  Land. 

Chicago. 

Lyon,  H.  L.  Observations  on  the  Embryogeny  Nelumbo  is  certainly  a  perplexing  form. 
ofNelumbo.  Minnesota  Bot.  Studies,  2:  -j^he  closed  bundles,  irregularly 
643-655,   1901.  °  ■' 

scattered,  present   a    distinctly    mono- 

cotyl  feature  ;  the  leaves  with  reticulate  venation  suggest  dicotyls,  while  the 
flowers  might  be  either  monocotyl  or  dicotyl.  The  earlier  observers,  dealing 
with  mature  seeds,  have  described  the  embryos  of  Nelumbo  and  of  other  mem- 
bers of  the  Nymphaeaceae  as  monocotyl.  Material  for  the  present  work  was 
collected  in  August,  1899,  and  August,  1900,  in  southeastern  Minnesota,  where 
acres  of  Nelumbo  lutea  grow  in  the  bayous  of  the  Mississippi  river.     A  study  of 


1576  Journal  of  Applied  Microscopy 

the  development  of  the  embryo  shows  that  it  retains  a  spherical  shape  until  it 
consists  of  several  hundred  cells.  The  single  cotyledon  then  appears  as  a 
crescent  shaped  organ  partly  surrounding  the  plumule.  The  single  cotyledon 
now  becomes  bilobed  by  a  localization  of  growth.  The  first  foliage  leaf  arises 
on  the  side  opposite  the  cotyledon.  The  radicle  is  transitory  and  does  not 
develop  into  a  primary  root,  but  the  work  is  done  by  secondary  roots  arising 
from  the  hypocotyl.  The  only  character  which  has  kept  the  Nymphseaceae  among 
the  Dicotyls  is  the  apparently  dicotyl  embryo.  Since  a  study  of  the  development 
shows  that  the  embryo  is  truely  monocotyl  and  since  the  anatomy  conforms  more 
closely  to  the  Monocotyls,  the  Nymphaeacece  should  be  classified  as  a  sub-series 
coordinate  with  the  Potamogetonace?e,  Alismaceae,  and  Butomaceae  in  the  series 
Helobiae. 

A  future  paper  will  deal  with  the  development    of    the   embryo-sac    and    fer- 
tilization, c.  J.  c. 


CYTOLOGY,  EMBRYOLOGY, 

AND 

MICROSCOPICAL  METHODS. 

Agnes  M.  Claypole,  Cornell  University. 

Separates  of  papers  and  books  on  animal  biology  should  be  sent  for  review  to 

Agnes  M.  Claypole,  125  N.  Marengo  avenue, 

Pasadena,  Cal. 

CURRENT  LITERATURE. 

Boveri.Th.    Die  Polaritat  von  Ovocyte,  Ei  und       ^^^^^-^   ^33    j^^je   the   interesting    dis- 
Larve  des  btrongylocentrotus  hvidus.  Zoolo-  ° 

gische    Jahrbiicher,   Abth.   f.    Anat.,     July,       covery   that  the    principal    axis   of   the 

'9°'-  Strongyloicntrotns  is  marked  out  in  the 

egg  before  fertilization,  and  that  it  probably  corresponds  to  the  axis  of  the 
ovocyte  of  the  germinal  epithelium.  The  unripe  egg  is  surrounded  by  a  gela- 
tinous envelope  which  is  so  transparent  that  it  is  invisible  under  ordinary  circum- 
stances. At  one  point  this  envelope  is  perforated  by  a  micropyle.  Into  this 
micropyle  the  polar  bodies  are  usually  extruded.  After  the  maturation  divisions, 
the  pigment  which  previously  was  scattered  uniformly  over  the  (t^g  collects  into 
a  broad  band,  surrounding  the  side  of  the  egg  opposite  the  micropyle.  The 
principal  axis  of  the  egg  is  thus  determined.  The  female  pronucleus  is  eccentric, 
but  stands  in  no  constant  relation  to  this  chief  axis.  Fertilization  is  usually 
effected  through  the  micropyle,  and  the  first  cleavage  spindle  lies  at  right  angles 
to  the  entrance  path  of  the  spermatozoon.  This  was  observed  when  the  sperm 
entered  near  the  equator  of  the  Qgg,  so  the  first  cleavage  plane  is  apparently 
determined  by  the  entrance  path  of  the  sperm.  No  trace  of  a  bilateral  organiza- 
tion of  the  egg  could  be  detected,  and  as  the  first  cleavage  plane  is  not  known  to 
stand  in  any  relation  to  the  bilateral  symmetry  of  the  larva,  the  entrance  path  of 
the  sperm  is  not  known  to  mark  out  the  plane  of  bilaterality,  as  it  has  been  said 
to  do  in  the  frog.  The  unpigmented  animal  pole  of  the  egg  forms  the  ectoderm, 
the  pigmented  belt,  which  lies  in  the  vegetal  half  of  the  egg,  forms  the  entoderm, 


and  Laboratory  Methods.  157 « 

while  the  unpigmented  area  at  the  vegetal  pole  gives  rise  to  the  primary  mesen- 
chym  and  larval  skeleton.  Boveri  holds  that  the  micropyle  represents  the  point 
of  attachment  of  the  ovocyte,  and  that  the  chief  axis  of  the  embryo  is  therefore 
traceable  back  to  the  germinal  epithelium  of  the  ovary.  s.  j.  h. 

Holmgren,  N.     Ueber  den  Bau  der  Hoden  und       The   seminal    tubules    of    the    testis   in 
die  Spermatogenese  von  Staphvlinus.    Anat.        „,      ,     ,.  ,.rr  i       •         i-rr 

Anz.  June  looi.  Staphyluius    diner    much    m    different 

seasons  of  the  year.  In  summer  the 
outer  zone  of  the  tubules  contains  spermatocytes  and  spermatids,  and  the  inner 
zone  is  filled  with  spermatozoa.  No  spermatogonia  are  present.  In  animals 
killed  in  the  winter  the  capsules  of  tubules  are  thickened  except  over  a  portion  on 
one  side  where  the  tubule  is  swollen  out.  This  swollen  part  contains,  besides  a 
Verson's  cell,  only  spermatogonia  ;  the  latter  differ  from  the  spermatogonia  of 
the  rest  of  the  tubule  in  size  and  structure,  and  become  arranged  radially  around 
the  Verson's  cell.  No  spermatocytes  of  the  second  order,  nor  spermatids,  are 
found  at  this  season,  but  there  are  a  few  spermatozoa  which  represent  remnants 
left  over  from  the  previous  summer.  The  thick  outer  capsule  of  the  testis  is  a 
syncytial  membrane  from  which  the  spermatogonia  arise.  The  spermatogonia 
become  enclosed  in  packets,  surrounded  by  a  capsule  derived  from  the  main 
capsule  of  the  tubule.  The  spermatozoa  that  are  not  discharged  are  finally 
ingested  by  the  capsule  and  resorbed.  This  is  apparently  a  normal  process  of 
utilizing  the  unused  spermatozoa.  The  spermatogonia  of  both  parts  of  the 
tubule,  although  they  differ  much  in  structure  and  history,  give  rise  to  sperma- 
tozoa, which,  so  far  as  could  be  determined,  are  all  alike.  It  is  a  fact  of  interest 
that  the  same  kind  of  specialized  cell  is  developed  by  two  different  routes. 

S.  J.  H. 

Conklin,  E.  G.     The  Individuality  of  the  Germ       In  Crepidula  platia  it  is  very   probable 
Nuclei  during  the   Cleavage  of   the  Egg  of         ,  ,  i    •    i  ,  1^1 

Crepidula.    Biol.  Bull.  II,  June,  1901.  that  the  germ  nuclei  do  not  completely 

fuse,  as  is  indicated  by  the  double 
character  of  the  nuclei,  which  can  be  traced  to  quite  an  advanced  period  of 
cleavage.  The  double  nature  of  the  nuclei  appears  most  clearly  during  the 
telophase  of  each  division,  but  may  sometimes  be  observed  throughout  the  entire 
resting  period.  The  chromosomal  vesicles,  when  the  daughter  nuclei  are  being 
formed,  fuse  into  two  separate  groups,  which,  for  a  time,  are  separated  by  a 
partition  wall.  In  the  early  cleavage  stages  two  nucleoli  are  present.  These 
may  represent  the  descendents  of  the  nucleoli  of  the  male  and  female  pronuclei, 
as  each  pronucleus  possesses  a  nucleolus.  It  is  probable  that  the  half  of  the 
nucleus  lying  nearest  the  animal  pole  of  the  egg  is  derived  from  the  female 
pronucleus,  and  the  other  half  from  the  sperm.  s.  j.  h. 

Petrunkewitsch  A.    Die  Richtungskorper  und  ^he  theory  that  the  unfertilized  eggs  of 

ihr    bchicksal    im    befruchteten    und  unbe-  •'                                                °° 

■  fruchteten    Bienenei.      Zoologische    Jahr-  bees  give  rise  to  drones  is  supported 

bucher,  Abth.  f.  Anat.,  July,  1901.  ^^  ^^e  observations  of  the  author,  who 

finds  that  the  eggs  laid  by  the  queen  in  the  drone  cells  are  always  unfertilized. 
In  both  the  fertilized  and  unfertilized  eggs  the  first  maturation  division  is  an 
"  equation  division."      The  first   polar  body  divides,  the  outer  portion  being 


1578  Journal  of  Applied  Microscopy 

extruded  from  the  egg,  while  the  inner  copulates  with  the  second  polar  body, 
forming  a  large  nucleus.  This  nucleus  soon  disintegrates  in  the  fertilized  eggs, 
but  in  the  parthenogenetic  eggs  divides  repeatedly,  taking  part  in  the  formation  of 
eight  cells,  which  wander  into  the  central  portion  of  the  egg,  where  their  ultimate 
fate  could  not  with  certainty  be  determined.  No  copulation  occurs  between  the 
female  pronucleus  and  either  of  the  polar  bodies.  How  the  normal  number  of 
chromosomes  is  regained  in  the  parthenogenetic  eggs,  repeated  efforts  failed  to 
discover.  s.  j.  h. 

Montgomery,  T.  H.    A  Study  of  the  Chromo-      This  paper  is  divided  into  two  portions. 

somes    of    the    Germ    Cells   of    Metazoa.       ^he  first  is  devoted  to  detailed  observa- 

Trans.  Am.  Philos.  boc,  1901. 

tions  on  the  spermatogenesis  of  forty- 
two  species  of  Hemiptera.  The  subjects  that  received  especial  attention  are 
the  changes  that  occur  during  the  synopsis  stage,  the  reduction  in  the  number  of 
chromosomes,  and  the  chromatin  nuclei.  The  interesting  discovery  is  recorded 
that  in  four  species  of  Hemiptera  the  normal  number  of  chromosomes  is  odd. 
In  the  second  portion  of  the  paper  there  is  a  discussion  of  subjects  of  a  general 
nature,  chief  of  which  are  the  following :  The  individuality  of  the  chromosomes, 
the  significance  of  the  chromatin  nucleoli,  the  relation  of  number  of  chromosomes 
to  genetic  affinity,  the  factors  which  determine  the  number  of  chromosomes, 
significance  of  the  uneven  number  of  chromosomes,  and  the  problem  of  reduction. 

S,  J.   H. 

^""^'i^-^'A    ^m'''%''    zur     Entwicklungsge-      Investigation    was   carried    on    mainly 
schichte  der  Musciden.  Zeit.wiss.  Zool.,1901.  *  ^ 

upon    CaJliphora    erythrocephala.     The 

following  subjects  are  treated  :  The  first  developmental  changes  in  the  fertilized 
egg,  formation  of  polar  bodies,  origin  of  the  yolk  cells,  forniation  of  the  blasto- 
derm and  germinal  layers,  development  of  the  alimentary  canal,  and  the  fate  of 
the  pole  cells.  In  regard  to  the  last  subject  the  author  finds  that  the  pole  cells 
wander  into  the  archenteron,  becoming  embedded  finally  in  the  entoderm,  where 
their  further  fate  could  not  be  followed.  s.  j.  h. 

The  author  uses  a  mixture  of  2  pts.  80 
Henninp,   C.    Dr      Depigmenting  the  Eyes  of  ^     ^i^oj^^i     ^^^    ^       ^     glycerin     to 

Arthropoda.     Zeit.     \\  iss.   Mick.     17,   1900.       ^     _  r       ts  j 

which  2  vols,  of  strong  sulphuric 
acid  are  added.  The  solution  acts  best  at  a  temperature  of  about  35°  C,  the 
time  required  varying  from  10  minutes  to  about  12  hours  according  to  the  kind 
of  pigment.  The  prolonged  action  of  the  fluid  is  not  injurious  to  the  eye 
tissues.  A.  M.  c. 


CURRENT  ZOOLOGICAL  LITERATURE. 

Charles  A.  Kofoid. 

Books  and  separates  of  papers  on  zoological  subjects  should  be  sent  for  review  to 
Charles  A.  Kofoid,  University  of  California,  Berkeley,  California. 

Harvey,  N.  A.    Introduction  to  the  Study  of  ^^  elementary  laboratory  manual,  the 
Zoology  for  the  use  of   High  Schools  and  ^  ■' 

Academies.     208  pp.  190 1.     The  Western  outgrowth  of  the  normal  school  method, 

Publishing  House,  Chicago.  p^^   fo^^h  as   a  guide   for    work  in   the 


and  Laboratory  Methods. 


1570 


first  grade  of  the  high  school.  The  objects  studied  are  ahnost  exclusively 
arthropods  and  vertebrates,  and  the  M'ork  is  planned  for  laboratories  not  equipped 
with  microscopes.  The  comparative  method  is  well  sustained  throughout  the 
book,  and  the  manner  of  presentation  is  one  well  adapted  to  cultivate  independ- 
ence in  the  pupil,  and  to  develop  the  scientific  method.  The  efi^ort  to  combine 
both  text-book  and  laboratory  guide  in  two  hundred  pages  has  been  less  suc- 
cessful, and  the  reputed  micro-photographs  which  are  offered  as  substitutes  for 
the  compound  microscope  will  be  of  little  value  even  to  schools  with  meager 
equipment.  c.  a.  k. 


Peters,  A.  W.  Some  Methods  for  Use  in  the 
Study  of  Infusoria.  Am.  Naturalist,  35 : 
553-559.  I90'- 


The  Yarn  Siphon  is  used  for  separating 
Infusoria  in  large  numbers  from  both 
the  culture  water  and  the  solid  debris 
which  it  contains.  Transfer  the  liquid  containing  the  Infusoria  from  the  culture 
jar  to  a  Stender  dish,  and  mix  thoroughly  to  obtain  a  uniform  distribution  of  the 
organisms.  Next,  several  pieces  of  woolen  yarn  are  laid  side  by  side  without 
twisting,  moistened,  and  placed  with  one  end  in  the  Stender  dish  and  the  other 
hanging  down  the  side  of  the  dish,  and  siphoning  into  a  collecting  vessel.  The 
yarn  filters  the  water  so  that  only  active  organisms  pass  over  into  the  lower 
vessel. 

The  Tube  Filter  is  an  apparatus  for  concentrating  the  Ciliata  contained  in 
a  large  amount  of  water,  and  changing  the  culture  medium.  The  ordinary 
methods  of  filtration  fail  in  this  case  because  so  many  organisms  stick  to  the 
filter  paper.  The  tube  filter  is  a  piece  of  large  glass  tubing,  over  one  end  of 
which  filter  paper  is  fastened  with  a  rubber  band.  This  tube  is  lowered  into 
the  culture  vessel,  and  the  filtration  takes  place  upward.  More  culture  fluid,  or 
any  desired  solution,  may  be  added  to  the  outer  vessel  from  time  to  time,  and 
the  filtered  water  removed  from  the  tube  by  means  of  a  siphon  with  its  lower 
end  bent  up  to  prevent  it  running  empty. 

The  U-Cell  (Fig.  1)  is  also  a  device  for  filtration,  but  on  a 
smaller  scale.  It  admits  of  microscopical  observation.  It  con- 
sists of  a  long  U  of  large,  close  fibered  darning  cotton,  which 
has  been  previously  moistened  and  placed  between  two  thin 
slides.  The  cotton  is  so  placed  that  the  U  is  two-thirds  the 
length  of  the  slide,  and  that  its  ends  barely  project  beyond  the 
parallel  ends  of  the  slides,  which  are  held  together  with  rubber 
bands.  If  it  is  desired  to  examine  with  a  higher  power  of  the 
microscope,  a  cover-glass  may  be  substituted  for  one  of  the 
slides,  but  in  this  case  narrow  strips  of  slides  should  be  laid 
over  the  cover-glass  where  the  rubbers  go  around  it.  The  cot- 
ton yarn  should  be  of  such  a  thickness  that  the  slides  will  be 
0.5  mm.  apart  when  the  cell  is  complete.  To  fill  the  cell  it 
should  be  held  nearly  vertical,  and  the  culture  fluid  and  In- 
fusoriaWnserted  through  the  open  end  of  the  U  with  a  small 
pipette.  Part  of  the  water  will  flow  out  through  the  cotton,  but  part  will  be 
retained  by  capillary  attraction.     By  repeated  use  of  the  pipette  any  desirable 


Fig.  1. 


1580 


Journal  of  Applied  Microscopy 


\V 


number  of  organisms  may  be  concentrated  in  the  same  cell.  The  cell  may 
also  be  filled  by  means  of  the  yarn  siphon,  a  single  strand  being  sufficient  for 
the  passage  of  many  Infusoria.  Such  preparations  may  be  kept  for  a  long  time 
by  placing  the  cells  in  a  larger  vessel  containing  water,  so  that  the  upper  open 
end  of  the  U  is  above  the  level  of  the  water,  while  a  part  of  the  U  is  immersed 
in  it. 

A  Circulation  may  be  kept  up  in  the  U 
cell  as  shown  in  Fig.  2.  The  cells  are  placed  in 
a  cylindrical  dish  with  their  inner  and  upper  ends 
resting  against  a  smaller  cylindrical  vessel  set  in 
the  center  of  the  first.  The  water  is  supplied  to 
the  cells  from  the  central  vessel  (the  edge  of 
which  should  be  but  5  mm.  above  the  open  end 
of  the  U's)  by  yarn  siphons  (S").  The  fluid  is 
removed  from  the  outer  vessel  by  means  of  a 
constant  level  siphon  (S').  The  inner  vessel  is 
supplied  with  water  from  an  elevated  bottle 
closed  with  a  two-hole  stopper.  Through  one 
of  these  holes  passes  the  siphon,  which  dips 
below  the  level  of  the  water  in  the  inner  vessel. 

Through  the  other 
the  air-tube  is  in- 
serted. This  passes 
nearly  to  the  bottom 
of  the  bottle,  and  by 
means  of  raising  and 
lowering  it  the  water 
in  the  inner  cylin- 
drical vessel  may  be  kept  at  any  desired  height.  In  siphoning,  woolen  yarn  pro- 
duces a  more  rapid  flow  than  cotton. 

Absorbent  Cotton  may  be  conveniently  used  for  making  temporary  or  per- 
manent microscopical'preparations.  A  very  small  amount  of  dry  absorbent  cotton 
is  spread  upon  the  slide,  a  few  drops  of  fluid  with  the  Infusoria  are  added,  and 
the  cover-glass  lowered  horizontally  so  that  the  animals  shall  be  caught  in  the 
meshes  of  the  cotton,  and  not  above  or  below  the  fibers.  The  cover-glass  is 
then  secured  with  rubber  bands,  and  the  preparation  treated  in  any  desired  way, 
subjected  to  the  action  of  killing,  dehydrating  agents,  etc.,  without  any  fear  of 
the  Infusoria  being  washed  away  or  misplaced.  F.  W.  Bancroft. 


Fig.  2. 


Looss,  A.    Zur  Sammel-  und  Conservierungs-      Directions  are  given  by  this   eminent 

technik    von    Helminthen.      Zool.  Anz.  24:       i     i     •    ^.u    \ •„<-   t^    t-u^   ^^u^^t-:^^    „„  i 

^„  ^^.  .^^  -,,«  T^^T  helminthologist  for  the  collection  and 

302-304,  309-3  I  C>,   I9OI.  o 

preservation  of  parasitic  worms  by 
methods  which  prevent  undue  contraction  and  distortion,  and  which  are  at  the 
same  time  available  for  field  work  or  in  the  absence  of  laboratory  facilities. 

lYcmatodes. — The  strongly  contractile  forms  are  not  readily  killed  in  extended 
state  except  by  the  old  method  of  washing  in  normal  salt  solution,  stretching 


and  Laboratory  Methods.  1581 

them  out  on  glass  plates  with  soft  brushes,  and  fixing  them  in  sublimate  solution 
while  held  in  this  position.  Other  trematodes,  large  and  small  alike,  are  very 
well  prepared  as  follows  :  One  or  two  cubic  centimeters  of  the  intestinal  contents 
are  put  in  a  test-tube  one-third  full  of  normal  salt  solution  and  shaken  vigor- 
ously for  one-half  to  one  minute.  Concentrated  sublimate  solution  is  then  added 
to  the  amount  of  one-half  the  quantity  of  salt  solution  in  the  test-tube,  and  the 
shaking  continued  for  one-half  minute  longer.  The  parasites  die  in  an  extended 
condition,  and,  if  need  be,  may  remain  four  to  six  weeks  in  the  sublimate  solu- 
tion without  injury. 

The  worms  may  be  freed  from  extraneous  material  by  repeated  shakings  and 
decantations,  and  put  through  alcohol  grades  and  iodine-alcohol.  When  numer- 
ous small  trematodes  are  amid  the  intestinal  villi  they  may  be  secured  by  cutting 
the  intestine  in  small  pieces  and  shaking  these  vigorously  in  normal  salt  solu- 
tion till  the  worms  are  freed,  when  the  intestinal  fragments  can  be  removed  and 
the  worms  treated  with  sublimate.  In  many  cases  the  duration  of  the  shaking 
must  be  adjusted  to  the  contractility  of  the  parasites  present.  Large  species 
should  be  placed  in  fiat-bottomed  dishes  till  hardened.  If  possible,  a  few  speci- 
mens of  each  species  should  be  killed  in  TO  per  cent,  alcohol  on  a  slide  beneath 
a  cover-glass,  supported  by  wax  feet.  The  alcohol  should  be  changed  from  time 
to  time,  and  when  the  killing  is  completed  the  whole  should  be  transferred  to 
90  per  cent,  alcohol,  and  freed  from  the  slide  and  cover-glass. 

Cestodes. — Small  species  with  chains  of  proglottids,  such  as  Taenia  echinococcus, 
are  preserved  by  the  above  described  shaking  method.  It  is  advisable  in  all 
cases  to  test  the  tenacity  of  the  proglottid  chains  before  submitting  them  to  the 
shaking  process,  for  some  forms  readily  break  up  when  shaken.  In  such  cases 
a  moderate  movement  of  the  preserving  fluid  suffices  to  preserve  the  worms  in 
an  extended  condition.  Larger  forms,  including  many  fish  and  reptile  tape- 
worms, become  tangled  if  shaken.  These  should  be  stretched  out  on  glass 
plates  to  harden  after  the  first  few  minutes  exposure  to  the  killing  fluid.  Tape- 
worms exceeding  5  cm.  in  length  should  not  be  subjected  to  the  shaking  process, 
but  may  be  gathered  in  shallow  dishes  in  normal  salt  solution.  From  this  they 
are  taken  singly,  by  grasping  the  posterior  proglottid  with  forceps,  and  are 
shaken  to  and  fro  in  a  one  to  two  per  cent,  solution  of  sublimate  in  normal  salt 
solution  until  contraction  ceases.  When  the  posterior  proglottids  are  too  readily 
detached,  the  worm  should  be  held  near  the  middle  of  the  chain.  Very  large 
worms,  such  as  Taenia  saginata  and  Motiiezia  expansa,  etc.,  are  not  easily  shaken 
out  in  the  ordinary  dish  of  killing  fluid.  Fine  specimens  may  be  secured,  how- 
ever, by  pouring  the  concentrated  sublimate  solution  over  the  worms,  suspended 
across  the  palm  of  the  left  hand  so  that  the  head  and  the  end  of  the  strobila 
hang  free. 

Nematodes. — Small  species,  such  as  Strongylus  subtilus,  when  present  in  con- 
siderable numbers,  may  be  collected  and  cleaned  by  the  shaking  and  decantation 
method  above  described,  treating,  however,  but  a  small  amount  of  the  material 
at  one  time  in  the  salt  solution.  Species  with  thin  cuticula  (Strongylus  from  the 
lungs,  Filaria  from  the  body-cavity)  should  be  placed  in  1-1.2  per  cent,  salt 
solution  to  avoid  the  swelling  which  attends  the  use  of  weaker  solutions.     The 


1582  Journal  of  Applied  Microscopy 

best  killing  fluid  for  nematodes  intended  for  collections  and  systematic  work  was 
determined  after  much  experiment  to  be  70  per  cent,  alcohol  heated  to  80-90°C. 
With  few  exceptions  the  worms  thus  killed  are  well  extended,  free  from  wrink- 
ling, and  give  excellent  histological  detail,  tissues  being  much  less  brittle  than 
when  killed  in  acids  or  metallic  salts.  For  subsequent  examination  the  worms 
are  transferred  to  a  mixture  of  glycerin  and  alcohol,  from'  which  the  latter  is 
evaporated  over  a  warming  oven  at  50-G0°C.  until  the  pure  glycerin  alone 
remains.  For  some  worms,  such  as  Scierostoma,  the  mixture  may  contain  as 
much  as  20  per  cent,  glycerin,  for  more  delicate  species  it  must  not  exceed  2-3 
per  cent,  before  evaporation.  The  evaporation  of  the  alcohol  must  be  very  slow 
in  the  case  of  those  forms  with  thick  cuticula.  Permanent  mounts  of  small 
species  may  be  made  in  glycerin  jelly.  Specimens  from  the  concentrated 
glycerin  may  be  transferred  to  96  per  cent,  alcohol  directly,  without  shrinkage, 
for  subsequent  sectioning.  All  attempts  to  bring  nematodes  from  formalin  into 
alcohol  without  collapse  failed  utterly. 

Acanthocephali. — Shaking  in  normal  salt  solution,  followed  by  similar  treat- 
ment in  sublimate,  leaves  the  specimen  fully  extended  with  proboscis  protruded. 

C.  A.   K. 

Marpmann.  G.       Eine  neue    Vorschrift  zum      The  author  recommends  the  following 

Konservireren  von  Zoologischen  mid  Anat-  ° 

omischen    Praparaten.      Zeitsch.  f.  angew.       fluids  for  use   in  fixing   and  preserving 

Mik.  7:  14,  1901.  zoological    specimens    and  anatomical 

material  where  it  is  necessary  or  desirable  that  alcohol  should  be  avoided.     The 

following  formula  is  given  for  the  fixing  fluid  : 

Sodium  fluoride,         -         -         -  50  gm. 

Formaldehyde  (40  per  cent.),  -         20  c.  c. 

Water, 1000  c.  c. 

From  this   fluid  the   preparations   are  passed  into  the  following  mixture  for 
preservation  : 

Glycerin  (28  °B)     -         -         .         -         500  c.  c. 

Water,       -----  1000  c.  c. 

Magnesium  chloride,     -         -         -         100  gm. 

Sodium  fluoride,         -         -         -  20  gm. 

This  fluid  is  said  to  have  preserved  the  natural  colors  of  anatomical  material 

and  of  reptiles.     Objects  for  sections  should  be  washed  three  or  four  times  in 

water,  and  then  treated  to  the  usual  grades  of  alcohol.     Glycerin  material  may, 

however,  be  passed  directly  to  the  embedding  soap.  c.  a.  k. 


and  Laboratory  Methods.  l5^-> 

NORMAL  AND  PATHOLOGICAL  HISTOLOGY. 

Joseph  H.  Pratt. 

Harvard  University  Medical  School,  Boston,  Mass.,  to  whom  all  books  and 
papers  on  these  subjects  should  be  sent  for  review. 

Wells,  H.  G.     Multiple     Primary      Malignant  yj^^^^  ^^s  collected  from  the    literature 
Tumors ;  Primary    Sarcocarrmoma   in    the 

Thyroid  of  a  Dog,  with  Mixed  Sarcomatous  Only  seventeen  cases  of  primary   multi- 
and  Carcinomatous  Metastases.     Journal  of  ]g    malignant    growths.        The     great 

Pathology  and    Bacteriology,    7:    357-366,  ^  °  "     .  .     ,.    ° 

igoi.  rarity  of  the  co-existence  of   different 

kinds  of  malignant  tumors  seems  to 
show  that  the  presence  of  one  variety  does  not  predispose  to  the  development  of 
another  type.  Their  occurrence  is  probably  fortuitous.  He  also  has  found  two 
cases  and  only  two  of  tumors  of  a  mixed  epithelial  and  mesoblastic  type,  and  to 
these  adds  a  case  of  his  own.  These  two  cases  are  not  complete,  but  are  prob- 
ably authentic.  In  his  case  there  was  a  primary  growth  showing  a  true  sarco- 
matous and  carcinomatous  type,  and  metastatic  nodules  were  found  in  the  lymph- 
nodes,  heart,  lungs,  and  kidneys,  showing  not  only  secondary  growths  of  both 
types,  but  in  several  nodules  there  were  both  sarcomatous  and  carcinomatous 
growths  in  the  same  nodule.  The  primary  tumor  was  in  the  thyroid  of  a  dog, 
but  in  his  plates  the  alveolar  carcinoma  and  the  sarcomatous  growths  are  not 
very  clearly  shown. 

He  discusses  further  the  bearing  of  these  cases  on  the  theories  of  the  etiol- 
ogy of  tumors  and  in  conclusion  states  that  the  very  great  rarity  of  these  sarco- 
carcinomatous  growths  offers  positive  proof  that  the  organism,  if  such  there  is, 
which  causes  malignant  tumors  is  not  the  same  for  sarcoma  and  carcinoma, 
otherwise  the  frequency  of  these  mixed  growths  would  be  greater.  He  thinks  it 
is  not  surprising  that  multiple  malignant  growths  should  occur  in  the  body,  and 
further  calls  attention  to  the  remarkable  fact  that  these  three  cases  of  mixed 
tumor  were  in  the  thyroid. 

We  do  not  think  the  author  is  fully  justified  in  claiming  such  a  great  rarity 
of  primary  multiple  malignant  growths.  His  list  does  not  include  all  the  pub- 
lished ones,  for,  looking  through  five  years'  records  of  1000  tumors,  microscop- 
ically examined,  we  find  a  number  of  multiple  primary  malignant  growths. 

H.  C.    Low. 

Sultan,  C.    Beitrag  zur  Kenntniss  der  Schild-      Katzenstein  stated  in  1899,  as  a  result 
driisen-Function.        Archiv     fiir      klinische 
Chirurgie,  63:  620-626,  1901.  of  his    experiments,    that  the    thyroid 

gland  is  not  an  organ  essential  to   the 

animal  economy,  and  that  it  can  be  removed  without  necessarily   destroying  the 

health    of    the    animal.     Sultan  has  followed  the  same  method  of    investigation, 

but  has  arrived  at  a  different  conclusion.     Total  extirpation  of  the  thyroid  glard 

in  dogs  and  cats  was  followed  by  severe,  characteristic  disease-phenomena  ending 

in  death  if  accessory  gland-tissue  was  not  present  in  sufficient    amount    to   take 

up  the  functions  of  the  thyroid.     Sultan  thinks  that    Katzenstein,    who    worked 

with  dogs,  overlooked  accessory  thyroids  and  was  thereby  led  into  drawing  false 

conclusions. 


1584  Journal  of  Applied  Microscopy 

Piana  found  accessory  thyroid  glands  in  sixty-six  per  cent,  of  all  the  dogs  he 
examined.  These  are  to  be  sought  for  in  the  neighborhood  of  the  aorta.  One 
of  Sultan's  cases  shows  how  readily  they  may  be  overlooked.  He  examined, 
microscopically,  the  lymph  nodes  from  the  arch  of  the  aorta,  and  not  until  he 
had  looked  over  many  sections  did  he  find  an  island  of  typical  thyroid  tissue.  It 
partially  surrounded  a  lymph-node  and  he  likened  it  to  a  skull-cap. 

In  cats,  accessory  thyroids  are  rarely  found.  j.  h.  p. 

Neumann,  E.      Das    Pigment     der     braunen       In  his  Studies  of  the  pigment  of  brown 
Luna;eninduration.     Virchow's    Archiv    fiir       .     ,        ^.  r   ^i      i  xt 

pathTAnat.,  161:  422-435,  1900.  induration  of  the  lung,  Neumann  never 

observed  the    formation    of  melanotic 

pigment  from  haemosiderin.     Granules  with  a   black    center    and    a    periphery 

more    or  less  the  color  of  haemosiderin  were  seen.     He  regards  these  bodies   as 

particles  of  carbon-dust  which  have  been  incrusted  with  haemosiderin.     He  also 

observed  bits    of  carbon    with  colorless  peripheries.     Both  the  colored  and  the 

colorless  borders  gave  the  iron  reaction  equally  well.     The  author  is    inclined  to 

regard  this  colorless  modification  as  the  last  stage  in  the  transformation    of  the 

blood  pigment  which  terminates  in  its  complete  disappearance  through  resorption. 

Neumann  found  these  peculiar  pigment  bodies  in  the  bronchial  lymph  nodes 

as  well  as  in  the  lungs. 

The  haemosiderin  is  formed  from  the  red  blood  corpuscles.     There  is    first   a 

diffusion    of   the    haemoglobin,    and  later  the  pigment  separates  itself  out  of  the 

solution.  J.  H.  p. 


GENERAL  PHYSIOLOGY. 

Raymond  Pearl. 

Books  and  papers  for  review  should  be  sent  to  Raymond  Pearl,  Zoological 
Laboratory,  University  of  Michigan,  Ann  Arbor,  Mich. 

Loeb,  J.  On  an  apparently  New  Form  of  The  author  found  that  when  the  gas- 
Muscular  Irritability  (Contact  Irritability?)  .  1  r  r  •  ^  • 
produced  by  Solutions  of  Salts  (preferably  trocnemius  muscle  of  a  frog  IS  put  in 
Sodium  Salts)  whose  Anions  are  Liable  to  certain  salt  solutions  in  a  Strength  of  1 
form  Insoluble  Calcium  Compounds.  Amer.  ,  ,  r  ^1  1.  •  o  ^  in 
Jour.  Physiol.  S:  362-373,  1901.  g^am  molecule  of  the  salt  in   b   to  10 

liters  of  water,  and,  after  being  sub- 
jected for  a  time  to  the  action  of  this  solution,  is  brought  back  into  air  or  certain 
other  substances,  it  goes  into  a  tetanus  or  performs  a  series  of  strong  contrac- 
tions. This  apparently  new  irritable  phenomenon  is  provisionally  considered  as 
"  contact  irritability."  The  substances  other  than  air  which  produce  the  con- 
traction when  the  muscle  is  passed  from  the  salt  solution  into  them  are  COo,  oil, 
2n  sugar  solution,  glycerine,  chloroform,  toluol  and  probably  mercury.  The 
salts  whose  solutions  bring  about  the  contact  irritability  are,  with  a  single  excep- 
tion, sodium  salts  ;  viz.,  sodium  fluoride,  sodium  carbonate,  Na2HP04,  sodium 
oxalate,  sodium  citrate  and  sodium  tartrate.  In  addition  to  these  (NH4)2  SO4 
has  the  same  effect.  The  anions  of  these  sodium  salts  produce  insoluble  calcium 
compounds  and  it  is  to  the  presence  of  these  in  the  surface  layers  of  the  muscle 


and  Laboratory  Methods.  1^^^ 

that  the  author  believes  the  effect  to  be  due.  This  view  is  supported  by  the  fact 
that  the  presence  of  solutions  of  calcium  salts  hinders  or  entirely  prevents  the 
contact  reaction.  This  contact  reaction  does  not  appear  in  curarised  muscle, 
indicating  that  the  action  of  the  solution  is  on  the  nerve  elements  in  the  muscle. 
If  the  nerve  of  one  of  these  nerve-muscle  preparations  be  put  into  the  solutions 
which  bring  about  the  contact  irritability,  the  muscle  will  begin  to  twitch  in  a 
few  minutes  and  tetanus  finally  ensues.  Removal  of  the  nerve  from  the  solu- 
tion stops  the  contractions,  which  will,  however,  begin  again  if  the  nerve  is 
brought  into  contact  with  any  solid  or  liquid  body.  Thus  apparently  the  ions 
do  not  stimulate  directly,  but  merely  increase  the  sensitivity  of  the  nerve  to  con- 
tact stimuli.  R.  P. 

Murbach,  L.    Physiology  in  the  High  School.      In  this  brief  note   the  author  gives   a 
Physician    and    Surgeon.     December  Num-         11^  ^i-  /■  ^i  ■        u      • 

ber   I  goo  skeleton  outhne  of  the  course  m  physi- 

ology taught  in  the  Detroit  Central 
High  School.  In  the  space  of  a  review,  mention  can  only  be  made  of  a  few  of 
the  more  particularly  noteworthy  features  of  a  wholly  excellent  course.  The 
standpoint  of  the  entire  course  is  experimentation  by  the  student,  and  independ- 
ence ("  forced,  if  necessary  ")  in  the  drawing  of  conclusions  from  experiments. 
The  experiments  and  laboratory  work  on  the  gross  anatomy  of  bodily  organs  are 
planned  so  as  to  bring  forcibly  to  the  student's  mind  nearly  all  the  fundamental 
principles  of  the  functional  activities  of  living  things.  Along  with  this  ground- 
work of  the  course  interesting  and  valuable  special  features  are  introduced. 
Food-stuffs  are  studied  experimentally  in  the  laboratory,  according  to  the  follow- 
ing plan  :  "  Simple  tests  are  made  for  starch,  sugar,  fat  and  proteid  in  known 
foods,  such  as  corn  starch,  glucose,  mutton  tallow  (extracted  with  chloroform), 
and  white  of  ^g^.  Then  students  are  given  yolk  of  ^gg,  milk,  beans,  castor 
beans,  peas,  flaxseed,  wheat  and  wheat  flour,  barley,  and  then  sprouted  barley — 
for  sugar^ — to  determine  the  principal  food  constituents.  The  uses  and  abuses 
of  alcohol  and  narcotics  are  discussed  from  a  scientific  and  moral  point  of  view, 
rather  than  from  a  dogmatic,  prejudiced,  emotional  one."'  Simple  experiments 
on  digestion  are  performed  by  the  students,  illustrating  the  principles  of  solution, 
emulsification,  osmosis,  ferment  action,  etc.  In  connection  with  the  study  of  the 
special  senses,  experiments  on  skin  sensations  are  introduced. 

These  examples  will  sufiice  to  indicate  something  of  the  originality  displayed 
in  the  whole  plan.  Such  a  course  as  this  of  Dr.  Murbach's  is  in  agreeable  con- 
trast with  those  mixtures  of  poor  anatomy,  poorer  physiology,  and  a  smattering 
of  hygiene  which  are  still  too  frequently  served  up  to  the  secondary  school 
student  as  "physiology."  R.  p. 

Stiles,  P.  G.    On  the  Rhythmic  Activity  of  the      This  paper  presents  the  results  of   ex- 
Oesophagus  and  the  Influence  upon  it  of      periments  on  the  effects  of  solutions  of 

Various    Media.     Amer.    Jour.    Physiol.    5 :  .  ,^  .  1         1.    ^l     •  ,. 

--g_-„      Q,  ^  various  salts  on  the  rhythmic  contrac- 

tions of  strips  of  the  oesophagus  of  the 
frog,  with  the  purpose  of  obtaining  light  on  the  general  question  of  the  activity 
of  plain  muscle  tissue.  The  oesophagus  was  chosen  for  the  work  because  its 
spontaneous  rhythm  was  found  to  be  more  regular  and    rapid    (4  to  6  beats  per 


1586  Journal  of  Applied  Microscopy 

minute)  than  that  of  any  other  portion  of  the  alimentary  tract.  It  was  found 
that  only  in  the  presence  of  Ca  and  K,  as  for  example  in  Ringer's  mixture,  would 
the  contractions  continue  regular  and  forcible  for  any  considerable  length  of 
time.  No  substance  was  found  which  could  be  substituted  for  the  Na  in  the. 
Ringer  mixture.  The  author  inclines  to  the  view  that  both  Ca  and  K  have 
specific  effects,  the  former  acting  as  a  stimulant  and  the  latter  having  an  inhibi- 
tory action,  instead  of  merely  serving  to  neutralize  the  toxic  effect  of  the  NaCl. 
The  Cl-ion  is  not  necessary  for  the  rhythmic  activity,  as  in  place  of  sodium 
chloride,  NaBr,  Nal  or  several  other  sodium  compounds  may  be  used.  Special 
attention  is  called  to  the  correspondence  between  the  oesophagus  and  the  venous 
end  of  the  heart,  which  manifests  itself  in  the  activity  and  relation  to  solutions 
of  the  two  organs.  R.  p. 

Levin,  I..    Physiological  Studies  on   the  Blood       The  purpose  of  this    investigation    was 
of  Animals  deprived  of  the  Adrenals.  Amer.       ,       i   ,  •  i     ^i        .1  ^-  r 

Jour.  Physiol.  5:  358-361,  1901.  ^o  determme  whether  the   secretion    of 

the  adrenal  bodies  serves  to  neutralize 
toxic  substances  formed  in  the  body  and  thus  prevent,  under  normal  conditions, 
auto-intoxication  ;  or,  on  the  other  hand,  merely  acts  on  the  nervous  system  to 
maintain  the  tonus  of  the  vasomotor  and  respiratory  centers  and  the  general 
muscle  tonus.  Both  of  these  views  as  to  the  function  of  these  organs  have  been 
held  by  physiologists.  It  was  found  that  when  the  blood  from  an  animal  (dog 
or  cat)  from  which  the  adrenals  had  been  removed  some  hours  previously  was 
injected  into  the  vascular  system  of  a  normal  animal  an  immediate  and  marked 
rise  in  the  blood  pressure  occurred.  This  result  was  uniformly  obtained  and 
indicates  that  the  blood  of  the  animals  without  adrenals  contains  some  active 
substance  which  is  under  normal  circumstances  neutralized.  The  author  con- 
cludes that  the  death  of  animals  deprived  of  the  adrenals  is  not  due  merely  to 
nervous  depression,  but  to  some  unfavorable  change  in  the  metabolism,  or,  as 
was  held  by  the  earliest  investigators  of  the  subject,  to  an  auto-intoxication  of 
some  sort.  '  r.  p. 

Qodlewski,    E.  (jun.)      O    wpywie  tlenu  na  The    author    finds  that    segmentation 
rozwoj  organismow  i  o    wymianie  gazow  w  •     t-u       u  f         <-  i 

pierwszychstadyachrozwojuzarodkauRana  ^^V    OCCur  m  the  absence  Ot    external 

temporaria.       (Ueber    die    Einwirkung    des  oxygen    supply,   but  that   development 
Sauerstoffs  auf  Entwickelung  und  liber  den  ,        ^,  ,.,.  ,  , 

Gaswechsel  in    den    ersten    Entvvickelungs-  ""^er   these   conditions   only   proceeds 

stadien  von  Rana  temporaria.)   Bull.  Internal.  to  a  certain  point.      CO. ,  has  a  specific 

Acad.    Sci.    de   Cracovie.      Pp.    1-24.  July,  ^      .  ^.  ,        , "  ,         t^i 

j„QQ  ^         -r   J    J,  toxic    action    on    development.       The 

amount  of  gaseous  metabolism  is  shown 
experimentally  to  increase  as  development  proceeds.  r.  p. 

Beer,  Th.    Ueber  primitive  Sehorgane.    Wie-      A  very  complete  summary  and   critical 
ner  klin.     Wochenschr.,    Jahrg.    iqoi.     Nr.        ,.  .  /-.it.        ^  ^1       1  • 

II   12  and  1 3  '   .'      o      /  discussion  of  the  literature  on  the  his- 

tology and  physiology  of  the  eyes  of 
lower  invertebrates.  The  "  objective  nomenclature  "  advocated  for  physiological 
work  by  Beer,  Bethe  and  von  Uexkiill  is  developed  for  the  physiology  of  light 
perception.  r.  p. 


and  Laboratory  Methods.  1587 

Wasmann,  E.     Nervenphysiologie  und  Tier-      A,  criticism  of  the  method  of  investiga- 
psychologie.      Biol.     Centralbl.  21:    21-ti,       ,.  ,         .        ,    ,     ,        .  ... 

jQQj       °  tion  of   animal  behavior  which  centers 

itself  in  the  study  of   the  physiology  of 

the  nervous  system.     The  author  favors  the  method  of  comparative  psychology, 

R.  p. 


CURRENT  BACTERIOLOGICAL  LITERATURE. 

H.  W.  Conn. 

Separates  of  papers  and  books  on  bacteriology  should  be  sent  for  review  to 
H.  W.  Conn,  Wesleyan  University,  Middletown,  Conn. 

Pierce,  Newton  B.    Walnut  Bacteriosis.    Bot.      Newton  B.  Pierce,  of  the  U.  S.  Depart- 
Gaz.  61  :  272. 

ment    of     Agriculture,     has     recently 

described  a  new  organism  as  the  cause  of  the  walnut  bacteriosis,  giving  to   it  the 

name  of  Fseudojnonas  juglandis.     The  organism  is  a  short  rod  with  rounded  ends, 

actively  motile,  bearing  a  single,  long,  polar,  unusually  wavy  flagellum,  occurring 

singly  or  in  pairs,  and   sometimes   in   short   or   long  chains.     This  organism  is 

very  strikingly  pathogenic  to  the  nuts,  leaves,  and  tender  branches  of  the  English 

walnut.     In  the  young  walnut,  the  epicarp    and    forming    shell   and    kernel    are 

destroyed.     The  author  has  produced  a  large  number  of    infections  by  spraying 

the  young  buds  with  a  pure  water  culture  of  the  organism.  L,  H.  Pammel. 

Eijkraann.     Ueber  Enzyme  der  Bakterien  und       ^^^^  ^^^^^^^^  ^as  experimented  upon  the 
Schimmelpilzen.     Cent.  f.  Bak.    u.    Far.    i,  _  ^  _       , 

29:  841,  1901.  formation  of  enzymes  by  bacteria    in  a 

somewhat  ingenious  and  extremely 
interesting  method.  The  method  consists,  in  brief,  in  using  agar  which  has  been 
mixed  with  a  certain  amount  of  material  upon  which  the  enzyme  to  be  tested  will 
have  a  solvent  or  other  noticeable  action.  For  example,  he  first  tested  the 
formation  of  an  enzyme  which  digested  casein.  To  do  this,  he  mixed  a  certain 
amount  of  fresh  skimmed  milk  with  agar,  making  a  white,  cloudy  liquid,  and 
then  inoculated  this  agar,  upon  an  ordinary  plate,  with  the  bacteria  to  be  tested. 
If  the  enzyme  is  produced,  it  diffuses  from  each  colony  and,  as  it  diffuses,  it 
digests  the  casein  so  that  it  dissolves  in  the  liquid  present.  The  result  is  that 
the  colonies  become,  in  a  few  days,  surrounded  by  clear  fields,  which  indicate 
the  digestion  of  the  casein.  On  the  other  hand,  bacteria  which  do  not  pro- 
duce this  enzyme  do  not  develop  such  clear  fields.  By  this  means  can  be  de- 
termined at  a  glance  whether  certain  bacteria  develop  the  enzyme  in  ques- 
tion. The  author  tested  a  large  number  of  bacteria,  finding  that  those  which 
liquefy  gelatin  produce  also  the  enzyme  which  digests  the  casein.  From  this  he 
concludes  that  the  casein  digesting  enzyme  and  the  liquefying  enzyme  are 
identical. 

The  author  tested  the  haemolytic  power  of  bacteria  by  adding  to  the  agar  a 
few  drops  of  blood,  thoroughly  shaking  the  mixture,  and  afterwards  inoculated  it 
with  the  bacteria  to  be  tested,  pouring  it  out  upon  plates  as  usual.     The  bacteria 


1588  Journal  of  Applied  Microscopy 

producing  this  haimolytic  enzyme  develop  colonies  whicii  become  surrounded  by 
a  clear  area,  due  to  the  disappearance  of  the  haimoglobin.  This  enzyme  does 
not  appear  to  be  the  same  as  the  casein  disgesting  enzyme,  since  some  bacteria 
produce  the  one  and  not  the  other.  In  a  similar  way  he  tested  the  production 
of  an  amylolytic  enzyme,  by  bacteria.  For  this  purpose  he  mixed  agar  with  a 
small  amount  of  boiled  starch,  and  inoculated  and  made  plates  as  usual.  After 
the  colonies  have  grown  there  can  be  detected  around  the  colonies  producing  a 
starch  digesting  ferment,  clear  fields  where  the  starch  has  disappeared,  or  the 
presence  of  the  enzyme  can  be  tested  by  throwing  a  weak  solution  of  iodine 
over  the  plate,  when  the  plate  will  turn  blue  in  all  regions  where  the  starch  has 
remained  undigested  ;  the  clear  fields  around  the  areas  are  not  rendered  blue, 
indicating  that  the  starch  has  been  converted  into  sugar.  This  production  of 
amylolytic  ferment  is  tested  for  many  bacteria.  Lastly,  he  tested  the  power  of 
micro-organisms  for  saponifying  fat.  The  method  in  this  case  was  to  mix 
certain  amounts  of  fat,  mutton  tallow  being  used,  with  the  agar  under  certain 
conditions,  and  then  inoculating  as  usual.  The  bacteria  which  produce  the 
saponifying  enzyme  become  surrounded  by  areas  in  which  the  appearance  of  the 
fat  has  been  greatly  changed,  due  to  its  saponification. 

The  method  is  quite  ingenious,  and  the  experiment  interesting  and  suggestive 
of  further  data  for  determining  physiological  properties  of  bacteria,  and  of  thus 
assisting  in  separating  different  species  from  each  other. 

H.    W.    C. 

Kreibich.     Ueber  bakterienfrei  Eiterung    beim       It     has     been     generally     assumed    in 
Menschen.   Wien.  klin.  Woch.  14 :  i;8i,  iqoi.  ,  .1     .  ,i       r  •  r 

•>  -^     ^  recent  years   that  the  formation  of  pus 

was  always  the  result  of  the  action  of  bacteria,  so  much  so  that  it  has  been  more 
or  less  a  dictum  of  bacteriologists  that  there  is  no  pus  formation  without  micro- 
organisms. It  has,  however,  been  recently  recognized  that  there  are  certain 
chemical  agents  which  are  certainly  capable  of  producing  pus,  totally  independ- 
ent of  micro-organisms.  For  example,  Croton  oil,  turpentine,  or  nitrate  of  silver, 
injected  subcutaneously  in  animals,  produce  pus  without  a  suspicion  of  bacterial 
action.  The  author  studies  the  question  whether  pus  is  ever  naturally  formed 
in  man  in  any  other  way  than  by  the  agency  of  bacteria.  That  pus  may  be 
formed  in  man  by  the  action  of  oil  of  turpentine  and  other  chemicals,  is  evident ; 
but  the  author  is  quite  convinced,  from  his  observations,  that  there  are  certain 
instances  when  pus  is  formed  naturally  in  the  human  body,  and  yet  independent 
of  the  action  of  bacteria.  He  mentions,  for  example,  some  cases  of  pus  forma- 
tion in  bubonic  pest,  in  which  the  most  careful  examination  has  failed  to  show 
any  trace  of  micro-organisms.  He  himself  studies  some  cases  of  eczema  in 
which  pus  is  formed.  The  most  careful  study  with  the  microscope  failed  to 
show  any  organisms  in  the  pus,  and  the  most  careful  investigation  by  culture 
methods  has  failed  to  reveal  their  presence.  A  long  series  of  studies  in  this 
direction  convinced  him  that  such  pus  is  sterile,  and  can  not  therefore  be  regarded 
as  due  to  the  action  of  bacteria.  Hence,  he  concludes  that  the  dictum,  no  pus 
without  micro-organisms,  is  totally  erroneous,  and  that  sometimes  pus  is  formed 
naturally  in  man  by  chemical  action.  h.  w.  c. 


and  Laboratory  Methods.  1589 

Neisser  and  Wechsberg,    Ueber  das  Staphylo-      -pj^g  authors  have  undertaken  the  study 

toxin.     Zeit.  f.   Hyg.   3o:   299,    1901.  / 

of  the  question  whether  the  bacteria 
going  under  the  name  of  Staphylococcus  produce  toxic  products.  The  pro- 
duction of  toxines  by  the  well  known  pathogenic  forms  of  Streptococcus  has 
previously  been  subject  to  much  experimentation,  but  the  Staphylococci  have 
been  generally  neglected.  The  author  studies  both  of  the  common  species  of 
this  genus,  the  Staph,  aurus  and  Staph,  albus.  His  method  is  to  cultivate  the 
organisms  in  proper  media  and  then,  after  filtration,  to  test  the  filtrate  for  the 
presence  of  toxic  products.  His  conclusion  is  that  both  of  these  species  produce 
toxines,  and  the  toxines  produced  are  identical  in  both  cases.  There  are  two 
such  toxines,  one  of  which  produces  a  dissolution  of  red  corpuscles,  called 
hcemolysin,  and  the  other  producing  a  dissolution  of  white  corpuscles,  and  called 
leukocidiii.  All  tests  indicate  that  the  toxic  products  produced  by  the  two  species 
are  identical,  inasmuch  as  they  always  answer  to  the  same  tests. 

The  author  is  further  of  the  opinion  that,  so  far  as  concerns  the  production 
of  these  toxic  products,  all  of  the  numerous  varieties  of  Staphylococcus  are 
essentially  the  same.  h.  w.  c. 


NOTES   ON   RECENT  MINERALOGICAL 
LITERATURE. 

Alfred  J.  Moses  and  Lea  McI.  Luquer. 

Books  and  reprints  for  review  should  be  sent  to  Alfred  J.  Moses,  Columbia  University, 

New  York.  N.  Y. 

Vernadsky    W.     Zur    Theorie    der    Silicate.  (Concluded from  November ^ 

Zeit.  f.  Kryst.,  34:  37-66,  1901.  \  j  j 

ALUMOSILICATES. 

Again  we  have  the  salts  and  "  derivatives,"  the  most  important  part  being 
that  of  the  salts  of  the  alumosilicic  acids,  for  which  the  general  formula  is 
m  MO  ^n  AlgOg  "/SiOg,  but  as  experience  shows,  ?n=Ji,  and  when  m=u^=\, 
then  p=\,  2,  4,  6,  and  rarely  8,  10,  or  12;  that  is,  only  the  following  alumo- 
silicates    have    been    observed:      MgAljSiOg,    M2Al2Si20g,     M2Al2Si40i2' 

M2Al2Si60i6,   M2Al2Sig02o,  M2Al2Siio024,    M2Al2Sii2028. 

The  first  of  these  may  be  called  the  group  with  the  chlorite  nucleus,  and  all 
the  others  the  group  with  the  mica  nucleus,  for  the  following  reasons  : 

(a)  There  is  no  known  reaction  by  which  alumosilicates  with  chlorite  nucleus 
can  pass  directly  into  alumosilicates  with  mica  nucleus,  or  the  converse.  It  can 
only  be  accomplished  by  the  introduction  of  "  derivatives." 

(Jt)  Alumosilicates,  with  mica  nucleus  readily  pass  from  one  division  to 
another. 

(<r)  By  weathering,  the  alumosilicates,  with  mica  nucleus,  yield  clay ;  those 
with  chlorite  nucleus  do  not. 

(cl)  Chrome  silicates,  with  chlorite  nucleus,  are  red  or  rose  colored,  but  with 
mica  nucleus  are  green,  indicating  different  structure. 


15^0  Journal  of  Applied  Microscopy 

It  is  probable  that  in  the  structure  formulae  the  hydroxyl  belongs  rather  with 
the  Al  than  the  Si,  because  the  relation  between  the  hydroxyl  group  and  alumina 
group  is  constant,  but  is  not  with  the  silica  group.  Also  because  in  reactions  this 
constancy  mentioned  remains,  while  the  silica  molecule  may  be  split,  and  finally 
because  on  destruction  of  the  alumosilicate  there  result  aluminates  and  not  silicates. 

The  following  may  be,  therefore,  taken  as  the  structure  formula;  of  the 
chlorite  nucleus  HoAljSiO,;,  and  the  mica  nucleus  HoAloSigOg. 


OH 


Al 

<> 

Si 

<> 

Al 

OH 

Chlorite  Nucleus. 

OH 

Al 

/     \ 
0          O 

0==Si          Si= 

=o 

0          O 

Al 

OH 

Mica  Nucleus. 

The  remaining  alumosilicates  readily  pass  into  the  second  type,  and  differ 
from  it  only  in  additional /(^r/rj-  of  SiOg  groups;  these  additional  pairs  differ 
from  the  first  pair  in  that  they  readily  separate  or  recombine.  From  the  two 
groups  represented,  however,  the  SiOj  can  only  be  separated  by  the  strongest 
methods,  involving  the  destruction  of  the  alumosilicate. 

Groups  with  the  Chlorite  Nucleus.- — -The  minerals  belonging  to  these 
groups  have  been  little  studied.  Very  few  simple  salts  are  known,  e.  g.,  marga- 
rite  CaAl2SiOg  and  some  chlorites.  They  form  very  complex  isomorphic 
mixtures  with  each  other,  and  also  isomorphic  mixtures  with  metasilicates.  In 
the  group  may  be  included  all  minerals  with  the  formulae  RoAl2SiO,j  or 
m  R2Al2SiOg  .  A,  and  include: 

1.  The    staurolite     and     clintonite     groups — usually    very    complicated 

formulae. 

2.  The  chlorite  group. 

3.  The  mellilite  group. 

As  yet  no  simple  formulae  will  express  their  composition,  and  there  is  there- 
fore no  substantial  basis  for  structure  formulae. 

Groups  with  Mica  Nucleus. — These  groups  are  much  more  important, 
and  have  been  more  thoroughly  studied.  As  before  stated,  characteristics  are  : 
I^asy  transformation  into  one  another ;  production,  on  earth's  surface,  of  clays 
by  weathering ;  constant  nucleus  in  structure  formula  little  affected  by  most 
reactions  ;  of  SiOg  atoms,  two  can  only  be  removed  by  destruction  of  the  com- 
pound, but  the  rest  can  be  easily  removed  or  added. 


and  Laboratory  Methods.  1591 

The  different  salts  of  mica  variety  readily  form  isomorphic  mixtures  with 
each  other,  and  also  with  metasilicates,  etc. 

All  "  derivatives  "  can  be  synthetically  obtained  at  definite  temperatures  by 
bringing  together  the  mica  nucleus  silicate  and  the  material  to  be  added,  and 
may  be  represented  by  the  general  formula  ;«R2Al2Si2+n02n  4  •  ■^-  The  sub- 
stance A  may  be  replaced  by  another  Aj  etc.,  without  alteration  of  the  nucleus. 

The  physical  characters  of  "  derivatives  "  are  peculiar.  For  instance,  the 
derivatives  of  many  colorless  alumosilicates  are  strongly  colored ;  for  instance  : 
/.  Na2Al2Si208  •  A,  rose  and  red  cancrinite,  yellow  cancrinite,  blue  lapis 
lazuli,  haiiynite,  sodalite. 

The  important  groups  are :  mica  group,  leucite  group,  feldspar  group, 
nephelite  group,  epidote  group,  garnet  group,  under  each  of  which  structure 
formulae  are  given. 

The  Clays. — The  Clays,  under  this  theory  of  the  silicates,  are  free  acids. 
No  clay  corresponding  to  the  chlorite  nucleus  has  been  found  in  nature,  nor 
is  any  known  corresponding  to  the  mica  nucleus,  though  by  heating  rectorite 
H2Al2Si20g  is  left.  The  clays  may  be  considered  as  natural  mechanical  mix- 
tures of  the  following  acids  and  their  derivatives,  and  possibly  others : 

(1)  Kaolin,         .  .  H2 Al2Si208  •  H2O. 

(2)  Halloysite,       .  .     H2Al2Si208  •  2H2O. 

(3)  Pyrophyllite,  .  H.AUSi^Ois- 

(4)  Montmorillonite,       .     HoAUSi^O^  2  •  ^H.O. 

(5)  Nontronite,  .  H2Fe2Si20g  •  HgO. 

A  classification  of  the  silicates  under  this  theory  is  given.  a.  j.  m. 


MEDICAL  NOTES. 

Year  by  year  the  importance  of  more  liberal  education  for  students  entering 
the  medical  college  is  more  clearly  recognized,  and  the  requirements  made  more 
rigid.  Prof.  Stanley  Coulter,  of  Purdue  University,  Lafayette,  Ind.,  recently 
read  a  paper  before  the  Indiana  State  Medical  Society  on  the  subject  of  Pre- 
medical  Education,  at  the  close  of  which  he  outlined  briefly  the  pre-medical 
course  as  given  at  Purdue  University : 

^'•Purpose  of  the  Pre-tnedical  Course. — The  pre-medical  course  of  Purdue  Uni- 
versity is  arranged  to  meet  a  three-fold  demand  : 

1.  To  furnish  a  broad  and  liberal  education. 

2.  To   give  special  and  extended  training  in  those  subjects  which 

underlie  the  strictly  professional  studies  of  the  medical  school. 

3.  By  this  co-relation  of  work  to  shorten  the  time  required  to  obtain 

the  university  and  professional  degrees. 

Graduates  of  Purdue  who  have  completed  this  course  are  admitted  to  the 
second  year  of  all  first-class  medical  schools,  thus  saving  a  full  year. 

The  preliminary  work  required  of  students  entering  this  course  is  that  of  the 
freshman  and   sophomore  years  of  the  general  course,  including  mathematics. 


1592  Journal  of  Applied  Microscopy 

English  literature,  history,  three  years  of  either  German  or  French,  and  one  year 
each  in  general  chemistry,  biology,  and  physics.  Additional  general  subjects 
taken  are  those  required  of  the  junior  and  senior  years  of  the  general  course, 
including  psychology,  literature  and  history,  geology,  and  German  or  French. 

Special  Pre-medical  Studies. — The  special  work  of  this  course  may  be  sum- 
marized as  follows : 

Quantitative  chemical  analysis,  lectures  (37  hrs.)  ;  laboratory  (2'i'2  hrs.). 
Organic  chemistry,  lectures  (52  hrs.);  organic  preparations  (108  hrs.). 
Physiological  chemistry,  lectures    (18  hrs.);    physiological  chemistry 

and  urine  analysis  (102  hrs,). 
Microscopical  technique,  lectures  (15  hrs.);  laboratory  (105  hrs.). 
Normal    and    pathological    histology,    lectures    (40    hrs.) ;    laboratory 

(140  hrs.). 
Vertebrate   anatomy  and  dissection,   recitation   (74  hrs.)  ;  laboratory 

(222  hrs.). 
Embryology,  lectures  and  laboratory  (88  hrs.). 
Bacteriology,  lectures  (87  hrs.)  ;  laboratory  (222  hrs.). 
Animal  physiology,  lectures  (37  hrs.)  ;  laboratory  (150  hrs.). 
Elective. — Technical  chemical  analysis,  eight  hours  weekly  through  the 
year. 

The  most  cursory  inspection  shows  that  in  such  a  course  none  of  the  pro- 
fessional studies  of  the  medical  course  are  undertaken.  There  is  no  attempt  to 
anticipate  them,  but  there  is  insistence  placed  upon  certain  subjects,  knowledge 
of  which  and  skill  in  which  are  evidently  of  the  highest  value  in  the  work  of  the 
medical  college.  Such  a  course  has  two  main  objects  :  First,  to  give  to  the  pre- 
medical  student  the  broad  vision  and  strong  mental  grasp  that  can  result  only 
from  a  broad  and  liberal  culture.  Second,  to  give  such  special  training  in  those 
subjects  which  underlie  the  science  of  medicine  that  he  may,  when  he  enters  the 
professional  school,  be  prepared  for  its  real  problems,  and  in  a  much  greater 
degree  than  at  present  understand  the  magnitude  of  the  work  he  has  undertaken 
when  he  has  chosen  to  enter  the  ranks  of  the  medical  profession.  Incidentally, 
this  method  saves  one  of  the  preparatory  years — no  small  matter  in  these  strenuous 
times." — Medical  Record,  10:  14.  -  c.  w.  j. 


Kaiserling  Method  for  the  Preservation  of  Pathological  Specimens. — 
It  is  held  that  by  employing  this  method  the  natural  colors  of  specimens  will  be 
preserved  almost  exactly,  and  apparently  for  an  indefinite  period  when  kept  in  a 
dark  place.  The  specimens  are  placed  as  soon  as  possible  in  the  following  solu- 
tion, in  which  they  remain  for  3  to  5  days  : 

No,  1.    Formalin,     ......         40  parts. 

Water, 200  parts. 

Potassium  nitrate,         ....  3  parts. 

Potassium  acetate,  ....       6  parts. 

Specimens  must  not  be  carefully  washed  out  in  running  water,  as  it  removes 
the  blood  on  which  the  color  depends.  Any  excess  of  blood,  etc.,  should  be 
simply  wiped  off  before   placing  specimens  in  solution  No.  1.     Specimens  lose 


and  Laboratory  Methods.  1593 

their  color  after  remaining  in  solution  No.  1  for  a  few  days,  but  the  color 
returns  after  the  specimens  are  transferred  to  the  next  solution,  which  consists 
of, 

No.  2.    Alcohol,  ...-.,         4  parts. 

Water, 1  part. 

and  in  which  they  remain  for  3  to  5  hours,  after  which  they  are  placed  in, 

No.  3.    Alcohol,  95  per  cent, 
for  1  to  2  hours,  and  finally  are  permanently  preserved  in, 

No.  4.    Potassium  acetate,  ....  1  part. 

Glycerin,    .......     2  parts. 

Water  distilled,      .....        10  parts. 

which,  before  use,  should  be  allowed  to  stand  for  48  hours,  and  then  filtered. 
Specimens  thus  prepared  and  preserved  should  be  kept  in  a  dark  place,  as 
light  produces  bleaching  in  the  course  of  time. — Texas  Med.  Ne^cs,  10:  10. 

c.  w.  J. 


Examination  of  Blood. —  Six  things  are  essential  in  order  to  make  satisfac- 
tory examination  of  the  blood  :  (1)  The  apparatus  must  be  absolutely  clean. 
(2)  The  various  stages  in  the  process  must  be  performed  rapidly,  because  the 
cell  coagulation  of  the  blood  will  interfere  with  any  of  the  tests.  (3)  The  work 
must  be  done  accurately.  (4)  Making  large  quantities  of  stain  and  keeping 
same  in  a  glass-stoppered  bottle  will  standardize  the  solution,  giving  mini- 
mum variations  in  intensity  of  stain.  (5)  Fixing  of  specimens,  by  continuous 
heat,  with  as  slight  a  degree  of  variation  in  distribution  of  heat  as  possible. 
(6)  Taking  of  blood  specimens  with  reference  to  time  of  day. 

In  order  to  more  accurately  fix  the  blood  count,  it  has  been  found  that  speci- 
mens taken  in  the  morning,  before  any  undue  excitement  is  indulged  in,  and 
previous  to  taking  of  liquids  or  solids,  will  give  a  more  uniform  blood  count  than 
those  taken  at  any  other  period  of  the  day. — /our.  Am.  Med.  Ass.  37 :  8. 

c.  w.  J. 


The  corner-stone  of  the  new  medical  building  of  the  University  of  Michigan 
was  laid  on  October  15th  by  Dr.  Leartus  Connor,  president  of  the  State  Medical 
Society.  The  building  will  contain  the  laboratories  and  class-rooms  of  the 
departments  of  hygiene,  bacteriology,  anatomy,  histology,  and  pathology.  The 
contracts  for  its  erection  call  for  an  expenditure  of  $88,000,  exclusive  of  what 
may  be  required  for  heating,  plumbing,  and  general  equipment. — Science,  14 :  356. 


1694 


Journal  of  Applied  Microscopy 


NEWS  AND  NOTES. 

An  Apparatus  and  Method  for  Rapidly  Staining  Large  Numbers  of 
Sputum  Specimens. — This  apparatus,  as  illustrated,  consists  of  a  long,  narrow 
copper  bath,  mounted  on  legs,  weighted  to  ensure  stability,  and  of  sufficient  height 
to  permit  the  use  of  a  Bunsen  burner  under  the  bath.  At  one  end  near  the  top 
are  two  inlets ;  the  upper  one  (A)  to  admit  the  stain,  the  lower  one  (B)  to  admit 
water.     In  the  bottom  of  the  bath,  at  the  same  end,  is  a  small  outlet  (C)  for  the 


Fig.  1. — Staining  Bath  for  Sputum  Specimens. 

Stain,  closed  by  a  rubber  tube  and  pinchcock.  At  the  other  end  of  the  bath, 
partitioned  off  by  a  false  wall,  is  a  one-half  inch  siphon,  the  inner  end  of  which 
is  about  three-eighths  inch  from  the  bottom,  and  the  top  about  on  a  level  with 
the  upper  inlet. 

Instead  of  ordinary  microscopical  slides,  a  thin  plate  of  glass,  etched  as  shown 
in  Fig.  2,  is  used.  The  etched  area  above  the  spacings  furnishes  a  surface  on 
which  desired  data  may  be  written.  This  large  slide  is  made  of  a  size  to  fit  the 
bath  and  is  held  upright  by  guides  in  the  ends  of  the  bath. 

To  use   this   apparatus,  proceed  as  fol- 
lows : 

Stain  is  admitted  through  A  until  the 
bath  is  about  two-thirds  full,  or  to  a  depth 
sufficient  to  cover  the  preparations  without 
starting  the  siphon.  The  specimens  are 
then  stained,  after  which  the  stain  is  drawn 
off  through  the  outlet  C.  Water  is  then  turned  into  the  bath  through  B.  The 
bath  fills  until  the  top  of  the   siphon   is  reached,  then  drains  rapidly  until  the 


Fig.  2. — Slide  for  Sputum  Specimens. 


and  Laboratory  Methods.  1595 

siphon  breaks,  when  the  bath  fills  again  and  empties  as  before,  and  so  on  until 
washing  is  completed.  A  glass  candy  tray,  a  trifle  over  nme  inches  long  and 
wide  enough  to  hold  two  slides  side  by  side,  is  recommended  for  decolorizing 
the  specimens.  Small  strips  of  glass  are  cemented  in  the  ends  of  the  tray  so 
that  when  the  slides  are  placed  in  it  specimen  side  down,  just  sufficient  space 
remains  for  the  required  amount  of  decolorizer.  After  decolorizing,  the  speci- 
mens may  again  be  washed  in  the  bath,  after  which  they  are  ready  for  examina- 
tion.— Jour.  Bost.  Soc.  Med.   Sd.,  5 :  8. 


A  Device  for  Paraffin  Embedding. — This  device  consists  of  a  printer's 

composing  stick,  six  inches  long  and 
two  and  one-fourth  inches  wide.  It  has 
an  adjustable  slide,  fastened  by  a  thumb 
set-screw,  and  set  at  a  distance  of  four 
inches  from  one  end,  giving  a  space 
four  inches  long  and  two  and  one-half  inches  wide.  This  space  may  be  divided 
by  nonpareil  slugs  and  quads  into  compartments  of  any  desired  size,  to  hold  the 
blocks  of  material.  When  the  tissue  is  ready  for  embedding,  a  small  amount  of 
melted  paraffin  is  poured  into  the  compartment,  the  tissue  properly  arranged 
therein,  and  the  compartment  filled  with  paraffin.  The  stick  is  then  cooled  on 
ice  or  in  cold  water,  so  that  it  requires  only  a  few  minutes  to  accomplish  the 
\vork. — Am.  Med.  1:1. 


RiNGiNc;  Slides. — Many  amateurs  are  unable  to  finish  well  made  mounts 
with  neatly  made  rings  of  cement.  This  is  often  caused  through  using  the  cement 
too  thick.  Professional  mounters  have  two  bottles,  one  containing  the  cement, 
the  other  the  solvent — generally  turpentine  or  methylated  spirits.  The  brush  is 
first  dipped  in  the  solvent,  then  in  the  cement,  and  a  thin  coat  is  deposited  on 
the  slide  as  it  is  rotated  on  the  turn-table.  Some  build  up  the  ring  at  once, 
others  allow  the  first  layer  to  dry  and  then  complete  the  process.  Each  time  a 
fresh  brushful  of  cement  is  taken,  it  should  be  preceded  by  a  dip  in  the  solvent. 
The  cement  can  then  be  deposited  with  cleanness  and  regularity. — Kno^vledgc, 
24:  186. 


The  annual  meeting  of  the  American  Society  of  Naturalists  and  Affiliated 
Societies  will  be  held  at  Chicago  on  Tuesday  and  Wednesday  of  Convocation 
Week,  that  is,  December  31st  and  January  1st.  The  discussion  before  the 
naturalists  will  be  on  Wednesday  afterpoon,  and  the  annual  dinner,  at  which  the 
president.  Prof.  Wm.  T.  Sedgewick,  will  give  the  address,  will  take  place  in  the 
evening.  The  subject  selected  for  the  discussion  is  "  The  Relations  of  the 
American  Society  of  Naturalists  to  Other  Scientific  Societies." — Science.,  14  :  356. 


A  new  science  building,  to  cost  $300,000,  is  being  constructed  at  Colorado 
College. 


1596  Journal  of  Applied  Microscopy 

A  catalogue  of  the  Marine  Invertebrata  of  Eastern  Canada,  by  Dr.  J.  F. 
Whiteaves,  has  been  published  by  the  Geological  Survey  of  Canada  (1901).  It 
consists  of  a  systematic  list  of  all  the  species  described  from  the  Bay  of  Fundy, 
the  Atlantic  coast  of  Nova  Scotia,  the  Gulf  and  Mouth  of  the  St.  Lawrence  river 
as  far  north  as  the  straits  of  Belle  Isle.  The  localities  at  which  some  of  the 
species  are  found  fossil  in  the  Pleistocene  deposits  are  also  briefly  indicated. — 
Nature,  64  :  1 666. 


Numerous  inquiries  have  been  received  asking  where  and  at  what  cost  the 
bulletin  of  photo-micrographs  issued  from  the  Biological  Laboratory  of  Earlham 
College,  and  noticed  in  our  August  number,  may  be  secured.  The  bulletin  is  pub- 
lished by  Nicholson  iv  Bro.  of  Richmond,  and  may  be  had  at  twenty-five  cents 
per  copy. 


QUESTION   BOX. 

Inquiries  will  be  printed  in  this  department  from  any  inquirer. 
The  replies  will  appear  as  received. 

REPLY  TO  QUESTION  No.   13. 

A  quantitative  test  for  the  bacteria  in  milk  can  be  made  with  very  low  magni- 
fying power.  To  do  this  a  known  quantity  of  milk  is  diluted  with  a  known 
quantity  of  sterilized  water.  This  is  evenly  spread  over  a  gelatine  culture  in  a 
Petri  dish.  Place  this  in  a  warm  (not  above  100°F)  and  dark  place  for  thirty-six 
hours,  and  it  will  then  be  ready  for  examination.  The  bacterial  colonies,  many 
of  which  can  be  seen  with  the  unaided  eye,  will  show  the  approximate  number  of 
bacteria  present  in  the  milk  used.  The  Petri  dish  may  be  placed,  without 
uncovering,  upon  the  stage  of  the  microscope,  and  the  colonies  examined  with 
the  low  power.  Objectives  ranging  from  the  2-inch  to  the  73-inch  may  be  used. 
Directions  for  making  the  gelatine  culture  may  be  found  in  almost  any  work  on 
bacteriology,  and  Petri  dishes  can  be  purchased  for  twenty-five  cents  apiece. 

C.  A.  Whiting. 
REPLY  TO  QUESTION  No.   14. 

In  response  to  the  inquiry  of  your  correspondent  this  month,  allow  me  to  say 
that  Tallqvist's  method  of  haimoglobin  estimation  consists  of  specially  prepared 
paper  upon  which  a  drop  of  blood  is  allowed  to  fall,  and  a  color  scale  for  com- 
parison. It  is  accurate  to  about  ten  per  cent.  It  costs  $1.25,  and  may  be 
obtained  of  the  Harvard  Cooperative  Society,  Boylston  street,  Boston. 

F.  W.  Hku-.ins,  M.  D. 

REPLY  TO  QUESTION  No.   15. 

I  have  frequently  transferred  alcoholic  specimens  of  animal  tissue  to  formalin 
without  bad  results.  In  fact,  tissues  originally  placed  in  dilute  alcohol  are  fre- 
quently improved  for  histological  purposes  by  being  changed  to  formalin. 

C.  A.  Whiting. 


For  Japan :     S.  Yoshizoe,  Itotanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  January,   t90t  No.  / 

LEADING  SUBJECTS 

Moses  C.  White. 

S.  H.  GAGE,  Cornell  University, 1109 

Fire  in  the  Veterinary  College  at  Cornell. 

S.  H.  GAGE, 1111 

Laboratory  Photography. 

Stereo  Photo-Micrography, 1113 

The  New  York  Botanical  Garden. 

D.  T.  MAC  DOUGAL, 1115 

Preliminary  Study  of  Mycetozoa. 

CLARA  LANGEN  BECK,  Wells  College, 1119 

Micro-Chemical  Analysis,  X.     Potassium. 

E.  M.  CHAMOT,  Cornell  University, 1121 

Easy  Method  of  Mounting  and  Preserving  Mosquitos. 

V.  A.  LATHAM,  M.  D., 1129 

Current  Botanical  Literature. 

CHAS.  J.  CHAMBERLAIN,  University  of  Chicago, 1131 

Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1133 

Normal  and  Pathological  Histology. 

RICHARD  M.  PEARCE,  M.  D.,  Harvard  College, 1136 

General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan,      1138 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1141 

Notes  on  Recent  Mineralogical  Literature, 

ALFRED  J.  MOSES,  LEA  McI.  LUQUER, 1144 

Medical  Notes, 1145 

News  and  Notes, 1147 

Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kntcred  at  the  Post  OIHuu  at  Rochester,  N.  Y.,  as  Secoort  Class  Matter 

tONBON :  Dawbarn  &  Ward,  Ltd.,  6  Farringdou  Avenue,  E.  C. 


PUBLISHER'S  ANNOUNCEMENTS. 


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CHICAGO, 

State  and  'WaBhineton  Sts. 


FIELD  GLASSES. 


SYNTHOL 


(ABSOLUTE) 


A  chemically  pure  synthetic  sub- 
stitute  for  absolute  ALCOHOL. 


PURER,      STRONGER,      MORE  STABLE,      CHEAPER. 

YNTHOL  may  be  used  for  every  purpose  for  which 
alcohol  is  used,  except  for  internal  consumption. 
Being  chemically  pure  it  does  not  have  as  much  odor 
as  absolute  alcohol  from  grain  or  wood.  It  is  perfectly  free  from 
color,  is  non-irritant  to  eyes  or  skin  and  has  from  ten  to  fifteen 
per  cent,  more  solvent  power  than  ordinary  alcohol. 


PRICES: 

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1  bbl.  of  50  gallons  per  gallon,  net,     2.25 


As  a  killing,  fixing,  or  hardening  agent  it  is  in  every  respect  equal 
to  the  best  absolute  alcohol  and  can  be  used  as  a  substitute  for  it  in  the 
preparation  of  stains,  reagents,  etc.  We  offer  a  special  low  price  on  Synthol 
in  barrel  lots  FOR  MUSEUM  PURPOSES.  As  a  preservative  it  is 
superior  to  any  alcohol.  Alcohol  becomes  tinged  with  color  on  expos- 
ure to  light.  Synthol  retains  its  absolute  colorlessness  under  all  condi- 
tions.   Sample  supplied  free  to  laboratories. 

MANUFACTURED  EXCLUSIVELY  FOR 

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For  Japan  ;     S.  Yosliizoo,  liotanical  Garden,  Imperial  University,  Toklo. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


Vol.  IV  February,   190t  No,  2 


LEADING  SUBJECTS 

The  New  Biological  Laboratories  of  Ripon  College. 

C.  DWIGHT  MARSH,  Ripon  College, 1149 

A  Combined  Condenser  and  Polarizer  for  Petrographical  Microscopes. 

W.  L.  PATTERSON,      1155 

A  Plan  for  a  Ureometer. 

J.  B.  NICHOLS,  M.  D.,  Washington,  D.  C, 1156 

A  Device  for  Supporting  Pasteur  Flasks. 

KATHERINE  E.  GOLDEN, 1157 

Laboratory  Photography.     High-Power  Photo-micrography. 

C.  E.  McCLUNG,  University  of  Kansas, 1158 

An  Improvised  Microtome. 

IRWIN  LA  VERNE  POWERS, 1162 

The  Study  of  Bacteria  in  the  Public  Schools. 

JAMES  E.  PEABODY, 1164 

Biology  Wall  Charts. 

ORSON  HOWARD, 1172 

Staining  in  Toto  with  Delafield's  Haematoxylin. 

NEWTON  EVANS,  M.  D.,  Am.  Med.  Missionary  College,  .  .  .  I  1  72 
Current  Botanical  Literature. 

CHAS.  J.  CHAMBERLAIN,  University  of  Chicago, 1174 

Cytology,  Embryology,  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1176 

Normal  and  Pathological  Histology. 

JOSEPH  H.PRATT,  Harvard  University  Medical  School,  .  .  1180 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1182 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1186 

Medical  Notes, 1188 

Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Knlt-rol  at  Ihe  Post  Oltici;  at  U.wh.stcr,  X.  V..  a^  Si-coii.l  Class  Matl.;r. 


LONDON :  Dawbarn  &  Ward,  Ltd.,  G  Farringdon  Avenue,  K.  C. 


LA  BORA  TOR  Y  APPLIANCES. 


For  Spring  botany 

Our  line  of  Botanical  Supplies  is  not  the  regu- 
lar stock  which  is  usually  found  on  the  market. 

Our 

Botanical  Mounting  Paper 

is  a  fine  quality  of  linen  ledger  paper  manufactured  especially  for 
us  after  our  own  specifications  with  a  view  to  producing  a  pure 
white  sheet  which  will  retain  its  whiteness  and  strength  with  age,  and 
have  that  much  desired  stiffness  which  makes  the  sheet  stand  out 
firmly  when  held  by  one  end  for  inspection  of  the  specimen.     Our 

Genus  Covers 

are  likewise  made  especially  for  us,  and  in  addition  to  being  made 
of  the  finest  fiber,  have  a  superior  writing  finish.  We  likev/ise  have 
our 


Driers 


made  to  order  to  secure  that  bibulousness  and  weight  which  add  to 
their  effectiveness.  Having  these  papers  made  in  quantity  enables 
us  to  quote  a  very  satisfactory  rate  on  large  orders  for  the  herbaria 
of  universities,  colleges,  and  other  institutions.  Our  prices  on  small 
lots  for  private  use  will  be  found  as  low  as  any,  and  you  get  an  extra 
quality  of  paper  besides.     In  the  matter  of  the 

fasGulum  aim  Plant  Press 

We  have  advantages  to  offer  in  the  convenient 
construction,  neat  workmanship,  durability,  and 
moderate  price.  Place  orders  early  and  do  not 
fail  to  get  estimates  from  us  before  ordering 
elsewhere. 

No.  1244  Mounting  Paper  (standard  size  used  in 

the  Asa  Gray  Herbarium  at  Harvard)  per 

ream  (500  sheets) $4.50 

No.  1242  Genus  Covers  for  above,  per  100  .  .  2.00 
No.  1240  Drying  Paper,  33x46  cm.,  ex.heavy.pr.lOO  1.00 
No.  1248  Vasculum,  with  strap  (size  13x20x40cm.)  1.50 
No-  1246  Portable  Plant  Press,  very  light,  with 

bands  to  prevent  disarrangement  of  speci^ 

mens  when  new  plants  are  added  in  the  field     2.00 

Bausch  &  Lomb  Optical  Co. 

ROCHESTER,  N.  Y. 

Chicago,  111.,  New  York, 

State  and  Washington  Sts.  25th  St.  and  Broadway. 

Goods  lu  Stock  at  Branch  Offices. 


MISCELLANEOUS  ADVERTISEMENTS. 


The  Thornton -Pickard 
Focal  Plane  Shutter 

is    the    only    Shutter    that    will    do 
full  justice   to   very  fast    Lenses 


A  complete  catalogue  of  Thornton- Pickard  Shutters  will 
be   sent   postpaid   to   any  address   upon   application  to 

ANDREW  J.   LLOYD   &   CO. 

323-325  \A/'ashington   Street,  -  -  BOSTON,   MASS. 


,"%/%/%/%/%/%■ 
(( 


A  Finger  Touch  Finds  the  Reference"         ^ 


IN  THE 


r 


"Y  &  E" 

gard  I  ndex 


4 


■^v  \  •    ^^^^^S^^t^^^^K^^^      Adapted  for  use  in  any  con- 

R^  ^1    ^^^^^Kk.  -P^--'  nection  for  any  subject.     If 

^^^^^^H|^  '  you   want   to   know   how   it 

.  -^^3:  vJ^^^H^^^  can  be  applied  to  your  special  require- 

"     A^^^^^  ments,  send  for  our  new  catalogue  No. 

27 -B.    It  illustrates   and   explains  35  different  forms  of  records  and 

gives  complete  information  on  the  subject  of  card  indexing. 


Principal  Branches :  Yawman  &  Erbe  Mfg.  Co., 

^        NEW  YORK,  360  Broadway. 

5        CHICAGO,   138  Wabash  Ave.  Fac.ories  and  Main  Offices  :   ROChCSter,   N.   Y. 

~        SAN  FRANCISCO,  29  New  Montgomery  St.  "^  "^  '  ^ 


PHOTOGRAPHIC  APPARATUS. 


THE 


REMO 


The  Best  Camera  Made 


Fitted  with  all  the  latest  improvements  and  sup- 
plied with  the  Victor  Lens  and  Shutter.  Our 
absolute  guarantee  goes  with  every  Premo  sold. 

The  Premo  Uses  Both  Glass  Plates  and  Film 


1900  Catalogue  sent  on  application. 


Rochester  Optical  Co.,n^?hester,  n.y. 


IP" 

W4 

M 


i 


m 


THE  PONY 

PREMO 


is  a  special  camera  for  wheelmen. 
So  compact  and  portable  that  it  can 
be  attached  to  a  bicycle  the  same  as  a 
tool  bag.     Price,  $10.00  and  upward. 


For  Japan  :    S.  Yoshizoe,  Botanical  Garden,  Imperial  University,  Toltlo. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


Vol.  IV  March,   i90t  No,  3 


LEADING  SUBJECTS 

Micro-Chemical  Analysis,  XI.    Ammonium. 

E.  M.  CHAMOT,  Cornell  University, 1189 

Staining  Sections  for  Class  Work. 

NEWTON  EVANS,  M.  D.,  Am.  Med.  Missionary  College,  ...  1  194 
Home  Made  Wall  Charts. 

CHARLES  E.  BESSEY,  University  of  Nebraska, 1195 

Flattening  and  Fixing  Paraffin  Sections  on  Slide. 

B.  M.  DAVIS,  State  Normal,  Los  Angeles,  Cal.,     1196 

A  Ventilated  Dish  for  Bacteria  Cultures. 

GEORGE  C.  WHIPPLE,  Mt.  Prospect  Lab.,  Brooklyn,  N.  Y.,  1  197 
Laboratory  Photography  : 

The  Processes  of  Photo-Micrography. 

C.  E.  McCLUNG,  University  of  Kansas, 1199 

A  Laboratory  Camera  Stand. 

P.  H.  ROLFS,  Clemson  Agri.  Coll.,  S.  C, 1202 

Current  Botanical  Literature. 

CHAS.  J.  CHAMBERLAIN,  University  of  Chicago, 1206 

Cytology,  Embryology,  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1208 

Current  Zo'dlogical  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1212 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  .  .  1214 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan,      12 IS 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1222 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES,  LEA  Mc  1.  LUQUER, 1224 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kntcre<l  at  the  Posl  Oflice  al  Rochcsler,  X.  Y.,  as  .Sicoii.l  ria->  .Mattir 


I.ONDOX:  Dawbarn  &  Ward,  Ltd.,  6  Farringdon  Avenue,  E.  C. 


CHEMICALS. 


SYNTHOL 


(ABSOLUTE) 


A  chemically  pure  synthetic  sub- 
stitute  for  absolute  ALCOHOL. 


PURER,      STRONGER,      MORE  STABLE,      CHEAPER. 

YNTHOL  may  be  used  for  every  purpose  for  which 
alcohol  is  used,  except  for  internal  consumption. 
Being  chemically  pure  it  does  not  have  as  much  odor 
as  absolute  alcohol  from  grain  or  wood.  It  is  perfectly  free  from 
color,  is  non-irritant  to  eyes  or  skin  and  has  from  ten  to  fifteen 
per  cent,  more  solvent  power  than  ordinary  alcohol. 


PRICES: 

1  pint,  net $    .40 

I  gallon,  net 2.50 

1  bbl.  of  50  gallons  per  gallon,  net,     2.25 


As  a  killing,  fixing,  or  hardening  agent  it  is  in  every  respect  equal 
to  the  best  absolute  alcohol  and  can  be  used  as  a  substitute  for  it  in  the 
preparation  of  stains,  reagents,  etc.  We  offer  a  special  low  price  on  Synthol 
in  barrel  lots  FOR  MUSEUM  PURPOSES.  As  a  preservative  it  is 
superior  to  any  alcohol.  Alcohol  becomes  tinged  with  color  on  expos- 
ure to  light.  Synthol  retains  its  absolute  colorlessness  under  all  condi- 
tions.    Sample  supplied  free  to  laboratories. 


MANUFACTURED  EXCLUSIVELY  FOR 

BAUSCH  &  LOMB  OPTICAL  CO., 


CHICAGO. 


ROCHESTER,  N.  Y. 


NEW  YORK. 


MISCELLANEOUS  ADVERTISEMENTS. 


The  Thornton -Pickard 
Focal  Plane  Shutter 

is    the    only    Shutter    that    will    do 
full  justice   to   very  fast    Lenses 


A  complete  catalogue  of  Thornton- Pickard  Shutters  will 
be   sent   postpaid   to   any  address   upon   application  to 


ANDREW  J.   LLOYD   8z:   CO. 

323-325  Washington  Street,  -  -  BOSTON,   MASS. 


A  Finger  Touch  Finds  the  Reference" 

IN  THE 

"Y  &  E" 

Qard  Jndex 


^ 


V 


Adapted  for  use  in  any  con- 
nection for  any  subject.  If 
you  want  to  know  how  it 
can  be  applied  to  your  special  require- 
ments, send  for  our  new  catalogue  No. 
27 -B.  It  illustrates  and  explains  35  different  forms  of  records  and 
?ives  complete  information  on  the  subject  of  card  indexing. 


■  'fncipal  Branches :  Yawman  &  Efbe  Mfg.  Co., 

\        NEW  YORK,  360  Broadway.  *  ' 

5        CHICAGO,   138  Wabash  Ave.  D        U  M     V 

J        SAN  FRANCISCO,  29  New  Montgomery  St.  Factories  and  Main  Offices  :   KOCttCSter,   IM  .    I  . 


PHOTOGRAPHIC  APPARATUS. 


THE 


REMO 


The  Best  Camera  Made 


Fitted  with  all  the  latest  improvements  and  sup- 
plied with  the  Victor  Lens  and  Shutter.  Our 
absolute  guarantee  goes  with  every  Premo  sold. 

The  Premo  Uses  Both  Glass  Plates  and  Film 


1900  Catalogue  sent  on  application. 


Rochester  Optical  Co.,'*'to?hester,N.Y. 


7m 

7M 
7M 


7M 

m 
m 

M 

0^ 


w 

Hi 
^^ 

i 

Is 


THE  PONY 

PREMO 


is  a  special  camera  for  wheelmen. 
So  compact  and  portable  that  it  can 
be  attached  to  a  bicycle  the  same  as  a 
tool  bag.     Price,  $10.00  and  upward. 


For  Japan :    S.  Yoshlzoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


Vol.  IV  April,  190t  No.  4 


LEADING  SUBJECTS 

The  Laboratory  Equipment  of  the  "Bahama  Expedition"   from  the  University 

of  Iowa. 

C.  C.  NUTTING,  University  of  Iowa,     1229 

Laboratory  Photography. 

The  Value  of  the  Telephoto-Lens. 

MORTON  J.  ELROD,  University  of  Montana, 1241 

Micro-Chemical  Analysis  XII.    The  Analytical  Reactions  of  Group  II. 

E.  M.  CHAMOT,  Cornell  University, 1242 

A  Description  of  the  New  Wing  of  the  Laboratory  of  Hygiene  at  the  University  of 

Pennsylvania, 1234 

Course  in  Biology  in  the  Horace  Mann  High  School, 1249 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   ....    1255 

Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1257 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1260 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  .   .   .    1262 

General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1264 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1266 

Medical  Notes, 1268 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kntereil  at  the  Past  Offlco  at  Rochester,  N.  Y.,  a.s  Second  Class  Matter. 


tiONDOX :  Dawbarn  &  Ward,  Ltd.,  6  Farringdon  Avenue,  E.  C. 


LABORATORY  APPARATUS. 


LABORATORY 
TABLE  DESK 

FINE  QUARTERED  OAK.     FULL  OFFICE  SIZE. 


THE  problem  of  the  Laboratory  Table  is  still 
far  from  settled  to  the  satisfaction. of  all, 
but  all  who  have  had  an  opportunity  to 
work  at  one  of  these  desks  will  be  loth  to  try 
any  other.  The  general  construction  is  the 
s  inie  as  of  a  fine  Roll  Top  Office  Desk,  but  modified  to  suit  the  requirements  of  the 
laboratory.  There  are  thirty-four  glass-stoppered  reagent  bottles  in  the  top  case,  covered 
by  a  roller  curtain  with  lock  and  key.  The  top  of  desk  is  plate  glass.  There  are 
seven  large  side  drawers  for  apparatus,  the  lower  right  deep  for  microscope  ;  the  upper 
left  with  receptacles  for  microscopic  preparations. 


PRICE,    $50.00. 


BAUSCH  &  LOMB  OPTICAL  CO. 


ROCHESTER,  NEW    YORK. 


BOTANY 
CLASSES 

puzzle  tlie  instructor  as  to  what  to  recommend  for  an 
all-round 

MAGNIFYING      GLASS      AND 
DISSECTING    MICROSCOPE. 


QR    MAGNIFIER. 


THE  QR  Magnifier  has  stood  the  test  of  time 
and  offers  many  advantages  where  a  low 
priced  lens  is  required.     It  costs  so  little 
every  student  can  afford  one,  and  being  adjusta- 
ble for  focus  and  having  a  large,  clear  field,  with  good  magnifying  power,  serves  the 
purpose  of  a  pocket  lens  (with  legs  removed)  and  dissecting  microscope. 


QR,  50  CENTS.     SPECIAL  PRICES    IN  QUANTITY. 


BAUSCH  &  LOMB  OPTICAL  CO. 


ROCHESTER,    NEW    YORK 


The  Post  Express  Printin?  Compaay, 
Rochester,  N.  Y. 


MISCELLANEOUS  ADVERTISEMENTS. 


Thornton-Pi  ckard 
^^^  Shutters  ^^^ 


THE  GREAT  THING  ABOUT  THESE  SHUTTERS  IS  THAT  THEY  ADMIT 
MORE  LIGHT,  EXPOSE  THE  EDGES  OF  THE  PLATE  AS  EVENLY  AS 
THE  CENTER,  AND  DO  THE  LENS  NO  INJURY  BY  JARRING.  THEY 
ARE  ROLLER  CURTAIN  SHUTTERS,  SIMPLE  IN  CONSTRUCTION, 
RELIABLE  IN  ACTION,  AND  NOT  LIKELY  TO  GET  OUT  OF  REPAIR. 

For  general  purposes,  the  Time  and  Instantaneous,  either  before  the  lens 
or  behind  the  lens,  is  the  best  shutter  made. 

For  very  rapid  work,  The  Thornton-Pickard  Focal  Plane  Shutter 

is  unapproached  ;  works  directly  in  front  of  the  plate,  passes  an  enormous  quantity 
of  light,  and  is  the  only  shutter  to  do  justice  to  the  full  powers  of  the  fastest  lenses. 
Very  simple,  reliable,  and  not  easily  put  out  of  order. 

Thornton-Pickard  Catalogue  free  on  application. 


Andre^v  J.  Lloyd  &  Co., 


323  'Wasbinsrton  Street, 
BOSXOI9. 


The  New  Edition  of  our  Encyclopaedia  will  be  ready  about  May  10th. 
Send  30  Cents  and  it  will  be  sent  promptly  when  issued. 


"A  Finger  Touch  Finds  the  Reference"         t 


IN  THE 


"Y  &  E" 

Qard  Index 


Adapted  for  use  in  any  con- 
nection for  any  subject.  If 
you  want  to  know  how  it 
can  be  applied  to  your  special  require- 
ments, send  for  our  new  catalogue  No. 
27 -B.  It  illustrates  and  explains  35  different  forms  of  records  and 
gives  complete  information  on  the  subject  of  card  indexing. 


Principal  Branches  :  Yawman   &  Erbe  Mfg.  Cc, 

^        NEW  YORK,  360  Broadway. 

5        CHICAGO,  138  Wabash  Ave.  ^  ^  ..   .    ^„.  D^^Uooto^     M     V 

f        SAN  FRANCISCO,  29  New  Montgomery  St.  F«tor.es  and  Mam  Offices  :   KOCtteStCr,   IN  .    I .  ^ 


PHOTOGRAPHIC  APPARATUS. 


PREMO  AND  POCO 
CAMERAS 


The  recognized  standard  high-grade 
Cameras  of  the  world,  have  been 
reduced  in  price  25  to  50  per  cent. 
Our  increased  faciUties  have  made 
this  reduction  possible. 

New  Ideas  and  New  Features 

Especially  adapted  for  scientific 
photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
Cameras  the  best  and  most  practical 
instruments  ever  constructed. 

Write  for  complete  Art  Catalogue,  describing  this  entire  line. 

Rochester  Optical  &  Camera  Co., 


Scientific  Department. 


Rochester,  N.  Y.,  u.  s.  a. 


For  .Tf-  p:  n  :     S.  Yoshizoe,  Rotanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VcL  IV  May,  190 1  No,  5 

LEADING  SUBJECTS 

The  University  of  Montana  Biological  Station. 

MORTON  J.  ELROD,  University  of  Montana, 1269 

The  Marine  Biological  Laboratory  at  Cold  Spring  Harbor,  L.  I. 

H.  S.  PRATT,  Haverford  College,     1279 

A  Method  for  Injecting  Small  Vessels. 

LEON  J.  COLE,  University  of  Michigan, 1282 

Laboratory  Photography.  The  Photo-Micrography  of  Tissues  with  Simple  Apparatus. 

W.  H.  WALMSLEY,     1283 

Combined  Ureometer  and  Saccharometer. 

A.  ROBIN,  M.  D., 1286 

Micro-Chemical  Analysis  XIII.     Strontium,  Barium. 

E.  M.  CHAMOT,  Cornell  University, 1289 

A  Simple  Washing  Device. 

ROBERT  G.  LEAVITT, 1297 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,  ....  1299 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1301 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  .  .  .  1304 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1307 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1310 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES,  LEA  McI.  LUQUER, 1312 

Medical  Notes,      1314 

News  and  Notes,   .         1315 

Question  Box, 1316 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kntcri'il  at  the  Post  Office  at  Roehi'.ster,  \.  Y.,  as  .'fecoaii  Class  .Matter. 


I^OXnOX :  Dawbarn  &  Ward,  Ltd.,  6  Farringdon  Avenue,  E.  C. 


LABORATORY  APPARATUS. 


LABORATORY 
TABLE  DESK 

FINE  QUARTERED  OAK.     FULL  OFFICE  SIZE. 


THE  problem  of  the  Laboratory  Table  is  still 
far  from  settled  to  the  satisfaction  of  all, 
but  all  who  have  had  an  opportunity  to 
work  at  one  of  these  desks  will  be  loth  to  try 
any  other.  The  general  construction  is  the 
same  as  of  a  fine  Roll  Top  Office  Desk,  but  modified  to  suit  the  requirements  of  the 
laboratory.  There  are  thirty-four  glass-stoppered  reagent  bottles  in  the  top  case,  covered 
by  a  roller  curtain  with  lock  and  key.  The  top  of  desk  is  plate  glass.  There  are 
seven  large  side  drawers  for  apparatus,  the  low^r  right  deep  for  microscope  ;  the  upper 
left  with  receptacles  for  microscopic  preparations. 


PRICE,    $50.00 


BAUSCH  &  LOMB  OPTICAL  CO. 


ROCHESTER,  NEW    YORK. 


BOTANY 
CLASSES 

puzzle  the  iiLStructor  as  to  what  to  recommend   for  an 
all-round 

MAGNIFYING      GLASS      AND 
DISSECTING    MICROSCOPE. 


T 


OR    MAGNIFIER. 


^HE  OR  Magnifier  has  stood  the  test  of  time 
and  offers  many  advantages  where  a  low 
priced  lens  is  required.     It  costs  so  little 
every  student  can  afford  one,  and  being  adjusta- 
ble for  focus  and  having  a  large,  clear  field,  with  good  magnifying  power,  serves  the 
purpose  of  a  pocket  lens  (with  legs  removed)  and  dissecting  microscope. 


QR,  50   CENTS.     SPECIAL  PRICES    IN  QUANTITY. 


BAUSCH  &   LOMB  OPTICAL  CO. 

ROCHESTER,    NEW    YORK. 


The  Post  Express  Printing  Company, 
Rochester.  N.  Y. 


MISCELLANEOUS  ADVERTISEMENTS. 


1  hornton-Pickard 
■ Shutters  ^^^ 


THE  GREAT  THING  ABOUT  THESE  SHUTTERS  IS  THAT  THEY  ADMIT 
MORE  LIGHT,  EXPOSE  THE  EDGES  OF  THE  PLATE  AS  EVENLY  AS 
THE  CENTER,  AND  DO  THE  LENS  NO  INJURY  BY  JARRING.  THEY 
ARE  ROLLER  CURTAIN  SHUTTERS,  SIMPLE  IN  CONSTRUCTION, 
RELIABLE  IN  ACTION,  AND  NOT  LIKELY  TO  GET  OUT  OF  REPAIR. 

For  general  purposes,  the  Time  and  Instantaneous,  either  before  the  lens 
or  behind  the  lens,  is  the  best  shutter  made. 

For  very  rapid  work,  The  Thornton-Pickard  Focal  Plane  Shutter 

is  unapproached  ;  works  directly  in  front  of  the  plate,  passes  an  enormous  quantity 
of  light,  and  is  the  only  shutter  to  do  justice  to  the  full  powers  of  the  fastest  lenses. 
Very  simple,  reliable,  and  not  easily  put  out  of  order. 

Thornton-Pickard  Catalogue  free  on  application. 

Andrew  J.  Lloyd  &  Co.,  ^"^  ^"«osxo:S. **'""'' 

The  New  Edition  of  our  Encyclopaedia  will  be  ready  about  May  loth. 
Send  20  Cents  and  it  will  be  sent  promptly  when  issued. 


A  Finger  Touch  Finds  the  Reference" 

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I'HOTOGRAPHIC  APPARATUS. 


PREMO  AND  POCO 
CAMERAS 


The  recognized  standard  high-grade 
Cameras  of  the  world,  have  been 
reduced  in  price  25  to  50  per  cent. 
Our  increased  faciHties  have  made 
this  reduction  possible. 

New  Ideas  and  New  Features 

Especially  adapted  for  scientific 
photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
Cameras  the  best  and  most  practical 
instruments  ever  constructed. 

Write  for  complete  Art  Catalogue,  describing  this  entire  line. 

Rochester  Optical  &  Camera  Co., 


Scientific  Department. 


Rochester,  N.  Y.,  u.  s.  a. 


For  Japan :    S.  Yoshizoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  June,    190 1  No.  6 

LEADING  SUBJECTS 

Improved  Automatic  Microtomes. 

CHARLES  S.  MINOT,  LL.  D.,  Harvard  University, 1317 

New  Freezing  Microtome  for  use  with  Carbon-Dioxide  Tanks. 

CHARLES  R.  BARDEEN,  Johns  Hopkins  University,    ....    1320 

Micro-Chemical  Analysis,  XIV.    Barium. 

E.  M.  CHAMOT,  Cornell  University, 1323 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   ....    1332 

Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1334 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1339 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  .    .   .   1341 

General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1343 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1347 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES,  LEA  Mel.  LUQUER, 1350 

Medical  Notes,      1353 

News  and  Notes, 1354 

Question  Box, 1356 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Entered  at  the  Post  Office  at  Rochester,  X.  Y.,  as  Second  Class  Matter. 


LABORATORY  APPARATUS. 


LABORATORY 
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FINE  QUARTERED  OAK.     FULL  OFFICE  SIZE. 


THE  problem  of  the  I/aboratory  Table  is  still 
far  from  settled  to  the  satisfaction  of  all, 
but  all  who  have  had  an  opportunity  to 
work  at  one  of  these  desks  will  be  loth  to  try 
any  other.  The  general  construction  is  the 
same  as  of  a  fine  Roll  Top  Office  Desk,  but  modified  to  suit  the  requirements  of  the 
laboratory.  There  are  thirty-four  glass-stoppered  reagent  bottles  in  the  top  case,  covered 
by  a  roller  curtain  with  lock  and  key.  The  top  of  desk  is  plate  glass.  There  are 
seven  large  side  drawers  for  apparatus,  the  lower  right  deep  for  microscope  ;  the  upper 
left  with  receptacles  for  microscopic  ^^reparations. 


PRICE,    $50.00. 


BAUSCH  &  LOMB  OPTICAL  CO. 


ROCHESTER,  NEW    YORK 


UR    MAGNIFIER. 


BOTANY 
CLASSES 

puzzle  the  instructor  as  to  what  to  recommend  for  an 
all-round 

MAGNIFYING      GLASS     AND 
DISSECTING    MICROSCOPE. 


THE  QR  Magnifier  has  stood  the  test  of  time 
and  offers  many  advantages  where  a  low 
priced  lens  is  required.     It  costs  so  little 
every  student  can  afford  one,  and  being  adjusta- 
ble for  focus  and  having  a  large,  clear  field,  with  good  magnifying  power,  serves  the 
purpose  of  a  pocket  lens  (with  legs  removed)  and  dissecting  microscope. 


QR,  50   CENTS.     SPECIAL  PRICES    IN  QUANTITY. 


BAUSCH  &  LOMB  OPTICAL  CO 

ROCHESTER,    NEW    YORK. 


PHOTOGRAPHIC  APPARATUS. 


PREMO  AND  POCO 
CAMERAS 


The  recognized  standard  high-grade 
Cameras  of  the  world,  have  been 
reduced  in  price  25  to  50  per  cent. 
Our  increased  faciHties  have  made 
this  reduction  possible. 

New  Ideas  and  New  Features 

Especially  adapted  for  scientific 
photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
Cameras  the  best  and  most  practical 
instruments  ever  constructed. 

Write  for  complete  Art  Catalogue,  describing  this  entire  line. 

Rochester  Optical  &  Camera  Co., 


Scientific  Department. 


Rochester,  N.  Y.,  u.  s.  a. 


TRAVEL. 


TOUR     OF    THE    GREAT     LAKES     ON      THE     FLOATING      PALACES     OF     THE 

NORTHERN   STEAMSHIP  COMPANY 


A  new  steamer  and  two  sailings  weekly  service  to  Chicago,  Milwaukee,         |\\ 

^         and  Harbor  Springs,  will  be  added  to  the  regular  Buffalo-Duluth  service, 

which  opens  early  in  June. 

I  . 

K      For  particulars  regarding  service  and  extended  tours  appiy  to 

I  W.  M.  LOWRIE, 

General  Passenger  Agent,   Buffalo. 


''^>f'i!^^'^:^'^y9!S^''!iifff^^jfS!ffy:^^ 


For  Japan :    S.  Yosliizoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  J^hf    t90i  No.  7 

LEADING  SUBJECTS 

The  Value  of  Methylen  Blue  as  an  Intravitam  Stain  in  the  Tunicata. 

GEORGE    WILLIAM    HUNTER,    JR.,   DeWitt   Clinton   High 

School, • 1357 

Spermatozoa  of  Man,  Domestic  Animals  and  Rodents. 

L.  NAPOLEON  BOSTON,  M.  D.,  Philadelphia  Hospital,  1360 

An  Improved  Photo-Micrographic  Apparatus. 

B.  H.  BUXTON,  Cornell  Medical  College, 1366 

Micro-Chemical  Analysis,  XV.     Magnesium  Group. 

E.  M.  CHAMOT,  Cornell  University, 1373 

Current  Botanical  Literaiure. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   ....    1381 

Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1382 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1385 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,   .    .    .    1387 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1391 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES,  and  LEA  McI.  LUQUER,  Columbia  Uni- 
versity,     1393 

Medical  Notes,  Methods  for  Staining  Tubercle  Bacilli, 1395 

News  and  Notes, 1395 

Question  Box, 1396 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kniurert  at  the  Post  Ottioe  at  Rochi'ster,  N  Y.,  as  Second  Class  Matter. 


LABORATORY  APPARATUS. 


LABORATORY 
TABLE  DESK 

FINE  QUARTERED  OAK.     FULL  OFFICE  SIZE. 


THE  problem  of  the  Laboratory  Table  is  still 
far  from  settled  to  the  satisfaction  of  all, 
but  all  who  have  had  an  opportunity  to 
work  at  one  of  these  desks  will  be  loth  to  try 
any  other.  The  general  construction  is  the 
same  as  of  a  fine  Roll  Top  Office  Desk,  but  modified  to  suit  the  requirements  of  the 
laboratory.  There  are  thirty-four  glass-stoppered  reagent  bottles  in  the  top  case,  covered 
by  a  roller  curtain  with  lock  and  key.  The  top  of  desk  is  plate  glass.  There  are 
seven  large  side  drawers  for  apparatus,  the  lower  right  deep  for  microscope  ;  the  upper 
left  with  receptacles  for  microscopic  preparations. 

PRICE,    $50.00. 

BAUSCH  &  LOMB  OPTICAL  CO. 

ROCHESTER,  NEW    YORK. 


BOTANY 
CLASSES 

puzzle  the  instructor  as  to  what  to  recommend  for  an 
all-round 

MAGNIFYING      GLASS      AND 
DISSECTING    MICROSCOPE. 


T 


QR    MAGNIFIER. 


VHE  QR  Magnifier  has  stood  the  test  of  time 
and  offers  many  advantages  where  a  low 
priced  lens  is  required.  It  costs  so  little 
every  student  can  afford  one,  and  being  adjusta- 
ble for  focus  and  having  a  large,  clear  field,  with  good  magnifying  power,  serves  the 
purpose  of  a  pocket  lens  (with  legs  removed)  and  dissecting  microscope. 


QR.  50  CENTS.     SPECIAL  PRICES    IN  QUANTITY. 


BAUSCH  &  LOMB  OPTICAL  CO. 

ROCHESTER,    NEW    YORK. 


PHOTOGRAPHIC  APPARATUS. 


PREMO  !!<£  POCO 

CAMERAS 


NEW  IDEAS  m  NEW  FEATURES 


i 


The  recogni^td  standard  high-grade  Cam- 
eras of  the  world,  and  the  world's  greatest 
Cameras,  especially  adapted  for  scien- 
tific photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
CAMERAS  the  best  and  most  practical 
instruments  ever  constructed.    •:•   •:•    •:•    :• 

WRITE  FOR  COMPLETE  ART  CATALOGUE, 
DESCRIBING    THIS     ENTIRE     LINE. 


i 


ROCHESTER    OPTICAL 
&  CAMERA  CO. 


SCIENTIFIC   DEPT. 


ROCHESTER,  N.  Y.,  U.  S.  A. 


TRAVEL. 


i?^!!SSZ:^;?;»32?'<"^5^'2:5Se55?2g3a!2S:22^^ 


For  Japan :    S.  Yoshizoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  August,    i90t  No.  8 


LEADING  SUBJECTS 

Table  of  Specific  Gravities  of  Saturated  Solutions  and  Solubilities  of  Anilin  Stains. 

LOUIS  LEROY,  Vanderbilt  University,      ,    .    1397 

Laboratory  Photography.     Photomicrography. 

D.  W.  DENNIS,  Earlham  College, 1399 

Notes  on  Testing  for  B.  Coli  in  Water. 

STEPHEN  DeM.  GAGE,  Lawrence  Experiment  Station,     .    .    .    1403 
The  Cone  Net. 

E.  A,  BIRGE,  University  of  Wisconsin, 1405 

A  Modification  of  the  Birge  Collecting  Net. 

ROBT.  H.  WOLCOTT,  University  of  Nebraska, 1407 

A  Method  of  Determining  the  Comparative  Gravity  of  Alcohol  when  Dehydrating 
by  Osmosis. 

R.  P.  WOODFORD, 1409 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   .    .    .    .    1411 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1414 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1418 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,   .    .    .    1421 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1423 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1427 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES  and  LEA  McI.  LUQUER,  Columbia  Uni- 
versity,     1431 

Medical  Notes, 1434 

News  and  Notes, 1435 

Question  Box, 1436 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

EaicTcf]  at  tbe  Post  Offici-  at  Rochister,  N.  Y.,  as  Second  Class  Mailer. 


LOKDON :  Dawbarn  &  Ward,  Ltd.,  6  Farringdon  Avenue,  E.  C. 


LA  BORA  TORY  APPARA  TUS. 


MI  NOT  ^^^"Ji'^'^ 

Microtome 


Cuts  Pai'affin  or 
Celloidin  Equally  Well.       ^^ 


The  Most  Accurate  Microtome  Made. 
For  Serial  Sectioning  Has  no  Superior. 


OUR  NEW  CATALOGUE  OF  MICROTOMES 
and  accessories  contains  full  descriptions  of  an 
entire  new  line  of  Microtomes  and  is  illustrated 
with  photographic  reproductions  of  the  instruments, 
showing  all  details.     Mailed  free. 


Bausch  &  Lomb  Optical  Co.,  Rochester,  N.  Y. 


NEW  YORK. 
CHICAGO. 


I'be  i'ost  Express  Printing  Company, 
Rochester,  N.  Y. 


PHOTOGRAPHIC  APPARATUS. 


PREMO  iiiB  POCO 

CAMERAS 


NEW  IDEAS  as  NEW  FEATURES 


i 


The  recognized  standard  high-grade  Cam- 
eras of  the  world,  and  the  world's  greatest 
Cameras,  especially  adapted  for  scien- 
tific photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
CAMERAS  the  best  and  most  practical 
instruments  ever  constructed,    v    -:•    -:•    •:• 


WRITE    FOR    COMPLETE    ART    CATALOGUE, 
DESCRIBING    THIS     ENTIRE     LINE. 


i 


ROCHESTER    OPTICAL 
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SCIENTIFIC   DEPT. 


ROCHESTER,  N.  Y.,  U.  S.  A. 


TRAVEL. 


I       TOUR     OF    THE    GREAT     LAKES     ON      THE     FLOATING      PALACES     OF     THE 

^  NORTHERN   STEAMSHIP  COMPANY 

A  new  steamer  and  two  sailings  weekly  service  to  Chicago,  Milwaukee, 
and  Harbor  Springs,  will  be  added  to  the  regular  Buffalo-Duluth  service, 
which  opens  early  in  June. 

;S      For  particulars  regarding  service  and  extended  tours  appiy  to 

W.  M.  LOWRIE, 

General  Passenger  Agent,  Buffalo. 


. 


,> 


i 


For  Japan ;    S.  Yoshizoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


Vol.  IV  September,  i901  No.  9 

LEADING  SUBJECTS 

Laboratory  Courses  by  Correspondence. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,     .    .    .    1437 
Laboratory  Photography : 

Photographing  Diatoms. 

P.  C.  MYERS,  University  of  Iowa, 1439 

The  5  mm.  Apochromat,  after  Prof.  Charles  S.  Hastings    in  the  Photog- 
raphy of  Diatoms, 1442 

The  New  Medical  Laboratories  of  the  University  of  Pennsylvania. 

SIMON  FLEXNER,  University  of  Pennsylvania, 1445 

Magnifiers. 

J.  DEARNESS,  London  Normal  School,  London,  Canada,     .    .    1448 
A  New  Thermo-Regulator. 

THOMAS   PALMER, 1449 

A  Rapid  Method  of  Making  Slides  of  Amoeba. 

M.  A.  WlLLCOX,Wellesley  College, 1450 

Micro-chemical  Analysis,  XVI,  Zinc,  Cadmium. 

E.  M.  CHAMOT,  Cornell  University, 1451 

Device  for  Leveling  the  Microscope. 

T.  O.  REYNOLDS, 1458 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   ....    1460 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1462 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1465 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,   .    .    .    1468 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1470 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1472 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES  and  LEA  McI.  LUQUER,  Columbia  Univ.   1474 
Medical  Notes .  ...    1476 

Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Kntere'l  at  the  Post  Office  at  Rochcsier,  X.  Y.,  as  Second  Class  Matter, 


LONI>OX:  Dawbarn  &  Ward,  Ltd.,  6  Farringdon  Avenue,  E.  C. 


LABORATORY  APPARATUS. 


Automatic 
Laboratory  Microtome 


A  PRACTICAL  LABORATORY  INSTRUMENT. 


Cuts    Paraffin    or    Celloidin    Material 
equally  well. 

Cuts  any  thickness  from  two  to  sixty 

Microns. 

1* 
Has    all    Adjustments    for    orienting 

large  and  small  objects  required. 

Moderate  in  price. 


OUR    NE^A^    CATALOGUE 

describes  a  complete  new  series  of  Mic= 
rotomes  of  several  different  styles,  all 
entirely  remodeled  and  representing  the 
highest  mechanical  attainments  in  con- 
struction. Jb  Photographic  illustrations 
show  each  in  detail.     Mailed  free. 


Bausch  &c  Lomb  Optical  Co., 


NEW  YORK. 


ROCHESTER,   N.  Y. 


CHICAGO. 


The  Post  Express  Printing  Company. 
Rochester,  N.  Y. 


LABORATORY  APPARATUS. 


MI  NOT  filllilif, 

Microtome 


Cuts  Paraffin  or 
Celloidin  Eqiially  Well.       ^1 


The  Most  Accurate  Microtome  Made. 
For  Serial  Sectioning  Has  no  Superior. 


OUR  NEW  CATALOGUE  OF  MICROTOMES 
and  accessories  contains  full  descriptions  of  an 
entire  new  line  of  Microtomes  and  is  illustrated 
with  photographic  reproductions  of  the  instruments, 
showing  all  details.     Mailed  free. 


NEW  YORK. 


Bausch  &  Lomb  Optical  Co.,  Rochester,  N.  Y.      Chicago. 


PHOTOGRAPHIC  APPARATUS. 


PREMO  its  POCO 

CAMERAS 


NEW  IDEAS  as  NEW  FEATURES 


i 


The  recognized  standard  high-grade  Cam- 
eras of  the  world,  and  the  world's  greatest 
Cameras,  especially  adapted  for  scien- 
tific photography  have  been  introduced, 
making  both  the  PREMO  and  POCO 
CAMERAS  the  best  and  most  practical 
instruments  ever  constructed.    •^    •:•    •:•   •:• 


WRITE    FOR    COMPLETE    ART    CATALOGUE, 
DESCRIBING    THIS     ENTIRE     LINE. 


i 


ROCHESTER    OPTICAL 
&  CAMERA  CO. 


SCIENTIFIC   DEPT. 


ROCHESTER,  N.  Y.,  U.  S.  A. 


For  Japan :    S.  Yoshizoe,  Botanical  Garden,  Imperial  University,  Tokio. 


Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  October,  1901  No,  to 


LEADING  SUBJECTS 

The   Botanical   Laboratory   and  the   Botanical  Garden    of  the    Tokyo    Imperial 
University,  Japan. 

KIICHI  MIYAKE,  Cornell  University, 1477 

The  Course  of  Study  in  Invertebrate  Zoology  in  the  Marine  Biological  Laboratory 
at  Wood's  Holl. 

CASWELL  GRAVE,  Johns  Hopkins  University, 1481 

Botany  at  the  Biological  Laboratory  at  Wood's  Holl. 

C.  H.  SHAW,  Temple  College, 1486 

Laboratory  Photography : 
The  Ortol  Developer. 

R.  P.WOODFORD, 1487 

Contributions  to  Our  Knowledge  of  Color  in  Photo-Micrography,  1489 

Kresylechtviolett. 

RALPH  L.  MORSE,  University  of  Michigan, 1492 

Micro-Chemical  Analysis,  XVII,  Magnesium  Group-Separations. 

E.  M.  CHAMOT,  Cornell  University, 1495 

A  Damp  Chamber  for  Use  on  the  Klinostat. 

HOWARD  S.  REED,  University  of  Michigan, 1499 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,   ...       1502 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1504 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1506 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,   .    .    .    1508 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1510 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES  and  LEA  McI.  LUQUER,  Columbia  Univ.   1513 

Medical  Notes,      1514 

News  and  Notes, 1516 

Question  Box, 1516 

Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Knlered  at  the  Post  Office  at  Rochester,  N.  Y..  as  Second  Class  Matter. 


LONDON :  Dawbarn  &  Ward,  L,td.,  6  Farringdon  Avenue,  E.  C. 


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A  POPULAR  description,  by  the  foremost  authority  in  this  coun- 
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Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


Vol  lY  November,  1901  No,  it 


LEADING  SUBJECTS 

Studying  and  Photographing  the  Wild  Bird. 

FRANCIS  H.  HERRICK,  Western  Reserve  University,  ....  1517 
A  Few  Remarks  en  the  Technic  of  Blood  Preparations. 

B.  L.  RAWLINS, 1524 

Laboratory  Photography : 

Photomicrography,  II.    An  Apparatus  Adapted  to  all  Kinds  of  Work. 

D.  W.  DENNIS,  Earlham  College, 1525 

Staining  Bacteria  in  the  Root  Tubercles  of  Leguminous  Plants. 

GEORGE  J.  PEIRCE,  Leland  Stanford  Jr.  University,  ....  1528 
Micro-Chemical  Analysis,  XVIII. 

E.  M.  CHAMOT,  Cornell  University, 1529 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,  ....  1535 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1537 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1541 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  ...  1544 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1546 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1548 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES  and  LEA  McI.  LUQUER,  Columbia  Univ.   1553 


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PHOTOGRAPHIC  APPARATUS. 


Nezu  Ideas  and  Neiv  Features 


PREMOandPOCO 


AMERA 


HE  recognized  high-grade  Cameras  of  the 
world,  and  the  world's  greatest  Cameras, 
especially  adapted  for  scientific  photog- 
raphy, making  both  the  PREMO  and  POCO 
CAMERAS  the  best  and  most  practical  instru- 
ments ever  constructed. 

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Cbe  Tnsect  Book 

By  DR.  L.  O.  HOWARD, 

Chief  of  Division  of  Entomology,  U.  S.  Department  of  Agriculture. 

16  colored  plates,  32  full-page  half-tones  and   300  text  cuts 
illustrating  hundreds  of  species. 

A  popular  description  by  the  foremost  authority  in  this  country  of 
the  Bees,  Wasps,  Ants,  Grasshoppers,  Flies,  and  other  North  American 
insects — exclusive  of  the  Butterflies,  Moths,  and  Beetles.  It  has  full  life 
histories,  giving  an  intimate  account  of  the  most  wonderful  facts  in  that 
insect  world  all  around  us,  which  is  so  incompletely  known,  even  by  scientists. 
A  man  may  go  into  a  city  park,  and  with  patience  and  industry  find 
insects  never  described  by  the  entomologists.  There  is  to-day  almost 
nothing  that  covers  authoritatively,  yet  popularly,  this  vast  field,  so  that 
this  volume  has  special  importance. 

The  INDEPENDENT  says  :   "It  is  refreshing  to  see  an  entomologist  of  such  eminent 

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Journal  of 

Applied  Microscopy 

and 

Laboratory  Methods 


VoL  IV  December,  1901  No.  t2 

LEADING  SUBJECTS 

General  Methods  for  the  Study  of  the  Nervous  System. 

GILBERT  L.  HOUSER,  University  of  Iowa, 1557 

Method  for  Rearing  Amoeba. 

MELT.  COOK,  DePauw  University, 1566 

A  Short  Method  for  the  Widal  Test,      1 565 

The  Arrangement  of  Cilia  on  Paramecium, 1566 

Immersion  Oil  in  Collapsible  Tubes 1567 

CHARLES  WRIGHT  DODGE,  University  of  Rochester. 
Laboratory  Photography : 

Further  Notes  on  the  Use  of  the  Telephoto  Lens. 

MORTON  J.  ELROD,  University  of  Montana, 1568 

Current  Botanical  Literature. 

CHARLES  J.  CHAMBERLAIN,  University  of  Chicago,  ....    1573 
Cytology,  Embryology  and  Microscopical  Methods. 

AGNES  M.  CLAYPOLE,  Cornell  University, 1576 

Current  Zoological  Literature. 

CHARLES  A.  KOFOID,  University  of  California, 1578 

Normal  and  Pathological  Histology. 

JOSEPH  H.  PRATT,  Harvard  University  Medical  School,  .   .   .    1583 
General  Physiology. 

RAYMOND  PEARL,  University  of  Michigan, 1584 

Current  Bacteriological  Literature. 

H.  W.  CONN,  Wesleyan  University, 1587 

Notes  on  Recent  Mineralogical  Literature. 

ALFRED  J.  MOSES  and  LEA  McI.  LUQUER,  Columbia  Univ.   1589 

Medical  Notes, 1591 

News  and  Notes, 1594 

Question  Box, 1596 


Publication  Department  BAUSCH  &  LOMB  OPTICAL  CO.,  Rochester,  N.  Y. 

Entered  at  the  Post  Office  at  Rochester,  N.  Y.,  as  Second  Class  Matter. 


PHOTOGRAPHIC  APPARATUS. 


Neiv  Ideas  and  Neiv  Features 


PREMO  AND  POCO 


AMERA 


HE  recognized  standard  high-grade  Cam- 
eras of  the  world,  and  the  world's  greatest 
Cameras,  especially  adapted  for  scientific 
photography,  making  both  the  PREMO  and 
POCO  CAMERAS  the  best  and  most  practical 
instruments  ever  constructed. 

WRITE  FOR  COMPLETE  ART  CATALOGUE  DESCRIBING  THIS  ENTIRE  LINE, 


ROCHESTER  OPTICAL 
AND  CAMERA  CO.  -«--«*- 


Scientific  Department. 


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Laboratory  §team  Sterilizersl 

f-       -p>  FOR  UNIVERSITIES  AND  COLLEGES.  I 


Improved  Style  with  Side  Door. 


The  Arnold  Sterilizer, 

illustrated  on  this  page,  is  adapted 
for  class  work.  It  is  inexpensive 
and  a  number  of  them  can  be  pur- 
chased for  a  small  outlay,  thus 
permitting  the  use  of  one  sterilizer 
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PUBLISHER'S  ANNOUNCEMENTS. 


ALASKA 


The  Results  of  the  Harri- 
man  Alaska  Expedition. 
Published  with  the  co- 
operation of  the  Washing- 
ton Academy  of  Sciences. 


ITS  NATIVES, 

BIRD  AND  ANIMAL  LIFE, 
TREES  AND  FLOWERS, 

AND  RESOURCES. 


iC^ 


WHAT  THE 

BOOK  IS. 


An  Epoch  Making 

Work  of 

Scientific  Travel 

with  40  Superb 

Colored  Plates, 

85  Photogravures, 

300  Text  Drawings 

and  5  Maps. 

2  Vols. 
$15.00  NET. 


This  work  contains,  in  popular  form,  the 
results  of  the  expedition  sent  in  1899,  by 
Mr.  Edward  H.  Harriman,  to  this  land  of 
fogs,  glaciers,  volcanoes,  seals,  and  gold. 

The  twenty-five  eminent  scientists  made 
extensive  collections  in  their  various 
branches  of  research  ;  no  less  than  twenty- 
six  new  species  of  mammals  were  dis- 
covered, and  the  results  in  other  branches 
of  zoology,  inbotany,  ornithology,  geology, 
etc.,  were  almost  as  important.  Many  new 
glaciers,  and  a  large  fjord,  hitherto  un- 
mapped, were  also  explored.  It  is  not  only 
the  final  authority  on  Alaska,  but  a  book  of 
fascinating  interest. 


^S^ 


John  Burroughs,  W.  H. 
Brewer,  William  Healey 
Dall,  Bernhard  E.  Fernow, 
Henry  Gannett,  Geo.  Bird 
Grinnell,  Charles  Keeler, 
C.  Hart  Merriam,  John 
Muir,  M.  L.  Washburn. 


WHAT  IS 

SAID  OF  IT. 


"  Sumptuous. 


-New  York  Tribune. 


"  The  finest  example  of  the  publisher's 
art  that  the  present  season  has  produced." 
— New  York  Telegram. 

"  Nothing  approaching  the  pictures,  in 
range,  variety,  and  beauty  has  ever  been 
obtained  before.  The  most  beautifully 
illustrated  work  of  travel  ever  issued  on 
this  side  of  the  Atlantic." 

—The  Nation, 

"  Chaste  and  elegant  in  design  and  exe- 
cution, artistic  from  every  point  of  view, 
lavishly  and  exquisitely  illustrated." 

—The  Dial,  Chicago. 


Size,  7  X  10  ;  pages,  about  500  ;  binding,  cloth,,  decorated  ;  illustra- 
tions, 40  in  color,  85  photogravures,  apd  300  drawings  from  photo- 
graphs and  paintings  by  Louis  Agassiz  Fuertes,  Charles  Knight,  R. 
Swain  Gifford,  F.  S.  Dellenbaugh,  etc.;  2  vols.;  price,  net,  $15.00. 


DOUBLEDAY,  PAGE  &  COMPANY, 

34  UNION  SQUARE,  EAST,  NEW  YORK. 


New  York  Botanical  Garden   Librar 


3  5185  00264  0504 


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