Transactions

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TRANSACTIONS

OF

lak AMERICAN

Microscopical Society,

ORGANIZED 1878. INCORPORATED 1891.

EDITED BY THE SECRETARY.

_ EIGHTEENTH ANNUAL MEETING

CORNELL UNIVERSITY, 1ITHACA,-N. Y.

August 21, 22 and 23, 1895.
atl —

oe L
VOLUME XVII. i,

BUFFALO, N. Y.:
THE WENBORNE-SUMNER CO., PRINTERS,
1895.

OFFICERS FOR 1895-96.

President: A. CLIFFORD MERCER, Me Oe : : Syracuse, N. WS < LY
Vice-Presidents: EDWARD PENNOCK, . é : Philadelphia, Pa. 2 O
Miss V. A. LATHAM, _ Chicago, Ill. |
Secretary: WiLL1aM C. Krauss, M. D., Buffalo, Ilo WE fia es .
Treasurer: MAGNUS PFLAUM, . 3 ; : . Pittsburg, Pa,
ELECTIVE MEMBERS OF THE EXECUTIVE COMMITTEE. V. [ %
O I >)
Pa ee a: BUIGENMANN 200 Sele les 'y on eee Bloomington, Ind. Op, a
HERMAN SCHRENKE, 5 : : , SR . St. Louis, Mo.

Miss M. A. Boots, . Longmeadow, Mass.

PAST PRESIDENTS.
Ex-officio members of the Executive Board.
i. R. BH. Wurp, M\D!/ 8, BR. MS., of Troy, N. Y.,
at Indianapolis, Ind., 1878.
_ BR. H. Warp, M.D F. R-M..8., of Troy, N. YS
at Buffato, N. Y¥., 1879:
H. L. Switu, DL... D., F: R. M. S., of Geneva, N. Y.,
at Detroit, Mich., 1880.
4. J. D. Hyatt, F. R. M.S. (absent), of New York City,
at Columbus, Ohio, 1881.
5. Gro. E. BLACKHAM, (M.D., F. R. M. S., of Dunkirk, N: Y.;
at. Elmira, N. Y., 1882.
6. ALBERT McCauua, PH. D., F. R. M. S., of California,
at Chicago, Ill., 1883.
i. J. D. Cox, LL. D., F. R. M.S., of Cincinnati; Ohio,
at Rochester, N. Y., 1884.
8. H. L. Stu, LL. D., F. BR. M.S., of Geneva, Nowy,
at Cleveland, Ohio, 1885.
9, T. J. BURRILL, PH. D., F. R. M. S., of Champaign, IIl.,
at Chautauqua, N. Y., 1886.
10. Wa. A. Rogers, A. M., F. R. M. S., of Waterville, Me.,
at Pittsburgh, Pa., 1887.
li. D. 8S. KeLuicoTtt, PH. D., F. R. M. S., of Columbus, Ohio,
at Columbus, Ohio, 1888.
12. WM. J. Lewis, M. D., F. R. M. S., of Hartford, Conn.,
at Buffalo, N. Y., 1889.
15. GEO. E. FELL, M. D., F. R. M. S., of Buffalo,.N. Y.,
at Detroit, Mich., 1890.
14. Frank L. JAMES, Pu. D., M. D., F. R. M. S., of St. Louis, Mo.,
at Washington, D. C., 1891.
0. MARSHALL D. EWELL, M. D., F. R. M. S., of Chicago, IIL,
at Rochester, N. Y., 1892.
16, Jacop D, Cox, LL. D., F. R. M: S., of Cincinnati, Ohio,
at Madison, Wis., 1893.
17. Lester Curtis, M. D., F. R. M. S., of Chicago, IIL,
at Brooklyn, N. Y., 1894.
18. Simon Henry Gace, B.S., of Ithaca, N. Y.,

vo

~

at Ithaca, N. Y., 1895.

The Society does not hold itself responsible for the opinions expressed by members in its
published Proceedings unless indorsed by a special vote,

PROCEEDINGS

or

The American Microscopical Society

ADDRESS OF THE PRESIDENT.

’ The Processes of Life Revealed by the Microscope ; a Plea for
Physiological Histology.

Simon Henry Gace, B. S., Ithaca, N. Y.

It is chatacteristic of the races of men that almost at the
dawn of reflection the first question that presses for solution is
this one of Life; life as manifested in men and in the animals
and plants around them. What and whence is it and whither
does it tend? Then the sky with its stars, the earth with its
sunshine and storm, light and darkness, stand out like great
mountain peaks demanding explanation. So in the life of every
human being, repeating the history of his race, as the evolution-.
ists are so fond of saying, the fundamental questions are first. to
obtrude themselves upon the growing intelligence. There is no
waiting, no delay for trifling with the simpler problems, the most
fundamental and most comprehensive come immediately to the
fore and alone seem worthy of consideration. But as age ad-
vances most men learn to ignore the fundamental questions and
to satisfy themselves with simpler and more secondary matters, as
if the great realities were all understood or non-existent. No
doubt to many a parent engaged in the affairs of society, politics,
finance, science or art, the questions that their children put,
like drawing aside a thick curtain, bring into view the funda-
mental questions, the great realities; and we know again that
what is absorbing the power and attention of our mature intel-
lect, what perhaps in pride we feel a mastery over, are only sec-
ondary matters after all, and to the great questions of our own

4 PROCEEDINGS OF THE

youth, repeated with such earnestness by our children, we must
confess with humility that we still have no certain answers. It
behooves us then, if the main questions of philosophy and science
cannot be answered at once, to attempt a more modest task, and
by studying the individual factors of the problem to hope ulti-
mately to put these together and thus gain some just compre-
hension of the entire problem.

This address is, therefore, to deal, not with life itself, but with
some of the processes or phenomena which accompany its mani-
festations.- But it is practically impossible to do fruitful work
according to the Baconian guide of piling observation on obser-
vation. This is very liable to be a dead mass, devoid of the
breath of life. It is a well-known fact that the author of the
Novum Organon, the key which Bacon supposed would serve as
the open-sesame of all difficulties and yield certain knowledge,
this potent key did not unlock many of the mysteries of science
for its inventor. Every truly scientific man since the world
began has recognized the necessity of accurate observation, and
no scientific principle has ever yet been discovered simply by
speculation ; but every one who has really unlocked any of the
mysteries of nature, has inspired, made alive his observations by
the imagination; he has, as Tyndall so well put it, made a
scientific use of the imagination and created for himself what is
known as the “ working hypothesis.” It must be confessed that
for some investigators the ‘‘ hypothesis” becomes so dear that
if the facts of nature do not conform to the hypothesis, ‘‘ so
much the worse for the facts.’’ But for the truly scientific man,
the hypothesis is destined solely to enable him to get the facts of
nature in some definite order, an order which shall make appar-
ent their connection with the great order and harmony which is
believed to be present in the universe.

If the working hypothesis fails in any essential particular he is
ready to modify or discard it. For the truly inspired investi-
gator, one undoubted fact weighs more in the balance than a thou-
sand theories.

At the very threshold of any working hypothesis for the Biol-

AMERICAN MICROSCOPICAL SOCIETY. 5

ogist, this question as to the nature of the energy we call life
must be considered. The great problem must receive some kind
of a hypothetical solution. What is its relation to the energies
of light, heat, electricity, chemism and the other forms discussed
by the physicist? Are its complex manifestations due only to
these, or does it have a character and individuality of its own?
If we accept the ordinarily received view of the evolution of our
solar system, the original fiery nebula in which heat reigned su-
preme, slowly dissipated part of its heat, and hurled into space
the planets, themselves flaming vapors, only the protons of the
solid planets. As the heat became further dissipated there ap-
peared in the cooling mass manifestations of chemical attraction,
compounds, at first gases, then liquids, and finally, on the cooling
planets, solids appeared. Lastly upon our own planet, the earth,
when the solid crust was formed and the temperature had fallen
below the boiling point of water, the seas were formed and then
Life appeared. Who could see, in the incandescert nebula, the
liquids and solids of our planet and the play upon them of chem-
ism, of light, heat, electricity, cohesion, tension, and the other
manifestations so familiar to all? And yet, who is there that for
a moment believes that aught of matter or energy was created in
the different stages of the evolution? They appeared or were
manifested just as soon as the conditions made it possible. So
it seems to me that the energy called Zzfe manifested itself upon
this planet when the conditions made it possible, and it will cease
to manifest itself just as soon as the conditions become suffi-
ciently unfavorable. It was the last of the forms of energy to ap-
pear upon this planet and it will be the first to disappear.

In brief, it seems to me that the present state of physical and
physiological knowledge warrants the assumption, the working
hypothesis, that Z2fe is a form of energy different from those con-
sidered in the domain of physics and chemistry. This form of
energy is the last to appear, last because more conditions were
necessary for its manifestations. It, like the other forms of energy, -
requires a material vehicle through which to act, but the results
produced by it are vastly more complex. Like the other ener-

6 PROCEEDINGS OF THE

gies of nature it does not act alone. It acts with the energies of
the physicist, but as the master; and under its influence the mani-
festations pass infinitely beyond the point where for the ordinary
energies of nature it is written ‘‘thus far and no farther.”

It can be stated without fear of refutation that every physiolog-
ical investigation shows with accumulating emphasis that the
manifestations of Zvzwg matter are not explicable with only the
forces of dead matter, and the more profound the knowledge of
the investigator the more certain is the testimony that the Life
energy is nota mere name. And, strange to say, the physicist
and the chemist are most emphatic in declaring that life is an
energy outside their domain.

The statements of a chemist, a physicist and a biologist are
added. From the character and attainments of these men, their
testimony, given after years of the most earnest investigation and
reflection, is worthy of consideration:—

When Liebig was asked if he believed that a leaf or a flower
could be formed or could grow by chemical forces, he answered:
“T would more readily believe that a book on chemistry or on
botany could grow out of dead matter by chemical processes.”’

“The influence of animal or vegetable life on matter is infin-
itely beyond the range of any scientific inquiry hitherto entered
on. Its power of directing the motions of moving particles, in
the demonstrated daily miracle of our human free will, and in the
growth of generation after generation of plants from a single seed,
are infinitely different from any possible result of the fortuitous
concourse of atoms; and the fortuitous concourse of atoms is the
sole foundation in philosophy on which can be founded the doc-
' trine that it is impossible to derive mechanical effect from heat
otherwise than by taking heat from a body at a higher tempera-
ture, converting at most a definite proportion of it into mechan-
ical effect, and giving out the whole residue to matter at a lower
~temperature.”—Sir William Thomson (Lord Kelvin).

“The anagenetic [vital] energy transforms the face of nature by
its power of assimilating and recompounding inorganic matter,
and by its capacity for multiplying its individuals. In spite of

AMERICAN MICROSCOPICAL SOCIETY.

~I

the mechanical destructibility of its physical basis (protoplasm)
and the ease with which its mechanisms are destroyed, it success-
fully resists, controls and remodels the catagenetic [physical anc
chemical] energies for its purpose.’”’—Cofe.

What, then, are the manifestations of the life energy ? and what
are the processes which are discernible? All of us in whatever
walk of life will recognize the saying of Gould :—‘‘ Now, when
one looks about him the plainest, largest fact he sees is that of
the distinction between living and lifeless things.”’

As life goes on and works with power where the unaided eye
fails to detect it, the microscope—marvelous product of the life
energy in the brain of man, shows some of these hidden proc-
esses. It has done for the infinitely little on the earth what the
telescope has done for the infinitely great in the sky.

Let us commence with the little and the simple. If a drop of
water from an aquarium, stream or pool is put under the micro-
scope many things appear. It is a little world that one looks
into, and like the greater one that meets our eye on the streets,
some things seem alive and some lifeless. As we look we shall
probably find, as in the great world, that the most showy is
liable in the end to be the least interesting. In the microscopic
world there will probably appear one or more small rounded
masses which are almost colorless. If one of these is watched,
lo! it moves, not by walking or swimming, but by streaming
itself in the direction. First a slender or blunt knob appears,
then into it all of the rest of the mass moves, and thus it has
changed its position. If the observation is continued this living
speck, which is called an amceba, will be seen to approach some
object and retreat, indeed, it comports itself as if sensitive, with
likes and dislikes. If any object suitable for food is met in its
wanderings the living substance flows around it, engulfs it and
dissolves the nutrient portions and turns them into its own living
substance ; the lifeless has been rendered alive. If the eye fol-
lows the speck of living matter, the marvels do not cease. After
it has grown to a certain size, as if by an invisible string, it con-

>?
stricts itself in the middle and finally cuts itself in two. The

8 PROCEEDINGS OF THE

original amoeba is no more, in its place there are two. Thus
nearly at the bottom of the scale of life are manifested all of the
fundamental features—the living substance moves itself, takes
nourishment, digests it and changes non-living into living sub-

, Stance and increases in size; it seems to feel and to avoid the dis-

, agreeable and choose the agreeable, and finally it performs the
miracle of reproducing its kind, of giving out its life and sub-
stance to form other beings, its offspring.

I 200 Vinsytlon
’

|

Choice

oe

Nutrition
, 4
4

_. Big 1. The Amoeba in its various phases of activity—Locomotion,
Ghoice (irritability), Nutrition and Reproduction. The figures should be
read from left to right, as with words in a book. (p.) Pseudopod ; (¢.)
Crystal of substance distasteful to the amoeba, hence the amoeba withdraws
from it ; (f.) Food ingested and digested by the amoeba for its nourishment.

AMERICAN MICROSCOPICAL SOCIETY. 9

The indigestible matter (w.) is extruded from the body and left behind.
(n.) Nucleus. This is seen to divide first in reproduction, then the division
of the cell body is completed, thus giving rise to two individuals.*

It is the belief of many biologists that the larger and complex
forms even up to man himself may be considered an aggregation
of structural elements originally more or less like the amceba just
described; but instead of each member of the colony, each indi-
vidual itself carrying on all the processes of life independently, as
with the amceba, there is a division of labor. Some move, some
digest, some feel, think and choose, some give rise to new beings,
all change lifeless matter into their own living substance,

The processes and phenomena by which a new individual is
produced are included under the comprehensive term, Améry-
ology.

All organisms great or small are but developments of minute
germs budded off by the parent or parents, and the way in which
these minute beginnings develop into perfect forms like their par-
ents can only be followed by the aid of a microscope. Indeed in
no field of biology has the microscope done such signal service
in revealing the processes of life.

The method of the production of a new being with the amceba,
as we have just seen, is for the parent to give itself entire to its
offspring—the parent ceasing to be in producing its offspring.
With some other lowly forms a part of the body of the parent buds
out, grows and finally falls off as an independent organism, or re-
mains connected with the parent to form a colony. In the vege-
table world a familiar example of a colony is represented by the
plant that the children call “Old Hen and Chickens.”

In the higher animals, however, where specialization has been
carried to its extreme limit, some myriads of cells forming the
body are set apart to produce motion, others digest food, still
others think and feel, while comparatively few, the germ cells
are destined for the continuation of the race. In the higher and
highest forms especially, all observation goes to show that the
life energy, not satisfied with the mere vitilization of matter and a

*The figures illustrating this address were drawn by Mrs. Gage.

10 PROCEEDINGS OF THE

dead level of excellence, is aiming at perpetual ascent, greater
mastery over matter and its physical forces. For the more cer-
tain attainment of this end, the production of offspring is no
longer possible for one individual ; two wholly separate individ-
uals must join, each contributing its share of the living matter
which is to develop into a new being. In this way the accumu-
lated acquirements of two are united with the consequent
increase in the tendencies and impulses for modification, and
nearly double the protection for the offspring. Thus, in striking
contrast with the amceba, where the single parent gives all of
itself to form offspring and in so doing disappears and loses its
identity, in the higher forms, while two must unite to form the
offspring, the parents remain and retain their individuality and
the ability to produce still other offspring. The process by
which this is accomplished may be traced step by step with the
microscope. A germ cell of the father and one of the mother fuse
together, and from this new procreative cell formed by the fusion
of two with all their possibilities combined, the new individual
arises. This certain knowledge is the result of the profound
investigation of the last few years and shows the literalness of the
scriptural statement, ‘‘ they shall be one flesh.”’

After this fusion of the father and mother germ cells, the
single cell thus formed, like the amoeba, divides into two, and
these into four and so on, but unlike the amceba all the cells
remain together. Within this celular mass, as if by an unseen
builder, the cells are deftly arranged in their place, some to form
brain, some heart, some the digestive tract, others for movement ;
so that finally from the simple mass of cells, originally so alike,
arises the complex organism, fish or bird, beast or man. How _
perfectly the word offspring describes the life process in the pro-
duction of this new being! That the child should resemble both
father and mother is thus made intelligible, for it is a part of
both. Yes, further, it may resemble grandfather or great grand-
‘father or mother, for truly it is a part of them, their life con-
served and continued. ‘There is no new life, it is only a continu-
ation of the old: ‘ Omne vidum ex vivo,’ all life from life. But

AMERICAN MICROSCOPICAL SOCIETY. II

S52 SS

Tone

: slain ay
gc ri I:

§ pnteron CORT IRETR EDGE Bip =

D> = |
Bs

J) o)
5 Bases sae

S2E eSsese Be oOSS,

Fig. 2. Various phases in the reproduction of one of the higher animals.
In the upper series is shown the fusion of the father and mother germ-
cells, in the middle series appear some of the earlier phases of segmenta-
tion of the fertilized ovum. The lower figure (modified from Marshall)
represents a medisection of an amphibian embryo sufficiently far advanced
to show that the original cells into which the ovum divided, have differ-
entiated and arranged themselves in such a manner as to form the
beginnings or protons of the great systems of organs—Brain, Enteron and
Heart.

O. Ovum, (n) Nucleus of the ovum. ¢ This sign indicates that the ovum
is a mother or female germ-cell. Z. Zotsperm. ¢ Sign indicating that
the zodsperm is a father or male germ-cell. (f. pn.) Female pronucleus.
(m. pn.) Male pronucleus. These two pronuclei fuse and form the nucleus
of the true reproductive cell, the fertilized ovum. In the two figures at
the right both signs (2 ¢) are used to indicate that both germ cells are rep-
resented in each figure. F. O. Fertilized Ovum. That is the true repro-
ductive cell composed of a father or male, and a mother or female germ-
cell fused. The steps of the fusion are shown in the upper series.

s.n. Segmentation nucleus. 2c. s. Two-cell stage, that is the fertilized

12 PROCEEDINGS OF THE

ovum has divided once forming two (compare the reproduction of the
amoeba). 4¢.s. Four-cell stage ; bl. blastula stage in which the fer need
ovum has divided into very many cells, all remaining together.

Embryo. The division of the ovum has proceeded very far, and the
cells have begun to differentiate and form organs. Brain, ch. Body axis
or notochord, Hnteron or alimentary canal and Heart.
the demonstration of this prime fact required a microscope, and
it is an achievement of the last half of this century. How
counter this statement still is to the common belief of mankind
we may perhaps better appreciate if we recall our own youth,
and remember with what absolute confidence we expected the
stray horse hairs we had collected and placed in water to turn
into living snakes.* The belief that it is an every-day occurrence
or living beings to arise from lifeless matter was not by any
means confined to those uneducated in biology. It was held by
many scientific men within the memory of most of us. Indeed
this goblin of spontaneous generation, even for the scientific world,
has been laid low so recently that the smoke of battle has
scarcely yet cleared from the horizon.

In the complex body of animals, as stated above, the constitu-
ent elements perform different functions. Is there any hint of the
way in which the action is accomplished? Let us glance at two
systems, the nervous and the glandular,widely different in struct-
ure and function. All know how constantly the glands are
called into requisition, the salivary glands for saliva, those of the
stomach and the pancreas for their digestive juices, etc. If we
take now the pancreas as an example, and that of a living, fast-

“Reference is here made to the nematoid worm Gordius. This worm
lives a part of its life as a parasite in the larvee of aquatic insects and in
some fish. In the adult free condition it differs markedly from the larval,
parasitic stage, and is very slender and much elongated, often reaching a
length of 20 to 80 centimeters (8to 10 inches), and has the general appearance
of a coarse hair like that from the tail of a horse. It lives in water and in
wet places and frequently appears in horse troughs and the wet places
where the trough overflows. From the hair-like appearance it was and
Still is believed that a hair from the horse’s tail or mane had directly trans-
formed into a living creature. By many persons itis called a hair-snake,
by others a hair-worm. Often one or several become tangled in an al-
most inextricable knot, whence the name from the famous “‘Gordian Knot.”

AMERICAN MICROSCOPICAL SOCIETY. 13

ing animal is put under the microscope so that its constituent
cells can be observed, it will be seen that they are clouded, their
outlines and that of their nuclei being vague and _ indistinct.
The cell is apparently full of coarse grains. If now the animal is
fed, as the digestion proceeds. the pancreas pours out its juice.
At the same time the granules, and with them the cloudiness,
gradually disappear, the cells become clear and both they and their
nuclei are sharply outlined. That is, the substance which is to
form the pancreatic juice is stored in the cells in the form of
granules during the periods of rest, and held until the digestive
agent is demanded, and if the demand is great all the granules
may be used up. But as soon as the demand ceases the cells
begin again their special vital action and again the granules begin
to appear and increase in number until finally the cells become so
full that they are fully charged and again ready to pour forth the
digestive fluid. This is a daily, almost an hourly process.

Let us take another example in which it would almost appear
that there is organic memory on the part of the gland cells.
No doubt all have seen the clear jelly-like masses surrounding
the eggs of frogs and salamanders. Whence comes this jelly
that is so resistent to the agents that work so quickly the de-
struction of ordinary organic matter? As spring advances the cells
of the oviduct increase enormously in size. The microscope
shows this increase to be due to a multitude of clear granules.
As the eggs move along, the ova are coated with the jelly formed
from the granules given out by the cells. As this material for
the jelly is poured out the cells gradually shrink to their original
size and then wait another twelve months before doing their des-
tined work.

If one can thus catch a glimpse of some of the finer processes
taking place in gland action, how is it with nervous action, the
highest function of which living matter is capable? While it
has been known for a long time that the nervous system is the
organ of thought and feeling and the director and co-ordinator of
the motions of the body, and many speculations had been made
concerning the processes through which the nervous tissue passes

14 PROCEEDINGS OF THE ;

Charged Discharging Discharged ¥%

Fig. 3. Sections of a gland in various phases of activity. The upper
series represents the gland as in longisection or lengthwise and the lower
series in transection or cut across. In the longisections, (lJ) in the right
hand figure represents the cavity or lumen of the gland into which the
secretion of the gland is poured. Thearrows at the top represent the direc-
tion taken by the secretion when it is poured out.

In the lower right hand figure, (1) represents the lumen, (n) the nucleus
of one of the cells, and (el. b.) the cell body of the same cell. The words
Charged, Discharging and Discharged designate the various phases of the
gland activity. The process of becoming recharged is not shown.

in performing its functions, it was left to an American student,
Dr. Hodge, to first successfully show that there were visible
changes through which the nervous system passes in its work.

AMERICAN MICROSCOPICAL SOCIETY. 15

The question is, can the activity of the nervous system be traced
as surely by changes occurring in the living matter forming its
basis, as the action of a gland can be seen by the study of the
gland cells ?

The demonstration is simple now that the method has been
shown. No doubt everyone has had the experience of failing to
perform some difficult muscular action at one time and then at
another of doing it with ease, or of finding true the reverse of
the adage, “ practice makes perfect.” For example, in a trial of
skill, as in learning to ride a bicycle, all the complicated action
may be performed with considerable ease and certainty at the
beginning of a lesson, when one is fresh, but as the practice con-

tinues the results become progressively less and less successful,
and finally with increasing weariness there is only failure, and one
must rest. We say the muscles are tired ; this is true in part,
but of much greater importance is the fatigue of the nervous
system, as this furnishes the impulses for the action and co-
ordination of the muscles. Now, as muscular action can be
seen and the amount can be carefully controlled, here was
an exact indicator of the time and amount of the nervous
activity. Furthermore, as animals have two similar sides, one
arm or leg may work and the other remain at rest, and con-
sequently corresponding sides of the nervous system may be
active and at rest. By means of electrical irritation one arm
of a cat or other animal was caused to move vigorously for
a considerable time, the other arm remaining at rest. Then the
two sides of the nervous system, that is, the pairs of nerves to
the arms with their ganglia and a segment of the myel (spinal
cord) were removed and treated with fixing agents, and ‘carried
through all the processes necessary to get thin sections capable
of accurate study with the microscope. Finally upon the same
glass slide are parts of the nervous system fatigued even to ex-
haustion, and corresponding parts of the same animal which had
been at rest. Certainly if the nervous substance shows the re-
sult or processes of its action the conditions are here perfect.
Fatigued nerve cells are side by side with those in a state of rest.

* 56 PROCEEDINGS OF THE

The appearances are clear and unmistakable; the nucleus
markedly decreased in size in the fatigued cells and posse
jagged, irregular outline in place of the smooth, rounded for
the resting cells. The cell substance is shrunken in size a
possesses clear scattered spaces, or a large clear spa rout

—Santel Bal

the nucleus.

Rest

Honey Bee

45 Rest Atos Fatigue
\ Fig. 4. Figures from Hodge (Jour. Morphology, Vol. VIL), showi
changes in the nerve cells of the spinal ganglia, in the cat and of the brs
in the honey-bee. The words Rest and Fatigue indicate the appearance
the cells in these two conditions. (n) Nucleus, (cl. b.), cell body,

clear space around the shrunken nucleus in the fatigued cells. °

AMERICAN MICROSCOPICAL SOCIETY. 17

If the nervous substance was not fixed at once but remained in
the living animal for 12 to 24 hours in a state of repose, the signs
of exhaustion disappeared and the two sides appeared alike. By
studying preparations made after various periods of repose all the
stages of recovery from exhaustion could be followed.

For possible changes in normal fatigue, sparrows, pigeons and
swallows and also honey-bees were used. For example, if two
sparrows or two honey-bees as nearly alike as possible were se-
lected, the nervous system of one being fixed in the morning after
the night’s rest and that of the other after a day of toil, the changes
in the cells of the brain of the honey-bee or sparrow and in the
spinal ganglia of the sparrow were as marked as in case of artifi-
cial fatigue. After prolonged rest, then, the nerve cells are, so to
speak, charged, they are full and ready for labor, but after a hard
day’s work they are discharged, shrunken and exhausted.

There is one more step in this brilliant investigation. If in the
morning, after sleep and rest, animals and men are full of vigor,
and in the evening are weary and exhausted, how like is it to the
beginning and end of life? In youth so overflowing with vigor
that to move, to act, is pleasure and continued rest a pain; but
in the evening of life a warm corner and repose are what we try
to furnish those whose work is done. How is this correlated in
the cells of the nervous system with the states of rest and fatigue ?
With a well-nourished child which died from one of the accidents
of birth the nerve cells showed all the characters of cells at rest
and fully charged. Ina man dying naturally of old age the cells
showed the shrunken nuclei and all the appearances of exhaust-
ing fatigue. In the one was the potentiality of a life of vigorous
action, the other showed, the fizal fatigue, the store of life energy
had been dissipated and there was no recovery possible.

For the animals that possess an undoubted nervous system,
probably all would admit that there is some sort of nervous action
corresponding to sensation; but what of living matter in the hum-
bler forms where no nervous system can be found? That these
have vital motion, that they breathe, nourish themselves, grow
and produce offspring none can deny. Do they have anything

18 ; PROCEEDINGS OF THE

comparable with sensation? As most of the lowest forms are
minute, the microscope comes to our aid again, and in watching
these lowliest living beings, it is found that they discriminate and
choose, going freely into some portions of their liquid world and
withdrawing from other portions. If some drug which is unusal
or we must believe disagreeable is added to a part of the water,
they withdraw from that part. It seems to have the same effect
as disagreeable odors on men and animals. On the other hand
there are substances which attract and into the water containing
these they enter with eagerness. Strange is it, too, that, as proved
by experiment, if an unattractive substance is used and also one
on the other side that has been found still more unattractive, the
less disagreeable is selected, the less of. the two evils is
chosen.

As man, the horse, dog and many other animals adapt them-
selves gradually to temperatures either very cold or very warm,
and that, too, by a change in their heat-regulating power rather
than by a change of hairy or other clothing, so these lowly or-
ganisms are found in nature in water at temperatures from near
freezing up to 60 or 80 degrees centigrade, a point approaching
that of boiling water. It may be answered that each was created
for its place, but by means of a microscope and a delicate ther-
mostat, to be certain of every step and to see all the results, Dr.
Dallinger, through a period of seven years, accustomed the same
unicellular organism and its progeny to variations of temperature
from 15 to 20 degrees centigrade, z. e., about the temperature of a
comfortable sitting-room, up to 70 C.. For those at the cooler
temperature it was death to increase rapidly the heat 1o degrees,
and for those at the higher temperature it was equally fatal to
lower it to the original temperature of 15 to 20 degrees. These
examples seem to show that it is one of the fundamental char-
acteristics of living substance, whether in complex or simple
forms, to adapt itself to its environment.

There is another fact in nature that the microscope has revealed
and that fills the contemplative mind with wonder and an aspira-
tion to see a little farther into the living substance and so per-

AMERICAN MICROSCOPICAL SOCIETY. 19

chance discover the hidden springs of action. This fact may be
called cellular altruism. In human society the philanthropist and -
soldier are ready at any time to sacrifice themselves for the race
or the nation. With the animals, the guards of the flock or
herd are equally ready to die in its defense.

So within each of the higher organisms the microscope has
shown a guarding host, the leucocytes or white-blood corpuscles.
The brilliant discoveries in the processes. of life with higher forms
have shown that not only is there a struggle for existence with
dead nature and against forms as large or larger than themselves,
but each organism is liable to be undermined by living forms,
animal and vegetable, infinitely smaller than themselves, insig-
nificant and insiduous, but deadly. Now to guard the body
against these living particles and the particles of dust that would
tend to clog the system there isa vast army of amceba-like cells,
the leucocytes, that go wherever the body is attacked and do
_battle. If the guards succeed the organism lives and flourishes,
otherwise it dies or becomes weakened and hampered. This
much was common scientific property three years ago, when one
of our members (Miss Edith J. Claypole) came to my laboratory
for advanced work. I discussed with her what has just been
given and told her that there still remained to be solved the
problem, what becomes of the clogging or deleterious ‘material
which the leucocytes have taken up? These body guards are, after
all,a part of the organism, and for them simply to engulf the
material would not rid the body entirely of it, and finally an in-
evitable clogging of the system would result. The problem is
simple and definite ; what becomes of the deleterious substances,
bacteria and dust. particles that get into the body and become
engulfed by the leucocytes? Fortunately, for the solution of
this problem, in our beautiful Cayuga Lake, there is an animal,
the Necturus, with external gills through which the blood cir-
culates for its purification. So thin and transparent is the cover-
ing tissue in these gills that one can see into the blood stream
almost as easily as if it were uncovered. Every solid constituent
of the blood, whether red corpuscle, white corpuscle, microbe or

20 PROCEEDINGS OF THE

particle of dust, can be seen almost as clearly as if mounted on
a microscopic slide.

Into the veins of this animal was injected some lampblack
mixed with water, a little gum arabic and ordinary salt, an
entirely non-poisonous mixture. Thousands of particles of car-
bon were thus introduced into the blood and could be seen circu-
lating with it through the transparent gills. True to their duty
the white corpuscles in a day or two engulfed the carbon par-
ticles, but for several days more the ieucocytes could be seen cir-
culating with the blood stream and carrying their load of coal
with them. Gradually the carbon-laden corpuscles disappeared
and only the ordinary carbon-free ones remained. Where had
the carbon been left? Had it been simply deposited somewhere
in the system? The tissues were fixed and serial sections made.
The natural pigment was bleached with hydrogen dioxid so that
if any carbon was present it would show unmistakably. With
the exception of the spleen no carbon appeared in the tissues, but
in many places the carbon-laden leucocytes were found. In
mucous cavities and on mucous surfaces and on the surface of the
skin were many of them; in the walls of organs were many more
apparently on their way to the surface with their load; that is
the carbon is actually carried out of the tissues upon the free sur-
faces of the skin and mucous membranes where, being outside of
the body, it could no more interfere in any way with it. But
what is the fate of the leucocytes that carry the lampblack
out of the tissues? They carry their load out and free the body,
but they themselves perish. They sacrifice themselves for the
rest of the body as surely as ever did soldier or philanthropist
for the betterment or the preservation of the state.

Thus I have tried to sketch in briefest outline some of the phe-
nomena or processes of life revealed by the microscope. Most
of those discussed have come under my own personal observa-
tion and are therefore to me particularly real and instructive.
But to every one long familiar with the microscope and with the
literature of biology many other examples will occur, some of
them even more striking. The discussion has been confined to

AMERICAN MICROSCOPICAL SOCIETY. 21

Gill Filament of Necturus

Leucocytes. Emigrating

Fig. 5. These figures represent various steps in the removal of foreign
matter from the blood of Necturus.

Gill Filament of Necturus. Part of a single gill filament greatly magni-
fied to show the blood vessels containing the red blood corpuscles (7. be.)
and the leucocytes (/.) or white-blood corpuscles. The black dots (c.) within
the blood vessels represent carbon particles which had been injected into
the veins. In many of the leucocytes are several carbon particles, there are
also several shown free in the blood plasma. (g. ¢.) The tissue of the gill
filament between the bluod vessels.

Leucocytes Emigrating. This, the lower figure, represents a section of
the skin with its covering epithelium (ep.) and the corium (cor.) or true
skin. The leucocytes containing carbon particles (c.) are seen in the corium
and penetrating the epithelium and finally free outside the epithelium.
The arrows indicate that the leucocytes emigrate from the body through
the corium and the epithelium, and finally into the space outside the
epithelium.

22 PROCEEDINGS OF THE

the above also because it seems to me to show with great clear-
ness the way in which we can justifiably hope to do fruitful work
in the future. This sure way, it seems to me, is the study of
structure and function together ; the function or activity serving
as a clue and stimulus to the investigator for finding the mechan-
ism through which function is manifested and thus give due sig-
nificance to structural details which, without the hint from the
function, might pass unnoticed.

This kind of microscopical study may be well designated as
Physiological Histology. t is in sharp contrast with ordi-
nary histology in which too often the investigator knows
nothing of the age, state of digestion or of fasting, nervous activ-
ity, rest or exhaustion. Indeed, in many cases it is a source of
congratulation if he knows even the name of the animal from
which the tissue is derived. Such haphazard observation has not
in the past, and is not likely in the future to lead to splendid
results. If structure, as I most firmly believe, is the material
expression of function and the sole purpose of the structure is to
form the vehicle of some physiological action, then the structure
can be truly understood only when studied in action or fixed and
studied in the various phases of action. :

Indeed if one looks only for form or morphology in the study
of histology the very pith and marrow is more than likely to be
lost.*

For example, if one wished to study the comparative histology

*Although in a different field, the words of Osborn in discussing the un-
known factors of evolution are so pertinent that they may well be quoted:
“My last word is, that we are entering the threshold of the Evolution
problem, instead of standing within the portals. The hardest task lies be-
fore us, not behind us. We are far from finally testing or dismissing these
old factors [of evolution], but the reaction from speculation upon them is
in itself a silent admission that we must reach out for some unknown
quantity. If such does exist there is little hope that we shall discover it
except by the most laborious research ; and while we may predict that con-
clusive evidence of its existence will be found in morphology, it is safe to
add that the fortunate discoverer will bea physiologist” [armed with a mi-

croscope|. I would like to add the last four words. S. H. G.. Am. Nat.,
May, 1895.

. Par
ares
—————————— PT

\

o
AMERICAN MICROSCOPICAL SOCIETY. 23

of the pancreas and were to take pieces from various animals to
be compared without regard to their condition of fasting or di-
gestion, he might find the coarser anatomical peculiarities in each.
In all probability he would also find two distinct structural types.
One type with clearly-defined cells and nuclei, the other with the
cells clouded, filled with granules and with the outlines of cells
and their nuclei almost indiscernible. Between these there might
be various gradations in the different forms. And yet, from
what has been stated above it is plain that all these different
structural appearances represent phases of activity, and all might
have come from the self-same animal. In like manner, if certain
parts of the nervous system were to be studied comparatively and
the tissue taken from one animal after refreshing sleep and rest,
from another after exhausting labor, another in infancy and another
from an animal decrepit with years, the difference in general ap-
pearance and in structural details would be striking enough to
satisfy any morphologist that, as with the structure of the pan-
creatic cells, there were two or more distinct types; but the physiol-
ogical histologist would recognize at once that the differences so
much insisted upon represented different phases of activity, and,
as with the pancreatic cells, might be all represented in the same
animal at different times.

I would be far from saying that there are no structural differ-
ences in the different animals independent of any particular phase
of functional activity ; but if these only are sought and the others
neglected the physiological appearances will often obtrude and
confuse if they do not utterly confound.

I have, therefore, for the last 10 years urged my students, and
mean to go on advocating with all the earnestness of which I am
capable, that’in studying an organism or its tissues, the investi-
gator, to gain certain knowledge,must know all that it is possible
to learn concerning the age, health, state of nervous, muscular,
and digestive activity ; in fact, all that it is possible to find out
about the processes of life that are going on and have gone on
when the study is made.

There are some microscopic forms in which the entire study

24 PROCEEDINGS OF THE

can be made while the creature is alive. With the higher organ-
isms, also, some of the living elements, as the white-blood cor-
puscles and ciliated cells, can be studied, and their various actions
and structural changes observed for a considerable time.

The white-blood corpuscles or leucocytes resemble the amoeba
very closely in their actions and powers, as we have seen in dis-
cussing the way in which the body is freed from foreign particles.
The ciliated cells are among the most striking of all the constitu-
ent elements of the body. One end is fixed firmly to the tissues,
the sides are in contact with their fellow cells, but the other end
is free and bears great numbers of hair-like processes, the cilia,
which project freely into some cavity or upon some surface.
What histologist would be able for a moment to suggest the power
of these hair-like processes if he studied the dead cells alone?
Yet the moment these cells are studied alive under the micro-
scope it is seen that for the service of the body all the powers of
these cells are concentrated into one, that of motion, and all the mo-
tion is manifested by the little cilia. These sweep with almost
incredible rapidity in one direction and more slowly on their re-
turn, thus producing a current in the direction of most rapid mo-
tion. This motion with the resulting current ceases only with
life. Each individual cilium is weakness itself, but with their com-
bined action the untold millions covering the cells, in the air pass-
ages for example, make a strong current in the liquid covering
them. This current is from the interior of the lungs toward the
throat and carries along with it particles of dust inhaled into the
lungs. In this way the delicate breathing organs are swept clean
and left unincumbered for their work of receiving oxygen and get-
ting rid of carbon dioxide.

If now one puts under the microscope some cells from the small
intestine of almost any animal from the lamprey eel to man, the
cells appear almost identical with those just described. The end
projecting to the free surface of the intestine seems to have a sim-
ilar brush of fine hairs, with a clear line along their base. Ifa
striated and a dead ciliated cell are under the same microscope

ide by side it is almost impossible to distinguish them. Indeed

AMERICAN MICROSCOPICAL SOCIETY. 25

Ciliated Epithelium
yn
z fia

EPEC PSTN es

Absorbing saree

TE

w

i

it

PETE

Maga iper aval

ij

ae

Fig. 6. Figures showing the similarity in appearance of the absorbing
epithelium of the intestine and of a ciliated epithelium. The free ends of
the cells point upward toward the top of the page and the attached ends to-
ward the bottom of the page. (cilia). The minute hair-like processes project-
ing from the free end of the cells and constantly swinging rapidly in one
direction and returning less rapidly to the starting point. In this way a
current is made in the direction of the most rapid motion (indicated by the
arrow). At the base of the cilia is a clear plate or segment (c. s.)

In the absorbing epithelium the segment appearing like the cilia is called
the striated border or segment (st. b.) and rests on a clear segment (Cc. 8.)
comparable with that on which the cilia rest. In the absorbing epithelium
food particles (f.) are represented as passing through the cell from the free
end toward the base, as indicated by the arrow.

so difficult is it that those from the intestine have been described
as ciliated more than once. If both cells are living no one could
confuse them. The striated end of one is motionless, the lines or
cilia of the other are in constant motion. One serves for produc-

26 PROCEEDINGS OF THE

ing currents, always in the same direction, the other is for the pur-
pose of absorbing and passing into the tissues the products of di-
gestion. One is a moving the other an absorbing cell.

Most of the tissue elements of the higher forms cannot be thus
studied alive, however, and the best that can be done is to fix the
different phases of action, as by a series of instantaneous photo-
graphs, then with a kind of mental kinetoscope put these together
and try to comprehend the whole cycle. .

Fortunately for the histologist the incessant experimentation
of the last twenty-five years has brought to knowledge chemical
substances which do for the tissues the wonder that was ascribed
to the mythical Gorgon’s Head,—to kill instantly and to harden
into changeless permanence all that gazed upon it. So the
tissues may be fixed in any phase and then studied at length.
If then the investigator observes and keeps record of every point
that may have an influence on the struétural appearances,whether
shown by experience or suggested by insight, and this rec-
ord always accompanies the specimen, thus and thus only, it
seems to me, can he feel confident that he is liable to gain real
knowledge from the study, knowledge that represents actuality
and which will serve as the basis for a newer and more complete
unraveling of the intricacies of structure, an approximate insight
into the mechanism through which the -life energy manifests
itself,

And so, with all the light that physics and chemistry can give,
commencing with the simplest problems and being careful that
every factor that can influence the result is being duly considered,
the microscopist can go forwaid with enthusiasm and with hope,
not with the hope that the great central question can be answered
in one generation, perhaps not: in a thousand, but confident that
if each one adds his little to the certain knowledge of the world,
then in the fullness of time the knowledge of living substance and
the life processes will be so full and deep that what Life Js,
though unanswered, may cease to be the supreme question.

AMERICAN MICROSCOPICAL SOCIETY. . 27

BIBLIOGRAPHY.

The following are a few of the works used in the preparation

of the foregoing address :

For general discussions of the problems treated, the works of

Herbert Spencer and other philosophers may be consulted with

profit. For extended references, the Index Catalog of the
Library of the Surgeon General’s Office, the Index Medicus, the
Physiologisches Centralblatt and the Anatomischer Anzeiger will

put the reader on the track of most of the books and papers that
have appeared.

"7879. Bernard, Claude—Legons sur les phénoménes de la vie commune

"95.

So.

aux animaux et aux végétaux. ‘lwo volumes, Paris, (1878-79).
These volumes show, in the way only a master like Bernard could
show, the essential unity of the life processes in animals and plants.
Chittenden, R. H.—On digestive proteolysis, being the Cartwright
lectures for 1894. New Haven, 1895, p. 187. He makes very clear
that in absorption the vital activity of the epithelium is necessary,
and that it is not a mere matter of physical diffusion. See p. 116, etc.

. Claypole, Agnes M.—The enteron of the Cayuga Lake Lamprey.

Proc. Amer. Micr. Soc., Vol. XVI. (1894), pp. 125-164, eight plates.

Besides the structural changes in the physiological process of trans-
formation from the larval to the adult condition, the enteric
epithelium and its structural features in action are shown; also the
ciliated and striated border of the enteric epithelial cells.

. Claypole, Edith J.—An investigation of the blood of Necturus and

Cryptobranchus. Proc. Amer. Micr. Soc., Vol. XV. (1893), pp. 39-76,
six plates. This is the investigation referred to in discussing
cellular altruism, p. 17 of the address.

. Cope, E. D.—The Energy of Evolution. American Naturalist, Vol.

XXVIII. (1894), pp. 205-219. From this paper is taken the quotation
on p. 0 of the address.

Dallinger, W. H.—On a series of experiments made to determine
the thermal déath-point of known monad germs when the heat is
endured in a fluid. Journal of the Royal Microscopical Society, Vol.
III. (1880), pp. 1-16. See also his Presidential] Address published in
the same Journal, pp. 185-199 (1887).

This investigation was carried on for nearly seven years, and
organisms living normally. at a temperature of 15° to 20° centigrade
were enured to a temperature of 70°C. Dr. Dallinger pointed out
some of the physical appearances passed through by the organisms
in their acclimatization. See also Davenport and Castle.

See p. 16 of the address.

i)
(72)

88.

"93

PROCEEDINGS OF THE

. Davenport, C. B. and Castle, W. E.—On the acclimatization of

organisms to high temperatures. Archiv fiir Entwickelungsmechanik
der Organismen. II. Band, 2 Heft. pp. 227-249. This paper gives |
in tabular form a history of the observations of various authors on
acclimatization of various living forms naturally, as in the waters of
hot springs, and artificially. Their own experiments on tad-poles
are highly suggestive and they point out some of the chemico-
physical changes occurring in the adaptation of the living substance
to the unusual environments. See also Dallinger.

Foster, M.—Text book of physiology. (New York and London, 1895).
In this work there is stated very clearly what is known and what is
not known concerning the processes of life.

Gage, Simon H.—The limitations and value of histological investi-
gation. Proceedings Amer. Assoc. Adv. Sci., Vol. XXXIV. (1885),
pp. 345-349. In this paper is pointed out the necessity of study-
ing function as well as structure in histological investigations if any
thing like a complete understanding of a tissue or organ is obtained.

. Gould, George M.—The meaning and method of life. 297 pages,

New York (1893). This is a most stimulating and inspiring work.
The quotation on p. 5 of the address is from it.

Hertwig, Oscar.—The cell, outlines of general anatomy and physi-
ology, translated by M. and edited by H. J. Campbell. P. 368, 168
illustrations. (London and New York, 1895) Dr. Hertwig lays
special stress on the function of the structural elements.

Hodge, C. F.—A microscopical study of changes due to functional
activity in nerve cells. Journal of Morphology, Vol. VII. (1892), pp.
45-168. Two plates. In this paper and the next are given the facts
on which the statements concerning the changes in nerve cells
mentioned in this address are based. There is also in this an
excellent resumé of what is known of structural appearances due to
vital activity in gland cells.

Hodge, C. F.—Changes in ganglion cells from birth to senile death.
Observations on man and the honey bee. Journal of Physiology,
Vol. XVIL., pp. 129-134, one plate.

Howell, W. H.—The Physiology of Secretion. The Reference Hand-
Book of the Medical Sciences, (N. Y., 1888), pp. 363-379.

In this article Dr. Howell gives a very admirable account of
secretion; and bearing upon the dissimilarity of living and lifeless
things says that something more than simple pbysical law is neces-
sary to explain the differences.

Kingsbury, B. F.—The histological structure of the enteron of
Necturus maculatus. Proceedings of the American Microscopical
Society, Vol. XVI. (1894), pp. 21-64, eight plates.

In this paper the structural appearances accompanying activity in
the enteric epithelium are described and figured.

Langley, J. N.—On the histology and physiology of the pepsin form-
ing glands. Philos. Trans., pp. 663-711 (1881).

Metchnikoff, Elias.—Lectures on the comparative pathology of
inflammation delivered at the Pasteur Institute in 1891. Translated
from the French by F. A. and E. H. Starling. P. 218; three colored
plates and 65 figures in the text. (London, 1893).

‘My principal object in writing this book is to show the intimate
connection that exists between pathology and biology properly so
called Author's preface. For the purposes of the preceding address

AMERICAN MICROSCOPICAL SOCIETY. 29

the parts of the book showing the activities of unicellular organisms,
their attraction and repulsion by various agents and the action of
the leucocytes in ridding the body of hurtful or clogging matter are
of especial importance.

’95. Sedgwick, Wm. T. and Wilson E. B.—An introduction to general
biology, p. 231, 105 Figs. 2d edition (N. Y., 1895.)

This work emphasizes the physiological side of the organism, and
the first chapters discuss with clearnesss and force the characters of
living things.

’g2. Thomson, Sir Wm. (Lord Kelvin).—On the dissipation of energy.
Fortnightly Review, Vol. 57 (1892), pp. 313 to 321. In this paper
may be found the quotation given on p. 4 of the address and also
the statement of Liebig. For this see the foot note to the article of
Thomson, p. 317.

’85. Tait, P. G—Properties of matter with an appendix on hypotheses as
to the eg ueupulien of matter by Prof. Flint, D. D. (Edinburgh, 1885).
P. p. 320.

794.95. Thurston, R. H.—The animal as a machine and a prime
motor. (N. Y., 1894).

See also Science, April 5, 95, and Journal of the Franklin Institute,
January-March, 95. It is shown that the animal machine is the
most efficient of all known machines, and the sentiment is expressed
that a comprehension of the processes of life is of as much interest to
the engineer as to the physiologist.

’95. Whitman, C. O.—Evolution and epigenesis. In Biological lectures
delivered at the Marine Biological Laboratory at Wood’s Holl, in
1894. In the prefatory note is given a discussion relating to matter
and energy.

See also his articles in the Journal of Morphology, Vol. I., pp.
227-252; Vol. IL, pp. 27-49; Vol. VIII., pp. 639-658.

PROCEEDINGS

OF

The American Microscopical Society

MINUTES OF THE EIGHTEENTH ANNUAL MEETING,

HELD AT

Cornell University, Ithaca, N. Y., August 21st, 22d and 23d, 1895.

WEDNEsDAY, August 21, 1895.
The members assembled in McGraw Hall at to minutes before
10 o'clock ; about 60 persons present.
The meeting opened with the following address of welcome by
the Hon. D. F. Van Vleet, of Ithaca:

Mr. President, Ladies and Gentlemen of the Society :

When I was asked, a few days ago, to makea formal address of
welcome to you, I suggested to one of my good friends that,
while it was something not new to my line, it would be the first
time that I had submitted my humble remarks to 75 or too
microscopists. I assure you that I feel rather diffident about talking
to you; and yet I have words of welcome for QO which are of
the best which I can give you.

_ IT realize the fact that there is no body of scientists in the world
who are such close observers, who have done so much and are
to do so much for humanity at large, as your own society. In
looking back over the history of microscopic investigation, I am
amazed, upon a hasty examination, to see the great improvements
which have been made in the microscope. There is, I take it, as
much difference between the magnificent instruments which you
have to-day, and which you are using, and the old instruments of
20 or 30 years ago, as there is between the old-fashioned churn

3? PROCEEDINGS OF THE

of our grandmothers and the magnificent apparatus for separating
cream and butter from milk which you may see in an adjacent build-
ing. There is nothing manufactured, to my mind, where such
enormous strides have been made in development, as with the
microscope. I looked over hastily last evening one of your ear-
lier reports. I found there that a great deal of time was given to
the discussion of the proper forms of the microscope and its mech-
anism. Then out of curiosity I turned to your last report, and
I found there that you are discussing questions of the greatest
importance to the human race; so that I take it that you have
now, ladies and gentlemen, a perfect piece of mechanism; and
your usefulness as a society, it seemed to me—as a layman in-
tensely interested in the work which you are doing—your useful-
ness to humanity at large is but just beginning.

Already, if you wish to see an object lesson, let me
state to you that this little city where you are welcomed as
guests, is expending something like $500,000—just beginning
the work. And why ?—because you, ladies and gentlemen of
the microscope, have shown to the people that there is that in
sewage which is an enemy to the human race; and so all over
this country, not only in the little city of Ithaca, but every-
where, the results of your investigations are bearing fruit for the
bettering of mankind. :

Therefore, while your members are increasing, while the work
that you have done in the past has been of the greatest import-
ance, I predict that the next 10 years of your existence as a So-
ciety will bring about far greater results than ever before. I do
not believe that there is a body of men in the United States who
have it within their grasp to do more for humanity, to do more
for the world at large, than your association.

Therefore I hope and trust that your deliberations will con-
tinue year after year, that you will continue to do the good work
already started and which is as yet in its infancy.

Now, in behalf of the mayor of this, our beautiful city, it is my
function to welcome you. Ithaca is one of the most beautiful,
one of the proudest little cities on earth. It is proud of its hills

AMERICAN MICROSCOPICAL SOCIETY. 33

and its valleys, proud of its lake, its streams, its waterfalls, proud
of its university, and proud to be the host of guests like you.
In behalf of the mayor, therefore, of the city of Ithaca, I extend
to you a most cordial welcome. We welcome every year here
hundreds of guests ; we were never so proud to welcome a body
of people as we are to welcome you, the leading scientists of
America, who have seen fit to honor us and the great university
which we love, by your presence.

In behalf of the 3,000 alumni who have graduated from the
halls of this university, and who love it as they love nothing else
on earth except their families, in behalf of that body of alumni I
welcome you. There have gone out from these halls men well
equipped to grapple with all the problems of this life. Some of
them are of your number ; and the alumni of Cornell University,
young though they are in years, stand before you, representative
scientists, as men who have taken the front rank. Cornell Uni-
versity has graduateda Jordan, a Comstock, a Gage, your honored
President ; and I might stand here an hour and tell you, ladies
and gentlemen, what the alumni of this university have done.
Therefore, in behalf of them, and I am sure every single one of
them would like to be present here and greet you and take you
by the hand—in behalf of them, as their present executive
officer, I greet you and welcome you, and say to you that we
desire, when you leave this, our alma mater, that you will carry
with you the idea that Cornell University is well equipped to do
the great work that it is doing, and that it is doing much to further
the interests represented by the society to which you belong.

Then, too, in behalf of the President of the university, who is
absent and who has delegated to me the task of welcoming you ;
in behalf of the university I would state to you that the univer-
sity is open to you, every department of this great university
greets and welcomes you. I doubt—of course I am prejudiced,
but I doubt if anywhere throughout the length and breadth of
this great land there is such another institution of learning as
this institution whose guests you are. Each of you should con-
sider yourself a guest of this university. You are here once

34 PROCEEDINGS OF THE

out of 18 years—I think this is your 18th annual meeting. I
am sure that I voice the sentiment of Cornell University when I
say to you that it would be the greatest pride to the university
if every meeting of your association might be here. I am sure
I voice the thought of the President of the university and of its
professors and of its alumni when I say that if this association
could meet here every year, and make here every summer a
creat department of a great university, it would further the ends
for which you are working, that it would be of greatest advantage
to you, that it would be of greatest honor to Cornell University.

In behalf of the university, therefore, in behalf of every citizen
of this little city, whether great or small, I desire to welcome you,
to say to you that our latch-strings are all hanging out, that we
desire among other things most of all to greet you, to meet you,
to become friends, to know you, to feel that we are honored in
doing so. Your time here is limited. We regret it. We wish
that instead of three days your stay could be extended to three
weeks. We wish that all the beauty of this most beautiful coun-
try, that all the advantages of this, one of the greatest universities,
all of its departments, everything which it has stored here in its
laboratories and in its museums, might be yours to inspect and
enjoy for weeks, aye, months.

Again expressing to you for every citizen, for every alumnus,
for every person connected with Cornell University, a most hearty
welcome, I venture to say that it is our belief that when you leave
us to go away you will come back to us again, and you can not
come back too often, you can not stay too long.

The President, Professor S. H. Gage, then made the following
' response to the address of welcome.

Mr. Van Vleet:

In behalf of this society I want to thank you most cordially for
this welcome. Last year, owing to circumstances over which we
had no control, one of the accidents that it is impossible to fore-
see, our meeting was not as successful as in some years. There
was a feeling on the part of the American Association for the Ad-
vancement of Science, that great association in this country which

AMERICAN MICROSCOPICAL SOCIETY. 35

covers all knowledge, that it was time that it carried its benefits
out on the Pacific Slope. You know, sir, that we have sent some
of our good men out there to carry the good tidings ; they have
been welcomed, and the whole association was going there.
This society was going asa feeble part of it. Owing to hard
times and various other things, that desire was not consum-
mated, and the American Association is now to meet in Spring-
field. The question then arose, where shall this society meet ?
It seemed tome, sir, that as so many members of the society whom
I had met pleasantly had asked about the university, that they
would like to come here to it; and I wanted them to come here
to see what our laboratories are like, to see our beautiful campus
-and city. I went to the president of the university and asked
him concerning it, and I was surprised at the heartiness with
which he acceded to every suggestion and the earnestness with
which he said that of all things which could come to the univer-
sity such meetings would give him the greatest pleasure. I
went then from him with his assurance as head of the university,
to the various professors, and everywhere was the same cordiality,
the same readiness to do everything that was possible to help in
making the meeting a success and in greeting its members. Then
some of us went down town; and with some hesitation, perhaps
you remember, I went to you, and asked you to be a member of
the local committee.

At your hearty consent to help us I felt encouraged. The
chairman of the local committee, Dr. Rowlee, of whom I can not
speak too highly for the efforts he has made, went to other
men in the town. The same cordiality was met everywhere ;
and so instead of there being town and gown in this welcoming
of us, there is only one, either all town or all gown, whichever
you wish to call it.

Now, then, to come back to the serious question, that is,
whether this society, whether we are worthy of this confidence,
this cordiality. In the past, as you have said in your remarks,
there has been work done in the society which I think has gone
for the advancement of knowledge. The question is, the main

306 PROCEEDINGS OF THE

question for us as a society is, whether in the work which we are
still to do are we going to add to human knowledge? If it
does add to human knowledge, and therefore to the possibility
of human happiness, it will add also to the security and honor
of the nation. I feel sure, sir, that I voice the sentiment of every
member of this society in saying that we will do everything we
can to be worthy of your confidence, of your cordiality. And
thanking you again for this greeting to our society, I now
declare the meeting open for the business of the day.

The names of a number of new members were then read as
recommended by the executive committee, and by resolution the
secretary cast the ballot of the society for them and they were
declared elected. They will be found in the list of members at
the end of the volume.

Mr. Pflaum, of Pittsburg, then read a paper on “‘ Alleged Me-
teoric Dust.”’

After the reading Prof. Kellicott said: I would like to inquire
the relative size of the particles found in Calcutta and at Pittsburg.

Mr. Pflaum: The size given for those there was from z75¢ of
an inch to gs}; ofan inch. But I found them far smaller. The
very smallest were but sa¢¢ of an inch. I do not believe that the
slide I have here gives the smallest, because I tried to seperate as
closely as possible the dross and dust from the shot itself. Of
course there were larger ones too. But the size seems immate-
rial. Under what conditions these grains are found I do not
know. But I imagine this: The iron and the silicious matter
in the iron becomes liquefied by the heat. When it rises from
the chimney it is cooled a little and under pressure from all
directions takes on a spherical form.

Mr. Seaman: There is only one question regarding this paper,
which seems to be of peculiar interest, and that is the way these
particles may be transported, not necessarily from the Pittsburg
blast furnaces, but even from those somewhere nearer by. Now
some of you may remember a few years ago there was an erup-
tion in the Indian Ocean that is known by the name of the erup-
tion of Krakatoa. I think very few in this country have any

AMERICAN MICROSCOPICAL SOCIETY. 37

idea of the tremendous character of that eruption, which threw
such volumes of dust into the atmosphere that it was supposed
fully one half of the surface of the earth had the sun somewhat
obscured and a yellowish haze in the atmosphere from the dust
with which the air was filled by that eruption. I am not pre-
pared to say that this was satisfactorily demonstrated, but it cer-
tainly is the best explanation which has been given of certain
atmospheric phenomena observed at that time. Of course that
eruption .was upon an enormous scale—a scale that has not been
paralleled since the destruction of Pompeii. Those of you who
wish to look into that matter will find in the Cosmopolitan for
April, 1895, a description of that scene by an eye-witness, which
certainly surpasses anything I know in literature since the
description of the destruction of Pompeii by Pliny. It will give
you an idea of the immensity of that phenomenon by which the
air was filled with this fine dust.

Mr. Hermann Schrenk then read a paper on the ‘ Corky Out-
growth of Roots, and their Connection with Respiration.”

Professor Gage said at the end of the paper: I imagine some
of you who have been listening to Mr. Schrenk’s paper have been
somewhat astonished by his speaking about plants using oxygen.
I was brought up on the pleasing theory that plants do not
breathe oxygen, but carbonic acid gas, and only animals oxygen.
Mr. Schrenk has been talking to you about organisms developed
by plants for the purpose of breathing oxygen. If you think for
a moment that the use of carbonic acid gas for a plant is simply
like our use of beaf steak, if you remember that plants, just as we
do, require oxygen to breathe, you will not be disturbed by
thinking that Mr. Schrenk, who used to be a student with us here,
has become unsound in his views by going to Harvard.

Professor Rowleé: I have been very much interested in the
paper by Mr. Schrenk. It will, be remembered by some members
of the society that I presented a paper upon this general topic
years ago, and at that time I looked up the literature relating
to the subject somewhat carefully. I did not then, nor till Mr.
Schrenk wrote me, anticipate that these structures would be

38 PROCEEDINGS OF THE

found in so many plants. There is a great deal yet, I think, to
be learned about the life histories of our marsh plants—it is one
of the most interesting fields for study.

The position that Mr. Schrenk takes with reference to these
organs, these peculiar developments, may be strengthened, I
think, by the general statement that these cells show gréat activ-
ity. They are cells that are highly charged with protoplasm—
not devoid of protoplasm as are some cells of the stem structure.
Now if this development were for floating purposes, as has been
urged, for the purpose of keeping the stem at the top of the
water, there would be certainly no occasion for unusual retention
of protoplasm and activity in it.

I may add, however, by way of contribution to the subject that,
being considerably interested in the matter of this development
of tissue, I thought that it might possibly be developed by
any plant, that any plant if submerged in a flood might develop
it. We frequently have floods in Ithaca, and when we do there
is an exellent opportunity to study the effects of submergence
upon the lowlands here. I think that it was perhaps two years
ago, after the vegetation was well started upon the alluvial bot-
tom toward the lake that there came a very heavy rain so that
the plants, many of them, were nearly submerged ; I presume the
water was nearly eight inches deep. I visited that region repeat-
edly with the special object of finding plants adapting themselves
to that submergence. The upland plants had started to grow in
normal terrestrial conditions, and they were submerged. To my
surprise nothing happened such as I had anticipated. In the
two weeks that the flood lasted those plants withstood it, appar-
ently with some injury, but without in any way adapting them-
selves to those aquatic conditions, as I expected they would.
It looks then as though certain plants had this method of stand-
ing submergence, and other plants had not. It is not possible
for many plants to respond.

Professor Rogers’ paper on ‘A Practical Method of Referring
Units of Length to the Wave Length of Sodium Light,” was then
read by Professor Moler.

AMERICAN MICROSCOPICAL SOCIETY. 39

Professor Gage: The question may arise to the minds of some
of you, What is the good of all this, anyway ? I remember that
back in 1882, at our meeting then in Elmira, Professor Rogers
was talking about metrology, the same subject that is being
discussed in this paper to-day. On that subject I believed most
thoroughly that the way to get the length of anything was to buy
a two-foot square in town at the hardware store, and measure it.
That was the same opinion as Professor Rogers has told me (1
know some of you have heard the story but some have not) was
held by a carpenter whom he wanted to level up a table. For
this purpose Professor Rogers gave him a good level—that is,
what Professor Rogers would call a good level. The carpenter
took it and tried to level up the table, but finally he gave it up,
declaring that the thing was no good, it was bobbing around all
the time. So he went home and gota 50-cent level, and had
not the slightest trouble in getting the table level. I think you
can see from this paper that if we are to get anything like abso-
lute measurements, not merely approximate ones, the best skill
of the most skillful men is necessary. We see from the paper
the difficulties that Professor Rogers had—after he got his brass
box, instead of its holding the air out it went in. All these diffi-
culties arise constantly in experiments where we reach for some-
thing absolute. Of course where it is simply an approximation,
a rough sort of approximation, there is no trouble; then we can
use the ordinary carpenter’s level or the two-foot measure that
we buy at the store. But Professor Rogers is trying to do some-
thing very different ; and he is, as many of us know, not exceeded
in the world as a metrologist ; he is recognized the world over,
in his determination of exact measurements as a master. It is
therefore with the greatest gratification that we have heard sucha
paper as that brought before us. To some of us perhaps there is
a good deal that is new and strange in this, but this is exactly, as
our worthy friend who welcomed us to the city and the univer-
sity has suggested, this is just what we are after—getting at things
that everybody does not know about, making additions to human
knowledge.

40 PROCEEDINGS OF THE

Mr. Seaman: I suppose that to many people who have not
thought upon the subject, the matter of weights and measures
appears extremely simple. They suppose every foot rule is ex-
actly like every other foot rule and every pound weight exactly
like every other pound weight. Now 1 think thata little thought
will show you how very far from a correct statement this would bein
the ordinary methods of business. If you go back in the history
of weights and measures you find that first of all people used as
standards, parts of the human body. We have relics of that yet 5
the hand with which we measure horses is the width of the human
hand; the unit of length known as a foot is derived originally
from the human foot. If you go back to classic days, the land
was measured by the number of hides of oxen that it took to
cover it, and we find traces of that in our language at the present
day. In England, in the time of Henry III., the grain, which we
now know as an apothecary’s measure, was actually a grain of
wheat ; and in order to establish some sort of uniformity they
attempted to prescribe by law that the grain should be taken
from the middle of the head of wheat, so that there might be a
uniform size, because the grains at the end and bottom of the
head were smaller than the other grains.*

Now it resulted from this attempt to furnish measures by natural
objects that weights and measures were of allsorts. If you take
a foot rule which you say is correct, and somebody else makes a
foot rule by it and that one goes on to some other part of the
country perhaps and somebody makes a foot rule by it, you will
find that the little-accumulated errors will soon amount to an
enormous variation from the standard. When the surveyor in
the city of New York attempted to compare and verify some of
the old surveys there, he found that it was impossible to do so,
often within five or six inches of the original lines. In New
York, where land is worth $250 a square foot, in many cases six
inches amounts to an enormous value. The reason was that the
old surveyors had never had any mode of verifying their chains.
The way it was actually done is described in one of the old

*See Remington’s Pharmacy, Art. Weights and Measures.

m P hee reas
SS —EEe eee

AMERICAN MICROSCOPICAL SOCIETY. 41

mathematical books, where the author prescribes that the surveyor
shall go to a church at the time when the congregation is coming
out, and call upon the first 16 people coming from the church to
stand together heel and toe, and he shall measure the length of
the line of feet for two rods; and thereby measured his chain.
You can see that under such conditions as that accuracy of
weights and measures was almost impossible. You could see in
the wall of an old building in the city of New York a few years
ago two iron staples or spikes driven in, in order that people
might fit their yard-sticks between them to verify their length.
Of course in a few years the spikes would become worn off so
that the yard-sticks would become longer all the time.

I suppose hardly any of you know that in this country to-
day there is but one, strictly speaking, legal system of weights
and measures, and that that system is not the one in customary
use. It is a very curious fact that the system of weights and
measures which we derive from England has never been legally
authorized by Congress. The only legislation upon the subject
of weights and measures which Congress has ever made consists
of two primary acts. One allowed the customs officers to use
the English weights and measures for the purpose of ascertaining
the customs to be paid to the government. The other act, which
was passed in 1866, established the metric system as the legal
system of weights and measures in this country. So we have
the curious anomaly that the great majority of the people in this
country are using a system of weights and measures which is,
strictly speaking, not according to law. Now in connection
with this I will state that when I first came to Washington 20
years ago and began to attend the meetings of scientific societies
there, I rarely heard of anything but the English system of
weights and measures. To-day you rarely hear anything but
the metric system. Measurements of objects of natural history
are mostly given in the metric system, so that that system has
actually come into use by a large number of people in this country.

There is one other thought that I may mention—I am simply
suggesting a few leading ideas. I stated that if you make a

42 PROCEEDINGS OF THE

measure and then compare another measure with that, and from
the second make a third, etc., there is very soon introduced a
great variation between the measures last made and the original
one. There are but two standards of measure in the civilized
world. The one is the metre of the archives in Paris; the other
is the platinum yard made by order of Parliament in England, after
the burning of the Houses of Parliament. If we take every meas-
ure in the world and refer them to those two standards, they will
all be alike ; and that is how this possibity of accumulating errors
is avoided. It is in order to make accurate comparisons with
these standards that all this work of Professor Rogers has been
done ; and it has been done with a perfection that, as Professor
Gage has stated, has been equalled nowhere else in the world.

Prof. Hyatt then read his paper on “The Mouth Parts and
Ovipositor of Cicada septendecim.”

Mr. Seaman: One of the privileges of science is to dispel
fictitious fears among people—the dread of that which is really
harmless. In connection with Professor Hyatt’s statement re-
specting the non-poisonous character of the Cicada septendecim,
I am reminded of a very extended series of experiments carried
out by Dr. Marx, a member of the Biological Society at Wash-
ington, in relation to the poisonous character of spiders, which
are usually supposed to be dangerous creatures. He took a num-
ber of the spiders of the kinds which have been reported in the
newspapers as having killed people, or having caused serious
illness, or at least having produced disagreeable swellings ; and he’
commenced by causing them to bite mice, and from mice he went to
guinea pigs, and from guinea pigs he went to rabbits, and from
rabbits to himself and other people. He tried many hundreds
of experiments ; he traced up, moreover, the histories of many
of the newspaper statements, and he proved, beyond the shadow
of doubt, that there is no spider belonging to this country, and
perhaps in no part of the world—because he tried many of the
large bird spiders of Brazil and other southern and tropical speci-
mens—that there is no spider in the United States whose bite
may be considered in any respect dangerous to life, or even

AMERICAN MICROSCOPICAL SOCIETY. 43

capable of producing in the majority of cases more than the
swelling which arises from a common mosquito bite. There
have been some cases—I think only two or three that he was
able to run down—where illness, perhaps in one case death, was
supposed to be connected with the bite of a spider; but a care-
ful examination of the evidence shows that it was not, strictly
speaking, due to the bite of the spider at all, but to that kind of
inflammation and blood poisoning which might result from the
scratch of a pin. Not but what spiders do have some poison,
but it is so feeble and in such small quantity that it is absolutely
without danger to human life.

Dr. Kingsbury read his paper on “‘ The Lateral Line System of
Sense Organs in Amphibia.” é

Professor Gage: I suppose we have all wondered in our
youth when we caught fish what that streak was along the side
of the body ; and it has been the purpose of Dr. Kingsbury in
giving us this paper not merely to speak of that in fishes but in
the Amphibia. Ithaca,as you know, is peculiarly rich in Amphi-
bia. There is a large variety of forms by which to test the
theory, whether it is due simply to the water or to something
else that we have that very interesting series, which Dr. Kings-
bury has shown to us. He has been trying to get behind the
appearances and find the reality, the true significance of things
which on the face of them may not seem to have much
significance—exactly the kind of work for which the society
stands, it seems to me.

A paper on ‘“‘A Comparison of methods of determining Hemo-
globin,” by F. C. Busch and A. T. Kerr, Jr., was then read.

Professor Gage: I may begin the questioning by asking the
reader if he will tell us how these tests correspond, for instance,
to determining the number of blood corpuscles in the blood
of the patient. Suppose we have a normal number of blood
corpuscles—let us suppose 5,000,000 as the normal number.
If now in your tests you found the hemoglobin was too much
or too little, would there be a corresponding increase or decrease
of the blood corpuscles ?

Mr. Busch: It has been found by observations made by @

44 PROCEEDINGS OF THE

number of observers that the hemoglobin does not necessarily
correspond to the number of corpuscles. It has been found, for
instance, as regards specific gravity that it has some constant
relation to the hamoglobin, but does not depend at all upon the
number of blood corpuscles.

Professor Gage: With a smaller number of blood corpuscles
then you might have more hemoglobin ?

Mr. Busch: Yes, you might have more hemoglobin.

Professor Gage: I believe there is at least an exceedingly
marked relationship between the haemoglobin and the number of
blood corpuscles if the variation is exceedingly large. If the
change in the number of blood corpuscles were smaller, say three
or four hundred thousand, it might not be a very important
factor. Dr. Moore, you have had expericnce in determining the
amount of hemoglobin and of blood copuscles ; can you say a
few words to us on the subject ?

Dr. Moore: I think not. The work I have had experience
in was not concerned with hemoglobin. But the question I was
just thinking about was this: In cases of anaemia, where you
have a large loss of haemoglobin, and in the organs such as the
spleen, the liver and the kidneys a deposition of blood pigment,
it seems to me that there must be a decided variation in the
specific gravity of the blood, if, as is supposed, the specific gravity
depends largely upon the amount of hemoglobin. Now whether
this hamoglobin is free in the plasma or still confined in the cor-
puscles is a question I am not prepared to speak about specif-
ically; but on general principles it seems that where blood cor-
puscles are destroyed in cases of anemia, as I think has been
found in certain of the lower animals—that in these cases the
hamoglobin is deposited and the specific gravity must vary. I
think there must be a correspondence in the lower specific gravity
of the blood and the diminution of the red corpuscles. What
effect an increase of the white corpuscles would have on this re-
lation of hamoglobin I cannot say. But in those cases we have
to deal with the fact that there is an unusual, an enormous in-
crease in the leucocytes of various forms.

Professor Gage: We have here one who has made a great

,

AMERICAN MIVROSCOPICAL SOCIETY. 45

study of the blood by aid of the micro-spectroscope. I would
like to ask, Dr. White, if in your spectroscopic study you can
make any correlation between experiments like those on the
number of blood corpuscles and the indications that would be
given by the spectroscope.

Dr. White: Ido not think I can answer the question satis-
factorily. I have noticed, however, that in examining specimens
of blood some specimens have very little hemoglobin, while others
have a great deal. In some cases there is a very great difference
in the corpuscles.

Just in that connection I would like to say one word in the way
of asking a question, which I shall not pretend to answer myself.
I have noticed in examining the blood of the dead in post-mortem
examinations, that there are bodies, sometimes as small as red
corpuscles, sometimes twice as large, that are colored almost as
much as the red corpuscles. I have even seen these in regard to
the living as well as in regard to the dead, and I am free to say
I do not know what those bodies are.

Mr. Moody: It seems to me that while there is a definite re-
lation or a somewhat definite relation, between the hemoglobin
and specific gravity, there are other things which enter into the
specific gravity of the blood, that may cause as great errors as
those arising in the experiments.

Dr. Moore: There is another question I would like to ask in
connection with the specific gravity of the blood—the time of day
of the experiments, and in what relation the taking of the blood
stood to the taking of food by the patient? I should think if
observations were made at different periods of the day in the same
individual, there might be changes due to the different relation to
the time of taking nourishment.

Mr. Busch: Our observations as a rule were made about mid-
way between meals. At that time the blood is more likely to be
in a normal condition. Some researches have been made by Mr.
Jones, of Cambridge, showing that the specific gravity varied during
the day under different conditions, as for instance, according to
the amount of liquid taken in. I know that one German ob-

46 PROCEEDINGS OF THE

server says that upon taking one and one-half mugs of beer the
specific gravity was diminished considerably.

Professor Gage: Have your investigations gone far enough
to find out whether this is really a practically useful clinical
method? I suppose that is a question we want to face and
answer. 1

Mr. Busch: I will not say absolutely just at this time, because
our observations are not completed yet; but there is no doubt
that this specific gravity method also is liable to considerable error.
We think that the other methods are liable to as much and perhaps
more error. But there is an advantage in having a method which:
is not a color test. People’s eyes vary so considerably. The
idea is this—that if we can get a method in which the error is
constant, then from day to day we can use it upon the same
patient and observe his improvement.

Dr. Moore closing the discussion: I would like to say just a
word or two in connection with this change in the blood after
food. In my experience when we have taken the blood of dogs
after death, if they were killed soon after they had been fed with
milk or with a light sort of mush, we found as a result that the
serum would contain such an enormous number of leucocytes,
that when the serum was set the surface would be completely
covered with a substance almost resembling cream in appearance—
in structure if not in color, for it is white—a substance that
really makes it useless for work. It seems to me that in taking
blood in such large quantities as that you would have great
variations. I should think that in determining the practical value
of these investigations, they would have to be taken in connection
with observation of the condition of the corpuscles, both red
and white. ;

Professor C. H. Eigenmann them made a few remarks prelimi-
nary to his paper on “ The history of the sex-cells from the time
of segregation to sexual differentiation in Cymtogaster,” and
at the conclusion of the paper the society adjourned.

Wednesday afternoon, August 21, about two o'clock, the
society assembled in the Physical Laboratory, at Franklin Hall,

AMERICAN MICROSCOPICAL SOCIETY. 47

and examined the ruling engine and comparator made by Wm.
A. Rogers. From the laboratory we went to the Entomological
department and inspected the excellent arrangement of insects
made by Professor Comstock, and also the beautiful engravings
made by Mrs. Comstock. Then the library was visited and
those who wished ascended the tower to enjoy the view of the
lake and country, while others examined the rare old books
Professor Mr. Burr exhibited in the White Historical Library.
The beginning of Cornell University was the Morrill land
grant of Congress to each State in 1862. After various struggles
in the State legislature over the disposition of the grant to New
York, during which Andrew D. White, of Syracuse, and Ezra
Cornell, of Ithaca, became closely associated, on October 7,
1868, the university was formally opened. The distinguishing
features of Cornell as a University are that it puts all truth on
a level, and gives to the scientific or technical student the same
standing as the student of literature and the classics. Hence the
spirit of the whole institution is preéminently American and scien-
tific as distinguished from those institutions where the dominating
influences have been those of tradition and caste. Hence the
young man who goes te Cornell receives a training that fits him
for the real life of the present age in which he is to play a part.
This year the university expects to welcome to her halls 2,000
students, including the women who find here precisely the same
opportunities as the men, and how well they make use of them
the pages of these proceedings bear witness. The library, while
it is one of the most recent, is undoubtedly one of the most
beautiful and interesting buildings on the grounds. Many of
the members wandered off to the other buildings near by, in-
cluding Sage college, the dormitory for women, adjoining which is
the Botanical department, and also to the Young Men’s Christian
Association building and the chapel, both near by. Attached to
the latter is a memorial chapel in which lie the bodies of Ezra
Cornell and Mrs. Andrew D. White, whose forms sculptured
in marble, rest on their marble couches as if they, too, were
an imperishable part of the institution which is for one of

48 PROCEEDINGS OF THE

them one of the proudest monuments ever reared for any man.
In connection with this visit to the collections of the University
the following letter from Dr. Wilder is appropriate for insertion
here:
THE UNIVERSITY MUSEUM OF VERTEBRATES.
To Dr. W. H. Seaman, Secretary of the American Microscopical Society :

Dear Sir— I regret my absence from Ithaca during the coming session
of the society upon both personal and official grounds. It would give me
pleasure to attend the sessions and meet the members.

As Curator of the Vertebrate Division of the Natural History collection
of Cornell University, I should invite inspection of it. In so doing I should
endeavor to state four points as follows: 1. The objects of the vertebrate
museum. 2. The principles upon which it has been formed and main-
tained. 38. Its special merits. 4. Its chief deficiencies.

1. The Museum is primarily and mainly educational. That is, it is
intended to illustrate the instruction of the classes in Physiology, Verte-
brate Zoology, Neurology, and Embryology. At each lecture or practicum
from 10 to 50 specimens are brought into the class room, and not only
exhibited and explained, but left there for one or more days for fuller
examination.

In accordance, however, with the doctrine that ‘‘none are so well fitted
to impart knowledge as those who are engaged in reviewing its methods
and extending its boundaries,” the collections have been made the basis of
numerous researches and publications by the staff of the anatomical depart-
ment, and by specialists elsewhere to whom specimens have been loaned.
There are also in rooms not open to the public considerable stores of mate-
rial for investigation or for preparation as museum specimens.

On the other hand the wants of the public have not been ignored. The
only division of the collection at present labeled with approximate explicit-
ness comprises the animals of Tompkin county, whose names, life-histories
and habits are of special interest to the farmers and fishermen of this
vicinity. From the beginning the museum has been freely accessible during
the whole of all week-days. The janitor is often at work there or within
call, and when funds have been available for the purpose a student-guide
has been in attendance on Saturdays and at commencement. The museum
is visited by schools from out of town and by excursion parties numbering
hundreds at a time. So far as lam aware no officer or employe has ever
received from visitors any pecuniary recompense for attendance upon
them, for the explanation of specimens or for the exhibition of living
animals.

2. The fundamental principle of the museum is the illustration of
important facts and ideas by means of specimens carefully selected and
well prepared. The wall-cases at the north end of the main floor contain
a synopsis of the vertebrate branch, and of the larger divisions commonly
regarded as classes. As an example may be taken the two cases at the east

AMERICAN MICROSCOPICAL SOCIETY. 49

end of the series. Here are about 15 mammals fairly representing the
range of difference in respect to form and mode of locomotion. Also some
dissections displaying the structural features which are both constant
throughout the class and peculiar thereto, e. g., the complete diaphragm,
the left aortic arch, etc.

In a case at the west side of the same floor may be seen a series of a very
different kind, viz., of the venomous vertebrates so for as obtainable.

Otherseries begun or contemplated are enumerated in my address on ‘‘Edu-
cational Museums of Vertebrates” in the American Association Proceedings
for 1885: Abstract in Science, Vol. VI., 1885, pp, 222-24.

The most significant of these relate to the exposition of the general
doctrine of evolution.

In the formation of these series mere number has not been considered,
nor has the cost or the rarity of a specimen determined its real value.
So far from taking all that is offered merely because it is cheap or alto-
gether without cost, I have held that a wise economy would be practiced
by paying for what was really needed, rather than in accepting less desir-
able objects as gifts. In an educational museum of vertebrates one flying
squirrel is more desirable than a dozen other kinds. It would be wiser to
pay ten dollars each for a Sphenodon, a Protopterus and an Apteryx than
to receive as gifts a hundred other lizards, fishes or birds. Indeed, con-
trary to the prevailing idea that the curator of a museum is mainly a
collector, I hold that one of his chief duties is to keep things out of it.

3. Among the desirable features of the museum may be enumerated the
following:

a. The large number of embryos, brains and hearts of all classes.

b. The numerous well-preserved human cerebrums especially of edu-
cated persons.

c. Thedissected preparations illustrating zoologic or physiologic facts and
ideas. Some of the best of these were made by your distinguished Presi-
dent.

d. The association of such dissections of soft parts in the same case with
the skins and skeletons of the same or allied forms.

‘e. The preservation of so many parts of one and the same individual.
For example, of a kangaroo there are the stuffed skin, the mounted skeleton,
the alinjected heart and brain, and some other viscera. This not only
exhibits correlations of structure but also in case of the detection of ane
error in the identification of the skin, renders it possible to extend the
rectification to all the other parts.

f. The designation of all parts of a single individual by one and the
same number, and the use of the same number upon all notes, photographs
and drawings relating thereto.

4. The most obvious defect of the collection is suggested by the last
paragraph; few of the specimens bear labels conveying adequate informa-
tion tv the visitor. But it must be remembered that the collection is
primarily for use in instruction and that although the tags may not be read
easily from without the case, they can be when the specimens are in the

50 PROCEEDINGS OF THE

lecture room. Also, the number refers to an Accession Book and to notes
that are accessible. Finally, among the reasons for the delay in labeling is
the intention to supplement the conventional label by a somewhat full
description and, in the case of anatomical preparations, by photographs,
or diagrams or published figures. The attainment of this “triple alliance
of object, drawing and description” requires much time and deliberation.

That many and serious gaps in the various seriesremain to be filled hardly
needs admission. For these desiderata we must await opportunities, the
thoughtfulness of our friends and the power of the trustees to supply the
means of purchase and preparation. .

The defects in the arrangement of the collection are due to three condi-
tions:

a. The present necessity of accommodating upon the main floor certain
ethnologic and archzologic collections which are incongruous and interrupt
the natural series.

b. The limitations of space in the alcove cases and in those at the north
end of theroom. The malposition most to be deprecated is the association of
the Dipnoans with the Ganoids, which constitutes in my mind, rank
zoologic heresy. ‘At present it is hardly to be avoided. ;

ec. A recent interchange of cases with another department necessitated
the transfer and storage of a considerable portion of the vertebrate series,
and the rearrangement has not yet been effected. Hence some unoccupied
spaces and some masses of undisplayed specimens. Should the American
Microscopical Society again honor Ithaca by meeting there I trust the verte-
brate division of the zoological collections may better represent my views
and intentions regarding it.

Very respectfully yours,
BURT G. WILDER.
Siasconset, Nantucket Island, Aug. 17, 95.

Wednesday evening, about eight o’clock the society gathered
in the Botanical lecture room to listen to the annual address
of the President, Professor Simon H. Gage, on The Processes
of Life Revealed by the Microscope; a Plea for Physiological
Histology. The address begins this Volume.

Norg.—Those who wish to know more of Ithaca and Cornell University,
in addition to the catalogs of the institution, which can be had.on applica-
tion, will find a little pamphlet of Andrus and Church, ‘‘ In and Out of
Ithaca,” very useful.

AMERICAN MICROSCOPICAL SOCIETY. 51

TuHurspay MorninG, August 22.

At 9.40 the society assembled, about 70 persons being present.

Professor Rowlee read his paper on ‘ The Chlorophyll Bodies
of Chara Coronata.”’ aa

The President then appointed Dr. Kellicott and Mr. Kihne as
a committee to audit the accounts of the Treasurer.

The following names were elected after free nominations to
serve as a nominating committee to select officers for the ensu-
ing year :

Messrs. W. W. Rowlee, D. S. Kellicott, W. H. Walmsley, G.
S. Hopkins and Wm. C. Krauss.

Miss M. A. Nichols then read her paper on “‘ Secondary Thick-
enings of the Root Stalks of Spathyema.”’

Professor Rowlee said when the paper was ended: It is due in
justice to Miss Nichols to say that she began what she and I both
believed to be a comparatively simple problem, and intended to
keep it within very definite limits, and it was her intention when
she undertook it to present it to this society. The discoveries
that she made as soon as she began to investigate, however, led into
such different lines that the paper has in it the possibility of a very
important contribution to our knowledge of the monocotyledon-
ous group. It is known probably to every one that the flowering
plants are divided into two great groups, the monocotyledons and
the dicotyledons, or, as they have been called, the exogens and the
endogens. The endogens or monocotyledons were believed once
not to have any secondary thickening, or, as the term implies,
increase by internal growth. As Miss Nichols has said, DeBary
found exceptions—others had found exceptions before him, and he
summarized the exceptions ; but they are comparatively few. There
are always exceptions in natural history, you know, and it is
fortunate if the exceptions do not come to be the rule.
Neither of us had a notion that we should find secondary thicken-
ing when the work was begun. It was with a view to the seed,
which Miss Nichols has described to us, that the study was made.
Not only is the discovery of the secondary thickening new, but
likewise the method of thickening is new; so that I feel we have

52 PROCEEDINGS OF THE

here a contribution to our knowledge of plants that is very im-
portant and which will be altogether creditable to our proceedings.

Dr. Hopkins: I would like to ask Miss Nichols if she has tried
any experiment to see whether or not the crystals would be re-
formed after being dissolved out. If they would be, I should
think it could be determined whether or not the irritation to the
taste was caused by the crystals. —

Miss Nichols: I did experiment with that in view, but the
crystals would not re-form voluntarily, for I let the solutions stand
for some time and examined them, but they did not re-form.
Whether they could be made to do so or not I do not know.

Dr. Holbrooks’ paper ona “‘ Fourth Study of the Blood, show-
ing the Relation of the Colorless Corpuscle to the Strength of the

by

Constitution,’ was read by title only, and the society then listened _
to Mr. K. M. Wiegand on “ Two Cases of Intercellular Spaces in
Vegetable Embryos.” The paper was discussed as follows :

Dr. Seaman: I would like to ask Mr. Wiegand as to the
shape of the limiting cells on these canals, as regards their
length. Are they longer than other cells, or not?

Mr. Wiegand: No sir, their length is about the same as their
diameter.

Dr. Seaman: They are mere altered parenchyma cells?

Mr. Wiegand: Yes, altered a little in form. J might say
that the arrangement of these spaces and of the cells around
them remind one very much of the arrangement of cells around
resin ducts in mature plants. The resin ducts in the pine family
of course have a sheath of cells arranged in this way.

The paper of Dr. E. J. Durand, on the “ Fruits of the Order
of Umbelliferae”” was then read by title, also that of Dr. P. A.
Fish, “‘ The Action of Strong Currents of Electricity upon Nerv-
ous Tissue.”’

The society then listened to Mrs. Gage’s paper on “ The
Morphology of the Brain of the Soft-shelled Turtle and the
English Sparrow compared.”’

Discussion on Mrs. Gage’s paper.

Dr. Krauss: I would like to ask if Mrs. Gage has made a

AMERICAN MICROSCOPICAL SOCIETY. 53

study of the cells of the cortex, especially of the frontal lobe, or
that part of the brain of the turtle corresponding to the frontal —
lobe in man; and also what the development of those cells is, as
compared with the development of the cells in the parietal lobe
or the occipital lobe.

Mrs. Gage: To answer Dr. Krauss’s question, I can simply
say that thus far in my investigation I have confined my work to
the general features of the brain, not atall to the histology ; I do
not know that I shall ever get to that point, there are so many
problems in the way which must be met in order to homologize
the parts and to be sure that the origin of the parts is the same,
before a comparison can be made of the cells.

Professor Ward: I should like to ask, in the light of this inves-
tigation of the three series of brains, to what extent Mrs. Gage
thinks the homology by fiber tracts can be followed out—are
they characteristic of certain groups and homologous in the
groups?

Mrs. Gage: Thus far only a few of these fiber tracts have been
considered. If I could find the cell nidz and the origin of them,
and find that they are perfectly comparable, then I should feel
that the fiber tracts which connect those nide are homolo-
gous. Otherwise I should feel that, simply because they go in the
same general direction and follow the same general course, it is
not necessary that they are homologous.

Professor Ward: Another word. To what extent have you
followed this out and found homology ?

Mrs. Gage: There are comparatively few of these which I
have already done. The one which I have mentioned in connec-
tion with this investigation in these two forms, seems to be exactly
comparable. There are certain tracts, like the great anterior and
posterior bundles, about which there seems to be no doubt at all.
There are these to be traced in all the forms of vertebrates and
they seem to perform in general the same function and take the
some direction; but some of these others, I should say, are
much more difficult to trace. There are several others which
are apparently, so far, quite easily determined. Some of the

54 PROCEEDINGS OF THE

more difficult ones, as those in the base of the cerebrum, I should
say, would take a great deal of time to decide finally that they
were homologous throughout the vertebrate series. 1

Dr. Humphrey: I was struck with regard to what Mrs. Gage
says concerning the size of the optic nerves of these turtles, and
I can state that these animals have a very acute sight. When
lying upon the stones or logs along the course of our rivers, they
will slide off when a person is sometimes at a distance of three or
four hundred yards. But on the other hand I do not consider
that their sense of hearing is very acute. A few weeks ago I was
walking along the banks of one of our smaller rivers with a rifle.
There were two turtles sitting on a log some sixty yards away.
As I was behind some bushes they did not see me. I assisted
one of them off the log with the rifle, but the other did not notice
it at all, so I assisted him off in the same way. Often I have
noticed that they pay very little attention to the crack of a gun,
but they pay a great deal of attention to one’s motion, and I believe
sometimes that they notice the puff of smoke from a gun.

Mrs. Gage: How about the sense of smell or the sense of
touch ?

Dr. Humphrey: I do not know about that, I presume though,
that it is very acute. I should infer that the sense of smell was
acute from the fact that they live mostly on dead fish. I may say
in regard to the habits of these, and of the large snapping turtles,
those whose heads are perhaps three inches across, very large
ones, I have found that their stomachs were filled with fresh-water
algze ; while, as far as I have been able to discover, the soft-shelled
turtles never take anything but flesh.

If Mrs. Gage will permit me I will say that I think a walk
along a stream would cause her to change one expression. She
said this animal led a sort of dreamy existence. It does in the
winter time, but in the summer time I can see no analogy to a
dreamy existence, unless because he is a nightmare to the fish-
erman.

Mrs. Gage: With reference to two points of which Mr. Hum-
phrey spoke: In observing a shoft-shelled turtle it is very evident

|

AMERICAN MICROSCOPICAL SOCIETY. 5

Un

that he keeps a very watchful eye upon you, but the only indica-
tion of motion whatever that he makes will be in the turning of
the eyeball. You know he is looking at you from that one fact.
There is no indication of nervousness on his part in any other
way.

As to the sense of hearing I have observed in a jar, where I
have studied them, that any loud sound near the jar will not pro-
duce any motion whatever in the turtle; wnile on shaking the jar
even in the slightest degree they will move. I conclude from
this that the sense of feeling was much more observable in this
turtle than the sense of hearing.

Professor Eigenmann: I may adda word as to the habits of
these turtles. We have had quite a number in the laboratory
this summer. They have a habit of burrowing under the sand
just sufficiently to cover themselves so that they cannot be seen.
One which we were trying to photograph, when we went to get
the plate, I think only across the table, was nicely covered when
we got back so that we could hardly see it. One very large
one that we got in a pen was missed after a few days. I thought
he was literally in the soup: But he afterwards reappeared.
These turtles lay their eggs in the sand. They lay quite a large
number of eggs in a nest—the largest number that we secured this
summer was thirty-two. The eggs are quite large—like good-sized
marbles. They are white, but the upper surface of the egg is
much whiter than the lower surface. I secured quite a number
of their little ones, but have not so far been able to get any eggs
to hatch. We still have a batch of eggs, however, and hope to
rear some.

Mrs. Gage: Where do they lay their eggs; how near the
water ?

Professor Eigenmann: They lay the eggs within a few fect
of the water’s edge. Those we secured were within, I should
say, ten feet of the edge of the water, right in the sand of one
of the lakes. The easiest way to get them is to take a hoe and
go toa likely place and just hoe over the whole of the region. You
may strike a nest and then by a little more careful hoeing you can

56 PROCEEDINGS OF THE

ro

get the eggs. We tried our fingers for quite a while, but found it
was wearing our nails and otherwise not very agreeable. A hoe
is the best thing

There is no sign on the surface. You can find marks where
the turtles have walked up, but on a beach a mile long the turtles
had walked up all along it so that the tracks do not help you
much. But once in a while you can find a particular track lead-
ing straight to a spot where there is a little depression in the sand,

a spade is not very good.

and there you can be pretty sure will bea nest. I may add
that right beside the nest of thirty-two we found another with
seventeen eggs. At any place where there was a point of land,
by using a hoe we could secure from 150 to 200 of the eggs in
a short time, perhaps hardly fifteen minutes.

Mrs. Gage: It seemed to me these turtles had the most self-
restraint of any animal I have ever watched. I sat by a tank
watching one for ten hours one day. I think Mr. Humphrey
would have been convinced that my conclusion about their
dreamy existence was correct. That turtle stayed at the bottom
of the tank for ten hours, and never once moved in that time.
He had filled up his lungs with oxygen and was breathing down
there by means of his gills situated in the throat.

Dr. V. A. Moore then read his paper on ‘‘ The Flagella of
Motile Bacteria.’ | 3

The President said: It is one of the sources of congratulation
for societies like this that we have problems presented to us for
investigation, and that as naturalists or investigators in science,
we have never to weep as Alexander did for new worlds to
conquer.

May I ask Dr. Moore if it is the supposition that these flagella
move, or have they been actually seen to move.

Dr. Moore: Some of the earlier observers have published
the statement that they have seen these flagella in the larger
forms, move in stagnant water. I will say that in examining a
water bacillus not long ago in a liquid culture, in the hanging-
drop preparation, as it is technically called, I saw these flagella,
a mass of them, rolling and twisting about, perhaps by means of

AMERICAN MICROSCOPICAL SOCIETY. 57

currents in the liquid; but I could not see them attached to the
bacilli themselves. But they have been observed and described.’
Dallinger and Drysdale watched a bacillus, as they called it at that
time, whatever it was I do not know, one of those organisms—for
ten or twelve hours, first one and then the other, until finally they
saw a curious vertical appearence of the liquid at the poles of the
organism ; they said they saw this fine hair-like projection move.
_ Those statements have been made, and I do not doubt that with
proper optical appliances they could be seen in the larger
forms.

Dr. White: I should like to make a statement in regard to
some of these bacteria that I think has not yet been published.
It may be of some interest on just this question of the motility.
Dr. Foote, of Yale, has prepared some cultures, both in plate
cultures and stab cultures in test tubes, and he asked me to
photograph them. Now these stab cultures in the test tubes in
glycerine after three days’ culture, the Cold communis appears as
a line or thread generally where the track of the wire was inserted ;
while a simple culture of the typhoid bacillus shows the culture
generally following the track of the needle, but also spread out
laterally so that it is funnel-shaped. At the top of the test tube
it will be very wide, perhaps half-an-inch wide, and tapering
down to the bottom. To my mind this different appearance is
one kind of proof that these bacilli move and travel in the
gelatin. Also in the plate cultures of the Coli communis, which is
generally considered to have very few flagella compared with the
typhoid bacillus—the colonies after three days’ culture in this
same material are clearly defined at the edges, sharply defined as
little dots; while the similiar cultures of the typhoid bacillus
have a distinct areola around them, about as wide as the colony
itself. If I had known that this paper was coming on this
morning I would have brought photographs to illustrate this
point.

Dr. Krauss: Just to show the importance of the flagella in
making a comparative examination of the bacilliabout a year and
a half ago we had an outbreak of typhoid fever in Buffalo. In

58 PROCEEDINGS OF THE

trying to detect the source we examined the water of different
localities. We suspected that the water commissioners were using
a source of water that had previously been used in Buffalo, but
not for the past few years. Now as to this old source of water
supply we know that certain parts of the city drain into it. It
was during a north-east storm when the water supply was rather
low that they used this old water supply. After a certain period
of incubation the typhoid fever broke out in all parts of the city. ,
We examined the bacilli in the old water supply and found
almost as much bacilli as water. In making comparative tests we
found that the typhoid bacillus was present and also other bacilli
resembling the typhoid so closely that it was almost impossible to
tell which was the typhoid and which was the other. It was only
‘on staining for flagella that we were able to state positively that
the typhoid was present and undoubtedly caused all the trouble.

Now as far as the function of the flagella is concerned, it occurs
to me, why cannot these flagella be weapons of defense for the
bacilli in their warfare against the phagocytes. We know that
almost all living bodies are supplied with weapons of defense of
some kind or other, and these may possibly be the creatures’ way of
fighting off the phagocytes as long as possible. The power of
resistance to disease in different persons is very different, as we
know, and certainly the resistance to antiseptic action of some of
these forms may be due perhaps to the flagella.

Professor H. B. Ward then read two papers, one on the
‘Primitive Source of Food Supply in the Great Lakes,” the other
on ‘ Some Experiments in Methods of Plankton Measurements.

Discussion on Professor Ward’s papers.

Professor Rowlee: I would like to ask Dr. Ward two ques-
tions. He mentioned three conditions which determined the pres-
ence of organisms more abundantly in the upper six feet of the
water than elsewhere. Those three conditions do not include one
that it occured to me might very considerably affect their presence
there, and that is the amount of air or oxygen that is in solution
in the water. Is it not true that there is much more of that
generally near the surface than in the deeper water ?

AMERICAN MICROSCOPICAL SOCIETY. 59

Professor Ward: That matter has never, so far as I know,
been investigated in fresh water. In salt water, Agassiz says in
“The Cruise of the Blake” that experiments made at a depth
of some one hundred fathoms show nearly the same amount of
oxygen as near the surface.

Dr. Eigenmann: I think that it has been shown that there is
more oxygen further down than on the surface.

Professor Rowlee: The other question was in regard to shoals
of fish visiting the mouths of rivers and streams. In Lake Ontario
the best fishing grounds are near the mouths of rivers ; and when-
ever there were great quantities of white fish taken by nets they
were usually, I do not know but universally, drawn where streams
flowed into the lake.

Professor Eigenmann: Was that in the fall of the year ?

Professor Rowlee: I think they drew them all summer, at all
times of the season. It occurred to me that that might be ex-
plained by some unevenness in the supply of food.

Professor Ward: I do not know how that is on Lake Ontario.
The best fishing grounds in ‘the northern lakes are not in such
situations. The best white-fish ground in the vicinity of Charle-
voix is on the west side of Beaver Island, which is a straight
sandy beach, so far as I remember, entirely without a break of the
smallest size. There is this much to be said with reference to the
white fish. There was evidence, I think, discovered last summer
to lead me to say that I believe the true white fish is a bottom
feeder, and in that respect differs from all the other species of
white fish in the lakes.

Dr. Fell: I wish to make a remark on some observations I
made in Lake Erie last summer. I noticed great quantities of
small fish cast up on the shore from Crystal Beach west and also
from there east, and I presume along the whole shore of Lake
Erie. There were millions of fish two or three inches long
thrown up on the sand and destroyed. Whether these were
products of the fish culturists or not I cannot say, but it seemed
to indicate that there was some great mistake made in supplying
the lakes with fish spawn, if such was the case.

60 PROCEEDINGS OF THE

But I have not seen any reference to this great destruction of
small fish before. I have not noticed it this year, but last year
the dead fish were plentiful, so that to walk along the shore of
the lake was very unpleasant indeed. You could not step with-
out putting your foot on two or three of them.

As to the question of food supply I recollect some years ago
we made an estimate of the character and quantity of diatoms
passing Niagara River; and we ascertained that some tons of
diatoms passed down there every day. Undoubtedly diatoms
are a great original source of food supply for the lakes.

Professor Eigenmann: Professor Ward’s paper was among
those which I especially was anxious to hear. The importance
of such stations has long suggested itself to me. I believe that
sometimes there is contemporaneous evolution in different regions,
and I think that something of that kind has occurred out our
way in regard to this matter of studying the conditions of fish
life. Illinois has established a station, Michigan has established
a station, Indiana has established one this year, and Ohio is.
going to establish one next year, to observe just such things
as Professor Ward has been telling us about.

I have made some observation on the plankton this summer, but
that was only as one of the elements of the environment of the fish.
We have attacked the fish problem in a little different way. We
have established a station on the continental divide between the
St. Lawrence and Mississippi basins, at a place where within five
or six miles we can get lakes of practically the same dimensions,
one belonging to one system and the other to the other system.
More than that, within a short distance we get lakes belonging
to Lake Erie, and other lakes belonging to Lake Michigan.

We are attempting there to study the environment of the fish
and the variation of the fish—the environment simply to give us
the unit for studying variation. We are catching as many
fishes as we think will suffice to get at the entire variation of a
given species in the water of these lakes—so many that if we
caught all the rest of them it would not make any difference in our
measurements. For instance, we have of one little fish, caught in

AMERICAN MICROSCOPICAL SOCIETY. 61

one of the lakes, something like 700 specimens now, in which
we will measure the variation. We have caught about as many ~
in a neighboring lake to measure the variation there. We are
measuring the environment and the variation in one place to get
a unit in order to measure the influence of change of environment
upon the same fishes in these different lakes to be found
around our present station.

To return to the turtle question for just a moment. Most of
these lakes have been raised in quite recent years by the building
of dams, and that has flooded some of the lowlands that were
covered with trees; the trees were afterward chopped down so
that we have along some of the margins many stumps. These
stumps have rotted so that there is a depression in the center.
These have been seized upon by the turtles as breeding places.
The little turtles crawl up these stumps and lay their eggs
inthem. We have got as many as 362 turtle eggs, I believe,
out of one of these little places. Rotten logs are frequently full!
of these eggs. There are places where in a cow track going
across a wheat field every depression that the cow made in
walking while the ground was soft contains turtles’ eggs.

Professor Kellicott: One or two questions have been raised
which I wish barely to mention. In regard to the distribution
of the plankton, I may say something in regard to the time which
it takes for a floating object to move from Lake Huron, say, to
the foot of Lake Erie. Those who have worked at Buffalo are
acquainted, I think, with the fact, that pine pollen is found float-

ing in the water past the city from the first of January to the first of
~ March. Early in the summer while the pine pollen is being
thrown into the lake there is none at all. After the date I have
mentioned there is practically none atall. Of course a storm will
sometimes stir it up from the bottom so that we find it in small
quantities throughout the year. But during the time from Jan-
uary to March a great deal of pine pollen comes through the
water to Buffalo. This would seem to show that the forces of
distribution are ample, even if these minute forms do move so
slowly of themselves.

62 PROCEEDINGS OF THE

In regard to the diatom Stephanodiscus Niagare it occurred also
in the winter, beginning earlier than I have mentioned, and late
in the fall, in great quantities, and living and continuing for two
or three months, when they ceased. Just where that diatom
occurs most luxuriantly Iam notaware. It has never been found,
I think, in shallow lakes, except Hemlock Lake. That fact would
also have, I think, some bearing.on this question. It must be, I
think, an inhabitant of deep water.

Mr. Ward speaks of crustacea being the food of a certain
fish. Perhaps he is aware that mysis sometimes makes its —
way into the water supply at Buffalo. We have taken it there,
always when the ice is going out in the spring.

In regard to the weighing of the plankton, we estimate the
quantity by weight. I suppose we were rather crude because we
did not dry the material absolutely. We dried it until the water
was just gone, and tried to estimate the exact amount of moisture
in that way. That method, when we weighed a large quantity,
could cause, I think, only a slight variation. It gave only.a re-
lative result, but I think that relative result was pretty good.
The method by ash I have some doubts about, because float-
ing in the waters of the lakes there is always a large quantity of
inorganic matter. If this matter were constant it could be disre-
garded, but it is very inconstant. .

Professor Ward: I think Professor Kellicott has a little mis-
understood my statement. We used the ash precisely for that —
reason, to get rid of that inorganic element. There is always, as
we found by microscopic examination, some sand floating in the
water. We can find individual grains of sand which in weighing
make considerable differences. This is especially important when
only a small amount of material is taken inahaul. For instance
in one haul we have taken only .95 c. c. of material. In estimat-
ing that, a small quantity of sand makes a great difference in the
weight. But by taking the weight air dried and taking the weight
of the ash and subtracting it, we eliminate the inorganic substance
in the water, eliminate everything except what is organic.

AMERICAN MICROSCOPICAL SOCIETY. 63

Tuurspay AFTERNOON,

On Thursday afternoon the society with their friends were °
given an excursion from the Campus to the Lake by the street
car.company and upon the Lake by the citizens of Ithaca. This
excursion was most enjoyable in every respect. The day was
perfect, and through the kindness of Professor Tarr and Professor
S. G. Williams, of the university, the interesting geological
features of the lake basin were pointed out to the members.

Fripay MornineG, August 23.

The society assembled at 10 o’clock in the McGraw building,
about 90 persons being present. The nominating committee
made its report as follows :

Officers of the society for 1895-06:

For President, Dr. A. Clifford Mercer, of Syracuse, N. Y.

For Vice-Presidents, Edward Pennock, of Philadelphia, Pa.;
Miss V. A. Latham, of Chicago, II.

For Secretary, Dr. William C. Krauss, of Buffalo, N. Y.

For Treasurer, Magnus Pflaum, of Pittsburg, Pa. ‘

For members of the Executive Committee, Professor C. H.
Eigenmann, of Bloomington, Ind.; Herman Schrenk, of St.
Louis, Mo., and Miss M. A. Booth, of Longmeadow, Mass.

Upon motion, the secretary was directed to cast a ballot for
the nominees as reported by the committee, and they were duly
elected.

Professor Conser then read his papers on “‘ Cocaine in ‘the
Study of Pond Life,’ and on “ Paraffin and Collodion Imbed-
ding.”

Discussion : _

Professor Eigenmann: I might add a word or two as to the
use of cocainێ. I have found it very useful in studying the habits
of fishes. In one case especially that I remember, where I secured
only one little larva with a net in sea-fishing, it was very valuable
and I wanted to make as many sketches as possible at successive
stages ; but it would not hold still, and of course J did not want
to kill it. So by judiciously adding a little of the cocaine, that
the druggist would let me have, I succeeded in keeping it still

64 PROCEEDINGS OF THE

quite long enough to draw; then by rapidly changing the sea-
water I succeeded in having it live another day. By repeat-
ing the operation in that way I got four or five sketches of it.
It thus proved very successful.

Dr. Mercer: Iam not very familiar with these methods as I
am not in the habit of doing much practical work now, but I
saw a “wrinkle” a year ago which I suppose some worker might
be glad to hear of. It was in Mr. Andrew Pringle’s laboratory
on the other side of the water. He was using the paraffin method
almost exclusively. He placed his specimen, when he wished
to saturate it with paraffin, in a chamber which he could exhaust.
The temperature of the chamber was regulated by a thermostat.
In a very short time, in comparison with the usual methods,
the paraffin had saturated the specimen, by reducing the atmos-
pheric pressure around the material. As a means of saving time
I dare say the busy worker will appreciate it.

Professor Conser: I have used the method of placing the
object under an air pump, which has reduced the time consider-
ably. It is a very practical method, though it is somewhat com-
plicated and troublesome.

Dr. Mercer: I might add that this was done in the case I
mentioned in a chamber which could be kept at any desired tem-
perature, which is a little addition to the air pump.

The President: I would like to ask Professor Conser why he
uses celloidin, which costs $1.25 an ounce, when gun-cotton can
be got for 25 or 20 cents an ounce; and I feel confident that any
one using it will find he gets just as good sections. This gun-
cotton is made in America, the celloidin in Germany. I had
occasion one time to buy eight or nine ounces of the German,
and I kept it very carefully and only used it as I thought we could
afford it, and in a little while it would not work at all. I tried it
and found it exceedingly acid. In despair I went down town
and bought an ounce of gun-cotton for 25 cents, and things com-
menced to work. After that I discarded the expensive for the
cheap.

Professor Ward: May Task the President if he has ever

AMERICAN MICROSCOPICAL SOCIETY. 65

had any difficulty in getting a quality of gun-cotton which is free
from little fibrous foreign bodies? All I have been able to find -
was quite dirty in that way.

The President: What I do in that case is to make up the
collodion and let it stand a little while; and all that settles to the
bottom, so that all the rest is perfectly clear. Or I filter it, but
that is not so good a way. If allowed to settle the upper part is
very excellent.

Professor Eigenmann: Is this gun-cotton precisely the same
thing as celloidin ?

Professor Gage: Yes.

Professor Conser: It won't go off?

The President: Yes, they will both go off if you want them
to. They are both of them nitro-cellulose, I believe.

Professor Conser: I want to add a word in regard to the
keeping qualities of celloidin. It must not be kept in air-tight
chambers. It can not be kept in glass-stoppered bottles, for
example. There will be some decoinposition in that case, especi-
ally if kept at the ordinary warmth of the laboratory. Though
I have used gun-cotton to a very small extent, I have preferred
the celloidin, from the fact that it was easier to dissolve, and more
ready at any time for operation. My experience with gun-cotton,
however, has not been very much ; it has only been experimental.

Dr. Krauss then read a paper: on “ Formalin as a Hardening
Agent for Nerve Tissues,’ and Dr. P. A. Fish one on “ The use
of Formalin in Neurology.”’

Discussion :

The President: Probably no reagent has been discovered and
applied to anatomical preparations, to microscopic work, which
is going to do so much, help us so much, as this one formol, and
every detailed account of experiments will be therefore of the
highest value to serve as a guide. I am particularly glad that
these two papers have come before the society to give us infor-
mation, and I hope that those among the members who have
made experiments will tell us of their experience. We then can
put all this testimony together and go on more successfully in

5

66 PROCEEDINGS OF THE

the future. I hope Professor Kellicott, who has experimented
with this substance, will tell us the results he has obtained.

Professor Kellicott: I have worked for about a year in pre-
serving by this method and I must. say that the result agrees so
closely with the results given in the papers that it seems hardly
worth while to take any time in stating my experience. I have
used the formalin in various solutions, and on a great variety of
tissues, in preparing them for museum and anatomical work,andthe
results have been most excellent. I think in our laboratory we
are going to harden them by one or two changes, then transfer
them to about half alcohol and half water, for preservation.
This method seems to give us better results than any other we
have tried, at least. Formalin hardens animal or other tissues
very quickly, and without much shrinkage, as has been said ; and
when transferred to alcohol they remain without shrinking, pro-
vided itis not too strong. The specimen will be preserved without
deposition of coloring matter or sediments, so that really sometimes
the formol does not need to be, filtered or changed. It preserves
the specimen clear and transparent, perfect in every way. I
have used it in preserving the brains of large animals as
well as small ones, for museum purposes, and have found nothing
like it. It requires less material, less time, and only a quarter
of the pains. You can put your specimen into the solution and
go off and leave it, and when you come back it will be all right,
just as you left it.

We have used it also for histological purposes to a limited
extent. In that regard we must work further before we are
ready to state any general conclusions.

As to the odor, I found that some students are unable to use
the formalin on that account without great annoyance to them-
selves. There was one student in my laboratory who could not
use at all a specimen that had been preserved in this way, the
irritation was so great.

| have had experience, I think, similiar to your own, in prepar-
ing anatomical materials for the ordinary dissecting work of the
laboratory. I think in that regard it is extremely valuable.

AMERICAN MICROSCOPICAL SOCIETY. 67

When the tissues are washed out in the usual way with a salt
solution and then injected witha formalin solution of two to four
per cent—I mean by volume—the animal will remain, if kept
cool, ready for use without any change or shrinkage. - We are
using it in that way constantly, and it is so valuable that I want
to recommend it to every one who has to prepare animals for the
dissecting laboratory.

I have had some experience in attemping to stain in bulk
before the tissue is hardened, with carmine, for example, and I
have had excellent results. When I have had time to experi-
ment so as to ascertain the right time and temperature for the
work, I am sure it will be of the highest value.*

The President: May I ask the writer of the first paper what
he had reference to in giving us the percentages? Here is going
to come in a real difficulty inthe use of formol. It is a forty-per-
cent. solution in water, and when we say we use a two-per-cent. ora
one-per-cent. or a four-per-cent. solution, the question immediately
arises—it did in my own mind when I came to use it—do we
mean that we take 97 or 98 c. c. of water and 2 or 3 c. c. of
this formalin solution, or do we mean that we take 95 c. c. of
water and 5 c. c. of the formalin solution, that is, to get a truly
two-per-cent. solution of formol. In some of the papers that I
have heard read the statement has not been distinct as to whether
it was by volume, as Professor Kellicott spoke of, where you
would use 2 c. c. of the forty-per-cent. solution formalin and ninety-
eight per cent. of water, or whether the absolute amount of formol
was meant. Dr. Krauss, what did you use?

Dr. Krauss: I used two per cent. by volume.

The President: That would be then really an eight-tenths-per-
cent. solution of the formol. I hope that will always be stated so
that there will be no mistake. The chances are if it is not stated
that some one who has had training as a chemist may use a really
two-per-cent. solution of the formol and that might be too strong.

*Professor Kellicott has written a detailed account of his experience
with formol, and the secretary has added it to the papers read at the
meeting.

68 PROCEEDINGS OF THE

Professor Ward: I want to make a few statements in regard
to this matter, because my experience, with perhaps a different
class of objects, is so absolutely different from what has been
reported this morning. If it were only on the basis of my per-
sonal experience, I should hesitate to emphasize the point as
strongly as I want to do. I know, however, by personal
correspondence of experiments carried on at the University of
Michigan, at Harvard, and at the Newport Marine Laboratory, and
they all agree with my own, that for certain purposes formol is
not only useless, but positively bad, worse than useless. Some of
you may remember noticing in the January Naturalist, if I re-
member correctly, quite an extensive article on the use of formol,
by a gentleman working in Professor Kingsley’s laboratory.
He was very enthusiastic in praise of formol. By personal cor-
respondence with one of our faculty at Lincoln, I have learned
that his preparations have all spoiled since that time. He recom-
mended formol very highly in the article on the basis of three
months’ experience. The same thing was true of my own ex-
periments, the same thing was true of experiments at the Newport
Laboratory and at Harvard. The preparations are good for a
limited time, but histologically at least they are useless after that
limited time. How long that time is has not yet, perhaps, been
determined. That of course is where the formol is used asa
preservative agent, and I think we ought to distinguish sharply
between its use as a hardening agent and as a preservative. As
a preservative I am convinced that for invertebrates at any rate
it is very bad. It gives distorted and incorrect histological
features every time. There may be a difference in the action on
vertebrate and invertebrate tissues, and I have not experimented
with it on vertebrates except for gross anatomy, but for preser-
vation of the invertebrates it is very bad.

In one of the papers, I think that of Dr. Krauss, I noticed a
little sentence that may have something to do with this. He said
that in some of his preparations the nerve cells seemed to be
slightly swollen, and the nucleus stained very deeply. Is that a
beginning of a change opposed to the ordinary shrinking, a

AMERICAN MICROSCOPICAL SOCIETY. 69

change by enlargement of the cell which, if it goes on long
enough, will result in serious damage to the tissue? Where we
are not concerned with a simple diagnosis of a certain kind of
cell, but are desirous of securing a correct histological image,
or making a careful physiological study of that structure, it is clear
that we must have some reagent which will preserve permanently
and as nearly as may be the actual character of the cell, neither
swollen nor shrunk, but in precisely its actual normal condition,

The statement quoted in one of these papers that the coloring
matter is preserved, I can distinctly negative for as widely-separ-
ated classes of invertebrates as the fresh water mites, crustacea,
worms and hydroids. In none of these is the color preserved
ordinarily beyond a few days. The color of planarian worms,
for instance, both fresh water and marine, while comparatively
well preserved up to a month, is entirely lost within a period
after that, so that at four months the color is entirely gone, and
at that time the worms are in poor condition.

In addition to these points let me say that formol is highly
volatile, and unless the bottles be very carefully closed the solu- -
tion deteriorates rapidly. So for instance we found that at the end
of four months an ordinary homceopathic vial corked as closely
as the best quality of A A * corks could close it, did not
contain a recognizable trace of formol. The specimens had
entirely deteriorated, and that not in a single vial nor due to
accident, but in a whole series. Whether this can be avoided by
the use of vaseline on the corks, I donotknow. Certainly under
ordinary conditions formol will disappear very rapidly.

Professor Eigenmann: The last point that Dr. Ward mentions
is a serious one, since so far as I know now we have no means of
testing the exact per cent. of formol that may be in the water ;
so that after we have used a solution once for hardening, for
instance, it must be thrown away or else we have to deal with a
solution whose per cent. we do not know. I have tried the
formol—there seems to be a perfect epidemic of formol experi-
ments in the country—I have tried it on various things. I tried
it on plankton and the plankton would not settle, so at least in

70 PROCEEDINGS OF THE

measuring it is useless. We tried it on crayfish and ali of them
spoiled, but I think that could be avoided by using a little stronger
solution. By injecting the crayfish with a strong solution and
then putting it in a weaker, it certainly becomes a beautiful object,
and looks as if it were going to crawl off. Frogs can be pre-
served so that they look very much alive indeed. On fishes I
have tried it and the color utterly disappears. I was in hopes I
could preserve the beautiful colors of our fresh-water fishes—and
they are certainly the most beautifully colored creatures alive. The
formol is also, I think, valuable in injecting into the muscles of
larger fishes that are to be preserved. Alcohol does not penetrate
quickly enough to preserve them. At Wood’s Holl last summer
I put a two-per-cent. solution of formalin in a small vial, and
crowded as many little fish into it as I could, fairly jammed them
in ; and I must say that a year afterward the fishes are just as hard,
just as firm, firmer in fact than when I put them in. [I tried it on
tad-poles. They are very hard to preserve. I tried a two-per-
cent. solution of formalin, in thirty-five per cent. alcohol, fifty. per
cent. alcohol, etc.,and also in simple formalin, and the formalin gave
the best results. The tad-poles are so firm that if you take them
by the tail and flop them they stand rigid, while, of course, in
alcohol preparations they are flabby and very disagreeable.

I would suggest that the difficulty regarding the percentage
could be avoided by using the name formol for the pure formol,
and formalin for the forty-per-cent. solution. I myself have used
it that way.

Miss Clara Harrison: I have tried some experiments with
formalin. Some three years ago, perhaps a little more than that,
[ had for sometime been trying to find some solution that would
preserve the color of fruits and flowers. I heard of formalin,
and as | could find no literature, that is, very meager literature,
my experiments with it were rather of a shot-gun kind. Ibegan
with five per cent. of the forty-per-cent. solution, and went down to
one per cent. I tried it on the Orchidacea, and I am sure their
coloring is quite as fine as that of our fishes. I found that the
yellows and the purples kept for a long time. I tried, as I

AMERICAN MICROSCOPICAL SOCIETY. mK

remember now, one of the Mexican Orchids and it kept clear
and perfect for about four months. I might say that I puta
grain or two of corrosive sublimate into the solution. For
about four months these kept perfectly in a little test tube
corked with a rubber cork. I was delighted; and one day I
thought I would take it down to the White House, where I had
obtained the specimen, and compare it with a fresh specimen. I
found the color identical. -But the next morning I was very
much disappointed to find the plant white and the solution a
beautiful red. So my experience is identical with that of Dr.
Ward. It does well for a short time, up to about four months,
in a one-per-cent. solution ; but after that I would not like to answer
for it, that is, on fruits and flowers.

The President: It seems to me that this discussion has
brought out the advantage of presenting a subject to a society
of people who are interested in the same thing, then by com-
paring notes it is possible to see wherein the thing is useful.
We have found, I suppose, all of us, that there is no panacea for
anything. Formalin is certainly admirable for certain things ;
but that does not say it is going to be admirable for everything.

A paper on “ New Points in Photo-micrographs and Cameras ”’
was then read by Mr. Walmsley.

The President: It was in 1882—before I had ever thought of
such a thing as making photographs with the microscope—that
I had the pleasure of hearing Mr. Walmsley speak upon this
subject. I think the society is to be congratulated that we have
with us, so to speak, a direct lineal descendant of Woodward.
As he has said, the work that Mr. Woodward did showed the
world what it was possible to do in the delineation of minute
things by photo-micrography.

Miss Latham then read a paper on ‘‘ The Question of the
Correct Naming and Use of Micro-Reagents.”’

The President: It is not often that we have so vigorous a
presentation of a thing, and I do not wonder that Dr. Latham
speaks with some feeling, because we have all gone through the
Same experiences, more or less, that she has.

72 PROCEEDINGS OF THE

Mr. Clark Bell then presented a communication on “ An
Inquiry Concerning the Possibility of Distinguishing Arsenic
from Different Packages by the Microscope.”

It is due to Dr. White that this question has been presented
to the Medico-legal Congress, and I thought it might be well
to ask the microscopists. of this ‘country if they would give us
some aid upon the subject. Any one who wishes to investigate
the question can address me and I will respond by mail and
send any literature upon the subject. The question arises out of
the celebrated murder trial in Connecticut. I will ask Dr. White
to explain the details of the case.

Dr. White: Inthe famous Hayden trial it was proved that
Mr. Hayden had purchased an ounce of arsenic in Middletown,
and it was claimed by the State that he had administered this to
Mary Stannard. He claimed that he had put the arsenic in his
barn, and an ounce of arsenic was found in a tin box in the barn.
The State asked whether it was possible to determime whether
that ounce of arsenic came from Middletown or not. The
arsenic which was found was examined with a microscope and
was found to consist of very many crystals with bright reflecting
faces, whereas the arsenic in the jar in Middletown had dull,
leaden faces. The arsenic found in the stomach also had dull
leaden faces. The question was whether these two parcels of
arsenic could be distinguished. Many parcels of arsenic were
bought from many different firms, to find, if possible, where this
arsenic came from. One expert in Chicago has stated that in his
opinion this dull, leaden color of the arsenic occurs by deoxidation,
but chemists tell us that cannot be. It is probably due to some
peculiarity in the grinding. What chemists know as glass
arsenic, when ground, will not have transparent reflecting faces.
The large crystals found in some places where arsenic is made,
if ground up, will also show rough faces; whereas the arsenic
found close to the furnace, if ground, will still show many
small bright faces.

If in this case the arsenic found in the barn was just the same
that Mr. Hayden bought in Middletown, it was clear that he might

AMERICAN MICROSCOPICAL SOCIETY. 73

be innocent. If it was from some other source it might be that
he was guilty.

This idea of distinguishing different parcels of arsenic by the
microscope was first brought to my attention in Medical Juris-
prudence some thirty or forty years ago. In 1860 a lot of
crystals were made for me by the chemist in Middletown. I have
had some of them preserved ever since ; and I think some of them
undergo a process of pitting. It is a question whether if kept for
some time they lose weight and form pits in the faces.

The paper by Dr. Krauss on ‘* A New Way of Marking Ob-
jectives’’ wasthen read. Thetwo following papers were read by
title only, viz.: ‘‘ Demonstration of Histological Preparations by
the Projection Microscope”’ by Drs. Krauss and Mallonee, and “‘Im-
provements in the Collodion Method ”’ by Professor S. H. Gage.

Dr. Mercer exhibited an improvement on the Syracuse solid
watch glass, having a peculiar shape so that they may be piled
together without slipping. |

Mr. Pflaum exhibited a metal block for centering slides which
he found very convenient. It is figured in his article in this
volume of the proceedings.

The society then adjourned to 3 P. M.

BUSINESS MEETING.

Fripay, August 23d.

The meeting was called to order at 3 P. M. in the McGraw
hall.

The Secretary: The Constitution requires that amendments
shall not be made except ona year’s notice. Article II provides
that membership may be acquired by application in writing,
recommendation by two members of the executive committee, and
election by the society. Notice is given that next year an
amendment will be proposed striking out the words from
“ nomination ” to ‘“‘society” inclusive, and so changing it that it
shall read ‘“‘ and election by the executive committee.”

74 PROCEEDINGS OF THE

The purpose of this is to prevent the long delay, often nearly
a year, before persons applying between the meetings could
become members, although in practice they were furnished the
proceedings like regular members.

The President: This notice of amendment then has been
duly given, and next year at our meeting it will be acted upon
and decided. .

The Secretary: A proposition has been submitted that there
be two classes of members, or two conditions of membership for
life, which would add to our income. The first is that persons
or organizations desiring to obtain the proceedings, by paying
$50 would be considered as subscribing members, without any
other rights of membership ; while those paying $100 would be
full-life members, and entitled to take part in the proceedings of
the society.

The President: This will be considered as due notice that
this amendment will be brought up for consideration and final
action next year. The proposition concerns membership and
requires an amendment to the Constitution.

The President: We have difficulty in storing the property of
the society, especially the plates. The drawings for these are
furnished by the writers, and they are made at the society's
expense. I propose that after an article has been published the
plates shall go to the writer.

I think the plates will do a great deal more good in that way
than in the hands of the society, where they are practically a
burden. Not one plate in fifty will ever be used again by the
society. A person interested enough to make the plate would
likely in future work find real use and convenience in having
these plates to show some other phase of the subject.

Professor Ward: Is this to be retroactive, or simply to deal
with the future ?

The President: As the stock is a burden to us, it seems that
it should refer also to the past as far as possible.

Professor Ward: J make a motion then to the effect that the
plates now in the possession of the society, or those to be used

AMERICAN MICROSCOPICAL SOCIETY. 75

in future publications, be given to the authors of the papers in
which they are used.

The society is supposed to get enough out of them to pay
for making them, otherwise they would never have been made.
Now they are so much dead timber on hand,

The Secretary: The society expects to keep some of its
publications, and there must be a place to store them. It is
very unfortunate that there cannot be a permanent place of
deposit for all our property. It is not merely a question of the
plates, but of back sets and also of donations of the proceedings
of other socities, of which we get a good many and would get
many more if we had a place to keep them permanently. There
are also some preparations and a number of other things. _A great
deal more has been lost than we now have on hand. The society
is expected to be national and to be permanent. The individual
is transitory, and his heirs do not take proper care of such
property as plates. Plates that have been prepared by students,
say in Cornell University, might be returned and they would be
kept properly by the University ; but that is not so with outside
contributors. To return to them will be throwing them away.
The society has paid for them, and they ought to remain its
property and a place provided for them. They take up actually
little space—all could be piled under one table. With the
exception of the plates for one number, all the plates made
before I became secretary have been practically lost.

The President: That isa ground on which I proposed that
they should be lost by the individual and not by the society.
Science is changing and the plates are of little value after a
short time, in most cases. Those making the plates may be
able to use them once or twice more before they have lost their
value.

Professor Rowlee: ‘There seems to be the larger question of
a permanent home involved. May it not be well to leave the
matter to a committee, of which the treasurer shall be chairman,
to provide a place for permanent housing of our property? I
am sure that the plates prepared in the botanical laboratory here

76 PROCEEDINGS OF THE

would be of more use to us in future publications than they
could be to the society. They may be used to illustrate some
new phase on the same general line.

The Secretary: It was stated in a previous meeting that the
Buffalo Academy of Arts and Sciences, in whose building the
Buffalo Microscopical Club has its home, would afford us a
place. I favor a committee to consider that subject. There is
much interest in microscopy at Buffalo, and it is conveniently
situated. I should think either there or Cornell University the
best place. .

Mr. Young: It is early in our history to begin giving away
things. We may think we must give away other things. I
think other members’ should be consulted, and move that the
matter be deferred to the next meeting. In the meantime I
favor a committee on the subject of a permanent home.

Dr. Mercer: I move that the whole matter be referred to a
committee of three or five, to consider also the larger question
of a home for the society, as well as its property. There we
could have a room, collect our books, etc., and meet there not
necessarily always, but when we had no other invitations.

This motion was seconded.

Mr. Milnor: If Pittsburg were made such a home I can
guarantee a place for storage that would be fire-proof.

Dr. Mercer: Dr. Krauss expressed the same thought to-day
as to Buffalo. The President stated the question to be on the
amendment to appoint a committee of three, as made by Dr.
Mercer.

Professor Rowlee: We must notice that we are about to
transfer all this material, with the change of office.

The President : No, the material is with the treasurer, not with
the secretary, according to the Constitution.

( It was explained that the plates had, by special arrangement,
been left with the secretary.)

Professor Ward: Plates to be useful at all must be used in a
very short time. The society will not print the same plate in a
volume within a year or two. After that it is of little use. But

AMERICAN MICROSCOPICAL SOCIETY. 77

this does not affect the question of considering a permanent
home—for the two questions must be distinguished clearly.

The Secretary: One paper in the journal this past year was
illustrated with a plate used in 1884, and last year. Dr. Stedman
applied to me for a plate used in 1882.

Dr. Mercer: Such plates would be very useful if in the
hands of the society for illustrating historical papers treating of
the development of some subject—as I have seen in articles
published by the Royal Microscopical Society.

The President: The question is on the amendment, that a
committee of three be appointed to investigate the whole matter
of the property of the society and a permanent home for the
society.

The President put the motion, which was carried.

The President: A proposition has been made to print the
Constitution and By-laws—four pages—every year in the pro-
ceedings, so that all new members may receive it and know
their rights and duties.

The Secretary: The proceedings are arranged to bind two
years, in a single cover. Every two years there is an index for
this double volume, there being only tables of contents with the
annual volumes. The Constitution has been put in only once in
this biennial volume, not in each annual volume.

After discussion, Professor Rowlee moved that the Constitution
be inserted in each annual volume.

Mr. Pflaum: I move as a substitute that the Constitution be
printed separately for distribution.

The President: And not bound up in the bound volume ?

Mr. Pflaum: No, because not necessary. We could send
these pamphlets to persons wishing to know the character of the
society.

President Gage: The American Association for the Advance-
ment of Science prints its constitution: in every volume of its
proceedings ; the constitution and list of members are also printed
separately for distribution, as suggested.

Mr. Pflaum: I then would make an amendment that they be

78 PROCEEDINGS OF THE

printed in the proceedings, and also separately for distribution.
I would include the list of members.

Professor Rowlee accepted the amendment and the motion
was carried.

The President: Our treasurer has had bound an official copy
of our proceedings. It is moved that an official copy be similarly
bound for the secretary.

Dr. Mercer: It would be well that this set, instead of look-
ing exactly like the other, should be marked as being the
secretary’s official copy.

This amendment was accepted by the mover and the motion
as amended was passed.

The President: The Spencer-Tolles fund is approximately
$400, and is invested in a building association or fund in Ohio,
netting six per cent. The treasurer proposes to transfer it to a
similar association in Pittsburg where it will net ten per cent., and
be safe. The executive committee has referred this to the
society for action.

Mr. Pflaum: This building and loan society in Pittsburg is
not a merely speculative and insecure one. It is national, and
its officers, whom I know personally, are well-known and per-
fectly trustworthy men. In seven years the fund will be at least
$1,000. The association has been paying fifteen per cent., but
interest is falling everywhere and they do not expect over ten per
cent, soon

The Secretary: This is an important matter and not of the
kind that we as a society are well fitted to decide. It depends
on judgment and investigation. I make a motion to refer the
matter to a committee of gentlemen residing in Pittsburg, three
members to be appointed by the President, with power to act if
in their judgment it is the best thing to do.

Mr. Pflaum: I amend the motion by saying that the full
membership of this society residing at Pittsburg be the committee.

The Secretary: I accept the amendment.

Mr. Milnor: I approve of what Professor Seaman has said.

[he society itself should not attempt to judge at a distance.

AMERICAN MICROSCOPICAL SOCIETY. 79

We should investigate thoroughly before the money is placed in
the hands of any society, especially a building and loan society.’

Dr. Mercer: I utterly oppose the investment of the money
in that way. Mr. Spencer and Mr. Tolles were men who would
have little sympathy with such a proposition. They would
prefer three per cent. in a safe place, to twenty per cent. in a building
society. If Mr. Spencer understood the principles on which
these societies are conducted he would oppose it. As I under-
stand it, they advertise to investors a high return—ten or twelve per
cent., and to borrowers remarkably cheap loans. But that is
impossible. It is the old story—the man who buys his coal by
the bushel pays a higher price. Mr. Spencer would be the last
to make him do that.

The Secretary: The object of the motion is that the com-
mittee shall do what in their judgment seems best—and I think
the society will be willing to trust these members. I think Dr.
Mercer must be under misapprehension as regards building
associations, for statistics of the United States show. that no other
investment is so safe. But there are different kinds of associations.
I should want to know the character of the men.

Mr. Milnor: I think if the matter is left to gentlemen in
Pittsburg it will be safe. I know this association to be first class.
But it has brought its rate of interest down now and _ will
eventually be much lower, so it may not bring over six per cent.;
and if that is the case I think bonds at six per cent. would be a
preferable investment.

Dr. Mercer: I do not think we ought to place this money in
any institution where any -person can even use the word
suspicious with reference to it.

This fund has been obtained with great difficulty and must not
be lost. I believe it should be taken from Ohio, and it would be
safer to put it into a savings bank at Pittsburg where it would
probably pay four per cent. Building associations are not the
place for trust funds. I should be afraid of an institution paying
ten per cent. when money is begging everywhere at two per cent.

Mr. Plaum: If the money had not already been in a build-

80 PROCEEDINGS OF THE

ing association I perhaps might not have suggested putting it in
this one. This one is better than that. A building society is
only a co-operative savings bank. The reason it pays such
large profits could be readily explained ; it depends on the com-
pounding of interest largely. The members get the difference
between six per cent, and ten per cent., taken by the proprietors
of savings banks. These associations have been of greatest
benefit to the poor men of Pennsylvania. The former opposition
to them—e. g. that of the courts, has been overcome by their
results. I think a committee could be trusted to treat this as a
sacred fund, as it Is. ;

Mr. Young: As I understand it the committee is not bound,
by the motion, to change the fund or to put it in a building
society, but as they see fit.

The Secretary: Yes,the resolution reads that the members of
the society residing in Pittsburg shall form a committee to invest
the Spencer-Tolles fund.

Mr. Phaum: (Answering a question.) There are about
seven active members in Pittsburg. It might be better that the
President appoint three or five out of these, as it would take
some time to sound all of them personally. I withdraw my
amendment and leave the original motion for a committee of
three or five. .

Mr. Milnor: I want the responsibility to come from the
Association—let the President appoint the committee.

The President: The motion then is that a committee of three
be appointed by the chair to re-invest the Spencer-Tolles fund.

Mr. Phaum: The change is necessary. The only member
at the time of the investment in Urbana, Ohio, has resigned, and
there is not a soul to look after the money there.

The motion as’stated by the chair was now put, and carried.

The Treasurer presented his report.

Dr. Mercer: I move that the report be accepted and adopted,
and that we thank the Treasurer for bringing to our attention
the character of the investment of this fund.

The President put the motion, which was carried, and then

AMERICAN MICROSCOPICAL SOCIETY. 81

stated that the executive committee recommend that 500 copies
of the proceedings be printed. .

Mr. Pflaum: This matter might perhaps best be deferred until
a permanent place of storage is decided on. As it is, our extra
copies take about 200 cubic yards and it costs $12 a year to store
them, without insurance. We have about 225 active members.

Dr. Moody: I move the society publish 300 copies.

The Treasurer stated the number of copies of each issue
actually on hand, viz.:

YEAR. YO COPIES. | YEAR. NO. COPIES.
LOG Se a ia: SO een pet UD 2: Hees ons, ad 57
iste ean he hae 30 SS Of eee ieee ae et SF) 246
TEST 2. . aL wee ee Ae F892 =SPartialitiyes ees cies. Le 185
Tio. Seng ae on os 43 Parties: SLA re
Poneer te ak. ok. ee eo Bantu rere cee: 76

UG ee ee Re 6 L893 —SParti lee ranrs aes ote en HB
ISIS. 2 Seg See 44 Parte il ce eee. 53
Llelol Gis Se RS I MMe tli te 35 arta teens oo. ehh 126
LSS ee PPS Se ee ee 236 Jedriniy Zine 8 We 152
LISIEISIG cas cok tS aa ee A eae 154. i [poh Aa A a a aa a a le
MSC chat i. co cclbbel cncrdvsieipis te 175

*Of these numbers the Secretary has several copies. If any members could supply
duplicates of 1884 to the Treasurer it would add to our complete sets,

Professor Ward: It is evident that even of the number in
1893, of which we have the most on hand, 350 copies were used.
We can never tell when one of the papers may find a consider-
able sale—they often do. If we get a permanent home it will
be no trouble to store even 250, while the lack of numbers at
any time when they are really desired is a very serious matter.
The cost of printing extra copies, not requiring type setting, is
comparatively very little.

The President: The question is on the amendment changing
the 500 to 300.

The amendment is lost.

Now we will proceed with the original motion, that 500 copies
of the proceedings this year be published, and also as many
copies of the Constitution, By-laws and list of members, as in the
wisdom of the executive committee is desirable.

The Secretary: The society furnishes to each author in the
proceedings 25 ‘‘ separates’ made by dividing up 25 of the 500
6

82 PROCEEDINGS OF THE

copies printed. As the Constitution, By-laws and list of members
are to be printed in each volume, this separation leaves 25 copies
of them, which can be used as desired. I have always furnished
lists of members, to people asking for them, out of these
copies. There has never been demand for more than that.

Thereupon the motion as above stated by the chairman was
carried.

The President: The next question is whether the proceed-
ings shall be printed as a single volume, as was done until the
Rochester meeting, or in four separate numbers, as has been
done since. The proposition is submitted by the executive com-
mittee without recommendation.

Professor Rowlee: I move the printing of the proceedings in
a single volume.

The secretary just elected stated to me, knowing that the
matter was coming up, that he thought it highly advisable that
the proceedings be in a single volume.. He did not believe he
would be able to bring out four separate numbers and do it
promptly. The work has been very hard for the present secre-
tary. It really requires four times the technical work and care.
It keeps one struggling the whole year instead of three months.
I believe the incoming secretary can keep his promise to get out
the entire proceedings of this meeting before the holidays. The
present method was an experiment. Dr. Seaman was willing to
give us the necessary time, as he has done; but he could not
keep it up permanently. In spite of Dr. Seaman’s best efforts the
numbers have at times been delayed, and the work has been
severe.

The President: I would prefer one number, and to look that
through, and not to have to tie the four up with a string before
they are bound. I have had a good deal to do with the printing
—of course only a fraction of what the secretary has done, and I
am convinced that to ask anyone to go over the task four times
is asking too much. Editing the whole proceedings need not be
much more work than one part. A person capable of being
secretary can hardly afford the time. The last number for last

AMERICAN MICROSCOPICAL SOCIETY. 83

year did not appear till after the meeting, and this year’s last
number will not come out until after this meeting, in spite of our
best efforts.

The question is that the proceedings for this year be published
in a single volume.

The motion is carried.

Professor Ward: There is to be started with the begin-
ning of the year a scientific undertaking in Europe that is
to be international and of such a character that I want to bring
it prominently before you, and urge action of the society in three
directions. Most of you know the great difficulty of tracing
bibliographical references, the extreme incompleteness of the
present system of Bibliography. There is to be established in
Zurich a bibliographical bureau for zoology, which will go into
operation January 1, 1896. It absorbs the Zoologischer
Anzeiger, and the /Jahresbericht, which is published at the
Naples station; it absorbs the Archiv fir Naturgeschichte, at
least the second volume, which publishes a résumé of the work
in the different portions of the animal kingdom for each year ;
it absorbs some other minor bibliographical publications ; and it
is hoped another year that it will include also the English Zoo-
logical Record, thus giving in the scope of a single enterprise
all the bibliographical notices of the world. In France a sub-
committee to push the matter is very thoroughly organized, and
the organization of the American side is thoroughly under way
In France the subcommittee is under the supportand encouragement
of the Zoological Society of Paris. Sub-committees have already
been organized in Switzerland, Italy, Hungary, Russia, Germany
and England. The members who furnish the bibliographical
records for the bureau give their services free. There are two or
three paid officials in connection with the bureau itself, one
of whom being the very well-known Professor Carus, of Leipsic,
editor of the Zoologischer Anzeiger, who is to give his entire
time for the munificent sum of $500 a year. This shows
you that the enterprise is a purely scientific one. The
men who are working in it are not doing it for the sake of

84 PROCEEDINGS OF THE

gain, but in the effort to lighten the labors of the workers in this
field throughout the world.

The bureau proposes to record all publications which touch
upon zoology, and to have the recording done not by librarians—
and you know that the cards sent out by library associa-
tions for scientific topics sometimes include Rotifera among the
Infusoria, and make all sorts of mistakes—but by zoologists who
know exactly what they are doing. The bureau is to have two
publications, a fortnightly bulletin and a card catalogue. The
bulletin is to be in exactly the same shape as the lists and short
reviews or résumés of contents now given by the ‘ Zoologischer
Anzeiger.’ The card catalogue is to have these same, but each
printed on a card after the fashion recommended by the Library
Association.

It is expected to receive subscriptions to both of these, and
also that the bureau will be able of offer specialists subscriptions
to that portion of the bulletins or cards which deals with their
special work—as for instance that part of the bulletin dealing
with entomology and the cards dealing with that subject.

There are three things that the bureau asks, and must have if
it is to succeed. It asks in the first place a certain subsidy,
for in the first two or three years it is practically certain that it
cannot pay expenses. The Swiss, French and German govern-
ments have already granted subsidies—the Swiss gave $400. I
had an interview in New York with the gentleman who is at the
head of the whole enterprise, who has put all of his time, and
of his fortune—not a great one—into the undertaking. He
told me the particulars of the votes by the several governments,
but I have forgotten some of them. The government in this
country can not take such a part as this. The matter can only
be pushed by bringing it before the learned societies. In this
country a subscription of $250 in addition to what has already
been raised is all that is asked. This is not an exorbitant share,
[am sure, The German Academy of Sciences, let me say, has
voted a subsidy to the undertaking. There are three societies
here that can be looked to for a subsidy—the Association for the

AMERICAN MICROSCOPICAL SOCIETY. 85

Advancement of Science, the American Society of Naturalists,
and our own society. ' What else is collected must be from
private persons or local societies. I know it is difficult to raise
even a small sum. The executive committee has discussed it,
and it is easy to see that the society is not rich. But I should like
to ask the society for $25—a very small sum. I ask this more
for the reason that I want to see the society in line with this
work, which is for the benefit of the whole world and of every
worker in this country.

Second, the bureau asks that we send a free copy of our
publications. And third it asks subscriptions for its publications,
but of course this concerns private persons rather than this society.

The President: If the chair may be allowed, I should like
most heartily to second this motion. I may add that since the
meeting of the executive committee, I have received a letter
from Dr. Kingsley saying that the combined societies in the
American Society of Naturalists have given a very cordial
endorsement of the movement, and that he counts on us for help.
“Cannot and will not you help us?” is the last sentence. It
seems to me that our society which publishes proceedings, wishes
them to be made known widely in just this way—we do not
want to be ina corner. I am happy to say that on talking with
some members of the society I can say that if this vote is passed
by the society it shall not come out of the treasury or from the

ordinary income of the society. It shall have the honor, if it is
such, of making the gift, but it shall not bear the expense.

All those in favor of giving our proceedings to this Bibliog-
raphical Bureau, and of granting a subsidy of $25, please say
aye. |

The motion is carried.

(It was stated by Professor Ward that this subsidy was for
one year only and did not bind the society after that.)

The President: I think that finishes the business of the
society, except for what we recognize as a pleasant function as
well as one of our duties, and that is our words of appreciation
for the kindness of the people of the place at which we meete

86 PROCEEDINGS OF THE

The first vote, I think, we all recognize, should be given to the
University for the welcome that has been given us, and for the
facilities offered for this meeting.

Professor Ward: We must all recognize that the success of
the meeting has been largely due to the place in which it has
been held. Entirely aside from the fact that Cornell University
has honored us with “A.M. S.,’ witness these buttons—and a
Master’s degree from Cornell University is a matter of congratu-
lation certainly for each of us—entirely aside from that, the cor-
dial reception which has been given us by the officers of the Uni-
versity, the way in which everyone has made us feel at home
within these walls, will cause all to carry away a feeling that
Cornell University keeps a unique place in our thought. To
those of us who are college men no affection can be quite like
that which one feels for his alma mater, but I must confess that
after the very pleasant time which I have had here, there is a
second place, comparatively near that, which will be given to
Cornell University ; and it is with a great deal pleasure that I
move that we extend to the President, the officers and faculty of
Cornell University our sincere thanks for the very cordial invita-
tion and reception which they have given us, and for the facilities
which they have placed in our hands, aiding. us in holding the
most successful meeting of the society which it has been my
privilege to attend.

The President: You have heard the motion, ladies and gentle-
men. All those in favor of this motion will rise—that is the
kind of voting one likes to see.

Next to the University, I think, we have to thank our local
committee. If anyone has ever beena member I feel quite sure he
would agree with me in that.

Mr. Seaman: Mr. President, having been secretary of the
Society several years, I think I am in a position to say some-
thing about the local committee. The first year in which my
active duties as secretary began was at Washington, and there we
had an active local committee. The next to the last year of my
duties was at Brooklyn, where there was no local committee and

AMERICAN MICROSCOPICAL SOCIETY. 87

nothing else. Now the difference between those two extremes
of conditions measures the difference between a successful meet-
ing of the society and a failure. There is nothing that can make
up for a want of interest in the place where the society meets, as
regards some at least of the conditions of a successful meeting.
Of course, if the society is strong in members, and those members
attend from all over the country, they can have a good meeting
where there is nothing but a house to cover them. But that is
not usually the case. There are certain requirements of attend-
ance, of providing for the comfort of members, of showing inter-
est in the meeting which contributes enthusiasm to those who
come from a distance; and more than that, if we look over the
meetings of the Microscopical Society, you will see that on
several occasions in the early days of the society the meetings
have been redeemed from failure by the activity of one or two
men in the place where the meeting was held. I think that I might
refer to the meeting at Elmira particularly as an instance. So
the meeting at Columbus, Ohio, which was one of the first that
I attended, was such that it seemed to me at that time that the
society had touched a very low ebb, and had it not been for the
efforts of one or two of the local members there would scarcely
have been any meeting. Now ina society that is in the early
days of what we hope will be a prosperous future, the local com-
mittee is one of the principal pillars upon which it must rest.
And in the case of our meeting here, I can say most heartily and
earnestly that the local committee has done the largest part of
the work, that to it is largely due the success of this meeting.
I therefore offer a resolution of thanks to the local committee from
the very bottom of my heart, that its members have lightened
my labors and made the meeting a success.

The President: I think that I will proceed with the original
method and not take a vote, because I am sure that is in the
heart of every one of us.

Next to the local committee we have to thank the city of
Ithaca. As I said in my remarks in answer to the address of

welcome, we had not hoped so very much from the city. That

88 PROCEEDINGS OF THE

was wrong in us. We misjudged the city. When we went to
them we found their hearts open, and they stayed open; and the
next time we have a meeting we shall not go with any doubt to
the men of the city, but with full confidence. I hope there will
be given in the heart of every one of us a hearty vote of thanks
for the city, and I would call upon Dr. Moody, who was a
student here for many years, and who knows the city better than
I do, to say a few words to us concerning it.

Dr. Moody: I must confess that I felt a little surprise when
our President announced that he approached the people of the
city with what we might call fear and trembling, with regard to
the reception of the society ; because it was my experience as a
student here that when anything was started on the campus the
town was always behind us. Surely in this case it was so. I
heard the chairman of the local committee the other day on the
street telling what great success he had had in the work he was
doing among the citizens. I am sure we all felt from the time
that we were welcomed by Mr. Van Vleet at the opening of the
meeting, that the hearty welcome he gave us, came from
his heart and from the hearts of the people—we felt that their
hearts were open to us. And Iam sure as we sailed down this
beautiful lake yesterday afternoon, amidst your delightful breezes,
we all felt then that the citizens had an interest in us, and that
the street-car company also, which carried us up and down these
hills, had an interest in us. I feel that each one of us will be
glad to come to Ithaca again, where we have received so warm
a welcome, to accept the invitation which has been extended to
us indefinitely for some future time. I therefore move a vote of
thanks to the citizens of Ithaca for their cordiality to us.

The President: I would like to put the motion that has just
been made, namely: that we render the citizens of Ithaca a
hearty vote of thanks for their courtesy to us, and their warm
welcome to the city,

The motion is carried.

The President: When the chairman of the local committee
and myself were discussing this meeting, the question of the press

AMERICAN MICROSCOPICAL SOCIETY. 89

arose immediately in our minds. What help could we get in.
giving information about the society, and for the encouraging
and interesting of the people here in Ithaca in the society? The
chairman of the local committee went to the papers in Ithaca, and
the welcome there was cordial. You who have seen the papers
since you have been here I know cannot help feeling that the
press of Ithaca has done everything that it possibly could to
make our meeting asuccess. They have acted, as the Honorable
Mr. Van Vleet said in the beginning, as though they felt honored
in the presence of the society in their midst, and they have done
for us, if one remembers the limited space of the great papers
nowadays, a great deal. I would ask some one to say a few
words on what the press has done for us in our meeting.

Mr. Plaum: A sure test of the grade of civilization of a
race, of a nation, lies in the short phrase, ‘‘ What’s going on?”
—both in the question and in the answer, and especially how
that answer is recorded, so that neither wind nor weather, nor
time, even, can ever blot it out. The question with the savage refers
to immediate bodily needs, and the answer dies with the acqui-
sition of his physical requirements. With advancing civilization
the question broadens in scope, and takes in friend and neighbor,
and gradually it leaps over territorial bounds till finally it takes
in all of society, and records its answer for future generations.
How painful is the attempt of our remote ancestors to tell us
what was going on. The invention of form writing, of hier-
oglyphics, to tell even a simple event, must have taken years of
the writer. With the invention of the alphabet the recording of
that answer was made more easy, but the process remained at
best a rude and slow one. It was the invention of printing that
gave the greatest aid. The art of printing gave an impulse for
more questioning and a wider answer. To-day the question and
answer to “‘ What is going on?” has become centralized in the
most civilized nations, and their standing is measured by the
quantity, quality and freedom of that grand modern organism
known as the press.

Our society most gratefully acknowledges that our work and

go PROCEEDINGS OF THE

aims have been recorded and spread broadcast as part of the
answer by the press to the question, ‘“‘ What is going on?” We
have been aided and benefited, and our labors lightened by the
help that is has afforded. The press of this city has nobly per-
formed its functions, it has faithfully recorded our endeavors to
add our part to human advancement. We appreciate the kindly
feeling and generous spirit of the press of Ithaca, and tender it
a most cordial au revoir.

The motion of thanks to the press of Ithaca was put and
carried.

Professor Rowlee: I should like to make a few remarks in
regard to the privileges that have been extended to us in another
relation, and I think I can do so without blushing. I think many
of the members do not understand how much we have to thank
the railroads and the street railroads of this city for the success
of this meeting. It was with a good deal of embarrassment
that I asked for the privilege of reduced rates to this meeting.
I feared that there might be some question as to whether we
were entitled to the privilege. I expected when I went to meet
the agent of the Trunk Line Association to find a strictly
business and railroad man, who would be very strict in his
treatment of us. But when I met the gentleman his first words
were: ‘We want to do for you people everything that we can.
We realize that you are working for the advancement of civilization
in this country, that you are doing it freely, and we want to do
everything for you that we can.” He has treated us in every
way most cordially. I am sure that any man who has to do
with this gentleman, or with the other railroad men of the
country, in regard to meetings for the furthering of human
knowledge, will have a very cordial reception, and he should not
be afraid to ask for anything.

In regard to the street railroad, the mere asking was the
having of the privilege of your riding free over their lines to the
lake. I did not ask them to carry you free all the time you
were here [am not sure but they would if I had asked them.
The President of this society has often reminded me that when

AMERICAN MICROSCOPICAL SOCIETY. gt

he wanted anything done that required a great deal of courage |
—he did not call it courage, however, he called it cheek, but I
think courage is a better word—he sent for me. But my
courage failed when I thought of asking free transportation
during the whole meeting.

I move that we tender a vote of thanks to the Trunk Line
Association of the Central States, and to the Street Railway
Company of Ithaca for the courtesies which they have extended
to us during our meeting.

The motion is carried.

The President: This completes our program, I believe, with
one exception, and that is the pleasant duty of the President to
resign his chair to the incoming President. I wish to say that
I have only sincere gratitude to the society for the cordiality
that I have received at their hands. I hope that the incoming
President will receive the same. I feel sure that he will, and I
think that if we go on with the society, each one of us trying to
do the very best he can, going in the spirit of true scientific
brotherhood, there won’t be any question ever arising as to
whether we are being treated properly or not. We won't think
anything about it. It is like the best kind of digestion ; we don’t
know that we have any stomach while we are all right. It is
only when we are dyspeptic that we know it—that we feel as the
little boy did after the Thanksgiving dinner, when he asked his
mother if the turkey was alive again. There will never be any
question of that kind with us. Weare helping our friends ; they
are helping us. I am frank to say that a great many of the
things that have been stated on this platform have given me the
greatest help, and real inspiration. It perhaps may not be
modest for me to say it, but it seemed to me as [| listened to
these papers, that in a great many of the societies I have attended
before, we have not had any that have exceeded these, to say
the least. If I may say in a general way, not to be egotistic, it
has done my heart good to see the genial look on the faces of
the people as I looked into them. Nobody seemed to feel that
he was being abused; nobody looked as if he was not enjoying

92 PROCEEDINGS OF THE

himself. Everybody seemed to think that things were going
rightly, and that look encourages the presiding officer and the
one who is reading the paper, and thus it makes the meeting a
success. Dr. Mercer—when acting as President, the best I can
say for you, the best wish I can have for you, is that you will be
accorded the same pleasant treatment.

Mr. Milnor: I think that one thing which always makes
a great deal, at least. depends upon it
—is the manner ia which the presiding officer discharges his

these gatherings pleasant

duties. And I think we all would like to express our apprecia-
tion to the President of the past year for the manner in which he
has discharged his duty, his courtesy and the way in which he
has made us all feel perfectly at home. I regret exceedingly
that this task of expressing to him our feelings, the most
pleasant one, I think, of all, has not been placed in the hands of
some one more eloquent than myself, one that could speak forth
the sentiments that I know ‘have filled the hearts of every one
who has attended these meetings. I therefore with the utmost
pleasure—it is one of the greatest pleasures of my life—move
that a standing vote of thanks be extended to Professor Gage
for the manner in which he has discharged his duty to this
meeting. :

( Applause and a standing vote of all the members.)

President Gage: I thank you gentlemen, most heartily, for
this vote of thanks.

Inaugural remarks of the new President.

President Mercer: /ellow members—I thank you for the
honor that you have conferred upon me by your choice. It is
an honor which I feel, and an honor at the same time which I
know perfectly well means something in the way of work. So
far as I am able to work for you I shall be happy to do so.
I hope that your wishes will be freely expressed in one and
another, so that I may know what to do.

If I could carry that face, which is beaming and smiling before
us, as our old President did, I have no doubt I should see beams
and smiles in the faces all around me. He forgot in wishing for

AMERICAN MICROSCOPICAL SOCIETY. 93

me the same cordial reception, that he but saw round about him
the reflection of his own face. I think it is a rather hard thing
for a successor of Professor Gage to get those smiles. However,
I am at your service and your pleasure.

The society then adjourned.

Friday evening, August 23, at eight o’clock, the society held a
soirée in the University gymnasium and armory, which was
attended by a large number of visitors from Ithaca. About sixty
microscopes were in use, and those present expressed a great
deal of pleasure at the opportunity afforded to examine objects
by the microscope. As one of the incidents of the occasion,
Professor Burr, who entertained the society so pleasantly
with his literary treasures, assured the writer that, although
connected with the university several years, he never before had
looked through a microscope. This entertainment closed one
of the most successful meetings the American Microscopical
Society has ever enjoyed.

[ The letter of Dr. Seaman resigning the position of editor and
secretary of the society, also the resolutions expressing the
society’s thanks and hearty approval of his labors, have been
misplaced aud could not be obtained by the present secretary to
insert in the proceedings —W. C. K.]

TREASURER'S” REPORT

FOR YEAR ENDING AUG. 21, 1895.

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MAGNUS PFLAUM,
Treasurer.

We hereby certify that we have examined the foregoing accounts, and
find the same correct with proper vouchers for expenditures.

F. W. KUEHNE.
Ithaca, N. Y., August 22, 1895. D. S. KELLICOTT.

SOME NOTES ON ALLEGED METEORIC DUST.

MacGnus PFLaum, Pittsburg, Pa.

The Spectroscope has made known that matter is universal ;
that elements constituting the earth are also contained in other
celestial bodies. It is, however, very human to endeavor phys-
ically to perceive what mentally we know, especially if the
object is from extra-terrestrial spheres.

This feeling of curious interest must have animated the
members of the Calcutta Microscopical Society, when, at one
of their meetings, a substance was presented to them described
as follows :

“The coarse-grained, dark-colored dust which drifts, as all
“Calcutta residents are aware, into the leeward corners of our
“flat house-tops, is found to contain, mixed with dried portions
‘of organic matter, certain magnetic and hyaline granules which
‘““present appearances highly indicative of previous fusion.
“These appear mostly as opaque or translucent spheroidal
‘bodies, sometimes single and isolated, and sometimes composed
“of more than one spherule fused together, varying in size from
“to'c0 tO 35 Of an inch in diameter. The opaque bodies vary
sail Pee from black to rusty red, whilst the transparent spheres,
“by transmitted light, are light brown or yellowish like colored
“glass. They frequently include bubbles, patches of granular
““matter, and more rarely crystals. The surfaces are usually
“smooth, and occasionally bubble-like protuberances bulge out
“on the sides. Composite grains are not uncommon in which a
‘“‘ glassy mass protrudes from an opaque body. Particles present-
‘“‘ing these characters have been obtained from house-tops widely
‘separated from one another and at considerable height, as on
“the tower of the High Court, facts which are of importance
“in considering any ‘conclusions as to the origin of this
“‘ material.’’*

The members inclined to assign to these particles a meteoric
origin.
# Am. Mo. Mic. J., Vol. XIV., page 72. _

96 PROCEEDINGS OF THE

It was no great leap of thought to cause the writer to speculate
upon the possible presence of such “meteoric matter” in the
Court House tower at Pittsburg, Pa., though it required more
“than a leap to carry that thought into action, by climbing the
steep ascent to the top floor of the tower. This floor is about
250 feet from the street, with open, narrow windows on all sides,
allowing winds to pass in and out in all directions. For this
reason comparatively little dust was found in the floor corners.
But what little was obtained was an ‘‘ omnium gatherum,” indeed.
What it did not contain might be easier to mention. But a
magnet separated the desirable matter from the dross, which,
upon being cleaned, showed the presence of the identical globules,
iron and hyaline, described above, as found in Calcutta.

But were these little strangers sent as visitors from some
celestial body ? Doubtful. Whether as solid or gas they could
not have left their former abode. The law of gravity would pre-
vent even heavier particles to leave a starry globe. Why could
they not be made artificially and thrown into the air by glass
houses and iron works? Pittsburg having both of these
industries in abundance, it seemed proper to make a practical
investigation. A visit to a glass house at once proved that no
glassy globules can be sent out of the ‘‘ Clay Pots” wherein
glass is melted. The pot is almost hermetically sealed. Fire
surrounds the vessel, merely melts, does not vaporize the con-
tents. The pot has no opening permitting glass, or any possible
vapor, to escape intoa chimney. However by way of precaution
some dust was taken from the opening in the pot out of which,
when ready, the glass is lifted. Upon examination nothing but
grains of sand and fine strings of glass were found. Next, a
blast furnace was visited, and a quantity of ‘ flue dust ” obtained ;
but in it nothing but ragged pieces of coke and iron found. The
Bessemer converter yielded better results. The cold blast
admitted causes a seething white heat in the metal and drives
out all impurities, which on reaching a colder stratum of air, are
solidified in the form of spherules, and the ground around is
covered with what looks like gunshot of all sizes, some

AMERICAN MICROSCOPICAL SOCIETY. 97

microscopical. These would answer our quest, were it not that.
no hyaline globules could be found in the gathering.

The next gathering was from the refining department of a
rolling mill. After three days of showers, the roof having
thereby been well washed, a quantity of fresh deposit was pro-
cured. This, upon being cleansed, produced, ’midst iron and
other particles of all shapes, the identical globules in large
quantity, exactly in every particular described as found in
Calcutta. There were not only reguar iron and hyaline spheres,
but all-the various zregu/ar particles, some even with both
hyaline and iron excrescences. There being no doubt that these
pretty little bodies are a product of iron mills the question
remains, how did they come to Calcutta? If there are no iron
works there, then they must be blown across from Europe.
The winds blow easterly. In iron works are high chimneys witli
strong drafts. It is therefore not at all unlikely that these
particles are sent up a comparatively high altitude, received by
easterly winds and deposited in regions beyond.

SOME MODIFICATIONS OF STEMS AND ROOTS FOR PURPOSES
OF RESPIRATION.

HERMANN VON SCHRENK,

The conditions surrounding many of our swamp and marsh
plants are such as to render the free absorption of oxygen a com-
paratively difficult matter. This applies particularly to the
organs growing in the mud and water, the roots, rhizomes and
bases of stems. In the majority of land plants oxygen is taken
up by these organs through the epidermis by diffusion, and is
carried from there to the cortical parts where it is needed. In
the water this free diffusion is rendered difficult and often nearly
impossible. In such cases we find the plant adapting itself by
varying and changing its structure, so as to enable it to grapple
with the new conditions.

Such variation may be in one of several directions (5), by
increasing the absorbing surface, by facilitating diffusion by means
of large intercellular spaces, or by producing organs or tissues
better fitted to absorb the necessary gases. Inthe extended root
system of Mikania and some of the palms we find an example
of the first change, the stem of the water lilies may represent
the second, and the tissues of Decodon Verticillatus ( Nese@a Ver-
ticillata) Jussi@a complete the series..

Whether these plants, z. ¢. species, which are so modified, have
shown such structure from the beginning of their existence is a
question. We may recall the explanation of Shaler (1) with
regard to the cypress knees. He considered the cypress a tree of
earlier geologic times than our own, crowded into the swamps at
the present time by more favored rivals. Here, under the
adverse conditions of respiration, it developed the knees. On
the hillsides, where the old conditions prevailed, no knees were

formed. In the case of Decodon, Lycopus and others, I believe,
we see something akin to this.

AMERICAN MICROSCOPICAL SOCIETY. 99

There are at present, then, a number of piants which have, in
their later stages, contrivances or organs enabling them to obtain
oxygen more easily than without such contrivances. The latter
have become, in the majority of cases, recognized characters of
their species, such as Decodon, Jusst@a, etc. The remaining
ones seem to have a greater plasticity of organization in this
respect. During the past year I had occasion to observe the
habits of several of this class. Lycopus sinuatus, Ell.is a labiate
growing ina variety of habitats, from dry borders of woods to
deeper swamps. If a specimen from a dry location be taken,
one will find the lower portion of the stem and the rhizome of
about the same size as the aérial stem. The characteristic
sclerenchyma fibres in the corners of the stem lie immediately
below the epidermis. Specimens from a swamp show a very
different structure. The lower portion of the stem is very much
swollen apparently, decreasing in size as one leaves the water
going upwards. Examination will show the epidermis ruptured
in many places, often entirely gone, exposing to the surrounding
air a mass of white, spongy tissue extending to the vascular
ring. The tissue consists of elongated cells linked by smaller
cells, forming a coarse network, as it appears, with large inter-
cellular spaces, cavities one might almost call them. The origin
of this tissue is similar to that found in Decodon (3). That the
formation of this tissue is due directly to the position of the plant
in the water, I am thoroughly convinced. Many specimens were
collected near Southold, Long Island, growing at the edge of a
pool in sphagnous ground. These had the “‘zerenchyma”’ tissue
developed to a large extent. On a tree stump in this sphagnous
ground other specimens of Lycopus flourished a foot or more above
the water level. These had zo sign of the tissue. In all there were
some 20 individuals on this stump. Other stumps near by and
in neighboring swamps gave the same result. A fact which I
have observed, but am at present unable to explain, is that @//
individuals of this species growing in damp places do not develop
the zrenchyma, as all individuals of Decodon do, for instance.
One will often find in the same swamp some with, others without,

100), PROCEEDINGS OF THE

this tissue. I believe in some parts of the country the Lycopus
never develops this tissue ; in Central New York I was unable to
find it.

The same may be asked of another plant, Ludwigia Sphero-
carpa. This plant grows luxuriantly in the muddy borders of
some ponds on Long Island. I collected it at Manor, L. lL.
The stems grow out from the mud with a layer of zrenchyma
nearly one-half inch in thickness around them, the layer extend-
ing nearly afoot above the surface of the water. Its appearance
is in every respect similar to the layer in Decodon, so similar that
I mistook the plant for Decodon several times. I have not col-
lected this plant sufficiently to say that it may grow without
zrenchyma, as does Lycopus sinuatus, but it evidently does, to
judge from such notes as ‘‘Bark below often spongy-thickened”’ *
(italics my own) and others of similar nature. Some individ-
uals on the bank in moist sand had less of the tissue than those
in deep water.

Before proceeding to another group of plants I would record
an instance of Decodon Verticillatus growing in a dried-up pond.
I had no means of telling absolutely how long the pond had
been dry, but it appeared to have been partially filled early in
spring. The clumps of Decodon grew a foot above the pond
bottom on tufts of Careces, many of the stems having bent over
in the characteristic manner. The bases of the stems showed a
very slight development of azrenchyma tissue, but every plant
had some of it. The tissue was shrunken and to all appearances
was dead, and had been so for some time. This is:what one
might have expected, the water gone, the need for aérating tissue
was no longer there, and it ceased to function. It will be of
interest to follow these plants in their next year’s growth.

Besides the plants noticed so far, we find modifications for
respiration in many shrubby plants, which may, as respects the
adaptation, be classed with those in Lycopus, etc. The most
striking example of this which I have been able to find, is the
common elder, Sambucus Canadensis, L. This is in the true

* Gray’s Manual, p. 188.

AMERICAN MICROSCOPICAL SOCIETY. IOI

sense of the word not a swamp plant in most places. Its habitat
is given by Gray as “rich soil in open places.” It grows along
fences and along stream banks. In Long Island its favorite
habitat is on the borders of inlets from the bays and marshy
banks of streams and ponds, oftentimes in water a foot deep.
Fig. 1 represents a small plant taken from a pond near Eastport.
One is struck by the great swelling which the stem has evidently
undergone below the water. (A-Z is the water line, the stem
having grown somewhat obliquely.) All over the surface are
snow-white excrescences of a warty nature, varying in size and
shape. These decrease in size and number as one goes upwards
from the water,and some inches above it one finds them merging
over into the ordinary lenticels (Fig. 1, Z.). The white spongy
tissue is at once proven to consist of cork cells (Fullzellen) pro-
truding from a structure of lenticellular character. These cells
are developed from a meristem at a surprising rate ; in many cases
the protuberances were two inches in length, three-eighths inch
wide, extending three-eighthsinch above the surface. Fig. 3 repre-
sents a cross section of a small one of these water lenticels (2).
It will at once become evident that we have here a modified
lenticel. A meristem (7) forms beneath the epidermis, before the
appearance of the phellogen layer, which gives rise to the ordi-
nary ‘‘ Fillzellen”’ (/'). These are formed in rapid succession
until the epidermis bursts. The activity of the meristem con-
tinues, pushing out large masses of these cells, which finally appear
as a column between the two lips of the Ienticel, the ruptured ends
of the epidermis. Meanwhile the phellogen layer (phe/) has
formed and has given rise to the periderm (gerd). Our figure
shows a series of periderm cells formed, which are being crowded
back by the rapidly increasing number of cork cells. The latter
are filled with air, giving the whole mass its white appearance.

The growth of these structures continues, several unite, and
then more, and we have at last long patches of this tissue of
loosely connected cells extending all over the stem, leaving but
little of the original bark (see Fig. 1) in position ; it would take
but little to form a continuous layer around such a stem, produc-

102 PROCEEDINGS OF THE

ing not peridermal cells, which resist the entrance ofair, but a tissue
exceptionally well fitted for the absorption of oxygen.

The water lenticels are by no means confined to the stem, but
are very often found on roots. Lenticels occur but rarely on
roots, and then only sparingly. But here we find them produced
abundantly. The submerged stems develop large numbers of long
fibrous roots, which branch sparingly. The roots grow out
horizontally, rarely into the mud below. On them the lenticels
are found. They differ in no respect from those found on the
stems, except perhaps in size. A noteworthy fact is that in many
cases the adventitious roots produced by the stems break through
these lenticels (Fig. 1, c), and sections show these roots growing
directly toward a lenticel from their very differentiation. This
directive influence of the lenticel on such roots has often been
noted, but is especially striking here where the lenticel has
assumed such large size.

In addition to the water lenticels the Sambucus stem presents a
modified cortex. The latter for some distance from the water
consists of round cells filled with active protoplasm and numer-
ous chlorophyll grains. The phellogen produces phelloderm
cells (f/d.) and thus increases the thickness of the cortex.
Intercellular spaces are few and comparatively small, even under
a lenticel. As one goes down the stem towards the water, these
spaces become larger. The cells of the cortex, still green and
full of protoplasm, become separated, and in the lower portions
they form a loose, spongy tissue, very different from the cortex
further up the stem. Here, too, the cells are more numerous, and
this, together with the air spaces, makes the cortex seem twice
as thick. This gives the whole stem the appearance of being of
so much greater diameter in the water.

Fig. 3 represents a section taken near D, Fig. 1. Here the
air spaces are quite marked (sf). They appear to originate near
the phellogen layer, the new phelloderm cells parting from one
another soon after their separation from the mother cell. It will
be seen that this system of canals filled with air, thus closely
surrounded by active protoplasmic cells, must insure to the fullest

AMERICAN MICROSCOPICAL SOCIETY. 103

extent the free diffusion of oxygen. Those passages are in close
connection with the outside through the water lenticels, which
offer little resistance to the available oxygen (Fig. 3). The
exact manner in which the lenticels act to absorb the oxygen of
the water is a problem yet to be solved. We have thus an
arrangement whereby the stem is enabled to obtain oxygen in a
manner both striking and efficient.

It may be asked, is it proven that these structures function as
respiratory organs? The fact does seem certain, as_ the
large quantities of air in the spaces and their connection with the
outside seem to indicate. Another proof is seen in the fact, that
they are entirely absent in plants not growing in the water. In
individuals of Sambucus Canadensis taken from dry soil, I was
unable to find any indications of the large intercellular spaces or
the large development of lenticels.

Other shrubs and trees show similar changes. Cephalanthus
Occidentalis is perhaps next in order. Its cortex shows spaces
similar to those of Samducus (Fig. 4). In these we find individ-
ual bast cells (4) torn away from the adjoining cells, some hang-
ing loosely in the air space. The water lenticels are present in
great numbers (Fig. 2), but differ in some respects from Sambucus.
They are long, very narrow and occur in patches along the stem.
The masses of ‘“ Fillzellen’’ extend further from the stem than
those of Sambucus, sometimes as far as one inch from the bark.
The roots of this plant were well supplied with lenticels.

Some of the larger trees showed the water lenticels in large
numbers. On the roots of Populus monilifera they protruded
from among the many fibrous roots. These lenticels showed
the lateral portions of the structure well formed. On the roots
of Acer rubrum, on which I first noticed these organs, they are very
Numerous, some 20 or 30 to the square inch. These are
perennial. At the end of. one year the cork column is perhaps
one-eighth inch high. A layer is then formed, similar to that in
the ordinary lenticel, closing the opening. This is pushed out
in spring, with the cork of the preceding year, and a new piece
is added to the column. This process may continue for many

104 PROCEEDINGS OF THE

years, so that an old root presents a peculiar appearance, especi-
ally in quiet water where the cork columns are not broken by
the flow of water against them.

How many other plants may show these modifications one can-
not tell, but I am confident that many more than we are at
present aware of, will, upon close -examination, prove to possess
to a greater or less extent adaptation furthering respiration. The
cases noted, especially Sambucus and Lycopus, show a plasticity
of organization which seems striking. It would seem that to
effect a change so marked upon a species, it would necessitate
the continued action of the environment upon individuals. But
here we have instances of plants responding to this action in the
short space of two months or less. Exactly how soon this
influence would make itself felt experimental evidence must bring
forth. Individuals should be grown under different conditions
of moisture, and hence of exclusion of oxygen, and the result
ought to explain some of the questions concerning which we are
stillin doubt. The function of respiration has often been under-
estimated, and it seems that modifications of this kind would
tend to emphasize the necessity of this function for the perform-
ance of the life activities. When plants of such different organi-
zations produce changes of this kind, so marked and so con-
stant, the importance of the end striven for must be recognized.

AMERICAN MICROSCOPICAL SOCIETY. 105

pIbLIOGRALTEY

Shaler, N. S.—Notes on Taxodium distichum. Science, 1883.

Schenck, H.—Uber das Aerenchyma, ein dem Kork homologes Gewebe
bei Sumpfpflanzen. Pringsh. Jahrb. f. Wiss. Bot., Bd. XX., 1889
p. 526,

Schrenk, J.—On the floating tissue of Nescea Verticillata. Bull. Torr.
Bot. Club, Vol. XVI., 1889, p. 315.

Wilson, W. P.—The production of aérating organs on the roots of
swamp and other plants. Proc. Am. Acad. Nat. Science, Philadelphia,
1889.
Rowlee, W. W.—The Aération of Organs and Tissues in Mikania and
other Phanerogams. Proc. Am. Micros. Soc., XV., p. 148.

(NotTe.—In this paper will be found a full list of papers relating to
this subject.)

EXPLANATION OF PLATES. —

G57) DURES eter
Fig. I. Sambucus Canadensis, Ts; base of stem with wa
A-B, water level; L, lenticels, C, root growing from a lentice x
Ss =

PEAT i

Av. 8. aD Nat DEL

’
‘ im
Af .
j
*
v= -
‘
a,
‘
|
_

108 | PROCEEDINGS.

PLATE II.

Fig. I. Cephalanthus Occidentalis, L., base of stem showing upturned
bark (B); at A some of the cork columns are seen projecting out from the
stem; C, root with water lenticels. x

PEATE, It:

sy

oP

— ai

a

bm xe) PROCEEDINGS.

PLATE Ill.

Fig. Ill. Sambucus Canadensis, L.—transverse section through a water
lenticel (about D, Fig.1). , ‘‘Fiillzellen;” m, lenticel meristem; perd,
periderm; p/el, phellogen; phd, phelloderm; sp, air spaces; 0, bast cells; ¢,
cambium; w, wood. can

Fig. IV. Cephalanthus Occidentalis.—Section taken immediately jinside
a water lenticel showing the air spaces, sp, with isolated bast cells, b.

Pia hia ie

l
4

If
i
i

al
pal
ooh)

Hy S. swat vex

SOME PECULIARITIES OF THE MOUTH PARTS AND OVIPOSITOR
OF CICADA SEPTENDECI/1.

Proressor J. D. Hyatt, New Rochelle, N. Y.

The year 1894, being the seventeenth since the last appearance
of this insect in the vicinity of New York, the 25th of May
seems to have been the day unanimously agreed upon by the
broods in this neighborhood to leave their subterranean abodes
and emerge to the light of day.

Although some skirmishers were seen in the woods a day or
two before, and many camp followers continued to appear above
ground after that date, yet the main army consisting of incalcu-
lable myriads came forth on the night preceding the 25th day
of May, and for several weeks they swarmed in certain localities,
over the trees, shrubs and foliage of all kinds.

It is not my purpose to describe this curious insect, or its
habits, as such description with its life history may be found in
any elementary work on entomology, but during its prevalence
alarming reports were frequently made in the newspapers of the
great damage done to crops, and to fruit and forest trees, by the
seventeen-year locust, and as nobody contradicted these state-
ments, the newspaper reporters manufactured ‘and _ published
circumstantial accounts of persons, preferably children, who had
lost their lives by being bitten or stung by these ‘“‘ venomous
insects.” While fully aware that these reports were purely
imaginary and sensational, my curiosity was stimulated to make
a microscopical examination of the “ biting and stinging ”’ organs
of Cicada, an additional motive being the opportunity, which
occurs at such long intervals, of obtaining abundant material for
the study.

It is hardly necessary to say that after handling and examining
hundreds of specimens, and carefully studying their habits, I
found no evidence that they ever bite or sting, or that after

DL PROCEEDINGS OF THE

reaching the winged state they ever do the slightest injury to
vegetation of any kind, but as I had never seen a description of
the mcroscopical appearance of the mouth parts and ovipositor,
I spent much time in making a careful examination of them, the
sections notably showing some curious features.

As this insect belongs to the natural order Hemiptera, the
mouth parts in a general way are typical of that order, the
mandibles and maxillz being drawn out into greatly lengthened
stylets or setee and enclosed in the labium.

Fig. I represents the extremity of the labium enclosing the
four slightly projecting stylets, x 120.

(To show the lengths of these parts under the same ampli-
fication, it would be necessary to make the drawing 30 inches
long.)*

The outer hooked pieces serve as anchors, which being inserted
into the tissues of a plant afford a leverage for forcing in the two
central stylets, which together form a sucking tube.

Fig. 2 is a transverse section through the labium and the four
stylets, showing the very curious manner in which a tube is
formed by holding in juxtaposition the two inner grooved pieces
and closing the laps by the outer pieces of the same sectional
form, all being held firmly together by the muscular labium
which is closely wrapped around them. This will be easily
understood by an inspection of sections of the four stylets as
seen in the lower figures.

While the mouth parts of Cicada are perfectly adapted to the
purpose of sucking the juices of plants, yet in the examination
of thousands that I have seen upon cherry, pear and other trees
I have never been able to discover one in the act of feeding,
either on the fruit or foliage.

The ovipositor is the instrument used by the female for making
incisions in the twigs of trees in which to deposit her eggs. It
consists of three parts, the two outer ones being tubular with an
opening near the extremity, through which the eggs are
extruded into the channel cut for their reception. They are

*(The original drawings were reduced one-third by the engraver.—ED.)

AMERICAN MICROSCOPICAL SOCIETY. 113

pointed at the ends, and a little anterior to the extremity are
somewhat enlarged and set with sharp teeth like a rasp, which —
extends spirally around the teeth pointing backwards.

Fig. 3 represents one of these saws, a being the outlet of the
oviduct which opens inward or toward the central piece.

The central piece which serves to hold the two Ovipositor
Saws in place and guide them in their movements, ends in two
extremely sharp points slightly curved outward. The insect in
cutting the channel for her eggs closes her legs around the twig and
forcing in the ovipositer saws beneath the bark and into the soft sap
wood, works them rapidly backward and forward cutting loose
but not removing the wood fiber. In doing this the outward
curves of the central piece causes the saws to spread so that she
cuts at the same time two channels, which diverge from the
entrance, leaving a ridge of solid wood between the two, nearly
a tenth of an inch wide at the extremity, while the channels
closely approximate at the entrance.

After finishing the cut, which is about three-tenths of an inch

in length, she withdraws the ovipositor, and forcing it in at the
8

114 PROCEEDINGS OF THE

first entrance to the greatest depth proceeds to deposit the eggs,
which are placed very symmetrically in pairs obliquely to the
middle partition, a little cavity being cut for each egg, into which
it exactly fits. The eggs are about 15 in number on each side.

As these insects are not at all timid, I was able to watch this
process with a large lens.

The extremely curious mechanism by which these processes
are accomplished will be easily understood by inspecting Fig. 4,
which is a section of the three pieces constituting the ovipositer,
in which aa is the central piece, having on each side, through-
out its length, a projecting rail or guide-bar which exactly fits
into a groove of similiar shape in the ovipositor; 0d are the
oviducts, which are bounded on the exterior side by a chitinous
frame, and have for their interior boundary the same material for a
short distance above the Z7-shaped groove, but this thins out into
what is evidently a muscular or contractile tissue above.

The two ovipositors are held together along their outer edges
by a peculiar pair of folds, which in section resemble a strong
hook on one side, grasped by a hand and thumb on the other.

As the insect, in cutting the groove for her eggs, sometimes
spreads her ovipositors widely apart, and sometimes uses them
close together, it is evident that she has the power at will to open
the hand seen on the left, or close it in the hook, as the particular
stage of the process may require.

‘THE LATERAL LINE SYSTEM OF SENSE ORGANS IN SOME
AMERICAN AMPHIBIA, AND COMPARISON
WITH THE DIPNOANS.

B. F. Kincssury, Pu. D., Defiance, O.,
Fellow in Vertebrate Zoilogy at Cornell University.

The general structure of the individual sense organs is ‘well
known, and their distribution has been worked out for a con-
siderable number of the Amphibia. The availability of many
Urodeles, some of them important and isolated forms, as
Amphiuma and Siren, led to a study of the distribution of the
organs in eight of the tailed Amphibia on which nothing had
been published, with the purpose of determining their presence
and the plan of distribution. A comparison with the condition
in the Dipnoans, Lepidosiren and Protopterus was made, and they
were added to the Amphibia first studied. Most of the specimens
studied were in the Museum of Vertebrate Zoology of Cornell
University, and for the privilege of examining them I am indebted
to Professor B. G. Wilder, Curator. Considerable material, also,
belonging to Professor S. H. Gage was placed at my disposal by
him, to whom also I am indebted for numerous suggestions and
kindly interest in my work.

To the distinctive features of the Ichthyopsida, or fish-like
vertebrates, enumerated by Huxley when he first pointed out
the natural provinces in which the Vertebrata. were grouped,
there might be added, since 1876, that system of sense organs
which, with the canals in which it is often enclosed, has been
variously spoken of in the literature of science as the muciparous
canals, lateral line system of sense organs, organs of a sixth
sense, branchial sense organs, etc. It is but lately that the
importance of these sense organs has begun to be fully estimated,
and more careful observations have been made upon certain of
the forms. The literature upon the subject is voluminous, but

116 PROCEEDINGS OF THE

despite the fact that so much has been written, except in the three
or four most recent contributions, especially those of Ewart, ’92,
and Allis, 89, there has been no account of this system at all
complete for any form, This is due largely to the fact that
attention has been mainly confined to a study of the canals.
and their branches rather than the structure, distribution and
innervation of the organs themselves—which is indeed very
much like attempting to understand the oyster or snail from a.
study of its shell alone.

The first-recorded observations upon the system were made.
by Stenonis, in 1664, upon the canals in a species of skate.
From that time the system attracted more or less attention.
Lorenzini and Monro secundus may be mentioned as two of the
more important early contributors. By most of the first writers.
the function of the system was regarded to be the secretion of
mucus and its distribution over the body of the fish. Jacobson;
in 1813 first arrived at the conclusion that the system of canals
constituted a sensory organ for the purpose of transmitting the
vibrations of the water to the nerves, as he believed. However,
it was not until Leydig in 1850 discovered the sense organs
themselves and made microscopic examination of them that
the step in the right direction was made, and a morphologic
basis given for the theory of the sensory function of the system.
His final and most complete account of the system appeared in.
1868, upon the ‘‘ Organs of a sixth sense” as he regarded them.
Since 1813, among others who worked upon this system in fishes
may be mentioned De Blainville, Robin, McDonnell, Schulze,
Bodenstein, Solger, Wright, Fritsch, Beard, Garman, Allis, Guitel,
Ewart, Pollard and Collinge; especially important are the
researches of Allis, Guitel, Ewart and Pollard, by whom the
distribution of the sense organs in the canals, their relation to
the pores by which the canals communicate with the exterior,
and the innervation, have been quite thoroughly worked up in
the forms investigated by them, namely, Amza, Lophius,
Laemargus and Raia; Pollard has also compared the distribution
of the organs in five Nematognaths with a view of determining

AMERICAN MICROSCOPICAL SOCIETY. 117

their taxonomic value. Garman has studied the canals in
many Elasmobranchs and in the Holocephala chiefly to ascertain
their value in classification ; for a more detailed account of the
early literature of the subject and the views and results of the
various writers Garman’s paper should be consulted ; upon this
the above account of the early history of the subject is based.

Much still remains to be done on the system, especially among
the Teleosts where theré is a considerable variation in its
development, which, in connection with the wide range in life-
habits, may afford some clue to the function and importance
of these sense organs. The modifications of the bones of the
skull due to the presence of the canals and the possible bearing
it may have on the origin of the vertebrate ear, demand that far
more exhaustive study be bestowed upon them than has been
up to the present. Particularly valuable will be careful and
thorough investigations upon the early appearance and develop-
ment of the system.

Merkel, in his monograph upon the nerve-terminations in the
skin, recognized two classes of related cutaneous sense organs
which he termed end-buds (Endknospen) and _ nerve-hillocks
( Nervenhigel), and to the latter belong the organs of the
lateral line system. For nerve-hillock, the monomym xeuromast
proposed by Wright is preferably employed.

The end-buds are found in fishes distributed in the skin, par-
ticularly of the head, and in the mouth cavity where, in the higher
vertebrates, they appear as the taste-bulbs. They are situated
upon papillz of the cutis and are formed of long rod-like cells
extending throughout the height of the bulb. In “ fishes” they
are always flush with the surface of the epithelium or even
project beyond it. When situated in the skin they appear to be
tactile organs; in the catfish the long barbels around the mouth
appear to be little else. than carriers of these sense organs
(Wright).

With the neuromasts, on the other hand, there is a differentia-
tion of certain of the cells, some of them being conical, pear-
shaped and short, but bearing a more or less distinct bristle ;

118 PROCEEDINGS OF THE

furthermore, the neuromasts evince a tendency to withdraw
themselves from the surface. This tendency is carried to an
extreme in the majority of “fishes” in which the lines of sense
organs, which constitute the lateral line system proper, sink
beneath the surface and occupy canals which open upon the skin,
by means of pores, generally between the sense organs. Com-
plications may be yet increased by enclosures of the canals in
bone, as if for protection, and the pores may become many times
divided, producing seemingly the effect of still further withdrawal
of the sense organs from the exterior. Examples of this last
occur in Ama and the Clupeide. Again, in other forms, the
organs of the lateral line system simply occupy pits in the epider-
mis, or are contained in an open groove; examples of each are
Lophius and Chimera.

In ‘“ fishes”’ the typical arrangement of the canals or lines is.
(1) one along the side of the trunk, causing the well-known
lateral line. When scales and a canal are present it perforates
each scale obliquely, and a sense organ and pore generally occur
in each. The organs of this group are innervated by the lateral
nerve, often spoken of as the lateral branch of the vagus. There
are four lines on each side upon the head; (1) one above the eye,,
the supra-orbital, innervated by the ophthalmicus superficialis
VII.; (2) below the eye, the infra-orbital, innervated by the
buccal branch of the seventh nerve; (3) the mandibular or hyo-
mandibular, upon the lower jaw, innervated by the hyomandi-
bular branch of the seventh, and (4) a transverse line in the occipi-
tal region which meets its opposite, thus uniting the system on
the two sides, and belongs to the lateral system. These lines
may either be independent of each other or connected. Accessory
lateral lines may be present, which in one genus (Mzgii,
McDonnell) reach the number of nine; lines of neuromasts
may also be present in the skin of the head and trunk in
addition to the typical lines, and a study of their distribution will
doubtless afford a clue to the origin of the accessory lateral
lines and explain departures from the type, which, because of
their greater development, were more readily observed.

AMERICAN MICROSCOPICAL SOCIETY. 119

In 1861, eleven years after the sense organs in the canals of |
fishes had been discovered, Schulze, 61, reported the presence
of homologous sense organs in branchiate Amphibia. In this
class (the Amphibia) the neuromasts retain their simple primitive
condition and remain in the skin, but slightly, if at all, withdrawn
beneath the surface. Since the first discovery of these organs in
Amphibia, there have been made, as far as I can ascertain, but
half a dozen or so communications upon the subject by six
investigators, namely, Leydig, Schulze, Langerhans, Bugnion,
Malbranc and Wiedersheim. Leydig’s more complete discussion
is given in his last article to which I have not had access. Of
the others Malbranc’s paper is by far the most exhaustive, being
a study of the structure and distributions of the sense organs in
Proteus, Menopoma, Triton, Salamandra and Salamandrina, and
the tadpoles of the anura Lombinator, Pipa and Rana. Wieders-
heim makes the important statement that in Aszdblystoma and
Salamandrina the sense organs become covered with epidermal
cells during the period of terrestrial existence and are again
uncovered when the life in the water is resumed. To the above
should be added Cope,* who, though he evidently did not
recognize the presence of this system of sense organs in the
Amphibia, speaks of the depressions which mark the location of
the organs as ‘“‘ mucous pores,’’ and describes their location for a
wide range of urodeles and has given figures showing the distri-
bution in a number, which, however, since he does not recognize
that they were more than ‘“‘ mucous pores,” are often inadequate.
There exist in the adult of Sa/amandra, which is a land form,
“pores” upon the dorsal side of the head in the region of the
so-called parotid gland of certain Amphibia, and upon the body
in two rows, occupying in general much the same position as two
of the lines in the sense organs did in the larva. They have
nothing to do with them, however, but are the openings of glands.
Apparently Cope regarded the “ pores” which he found in so
many forms as homologous with the gland openings in

* Of eleven general articles and books upon the Amphibia examined, in
five only was the presence of this system noted.

120 PROCEEDINGS OF THE

Salamandra, and therefore termed them ‘“ mucous pores.” * *
It is also possible he may have had in mind the pores by which
the canals in fishes opened upon the surface; the sensory nature
of the system had been too long known to admit of their being
spoken of as ‘‘mucous pores” were fore itself not inappropriate
in Amphibia if used in the same sense as in fishes.

The original matter here consists mainly in a presentation of
figures of the distribution of the organs in certain American
urodeles and comparisons of them with each other and forms in
which the distribution had been already investigated. Cope’s
descriptions will be used to supplement my own for forms in
which the system exists and which have not been accessible.
The purpose is to ascertain the presence of this system and
determine what may be the typical distribution of the organs for
Amphibia. |

In examining the distribution in Amphibia a comparison with
other Ichthyopsida is inevitable, and as far as possible the names
which have been applied to the lines in “fishes” will be employed
here. The only lines whose homology with those in teleosts,
ganoids and elasmobranchs is unmistakable are the lateral lines
and the lines above and below the eye, which are accordingly
spoken of as the supra and infra-orbital lines respectively. The
lines upon the side and venter of the head are not so easily com-
pared. They are doubtless innervated by the same branch of the
seventh (mental, Strong) and represent the hyomandibular canal

* * The close resemblance between the distribution of the sense organs in
the larva and the ‘‘pores” in the adult Salamandra, caused Leydig to
advance the rather remarkable theory that at transformation the sense
organs became changed into large glands. Undoubtedly, however, the
similarity in location is merely coincidence, and the sense organs in the
larva perish and are at the time succeeded independently by the glands,
Leydig’s words are: ‘‘Nachdem geschwanzte und ungeschwanzte
Batrachier aus Kiemenathmern zu Lungenathmern geworden sind, haben
sich die Organe der Larven zu den grossen Hautdriisen des Kopfes und der
Seitenlinien umgebildet, welche auch jetz noch durch die Art des Secretes
und dadurch, dass zahlreiche Nerven an die Gegenden, wo sie liegen,
herantreten, von gewéhnlichen Hautdriisen sich verschieden verhalten”
(Malbranc),

AMERICAN MICROSCOPICAL SOCIETY. Be

system of fishes, with any associated lines of free neuromasts
there may be. For convenience in description I shall use some >
locative terms employed by Garman in Elasmobranchs, even
though it is not demonstrated at least, that the lines in the two
classes are homologous, though occupying much the same
regions.* They are therefore only provisional.

Proteide. Of the two existing genera of this family, Proteus
of Europe has been studied by Bugnion and Malbranc; figures
I—3 and 16 show the distribution of the neuromasts in Wecturus.
Comparison of these two genera shows the conditions strikingly
alike, the chief difference seeming to be due to the more elon-
gated form of the head in Proteus. The organs in that genus
are described by Bugnion as occurring in groups, occupying
linear depressions in the epidermis. These groups again are
associated together in lines or series upon the body. Bugnion
recognized upon the lower jaw, “lignes divergentes,’”’ “series
marginales,’’ and between them “ groupes obliques ;’’ upon the
side of the head groups which converge toward the corner of the
mouth ; upon the dorsal side of the head a line above the eye
terminating in a cluster dorsad and cephalad of the prenares
(groupe nasal anterieur), and also a cluster of groups caudad and
ventrad of the nostril (groupe nasal et posterieur). In the
occipital region of the head is an aggregation of groups which
is continuous with the lateral line. A ventral line passes ventrad
of the arms and terminates in the region of the legs. A dorsal
corporal line of groups transverse to the long axis of the body
was also recognized by Malbranc. The number of individual
organs in each group varied from four to eight.

* The true homology of the lines, it is felt, is determinable by the nerve
supply. Tested in this way, Ewart finds for the lines in Teleosts the follow-
ing equivalents in Elasmobranchs, the canals of the latter being given as
named by Garman: Infra-orbital=orbital, sub-orbital, orbito-nasal, nasal,
half of median and prenasal canals; supra-orbital—cranial, rostral and
sub-rostral; hyomandibular=angular, jugular, oral, sub-pleural and
pleural. There exists the possibility that some of the lines in sharks repre-
sent lines of neuromasts existing free in the skin in the teleosts, which have
not been recognized.

122 PROCEEDINGS OF THE

Though his figures show a very close resemblance, the less.
elongated head of (Vecturus permits a better idea of the arrange-
ment. Upon the dorsum of the head in Wecturus there exists
caudad of each eye a cluster of groups which is divided into two
lines, one of which passes dorsad, the other ventrad of the eye,
the supra-orbital and infra-orbital lines. Fig. 2 requires but
little supplementary description ; many of the groups, especially
cephalad of the eye, are oblique, and upon the snout their
arrangement is as if radiating from a common center (Fig. 1). —
In the occipital region of the head is a triangle of groups con-
tinuous caudad with the lateral line; two or three groups form a
short transverse series suggesting an occipital line. Upon the
venter of the head (Fig. 3) are readily recognized the lines of
Buenion ; lines of groups diverging from the corner of the lower
lip toward the latero-caudal corner of the head, the gular lines,
a line along the margin of the lower lip, which approaches the
cular line at its cephalic end and will be spoken of as the oral
line. Associated with the groups of these lines whose direction
corresponds to that of the line are others which are transverse or
oblique (groupes obliques of Bugnion). Upon the side of the
head (Fig. 1) may be recognized (1) a line of somewhat oblique
groups from the infra-orbital caudad of the angle of the
mouth to the oral line; (2) a line of groups from the corner of
the mouth to the caudal end of the gular line, and (3) a trend of
groups from the throat where the gular line terminates caudally
toward the infra-orbital, communicating also with the occipital
group. These will be termed the angular, jugular and post-
orbital lines or series respectively.

Upon the trunk the three lines are present. The lateral line
is composed of longitudinal groups and extends from the occipi-
tal region nearly or quite to the end of the tail. The ventral
line curves around ventrad of the arms and extends to the region
of the vent ; it is not continuous with any of the groups on the
head. A dorsal line of transverse groups was present, though
apparently weak. However, the tendency of the skin to form
little transverse furrows and folds rendered it difficult to determine

AMERICAN MICROS COPI CAL SOCIETY. 123

the precise conditions existing ; doubtless other groups in addi-
tion to those shown in the figure exist and were not detected.
The organs in each group varied in number from six to eight.

Cryptobranchide. The distribution in Cryptobranchus alle-
ghaniensis has been heretofore described and figured by both
Malbrane and Cope, and my own observations can little more
than add confirmation. In this form the organs open by
circular or oblong pores upon elongated dermal papillz (Fig. 40),
which they thus divide into two, reminding one at once of the
“Spalt-Papillen” of Fritsch in the ray, containing free neuromasts
in the skin, and the papillze in the skin of the lamprey which are
similarly halved by a slit-like depression at the bottom of which
a sense organ is situated.* Though the orifices are rarely
oblong the tendency of the organ corresponding to the groups
in Vecturus is believed to be, as in the “ Spalt-Papillen,” trans-
verse to the papilla, and in the single diagram of the venter of
the head introduced for comparison, this is so indicated by a short
line (Fig. 40). This being the case, the tendency of the organs in
Cryptobranchus and the direction of the groups in Wecturus
correspond.

The three corporal lines occur as follows: The dorsal line is
weak and is represented by about ten or twelve transverse organs,
and extends to about the level of the end of the abdomen. The
ventral line contains about 36 organs ; it extends from just cephalad
of the legs, where it curves in toward the meson, to cephalad of
the arms, passing ventrad of them and curving dorsad to end in
a short transverse line. It occupies a position ventrad of the
lateral fold present in Cryptobranchus. The lateral line lies just

* The distribution of these organs in Petromyzon planeri has been
figured and described by Langerhans, who regards them, and correctly it is
thought, as homologous with the lateral line sense organs. They arearranged
in lines upon the head which are difficult to homologize with those in higher
forms. Upon the body Langerhans recognizes dorsal, lateral and ventral
lines, of which the first two are evident, the ventral is short and possibly
may not be a corporeal line. The histologic structure of the organs in the
Lamprey seems not to have received sufficiently accurate investigation,
and on the nerve supply practically nothing has been done; it would offer
an interesting and important, though rather difficult, research.

124 PROCEEDINGS OF THE

dorsad of this fold and extends to the tip of the tail. Upon the
head it seems to connect with the post-orbital series.

Upon the ventrad aspect of the head the gular and oral lines
are well developed, the latter covered for a portion of its dis-
tance by a labial fold. The former is accompanied by the usual
transverse organs and extends upon and over the lateral fold
upon the head. On the ventral side of this fold a double line of
twelve or fourteen organs extend from the gular to the corner of
the mouth (jugular). Upon the dorsal side of the fold there is
a trend of organs cephalad to the orbital group, which appears
to represent the post-orbital series. The eye is surrounded by
the customary series of organs, which cephalad of it are so
thickly placed that from the material at hand it was impossible
to reduce them to a system. Some were parallel and some
transverse or oblique to the assumed trend of the lines. An
angular passing around the corner of the mouth to the oral was
present. .

Microscopic examination shows the neuromasts in C7yffo-
branchus much larger than in WVecturus, though of the same
typical structure; compared with Wecturus they are also much
fewer, one organ in Cryptobranchus more nearly representing a
group in WVecturus.

Amblystomatide. Of the genus Ambdblystoma, species pune-
tatum, both larval and adult forms were examined and the dis-
tribution of the sense organs is given in Figs. 4-11, 17, 19, 33
and 36. <Azmblystoma appears in the relation of the groups to
the lines to form a single exception to the type presented in the
other urodeles examined, and for that reason and because the
abundance of larval material rendered it a convenient form in
which to illustrate it, the later development of the system in the
larva is represented by Figs. 33, 4,5,19,17, 10,11 andg. The
questions and difficulties which involve a study of the first
appearance and early development of the system render any
treatment impossible here. The Amphibia also do not seem
to afford as good opportunties for such an investigation as other
Ichthyopsida.

AMERICAN MICROSCOPICAL SOCIETY. 125

The presence of the sense organs could be detected in yet
unhatched larve. The distribution in a larva 16 mm. long is
shown in Figs. 4, 5 and 19. Upon the dorsum of the head the
supraorbital lines are readily seen and the groups in the occipital
region also. A yet simpler arrangement is shown in Fig. 33 of
a larva whose arms are just budding. Upon the venter of the
head the oral and gular series are recognized with tendency to a
transverse line just cephalad of the gular fold as in Necturus
and Cryptobranchus. There are small accessory, doubtless
developing, organs beside those forming the typical lines. Upon
the side of the head ( Fig. 19), are seen the infra-orbital, angular,
oral and jugular, each consisting of few organs simply arranged.
All three lines upon the body are present and presenting the
typical arrangement ; those in the lateral line were oblong as if
exhibiting a tendency to transverse fission.

In a larva 28.5 mm. long (Figs. 10, 11 and 17), the system
has reached a greater complexity and higher development, and
in place of single organs occur groups of two or three, as in
Necturus, though not marked by a depression. In the arrange-
ment, however, there is a difference. While in Mecturus and
also in the other urodeles examined the groups in the lateral and
ventral lines on the trunk, and generally the lines on the head, are
parallel or nearly so with the lines in which they occur and
transverse in the dorsal corporal line, in Amélystoma the con-
dition is reversed and the small groups are transverse to the
direction of the line in the ventral and lateral lines, and parallel
or nearly so in the dorsal lines, etc., agreeing in this with the
condition of the groups in the corporal lines in Anura larva
(Malbranc and Fig. 37). Herein lies the most puzzling
peculiarity of the distribution in Amélystoma. The general
regions of location recognized before in MVecturus are easily
traced in the plottings.

The dorsal view of the head of an older larva introduced for
comparison shows a yet greater development of organs and
groups. In an axolotl 15 cm. long, Malbranc states that in the
lateral lines the groups, which are as in Asudlystoma transverse,

126 - PROCEEDINGS OF THE

consist of as many as ten organs which are, however, oblong and
their long axis parallel with the direction of the line.

In the adult Amdlystoma the same arrangement occurs, though
the apparent number of organs is much reduced. Figures 6-8
are plottings of an adult 18 cm. long. In many individuals
taken in the Spring it is impossible to find more than a trace of
the system, and in one the lines upon the head and the lateral
and dorsal lines on the body were very distinct, but the ventral
line was apparently missing; this is doubtless due to the fact,
stated by Wiedersheim as quoted before, that in Azmdlystoma the
sense organs become covered by epidermal cells when the animal
assumes a terrestrial existence and again appear when it enters
the water, a fact I can confirm, since, when no organs can be
perceived from a superficial observation, sectioning reveals them
in the epidermis, but concealed by overlying epidermal cells. -
Irregularity in the reappearence of the organs explains the
apparent absence of certain lines, and though there is no positive
evidence, I am inclined to believe that none of the organs dis-
appear at transformation, but only a few reappear in the adult when
the life becomes aquatic at the breeding season, the rest remain-
ing concealed in the epidermis. In Amdlystoma the lateral line
terminates at about the level of the vent and a more dorsally
situated line succeeds upon the tail. Malbranc regarded this as
properly belonging to the lateral line rather than the dorsal, a
view which is supported by the condition in other urodeles,
notably Diemyctylus and Gyrinophilus.

Beginning at about the level of the vent there are upon the
dorsal surface of the tail on each side a row of some 60 or more
openings occupying the line of the sense organs and resembling
them greatly. These pores are oblong and transverse to the
axis of the body, and in some instances there are two openings
forming a transverse pair. Around each the pigment is absent
from the skin, thus making them more distinct. At first the
impression is strong that they are sense organs until dispelled by
closer examination and sectioning of the skin which shows them
to be gland openings. The sense organs may readily be detected

AMERICAN MICROSCOPICAL SOCIETY. 127

in their midst however. The condition illustrates how easily |
the two classes of openings may be confused and accounts for
Leydig’s wrong conclusions. | Cope also did not recognize the
difference. .

Cope figures, p. 60, the sense “ pores”” upon the head in A.
punctatum and describes them page 57. He also notes their
presence in A. conspersum, copeanum, jeffersonianum, and in the
allied genus Chondrotus.

Plethodontide. Cope describes and figures the distribution in
Stereochilus marginatus, p. 153. His words are ‘the mucous
pores on the head are distinct and large. They form a double
series along the canthus rostralis and a single one above the orbit
which turns round the latter behind and is continued below it and
along the side of the muzzle to the nostril. A series of similar
large pores extends along the middle of each side.”’

The system is distinct in Gyrinophilus. Figs. 18, 24-26 are
of a larva of Gyrinophilus porphyriticus 82 mm.long. The supra-
orbital line is but weakly developed, the organs being more
abundant toward the nostrils. The infra-orbital is well repre-
sented and presents the characteristic radiating arrangement
caudad of the prenares. Each organ is oblong and its location
readily marked by the absence of pigment from around it. The
oral line is well developed with 2—3 transverse organs accompany-
ing it, corresponding to the groups in Wecturus. The angular
line is present and the jugular represented by 4-5 organs. The
gular line is distinct though apparently weak near its middle.
The postorbital group is well developed. The lateral line
connects with a small occipital group on the head and extends
to the tip of the tail, where the organs are more crowded and
the line more dorsally located, as in Amédlystoma. The dorsal
line is represented by 12—15 transverse organs and does not reach
the level of the hind legs.. The ventral line is more strongly
developed, but yet weak. The position is typical as seen from
the figure (18), Gyrinophilus thus agrees closely with the general
scheme of arrangement exemplified in the forms examined
before. In the adult the neuromasts are present though the fact

128 PROCEEDINGS OF THE

is less easily recognized. Cope plots some of the organs in a
latva’ Tizem, long:).(Pl. XLII Bisse)

That the system is present in Sfelerpes ruber is evident from
the words of Cope on page 172: ‘“ The eye is encircled by a
series of pores. These extend anterior to those on the side of
the head to the nostrils and are more crowded. The lower edge
of the lower jaw is encircled by a single series of pores and there
are two other series nearly straight which start from the point of
the chin and diverge backwards.” The last two are clearly those
which I have designated as the gular lines; the orals of the two
sides would seem to meet at the meson.

Desmognathide. In Desmognathus the system is weakly
developed though present. An examination of the adult D.
fusca shows diverging gular lines consisting of seven or eight
organs each with accessory organs (2-3). The oral line is
represented by but two or three organs. The infra-orbital line
is the most developed and consists of about ten organs under
the eye and cephalad of it.. The supra-orbital is apparently
almost wanting, consisting of two or three organs only. Upon
the body the lateral and ventral lines are present, the organs
being about one for each segment. Of the dorsal line no organ
was detected. The conditions in Desmognathus render it often
difficult to determine their presence from superficial examination,
and no serial sections of the trunk have been made, so that
possibly a few organs may represent the dorsal corporal line
which were not detected.

Cope has recognized the presence of the “pores” in the
species of this genus. He says: ‘The pores in D. ochrophea
are very difficult to observe. In a few specimens I have seen a
few of those of the lower series; the upper I believe to be wanting.”
In D. fusca he finds ‘‘ one well and one little developed lateral
series of mucous pores.” Of Desmognathus nigra: “The
mucous pores are well developed, and the two lateral series are
often distinct in alcoholic specimens by their white color; when
they become dry they are difficult to observe. There are two
rather distant gular series within the mandibular rami on each

AMERICAN MICROSCOPICAL SOCIETY. 129

side, and one on each side extending inwards and forwards from:
the gular plica. The Superior lateral series extends from the
orbit to near the end of the tail ; the inferior round the humeri to
each side the pectoral region.” .

Pleurodelide. In the adult of Diemyctylus viridescens the
depressions marking the location of the organs are very distinct.
Upon the head the Supra-orbital and infra-orbital serjes are well
developed. In many the pores cephalad of the eye are oblong
and exhibit the same radiating arrangement shown by the groups
of organs in Mecturus. Upon the side of the head are distinct
the jugular and angular lines, and post-orbital Sroup, which, how-
ever, does not seem to have any marked trend. The gular line
starting from the caudal end of the jugular is formed of two
series of organs, the more lateral of which are often transversely
oblong, representing doubtless the transverse Sroups in Mecturus
and other forms. At its cephalic end it approaches the oral line
which consists of some half a dozen or more organs. Upon the
body the three lines are well developed, and in them the organs
are associated into groups of two or three ( Fig. 20). Of these
the lateral line is as usual best developed, and caudad of the
vent curves dorsad to Occupy a position along the base of the
caudal fin ; it is not closely connected with the occipital group
on the head. The dorsal line, in which the groups are transverse
to the long axis of the body, runs out just caudad of the vent
upon the caudal fin. The ventral line ( Figs. 20 and 21), curves
mesad beneath the humeri, cephalad upon the pectoral region,
and likewise at its caudal end just cephalad of the legs, having
thus its typical position and extent.

In the well advanced larva the conditions are very much as in
the adult. For comparison a figure showing the distribution
upon the dorsum of the head is given ( Fig. 23). The lateral
lines are very distinctly located by the absence of pigment from
around the sense organs.

Diemyctylus viridescens, whose life-history has been worked out
by Gage, passes through a period of terrestrial existence inter-

mediate between the aquatic larval and adult Stages, and there-
9

130 PROCEEDINGS OF THE

fore affords an excellent example of the forms, such as Salaman-
drina and Amblystoma, in which the sense organs persist and live
over the time of life on land by being covered with epidermal
cells, reappearing again when the aquatic life is resumed. The
skin during the land stage is of a red color and rough with
numerous horny papille. Examination of the skin or the exu-
vium with a lens fails to detect any of the orifices so conspicuous
in the olivaceous, smooth-skinned aquatic adult. Examination
of sections of the skin from the right regions demonstrates
their existence, and there is no doubt that the sense organs
persist from larva to adult covered, or nearly covered (as in the
one figured, Fig. 34), by epithelium cells.

Comparison of the distribution of the sense organs in this form
with European newts of this family, that have been examined by
Malbranc, shows a very close resemblance. Cope has noted the
presence and distribution of the pores in the other American
species of this genus, D. forosus (p. 204).

Amphiumide. An alcoholic specimen of Amphiuma means
was examined and this was supplemented by fragmentary obser-
vation of a living individual in which the location of the neuro-
masts was easily detected. The organs are in small groups of
two or three each, which are indicated upon the figures by short
lines. A comparison of the figures with those showing the
distribution in other genera indicates how closely Amphiuma
conforms to the typical amphibian arrangement. The supra-
orbital and infra-orbital lines are well developed, the former
extending some distance caudad of the eye. In connection with
each line are numerous groups which are transverse OF oblique
to the direction of the main lines, especially cephalad of the eye,
recalling the relations in Necturus. The lateral line extends
cephalad upon the head, curving ventrad toward the post-orbital
groups as in Necturus, and not connecting with the orbital lines.
Upon the venter of the head the arrangement of the lines is
almost diagrammatic. Both oral and gular lines are well
developed and in connection with each are the numerous trans-
verse groups which have been found so often before. Upon the

AMERICAN MICROSCOPICAL SOCIETY. Dar

‘side of the head are quite distinct jugular and post-orbital lines,
the former extending from the angle of the mouth to the caudal
termination of the gular; the latter, from the infra-orbital, of
which it appears as a caudal continuation, to the meeting of the
gular and jugular,

Although Cope states that no distinct rows of pores exist on
the body, the examination of the living specimen showed clearly
the presence of all three lines, the lateral extending throughout
the length of the elongated body, the ventral terminating in the
region of the vent, and the dorsal much more weakly developed.
The cephalic ends of all three lines are shown in Fig. 13. Cope’s
description of the mucous pores upon the head agrees in the
main with the figures.

It was not always possible to determine how many neuromasts
‘occurred in each group; where observed, the number was only
two or three, and uniformity in the size of the groups rendered it
probable that the latter number was not exceeded. Where the
organs were apparently single, a dot was employed to represent
their location.

Stremde. Four small alcoholic individuals of the genus
Stren were examined, and though the condition of the epidermis
made it impossible to give the system a thorough examination
and make complete plottings, Figs. 30 and 31 suffice to indicate
that the neuromasts exist and that they conform in their
distribution to the general scheme in Amphibia. Upon the
trunk three lateral lines occur and their position and extent are
entirely typical. Wherever the condition of the epidermis
rendered the number determinable, the small groups which were
found contained two organs each. They are longitudinal in the
lateral and ventral, and transverse in the dorsal line. The
relation of the cephalic and caudal ends of the ventral line to the
axilla and vent is shown by Fig. 31 and is typical, resembling
the condition with forms in which hind legs are present.

Upon the head it was difficult to determine more than the rich
abundance of the neuromasts, in the usual regions. Upon the
venter of the head the gula line is well developed ( Fig. 30 i

Liner) PROCEEDINGS OF THE

and the arrangement of groups complicated. The presence of
oral, angular and jugular series could not be determined because-
of the ill preservation of the epidermis.

Anura. But little attention has been paid to the system in
the tailless Amphibia. For comparison merely, the distribution.
in a species of Rana, probably catesbzana, is given ( Figs. 37 and
38). The existence of the neuromasts in many of the European
genera has been determined by Schulze, Leydig and Malbranc,*
though, as far as ascertained, only of Loméinator and Rana have
figures been published showing the distribution.

In Rana three lateral lines are readily seen, the dorsal meeting
the lateral at its cephalic end. The orbital lines are well
developed and encircle the prenares also. The ventral line
extends from the region of the vent making a curve dorsad
around the region where the arm is concealed and is continuous
apparently with a transverse line (Fig. 37). When in later
larval life the arm appears, the line passes dorsad of it instead of
ventrad as in the urodela, a rather noteworthy difference appar-
ently-unobserved hitherto. Another difference between the two
groups, constant for all forms examined except Ammblystoma, was
noted by Malbranc ; it is that while in the tailed Amphibia the
linear groups of neuromasts are longitudinal in the lateral and
ventral lines, and transverse in the dorsal, in the tailless forms
the relation of group to line is exactly reversed and the groups
are longitudinal in the dorsal, and transverse in the lateral and
ventral lines. In the orbital lines also the groups are transverse
or nearly so, especially near the eye.

It is interesting to note the existence (Malbranc) of the sense
organ in a larva of Pipa dorsigera, 2.5 cm. long, taken from the
back of the mother.

DIPNOANS.

When it was desired to compare the system in Amphibia and

* Schulze examined Bombinator igneus, Rana esculenta, and temporaria,
Pelobates fusca and Hyla arborea; Leydig, Bufo cinereus and calamita ;
Malbranc, Bombinator, Pipa and Rana. (Malbranc.)

AMERICAN MICROSCOPICAL SOCIETY, 133

in the Dipnoi, the only statements* upon the lateral line system
in that group that could be found were so unsatisfactory that an
examination of individual specimens was necessary. ©

Upon the system in Protopterus the most detailed account was
given by W.N. Parker, who is quoted by Wiedersheim. He
states that the organs are not confined to the lateral line, but
occur dorsad as well as ventrad of it; that they are especially
numerous upon the snout where they occupy grooves in the skin,
and occur always free in the skin as in Amphibia and the young
‘of many fishes. His rather general statement needs supple-
mentation and correction in several particulars. From the exami-
nation of five well preserved specimens, Figures 12, 27-29 were
constructed, and the following may be said in discussion of them.
As stated by Parker, the neuromasts are situated (in most of the
regions) free in the skin, and not in canals as in most “ fishes,”’
and occur in the lines on the trunk, in linear groups of from four
to ten organs each. Three lines are present upon the body as
in Amphibia—dorsal, ventral and lateral. The ventral line
arises ventrad of the gill slit, passes ventrad of the pectoral fin
to the pelvic fin where it is interrupted, turning slightly dorsad
instead of ventrad as in Amphibia, to be continued upon the tail
by a line of rather long groups. The dorsal line js more weakly

* For the opportunity of consulting the Dipnoan specimens and literature
I am indebted to Prof. B, G. Wilder,

+ ‘‘Diese (Hautsinnesorgane) weichen in ihrem Bau von dem gewohnlichen
Verhalten bei Fischen und Amphibien nicht ab. Sie sind nicht allein auf
die Seitenlinie beschrankt, sondern finden sich auch am iibrigen. K6rper
und zwar dorsal- wie ventralwiarts. Ob es sich aber an diesen Stellen um
eine regelmissige Anordnung handelt, vermag ich vorderhand nicht sicher
zu sagen. Am Kopf, wie namentlich an der Schnauze, sitzen sie ungleich
zahlreicher; dabei liegen sie auf dem Grund von grubigen Hauteinsen-

Es handelt sich aber nicht etwa um eine Betheiligung der Kopfknochen,
‘d. h. letzere bilden nirgends schiitzende F urchen und Caniile. Dasselbe
gilt auch fiir die im Bereich der Seitenlinie sitzenden Organe. ‘Auch sie
werden keineswegs von den Schuppen tiberlagert. Kurz, allerorts sitzen
die Hautsinnesorgane des Protopterus frei im Niveau der Epidermis, ein
Verhalten, welches mit dem der wasserlebenden Amphibien und den
-Jugendstadien sehr vieler Fische tibereinstimmt,” Parker, p. 7.

134 PROCEEDINGS OF THE

developed. The general direction of the groups composing it is:
longitudinal, although those in its cephalic part are somewhat
oblique. The lateral line is composed of linear groups closely
associated together. Upon the tail it either terminates some dis-.
tance from the end or shifts its position, as is more probable, and
is continued farther ventrad. Upon the head it is continuous.
with the orbital lines.

Cephalad of two transverse spurs in the occipital region, the-

continuation of the lateral line becomes enclosed within a canal

which opens by three pores, as indicated in the figures, and forks.

at the fourth. The dorsal branch is continued above the eye as.
the supra-orbital line. After a series of eight pores, the line
again occupies an open groove, succeeded by a short canal with,
three cephalad pores, of which the organs are again situated in.
a furrow in the skin.

The ventral branch passes through a canal with seven pores,
where the infra-orbital line arises as a line of organs situated in
a groove, the canal bending ventrad to be succeeded at the eighth
pore by a line of organs occupying a groove along the edge of
the lower lip, or the oral line. From its most caudal pore ex-

tends a line of neuromasts caudo-ventrad to meet another line-

upon the ventral surface of the head. They may be spoken of
as the jugular and gular, and their relations are seen in Figs. 27

and 29. In addition to these main lines the accessory lines occur

upon the head as shown in the figures.
As far as I can ascertain the only descriptions of the lateral
line system in Lepidosiren were those of its discoverer, Natterer*

* “ Diese Schleimkanile beginnen an der Spitze der Schnauze und bilden,
jederseits zwei wellenférmige, mehrere Zweige aussendende Linien, deren
sich eine oberhalb, die andere unterhalb des Auges, bis gegen das Hinter-
haupt hinzieht, wo sie sich wieder vereinigen, zwei gerade gegen das
Hinterhaupt aufsteigende Aestchen aussenden und von der Kiemenspalte
an in gerader Richtung lings den Seiten des Kérpers bis zum Schwanzende,
analog der Seitenlinie der Fische, verlaufen. Die untere dieser wellen-
firmigen Linien gibt vor ihrer Vereinigung am Mundwinkel einen Zweig
zum Unterkiefer ab, der den Kiefer umsiumet, sich von der Spitze des-
selben in einem doppelten Aste gegen die Kehle wendet, das Unterkinn
pegrenzet und von da in wellenférmigen Windungen die Kehle durchzieht.

,

AMERICAN MICROSCOPICAL SOCIETY. 135

and Hyrtl,* * when the system had not yet been discovered in
Amphibia and was regarded as a mucus-secreting organ in
“fishes.” However, the comparison of their descriptions and
the figure of Natterer, supplemented by the examination of a speci-
men in the Museum of Vertebrate Zoology of Cornell University,
from which figures 43 and 44 were made, suffices to show that
the distribution in Protopterus and Lepidosiren are very closely
comparable, almost identical ; that in Lepzdostven the neuromasts
occur free in the skin, and no portion of the system, as in Protop-
terus, occupies canals. From the description of Natterer the
presence of the ventral and lateral lines is evident, and I doubt
not that the dorsal line is also present. The poor state of pres-
ervation of the epidermis in Lefzdosiren did not permit my
ascertaining its existence.

In Ceratodus the system is enclosed in canals, and the relation
of the lines could not be determined from superficial examina-
tion. The lateral canal is the only one upon the trunk, per-
forating each scale and opening upon each by a trifid or quad-

und dieselbe in mehrere Felder theilt, sich dann aber in vollkommen
gerader Richtung zu beiden Seiten des Bauches dicht iiber die Hinterfiisse
hinweg, lings der Basis der unteren Schwanzflosse bis an’s Schwanzende,
erstreckt. Die obere sendet einen Zweig wellenférmig quer iiber den
Scheitel.” Natterer, °45, p. 5.

* * « Hin der Classe der Fische eigenthiimliches und bisher bei keinem
Amphibium beobachtetes System von Schleimcanilen findet sich unter
folgenden Verhiltnissen: Die Seitenlinie theilt sich, nachdem sie die ganze
Lange des Stammes durchlaufen und iiber der Kiemenéffnung zwei con-
vergirende Aeste gegen den Nacken abgegeben hat, hinter und iiber dem
Mundwinkel in zwei Zweige, welche schlangenférmig gewunden iiber und
unter dem Auge gegen die Schnauze ziehen, und am Lippensaum, zwei
Linien von einander entfernt, endigen. Der untere derselben schickt gleich
nach seinem Ursprunge drei Aeste zum Unterkinn, welche in der Mittel-
linie in einander iiberzugehen- scheinen, and durch mehrere gewundene
Zwischenschenkel mit einander communiciren, wodurch kleinere unregel- *
massige Facetten gebildet werden. Der obere haingt mit dem der anderen
Seite durch einen iiber den Scheitel weggehenden Verbindungsarm, und ein
Zoll hinter diesem, durch einen zweiten ahnlichen, zusammen... . . Die
Verbreitnng dieser Linien am Kopfe stimmt mit jener bei Chimcera voll-
kommen iiberein.” Hyrtl, 45, p. 6.

136 PROCEEDINGS OF THE

rifid pore. Whether the ventral and dorsal lines were present as
series of free neuromasts could not be determined.

Comparing the system in Proftopterus with that in urodeles,
resemblances appear in the presence of three corporal lines
occupying the same relative positions as in Amphibia, the pres-
ence of lines on the head apparently representing the oral, jugu-
lar and gular lines in Amphibia, connected with the infra-orbital.
Differences in detail in the two groups are manifest ; most con-
spicuous is the apparent fusion for a part of their length of the
infra-orbital and jugular, and the absence of an angular line.

In degrees of complexity of the system the three dipnoans
stand, Lepidosiren, Protopterus and Ceratodus. In the first the
entire system is superficial; in Cevatodus it is sunk in canals (except
perhaps dorsal and ventral lines and accessory groups) ; while in
Protopterus it is intermediate, a small portion only occupying
canals.

HISTOLOGIC STRUCTURE OF THE NEUROMASTS.

In structure, as has been suggested before, the neuromasts in
Amphibia are but little modified from the typical form. They
are situated in the epidermis, but little if at all withdrawn from
the surface and present throughout the class the same histo-
logic structure. The following description is based upon the
statements of Malbranc and my own observations:

The two kinds of cells are readily distinguished, the conical,
pear-shaped or so-called sensory cells, and the spindle or sus-
tentacular cells. The former occupy the center of the bulb
extending only partly through its height. In number they vary
from only a few to forty or so, according to the size of the
neuromast. The nucleus is generally large, round and clear,
possessing a small amount of chromatin. The cell-body,
further, blackens somewhat on the application of osmic acid,
though not so markedly as in the neuromasts of fishes. The
ectal end of the cell bears a refractive bristle which in the larva
is generally quite long ; in the adult itis short, reaching, however,
the free surface of the bulb. The ental or basal end which con-
tains the nucleus seems generally to terminate bluntly; often

AMERICAN MICROSCOPICAL SOCIETY. 137

however, delicate varicose processes are to be observed. ( Mal-
branc ).*

Surrounding these is an enveloping sheath of long spindle
cells which extend throughout the height of the bulb, their
form being modified according to the shape and position of the
surrounding cells.

The nucleus is generally .situated near the basal or ental end
which terminates in a number of delicate processes. Isolated
cells of the two kinds from Déemyctylus are shown in Figs. 42
and 43; for comparison also are figured the neuromasts in
Necturus, Diemyctylus and two developing (?) organs in Amblystoma
in which the typical histologic structure is readily seen. The
neuromasts in /Vecturus are the more withdrawn from the surface.
The precise mode of termination of the nerve fibers in the neuro-
masts still demands attention, possibly for the determination of
the value of certain theories.

GENERAL REMARKS.

The Amphibia afford in certain respects peculiar opportunities
for the study of a sensory system associated with existence in
the water. This is due to the fact that there are here included
forms purely aquatic and forms as purely terrestrial in their
habits of life, and yet others which spend a portion of their life in
the water and a portion of it on land. In every family of the tailed
Amphibia native in the United States the system has been found,
and in five families of the tailless Amphibia. Since Malbranc
has found the sense organs in a larval Pipa, and Leydig in a
larva of the viviparous Salamandra atra taken from the oviduct
of the mother, doubtless the system will be found in a more or
less perfect state of development in all Amphibia at some period
in their life-history. The genus Plethodon would be a good test
form for determining this, since it is said at no period of its

* Schulze has stated that these cells are directly continuous with nerve
fibers. This last is improbable; doubtless the application of modern
methods would show that such is not the case; it is important, however,
‘since in that case these would be true nerve cells in the second modified
epidermal cells.

138 PROCEEDINGS OF THE

development to live in the water. In order to ascertain it
possible the presence of the neuromasts in this genus, I examined
serial sections through the head of a well developed Plethodon
erythronotus embryo almost ready to hatch, but could detect no
certain trace of the sense organs. However, if larve just
hatched are examined, it is confidently expected that the organs.
will be found.

In the urodela the distribution may readily be reduced to the
following type: Upon the body, three lines, a /atera/ continuous
or not continuous with an occipital group, though not continuous
with the orbital lines; a ventral line extending from under the
arms. in the pectoral region to near the hind legs ; a dorsal line
somewhat closely connected with: the lateral at its cephalic end
and seldom extending as far as the level of the vent. Upon the
head, a series extending from behind the eye, above and below it
to the snout, the swpra- and infra-orbital lines; a line upon the
lower lip, the ora/, connected with the infra-orbital by the
angular; a line from the angle of the mouth to the lateral
corner of the head and there meeting a diverging line upon the
ventral side of the head, and, when this is sufficiently developed,
a line or trend of organs upon the side of the head; these the
jugular, gular and postorbital lines of the descriptions and figures.

Comparison with other Ichthyopsida may not be of much
value ; however, the distribution approaches most nearly that in
the Dipnoans, then in Elasmobranchs ; among the latter Ch/amy-
doselachus, apparently, in the greater extent of the gular line,
shows most resemblance to the Amphibia.

In the tailless forms there are three corporal lines, the lateral
and dorsal converging cephalad ; also well developed orbital lines,
and upon the ventral side of the head a line in the position of the
oral and jugular, possibly representing both, and two transverse
lines, with a possible gular. The changed form of body in the two
groups renders a comparison difficult; it should be based on —
nerve supply. The curious difference in the relations of the
groups in the lines upon the trunk in the anura and urodela has

been noted before.

AMERICAN MICROSCOPICAL SOCIETY. 139

The significance of the arrangement into groups is apparent
when the system is examined in the larva at different periods of
development. Evidently as has already been maintained by
Malbranc, each group sprang from a single organ by repeated
fission in the same plane. His figures and my own observations
clearly show that such is the case, as illustrated by Fig. 45.
Exactly how this takes place, however, is unknown. Whether
the sensory cells may arise from the supportive cells, or from
sensory cells alone, and supported from supporting cells or from
ordinary epidermal cells, yet awaits solution.

It is probable that the sensory cells alone determine the size,
shape and division of the neuromast, and multiply by the division
of previous sensory cells. In only one instance a karyokinetic
figure was observed in a nucleus apparently of a sensory cell.
The causes determining the plane in which fission takes place
must be closely connected with the function of the sense organs,
whatever it may be. Malbranc called attention to the often
recurring arrangement of groups upon two coordinates perpen-
dicular to each other, or nearly so (as in the gular line), pointing
out the physical advantage in such an arrangement in perceiving
the direction and strength of a vibration in the water, should
such be their function. In Ichthyopsida, in which the sense organs
are deeply sunken in canals, the pores often become many times
divided. In Amma, Allis found that the primitive pores divide
quite regularly in a certain plane for a number of times; these
secondary pores again often divide in a plane at an angle to the
first, generally a right angle or nearly so, reminding us of the
groups in Amphibia. In forms, then, in which the sense organs
are confined in canals, this division of the pores would seem to
represent a potential division of the sense organs, which in forms
in which the sense organs are freely situated, as in Amphibia,
can be actual. It does not appear that any physiological experi-
ments have ever been undertaken to attempt to ascertain the
function and value of the system, nor are there as yet data
for a comparison of the development of the system in different
Ichthyopsida with their habits and form, from which some idea

140 PROCEEDINGS OF THE

might be obtained. Of the theories advanced, the facts of
arrangement and development seems to render most reasonable
the one of Jacobson, viz., that it is the perception of vibrations of
the water, and that the distribution, and therefore multiplication of
the organs may, perhaps, be to a certain extent regulated by and
modified in accordance with mechanical advantages arising out
of the form and habits of the animal.

In connection with the development of the individual sense
organs should be mentioned the existence in Cryptobranchus, as
observed by Malbranc and by the writer, of small organs in the
skin which are covered with epidermal cells, and which he
regarded as developing organs, which later break through the
epidermis. This mode of development has been shown by Allis
to exist in Asma. However, I do not consider it necessarily the
proper explanation of these hidden organs in Cryptobranchus.
They were also observed in the skin from the parotid region of
an adult Asdlystoma (the only region examined); in a very
limited area, about five mm. square, were found two groups of
two each, and one of three of these small submerged neuromasts.
From the fact that the organs in the larva are more abundant
apparently than in the adult, it is suggested that these are but
supernumerary organs which do not appear in the adult, or do
so subsequently when needed.

This calls the attention to the necessity, imposed by the life
habits of certain urodeles, e. ¢., Diemyctylus, for the neuromasts
to live over a period of terrestrial existence, which is accomplished
by the protection of the organs by a growth of epidermal cells.
Doubtless this is also true for many other forms of semi-aquatic
habits of life, ¢..¢., Desmognathus. In certain other urodeles, ¢. g.,
Salamandra, and I believe Plethodon (if they exist at any time),
the system perishes entirely in the adult. This is also the case
apparently in all the Anura, though in Raza it persists until after
both legs and arms are well developed and the tail has begun to
be absorbed. There would seem, then, to be something other
than an aquatic existence necessary for the maintainance of the
neuromasts, since Raza catesbiana is more purely aquatic than

Ss ~€~

AMERICAN MICROSCOPICAL SOCIETY. 145

several of the Salamanders in which the system persists. Of the.
mode of final disappearance nothing is known.

The nerve supply of the system has never been worked out
for Amphibia with sufficient completeness so that the innervation
of all the various individual series is not definitely known. The
investigations of Ewart and Strong, the latter upon the cranial
nerves in the tadpole, have shown that in Amphibia and Elasmo-
branchs, at least, the nerves supplying this system arise just
cephalad and caudad of the eighth nerve. In Urodela, at the
level of the seventh, a nerve arises which divides, one branch
becoming associated with the Gasserian ganglion of the fifth,
and giving rise to the buccal and opthalmicus superficialis V/T.,
which, undoubtedly, supply the infra

and supra-orbital lines.
The second branch joins the seventh and forms the mental
(Strong) nerve which supplies the lines upon the ventral side of
the head, and probably also the jugular and post-orbital lines.
The lateral nerve arises just cephalad of the ninth and divides
into three branches, the more dorsal much the smaller, which
supply the group in the occipital region of the head and the
three lines upon the body. As far as it has been possible to make
comparative study the results sustain Strong. In Mecturus,
Amblystoma and Diemyctylus the roots described by him as
innervating the lateral line system above, were all recognized and
were in relative size quite proportional to the development of the
system. In Desmognathus the roots arose so close to the
seventh and ninth nerve that they were not readily distinguish-
able ; however, the customary branch to the Gasserian ganglion
was present, though very small, as it should be. In Plethodon
I was unable to detect it, and believe the lateral line roots
lacking. *

* Pinkus 95 has examined the larvee of Salamandra maculosa and atra,
Desmognathus fusca and Salamandrina and finds the lateral line roots as
described by Strong. In the adult Salamandra atra and Geotriton fuscus
the nerves are absent although in Salamandra asmall strand of fibers

_ passed from the seventh to the Gasserian ganglion, there to end without

any corresponding issuing nerves. In Protopterus he found the lateral line
nerves much as in the Amphibia. As would be expected from the distri-

142 PROCEEDINGS OF THE

Especially interesting and prominently advanced of late is the
theory of the origin of the ear from this system of sense organs.
In support of this are (1) the fact of the origin of the nerves
supplying the neuromasts from the immediate neighborhood of
the eighth, caudad and cephalad of it; (2) the general resem-
blance between the sensory cells and the hair cells of the ear ;
(3) the tendency of the neuromasts to sink below the surface in
development, as does the ear; and (4) the embryologic evidence
so far accumulated indicates that in fishes the ear and the lateral
line system develop from a common epibranchial thickening of
the ectoderm, which spreads caudad and cephalad to constitute
the lines of neuromasts.

Despite the plausibility and attractiveness of the theory, its
acceptance, it seems, should be held in abeyance until one or two
points are determined. Ayers, who is one of the most ardent
supporters of the theory, in the latest publication upon the
relation of the nerve fibers and hair cells in the ear, states that
cell and fiber are directly continuous and the nerve fiber does
not terminate freely among them as had been held hitherto. This
makes the hair cells true nerve cells, parts ( one-half or less ) of
nerve units, comparable therefore to the sensory cells of the nose.
On the other hand, the only application known to me of the
impregnation methods ( Retzius ’93 ) indicates that in the neuro-
masts the termination of the fiber is free in an end brush, as it is
in the end buds. This would make the sensory cells of the
neuromasts but modified epidermal cells, and not comparable ( if
both Retzius and Ayers are correct ) to the hair cells of the ear,
which would be the morphological representatives (in part ) of
the nerve cells of the ganglia in connection with the nerves of
the system. Therefore it is felt that the mode of termination
of the nerve fibers in both the macule of the ear and in the
neuromasts should be reinvestigated before the other facts are
accepted in support of the theory.

bution of the neuromasts, the lateral nerve divides into three superficial
branches, a ventral, lateral and dorsal, and a deep lateral one (R. lateralis ~
profundis), which he deseribes as innervating the caudal portion of the

tail (the caudal more ventral series of neuromasts).

AMERICAN MICROSCOPICAL SOCIETY. 143

METHODS.

The methods employed were of the simplest. Much of the
material was alcoholic which had been previously prepared for
museum purposes. Such forms as were available fresh, e. ¢.,
Diemyctylus, Necturus, etc., were carefully killed in weak ( one-
third per cent.) chromic acid with ether, and the epidermis was
then immediately examined with a lens, and the location and
arrangement of the neuromasts ascertained; in MVecturus they
were difficult to detect in specimens less carefully treated. The
larval Amblystomas were killed in either chromic acid ( one-third
per cent.) or platinum chlorid ( one-quarter per cent.) and pre-
served in alcohol, and examined by strong reflected light with
the compound microscope. Tissue for the histologic exami-
nation of the sense organs was fixed in Picro-aceto-sublimate.
(Formula: fifty percent. alcohol, 1o00cc.; mercuric chlorid, five
grams; picric acid, one gram; glacial acetic acid, 1occ.). The
stains employed were Gage’s hematoxylin and eosin, the last in
an alcoholic ( sixy-seven per cent.) solution. Collodion was used
in imbedding and the blocks were cleared in Fish’s Castor-thyme ©
oil mixture. (Fish, 3.)

144

89g.
185.

922.
82.

"73.

94.
18g.

192.

93.
784.

88.

ol.

"45.

"73:
715.

50.
67.
68.

PROCEEDINGS OF THE

LIST OF PAPERS REFERRED TO.

Allis, E. P., Jr.—The anatomy and development of the lateral line
system in Amiacalva. Journal of Morphology, Vol. II., pp. 463-566.
Beard, John—The system of branchial sense organs and their associ-
ated ganglia inIchthyopsida. A contribution to the ancestral history
of vertebrates. Studies from the Biol. Lab. of the Owen's College,
Vol. L., pp. 170-224; also Quart. Jour. Micr. Sez. (1885).

De Blainville—Principes d’anatomie comparée. 1822, I.

Bodenstein, E.—Der Seitenkanal von Cottus gobio. Zeitsch. f. Wiss.
Zool., Vol. XXXVIL., pp. 121-145.

Bugnion, E.—Recherches sur les organes sensitifs qui se trouve dans
Vepiderme du Proteé et de lAxolotl. Diss Inaug. de Zurich. Tiré
du Bull. Nr. 7. de la Soc. Vandoise de Sci. Nat., Vol. XII. (1878).
Collinge, W. E.—The sensory canal system of fishes. Part I, Gan-
oidei. Jour. Micr. Sci., New Ser., Vol. XXXVI. (1894), pp. 499-537.
Cope, E. D.—The batrachia of North America. Bulletin of the
National Museum, No. 34. Washington, D. C. (1889).

Ewart.—The lateral sense organs of elasmobranchs. I. The sensory
canals of Laemargus. IL. The sensory canals of common skate (Raia
batis). Roy. Soc. Edinb. Trans., Vol. XXXVI., Pt. I. (1892), pp.
59 and 87.

Fish, P. A.—A new clearer for collodionized objects. Proc. Amer.
Micr. Soc., Vol. XV., pp. 86-89 (1893).

Fritsch—Ueber den Bau und Bedeutung der Kanalsysteme unter der
Haut der Selachier, Sitzwngsber. d. Kénigl. Akad. d. Wiss. zu
Berlin, Halbbd. I. (1884).

Garman—On the lateral canal system of the Selachia and Holoce-
phala. Bull. Mus. Comp. Zool. Cambridge, Vol. XVII., No. 2, pp.
57-119 (1888).

Guitel, F.—Recherches sur la ligne laterale de la Baudroie (Lophius
piscatorius). Arch. de Zool. experimental et general, 2 ser., Vol. IX.,
pp. 125-190 (1881).

Hyrtl, J.—Lepidosiren paradoxa. Monographie. Trans. der Béhin.
Gesellsch. der Wiss., V. Folge, Bd. III.. pp. 1-64, Prag. (1845).
Langerhans, P.—Ueber die Haut der Larve von Salamandra maculosa,
Arch. f. mikr. Anat., Vol. IX., p. 744 (1878).

Langerhans, P.—Untersuchungen iiber Petromyzon Planeri. Ber.
der Natur. Gesellsch. zu Freiburg, Vol. VI. (1875), pp. 1-114.

Leydig, Fr.—Ueber die Schleimkanile der Knochenfische. Maller’s
Archiv (1850), p. 170.

Leydig, Fr.—Ueber die Molche (Salamandrina) der Wiirtembergischen
Fauna. Arch. f. Naturgesch., Vol. XXIIL. (1867).

Leydig, F.—Ueber Organe eines sechsten Sinnes. , Zugleich ein Bei-
trag zur Kenntniss des feineren Baues der Haut bei Amphibien und
Fischen. Nova Acta Acad, Caes. Leopold. Caroli. nat. curios.,
Dresden, 1868.

1678. Lorenzini.—Osservazioni intorno alle Torpedini, Firenzi, 1678;

London, 1705, Anlg.

62.
76.

So.

AMERICAN MICROSCOPICAL SOCIETY. 145

McDonnell, R.—On the system of the lateral line in fishes, Trans.
Roy. Irish Acad., Vol. XXTV.) Pee: pp. 161-187 (1862).

Malbranc, M.—Von der Seitenlinie und ihren Sinnesorganen bei Am
phibien. Zeitsch. f. Wiss. Zool., Vol. XXVI. (1876), pp. 24-86,
Merkel, Fr.— Ueber die Endigungen der sensiblen Nerven in der
Haut der Wirbelthiere. Rostock, 1880,

1785. Monro, A. (secundus)—The structure and physiology of Fishes.
1785.

°So.

. Natterer, Joh.—Lepidosiren paradoxa. Eine neue Gattung aus der

Familie der fischihnlichen. Reptilien. Wien, 1845

- Parker, W. N.—Zur Anatomie und Physiologie von Protopterus an-

nectens. Berichte der Naturforschenden Gesellschaft, Vol, IN 3
Heft, pp. 1-26 (1888),

Pinkus, F.—Die Hirnnerven des Protopterus annectens. Schwalbe's
Morph. Arbeiten, Vol. IV. (1895).

- Pollard, H. B.—The lateral line system in siluroids, Zool. Jahrbuch,

Vol. V., p. 525 (1893),

. Retzius—Die Nervenendigungen in den Endknospen, resp. Neven-

hiigeln der Fische und Amphibien. Bio], Unters., N. F. IV. (1893).
Robin C. Recherches Sur un appareil qui se trouve sur les poissons
du genre des Raies, Annales Sci. Nat. (8e sér.), Vol, VIL, pp. 193-204,
Schulze, F. E. —Ueber die Nervenendigung in den sogenannten
Schleimkanilen der Fische und iiber entsprechende Organe der durch
Kiemen athmenden Amphibien. Arch, Sf. Anat. u, Phys. (1861), p-
759.

Schulze, F. E.—Ueber die Sinnesorgane der Seitenlinie bei Fischen
und Amphibien. Arch, J. Mik. Anat., Bd. VI » Pp. 62 (1870),

. De Séde de Liéoux, P. —Recherches sur la ligne latérale des poissons

osseux. (1884,
La ligne latérale des poissons osseux. Rey, Scientif., Vol. XXXIV.,
467.

Mobo X.Vit. pp. 95-113 (1880). IL Die Seitenorgane der Selachier.
Arch. f. mikr. Anat., Vol. XVIL., pp. 458-478, III. Die Seitenorgane
der Knochenfische, Arch. f. mikr. Anat., Vol. XVIIL., pp. 364-390
(1880),

1664. Stenonis.—De muscalis et glandulis observationum specimen

10

cum duabus epistolis quarum una ad Guil. Pisonum de anatome
Rajae, etc. Amsterdam, 1664,

- Strong, O.—The cranial] nerves of Amphibia. A contribution to the

morphology of the vertebrate nervous system. Jour. Moph., Vol.
X., pp. 101-230 (1895).

. Wiedersheim, R.—Grundriss der vergleichenden Anatomie der Wir-

belthiere. 38te Auflage. Jena, 1893.

Wright, R. Ramsay—On the skin and cutaneous sense organs of
Amiurus. Proce. Canad. Inst,, Vol. II., Fasc. N 0. 8., p. 252. Toronto,
1884.

Wright R. Ramsay—Some preliminary notes on the anatomy of
fishes. Proc. Canad. Inst. Toronto, Feb, 7, 1885, p. 3-4,

146 PROCEEDINGS.

EXPLANATION OF THE PLATES.

The specimens from which most of the outline plottings were made
were alcoholic, 50 that in many Cases the shape and position of the
legs and arms are unnatural.

Figures 1-3, 6-8, 16, 18, 20-22, 94-26, 30 and 31 were outlined at the
required magnification by means of a vertical photographic camera. The
specimens, in water or alcohol, were placed in direct sunlight, and in place
of the ground-glass screen, a clear glass was used across which tissue paper
was stretched and fastened, whereon the outline and as many details as were
necessary were traced.

Figures 4, 9, 9-11, 17%, 19-23, 93 and 36 were drawn with a compound
microscope by the aid of an Abbe camera lucida.

From the curvature of the surface the relative position of sense organs
at the periphery of any aspect would be falsely shown, hence, in many of
the figures they were discarded and only the groups strictly belonging to
any one aspect shown. The approximate magnification is given for each
figure.

The names that have been applied in description to each line or series are
indicated in the figures by the following abbreviations:

30, — Supra-Orbital. O. = Oral.
10. — Infra-Orbital. A. = Angular.
PO. — Post-Orbital. Vv. = Ventral.
J. = Jugular. D. = Dorsal.
G. = Gular. L. = Lateral.

OC. = Occipital Group.

PLATE I.

Necturus maculatus.

Fic. 1, Lateral aspect of the head; the short lines represent linear
groups of 6-2 organs each, X 1.

Fic. 2. Dorsal aspect of head, X 1.

Fic. 3. Ventral aspect of head, X J.

Amblystoma punctaium.

Fic. 4. Larva 16 mm. long. Ventral aspect of head, < 8; each circle in
this and the following figures represents a sense organ.

Fic. 5. Larva 16 mm. long. Dorsal aspect of the head, X 8.

Fic. 6. Adult 18.5 cm. long. Ventral aspect of the head, X 1.

Fic. 7. Same. Dorsal aspect of the head, X 1.

Fia. 8. Same. Lateral aspect, X 4:

Fia. 9. Larva 52 mm. long. Dorsal aspect of the head, X 4.

—e SC

Amblystoma.

PEATE 1,

# {
yi {\ (\

Amblystoma.

Amblystoma.

PROCEEDING

PLATE Il.

SD:

Amblystoma punctatum.

Fig. 10. Larva 2
Fig. 11. Same.

8.5 mm. long. Dorsal asp
Ventral aspect of the head, xX 8.

ect of the head, X 8.

Protopterus annectens.

Fic. 12. Specimen 26 cm. long. Ventr
circles represent pores of the canals, the
indicated by dotted lines.

al aspect of the head, x 1. The
free lines of sense organs being.

Amphiuma means.

Fic. 13. Lateral aspect of the head,
Fic. 14. Ventral aspect of the head, X
Fic. 15. Dorsal aspect of the head, X 1

IN.
ie

PEATE IT,

cere ue
°
°
hn ° °

( : Q. G =

Amphiuma

Protopterus.

Amphiuma. ; Amphiuma.
VE

BLK del. ay

150 PROCEEDINGS.

PLATE Ill.

Fie. 16. Adult Necturus maculatus. Lateral aspect. of body. The
organs upon the head are omitted, X +:

Fic. 17. Larval Amblystoma punctatum, 28.5 mm. long. Lateral.
aspect of the body. X 4

Fig. 18. Larva of Gyrinophilus porphyriticus. Lateral aspect of the
body, X 1.

Fic. 19. Larva of Amblystoma punctatum 16 mm. long. Lateral
aspect, X 8.

Diemyctylus Virtaescens .

Fic. 20. Adult male. Lateral aspect, X 1.

Fig. 21. Same. Ventral aspect, X 1.

Fic. 22. Same. Dorsal aspect of the head, X Ibs

Fic. 23. Larva 32mm. long. Dorsal aspect of the head, X 8.
Gyrinophilus porphyriticus.

Fic. 24. Larva 82 mm. long. Dorsal aspect of the head, X 2.

Fig. 25. Same. Lateral aspect of the head, X 2.

Fic. 26. Same. Ventral aspect of the head X 2.

Protopterus annectens.

Fic. 27. Lateral aspect of the head, X 1. The circles in this and the two.
following figures represent pores, the free lines of sense organs being indi-
cated by a dotted line except in Fig. 29.

Fic. 28. Dorsal aspect of head, X 1.
Fic. 29. Lateral aspect of the body, X +:

Siren.

Fic. 30. Ventral aspect of the head, X 1. The short lines indicate.
groups.
Fic. 31. Lateral aspect of the body, X 1.

PLATE III.

Protopterus.

sate are

Protopterus.’

152 PROCEEDINGS.

PLATE IV.

Fic. 32. A neuromast from the skin of the snout. Necturus, < 325.
con.—conical or sensory cells.
sp.—=spindle cells.
Fic. 33. Larval Amblystoma punctatum, arms just budded. Dorsal
aspect of the head, 8.
Fig. 34. A neuromast from the skin of the throat (gular line). Diemyc-
tylus viridescens, red (land) form, 3825.
con.—conical cells.
sp.=spindle cells.
Fig. 35. A neuromast from the lateral line near the level of the hind
legs. Diemyctylus viridescens, adult viridescent (aquatic) form.
con.—conical cells, 325.
sp.—spindle cells.
Fig. 36. Larval Amblystoma punctatum, arms just budded. Ventral
aspect of the head, x 8.

Sows

Amblystoma.

PASE, SV

Amblystoms

BEKadd

154 PROCEEDINGS.

PLATE V.

Rana catesbiana.

Fig. 37. Latero-dorsal aspect of the body, x 2. Short lines indicate
groups.
Fic. 38. Ventral aspect of the head, x 2.

Cryptobranchus allegheniensts.

Fia. 39. Ventral aspect of the head, < 4.

Fie. 40. Diagram of the elongated papilla divided by the opening of the
sense organs, with a line beneath to show its representation in Fig. 39.

Fias. 41 and 42. Isolated cells of the neuromasts from the head of
Diemyctylus viridescens, X 615. Isolated by twenty hours’ maceration in
one-half Miiller’s fluid.

Lepidosiren paradoxa.

Fic. 43. Lateral aspect of the head, X 4.

Fia. 44. Dorsal aspect of the head, x 4

The lines or portions of lines seen are represented in the last two figures
by a succession of dots. From the condition of the specimen the plottings
were necessarily imperfect.

Fia.45. Two submerged (developing) organs from the skin of Amblystoma
parotid region, representing apparently an organ dividing by fission, 325.

con.=conical cells.

PEATE V:

Lepidosiren.

=

THE CHLOROPHYLL BODIES OF CHARA CORONATA.

W. W. RowLeE, Ithaca, N. Y.

No group of plants affords a better opportunity for studying
the contents of vegetable cells than the Characee. Of the two
principal genera in the group, WVetella is most often recom-
mended for histological studies. Chara coronata, however, is
without the cortical cells which interfere so seriously with
observation of cell-contents in other species of Chara. As this
particular species is more abundant in the vicinity of Ithaca than
any species of /Vite//a, it has been selected more often as a sub-
ject for laboratory investigation in vegetable histology, and one
of the interesting observations made in connection with these
studies has reference to the number of chlorophyll bodies in a
given area.

The plant body is made up of successive joints. At the
joints are lateral outgrowths frequently called “leaves.” The
plant simulates an ordinary terrestrial herb in having nodes and
internodes and in having leaves borne at the nodes. Each
internode consists of a single, large cylindrical cell. Inthe lower
portion of the axis the internodal cells are sometimes more than
an inch long and they grow successively shorter toward the
apex. The nodes consist of several cells. The internodal cells
are lined with an ectoplasm in which is imbedded the chloro-
plastids, to which, as is well known, is due the green of plants.

The chlorophyll bodies lie in longitudinal. rows and very close
together, but never so close as to overlap. The rows lie
obliquely in the cell, thereby giving the impression of being in a
spiral The number of chlorophyll bodies in a given area is
something surprising. They are larger and fewer in the older
parts of the plant axis, and in these parts number from 12,000
to 15,000 to the square millimeter. In the younger portions of

156 PROCEEDINGS OF THE

the axis and in the leaves their number rises to from 25,000 to
30,000 in each square millimeter, a number which may possibly
be more easily comprehended when we reduce this to inches, and
find that there are more than a million of these bodies to each
square inch of surface.

It is interesting to note that just as in terrestrial plants, the
chlorophyll is more abundant in the lateral organs.

CONTRIBUTIONS TO THE LIFE-HISTORY OF SYMPLOCARPUS
FCETIDUS.

W. W. RowLer and Mary A. NicHots, Ithaca, N. Y,

It seems that the characteristic Property which has given to
this plant its popular name and its malodorous reputation has
further served to render it immune from the attacks of Scientists.
Hence, although a prominent feature of our flora, a plant of wide
distribution and a member of a group which is especially interest-
ing in its position and relations, yet comparatively little concerning
its anatomy or life-history has found its way into the annals of
botany. The difficulties attendant upon its study are augmented
by its unattractive habitat, and the fact that many of its life
processes are carried on under a heavy blanket of snow. When,
late in February, Openings begin to appear at the summits of
cone-shaped, snow-walled cavities (note 1), to announce the ad-
vent of this earliest herald of plant life, one finds that he has
already flaunted his pollen within the warm, thick-hooded Spathe,
which nestles close to a sturdy leaf-roll. But the initia] stages
of growth may be studied through July and August. At this
time the next year’s spadix is already differentiated deep down in
the heart of the plant, usually six inches or more below the sur-
face of the ground. Now, too, one finds close to the perennial
rootstock, where they have fallen from the fleshy spadix of the
Previous autumn, a handful of brown, bulblet-like seeds, from one
to one and one-half centimetres in diameter. These seeds are
described by Gray in his “‘ Manual” as « filled by the large, glob-
ular and fleshy, corm-like embryo which bears one or several
plumules at the end next the base of the ovary ; albumen none.”
In the large number of seeds examined we have failed to find, in
any case, more than one plumule. The plumule consists ofa
central axis, upon which are born several rudimentary leaves,

158 PROCEEDINGS OF THE

forming successive over-lapping layers. As fast as these cap-like
layers push out into true leaves, new layers are cut off from the
tip of the axis. The axis contains a ring of well-defined vascular
tissue, which extends from the base of the plumule nearly to the
opposite end of the seed, and becomes more and more conspic-
uous as the xylem hardens with the growth of the seedling. The
radicle is entirely wanting, and hence no primary root is developed,
The plumule and axis are wholly surrounded by the fleshy scutel-
lum, which is so largely developed here as to fill the entire seed.
It is composed of loose-celled tissue of a bluish color, richly
stored with proteids, which take stain very readily. The axis of
the embryo gradually lengthens into rootstock, which contains a
central cylinder, limited by the ring of vascular tissue, and inter=
spersed with numerous concentric vascular bundles, irregularly
arranged, and much contorted. From these bundles secondary
roots develop adventitiously after the plumule has attained con-
siderable size.

The continuation of the bundles from the rootstock to the
petioles is not distinctly traceable and, moreover, the bundles of
the petioles and of the blades as they expand are collateral and
very regular.

A conspicuous feature in all the tissues of this plant is the ex-
traordinary number of sacs containing raphides. These occur in
seed, root, petioles, leaves, spathe and spadex, but are most nu-
merous in the fleshy rootstock, particularly in the regions of the
bundles. It has been suggested that these crystals may be the
cause of the painful irritation produced in the mouth by tasting of
the rootstock. The sensation is similar to that produced by the
corm of Indian turnip, with which many people are familiar. The
irritation is often pronounced mechanical. One may easily fancy
that he feels the needle-like crystals penetrating the tongue and
lips. When the crystals are dissolved out with H. Cl. the irritat-
ing property disappears. Whether, however, the irritation is
mechanical, or is due to the presence of some free acid (like the
formic acid of the stinging nettle), which is reduced by the H. Cl.,
has not been determined. Some discussion of the question rela-

AMERICAN MICROSCOPICAL SOCIETY. 159

tive to Ariseerma appeared in the Botanical Gazette for January,
1888, and September, 1889.

When the rootstock first pushes out from the seed, it has a
diameter of about one-half centimetre, while the old rootstocks
attain a size eight toten times greater. It has been established by
DeBary and others that stems and roots of monocotyledons in
general show no secondary. changes after the primary differentia-
tion of tissue. Increase in girth, which occurs after this, depends
upon “an increase in the volume of existing tissue elements, and
not upon a cambiogenetic secondary formation.” In exception
to this general rule, DeBary says: ‘“‘ Cambium and secondary
wood and bast appear, as far as at present known, only in the
more or less arborescent stems of A/vine@ (Alve, Lomatophyl-
lum, Yucca) ; of Beaucarnea and the Dracenee (Dracena, Cordy-
line, Aletris etc.); in the tubers of Dvzoscoreacee@ (Dioscora,
Tamus, Testudinaria) ; and the roots of Dvacene@. Vines adds
to these some shrubby /ridace@, such as Aristea and the stem of
Isoetes. To these exceptional monocotyls must now be added
our Symplocarpus fetidus. Entirely surrounding the central
cylinder, in which the vascular bundles are disposed, is a ring of
meristematic tissue. It consists of from two to five layers of
tabular cells. The outer layer conforms nfore or less closely to
the structure of the adjacent parenchymatous cortex, while within
the irregular radial and tangential divisions of the cells provide
for the formation of the much contorted and curiously disposed
xylem and pericycle. Vines, in his enumeration of the various
types of secondary thickening, gives the two following descrip-
tions :

First—‘ There is no primary cambium layer, the bundles
being all closed; secondary growth in thickness is effected by a
ring of meristem quite external to the primary bundles; this
occurs in the stems and roots of monocotyledons (arborescent
Liliacee, such as Yucca and Dracena ; and some shrubby /ridacee,
such as Aristea). The ring of meristem is usually developed in
the pericycle, but in the roots of Dracenew it is formed partly
from the pericycle and partly from the cortex. This meristem

160 PROCEEDINGS OF THE

ring is not termed a cambium ring, because it does not form wood
on one side and bast on the other; but it forms, centrifugally,
entire, closed, concentric (with external wood) bundles together
with intervening fundamental tissue.”

Second— There is no proper cambium layer, but the primary
bundles are invested by a pericyclic meristem ring, which gives
rise externally to a considerable amount of parenchymatous sec-
ondary cortex, and internally to a small amount of vascular
tissue ; stem of Isoetes.”’

Neither of the foregoing types conform strictly to the con-
ditions found in Symplocarpus. The structure here agrees with
the first type in that it forms centrifugally entire, closed, concen-
tric bundles and intervening fundamental tissue ; but differs from
this type in that it also forms, centripetally, parenchymatous
cortex. It differs from the second type in that it forms, internally,
not a small amount, but a large amount, of true vascular tissue ;
and also, as before stated, a considerable amount of intervening
or pericyclic tissue.

The results of our investigations so far may now be tabulated
as follows :

1. The embryo displays a peculiar individuality in the absence
of a radicle, the matked development of vascular tissue in the
axis and the conversion of the scutellum into a-fleshy bulb filling
the entire seed

2. There is a probability that the raphides have to do with the
irritating taste of the plant, and this probability needs confirming
or refuting by chemical analysis.

3. The rootstock contains a layer of meristematic tissue sep-
arating the cortex from a central cylinder in which the vascular
tissue is disposed, and this meristematic layer gives rise, centrifu-
gally, to complete closed bundles and inter-fascicular tissue and,
centripetally, to cortex.

Botanical Laboratory, Cornell University.

ee

162 PROCEEDINGS.

EXPLANATION OF PLATES,

PLATE I.

1. Drawn from nature, January 15,1895. (a) Tip of rootstock. (b) Per-
ennial rootstock. (c) Annual shoot, a leaf-roll only—no flower stalk
present.

2. (d) Spathe. (e) Leaf-roll. (f) Enveloping bract.

3. Young seedling, showing seed, leaf-roll and rootlets.

4. Flower cluster, the base of the spathe in longisection and the upper
part cut away to show spadix.

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164 PROCEEDINGS.

PLATE Il.

5. Cross section of rootstock. (c) Cortex. (m) Meristem. (p) Peri-
cycle. (x) Xylem.

6 and 7. Longisections of same.

8. Raphides-bearing cells.

NoTE 1.—These holes in the snow through which the young plants first
appeared were so conspicuous and so constant a character as to excite par-
ticular attention. They were usually from three to eight inches in diameter,
the smaller ones being larger at the base and forming a more or less arching
roof over the plant. The unusual regularity of appearance and the fact
that the plants were not yet on a level with the surface of the snow, sug-
gested the possibility that the melting of the snow around these plants is
induced by heat generated by plant activity. A series of careful experi-
ments was made to determine the difference between the temperature of
the plant at the base of the spadix within the spathe and that of the sur-
rounding atmosphere. The maximum difference observed was four degrees
C. This was when the temperature of the surrounding atmosphere was
4° C. and the temperature of the plant was 0°.

|) eo Be Co

; pos, L@. NiVe
ae =

SPECIFIC GRAVITY METHODS OF DETERMINING THE
PERCENTAGE OF HAEMOGLOBIN IN THE BLOOD
FOR CLINICAL PURPOSES.

—— eee
FC. Buscu. B. Siete Wel Kerr, Jr., B. S., Buffalo, N. Y.

Each year the importance of the clinical examination of the
blood is becoming better recognized. In this examination there
are two points to be ascertained which are generally acknow]-
edged. These are, the percentage of Hemoglobin and the num-
ber and kind of ted and white blood corpuscles,

For determining the hemoglobin there are several methods.
The hemometer of Fieischl, the hamoglobinometer of Gowers
and the spectroscopic method of Henocque, are fairly well known.
None of the above methods employ the microscope, but a deter-
mination of the hemoglobin is so intimately connected with a
microscopical examination of the corpuscles of the blood, that we

It is recognized that there js a relation between the specific
gravity of the blood and its percentage of hemoglobin: Ham-
merschlag has constructed a table giving the haemoglobin per-
centages corresponding to the different specific gravities of the

In these observations we have compared the specific gravity
method of Hammerschlag with the hemoglobinometer of Gowers
and the hamometer of Fleischl.

Fleischl’s hamometer consists of a colored wedge, with a
graduated scale attached ; a well with two compartments, one for

166 PROCEEDINGS OF THE

pure water and the other for diluted blood; and a capillary
pipette for measuring the blood. The blood obtained, by punct-
uring the finger, is drawn by capillarity into the pipette, from
which it is washed into one of the chambers of the well.

Here it is thoroughly mixed with the water. Both compart-
ments are then filled with water and the well is covered by a
glass plate. The well is placed upon the stand so that the com-
partment filled with distilled water is over the colored wedge.
This is moved by a screw until its color corresponds to that of
the diluted blood in the other compartment. The percentage of
hemoglobin is then read off from the attached scale. In using
the Fleischl, artificial light is necessary, daylight being excluded.

The hamoglobinometer of Gowers is usually manufactured
with but one colored tube, which is for use with daylight. There
is another form in which there are two tubes, one for use with
daylight and the other for artificial light. The one which we have
used is of the former kind. It consists of a sealed tube filled with
a glycerine-jelly solution of carmine and picro-carmine of the
color of an one-per-cent. solution of normal blood ; another tube of
the same diameter to hold the blood to be tested; a pipette
graduated to 20 cu. m.m. and a stand to hold the two tubes, side
by side. The blood measured in the pipette is mixed with a small
quantity of water in the graduated tube; water is then added
until the dilution corresponds in color to that of the standard
solution in the other tube. In making the comparison it is neces-
sary to hold the instrument against a white back ground, opposite
the source of light or directly between the eye and the window.

The method which we have used for determining the specific
gravity, and thus the haemoglobin of the blood, is not so well
known as the above and will therefore bear a more detailed
description. It is one used by Hammerschlag and depends upon
the well-known physical principle that a body which will float
indifferently in a liquid is of the same specific gravity as that
liquid. For this purpose, two liquids are taken, one of a higher
and the other of a lower specific gravity than that of the blood,
with neither of which it will mix. The necessary apparatus con-

AMERICAN MICROSCOPICAL SOCIETY, 167

sists of a hydrometer, hydrometer jar, chloroform and benzole.

In using this method, the finger is pricked and the blood thus
obtained is introduced into a mixture of chloroform and benzole
in the hydrometer jar. The drop of blood, since it will not mix
with either chloroform or benzole, retains its spherical form. If
the drop sinks the mixture is too light and must be made heavier
by adding chloroform. If it rises the mixture is too heavy and
must be made lighter by adding benzole. By carefully adding
one or the other a point is reached where the drop of blood will
neither rise nor sink, but will float indifferently in the mixture.
At this point the specific gravity of the blood is the same as that
of the mixture, By means of the hydrometer we can obtain the
specific gravity of the mixture and thus at the same time that of
the blood.

It is desirable to use a medium-sized drop of blood and it is
better not to divide this into several. Care must be taken, how-
ever, to mix the liquids thoroughly by stirring with the glass rod.
In order to facilitate Mixing, it is well, when the liquid is too heavy,
to add an excess of benzole and bring it back to the desired point
by adding chloroform. The latter being heavier, sinks and thus
mixes more readily with the mixture.

We have found it convenient to obtain the blood from the
palmar surface of the middle finger of the left hand, and have
used, for this purpose, an ordinary sharp-pointed steel pen with
one nib broken off. A new pen may be used for every test and
Should be sterilized by heat. The finger also should be washed
with some antiseptic, in order to take every precaution against
infection. This method of obtaining the blood was used by us
for the three instruments.

For introducing the blood into the chloroform-benzole mixture,
a pipette of fine calibre may be used. A sufficient quantity of
blood is drawn into this and expelled in the middle of the mixture.
Care should be taken that all of the blood is not blown out, but
that some remains in the tip of the pipette. That which has been
expelled will usually adhere to the pipette as a large drop and
must be shaken loose. By thus holding back a small portion of

168 PROCEEDINGS OF THE

blood, the liability of mixing air with the drop is avoided as much
as possible.

E. Lloyd Jones, of Cambridge University, uses a modification of
the method of Professor Roy. This, which depends upon the same
principle as the preceding, consists in the use of numerous solu-
tions of glycerine and water, the specific gravities of which are
known and which are successfully tried until one is obtained,
corresponding in specific gravity to that of the blood.

His apparatus consists of twenty to twenty-five one-ounce glass
bottles filled with standard solutions of glycerine and water, dif-
fering one from the other by .oo1 of specific gravity ; a number of
fine glass pipettes drawn out to a point and bent at right angles
near the tip; a cylindrical glass jar of about one dram capacity ;
and a number of clean, sharp suture needles. After puncturing
the finger on the dorsal aspect near the root of the nail, the blood
which exudes of itself or after the finger has been quickly
squeezed, is drawn into one of the pipettes. This is introduced
into one of the standard solutions and the blood gently blown
out. The solution chosen is of high or low specific gravity
according to the appearance of the patient. The bent point of
the pipette prevents the blood from being given an impetus up
or down when blown from the end,

According to whether the specific gravity of the blood is equal to,
greater, or less than that of the solution, it will pursue a horizontal
course, sink or rise. By trying a number of solutions one may
be found in which the blood neither rises nor sinks, or two are
found in one of which it rises and in the other sinks. In the last
case the specific gravity of the blood is between the two.

In our experience with the Gowers’ instrument, we have found
it very unsatisfactory. It is often quite impossible to get the tint
of the diluted blood to correspond to that of the standard one-
per-cent solution. Even when this is attained, a difference in
shade may be produced by looking at the instrument somewhat
from the side instead of straight from in front; by holding the
paper for reflection farther away from or nearer to the instrument ;
by holding the instrument between the eye and the window or by

AMERICAN MICROSCOPICAL SOCIETY, 169

moving farther away from the window. In the last case, in sev-_
eral instances, the differences produced by moving twenty feet
away from the source of light, was fifteen per cent., the blood
requiring to be more diluted when farther from the window and
thus giving a higher reading. These tests were made in a
hospital ward on a day of average brightness. Therefore it may
be seen that in addition to the other sources of error, the nature
of the day, whether it be bright or cloudy, will make an appreci-
able difference.

We have frequently disagreed in our readings of the same test
in both Fleischl and Gowers and others also have differed from
us as to when the proper shade was attained. In using the
Fleischl instruments, in comparison in the same cases, we have
generally found a difference in reading between the two. In
thirty per cent. of these comparisons the difference was as much
as ten per cent. We have also found that in one-fifth of our cases
we disagree in our readings of the same instrument.

We have found it a great inconvenience in making bed-side
tests in a hospital ward, to run to some other part of the ward or
building to a dark room. In order to obviate this difficulty we
have adopted the following device: This consists in our instru-
ment bag fitted with a cardboard cover; at one end of this a
hole is cut for the passage of a lamp chimney ; at the other end
a small hole for looking through the well of the instrument, and
at one side of this a window with a flap for inserting the hand to
move the wedge.

Hammerschlag’s method has the advantage that there is no
color test. Every one must agree as to whether the drop rises
or sinks or stays where placed. It is also very inexpensive, all
that is necessary being a hydrometer jar, chloroform and benzole.
The method of Roy and Jones necessitates keeping on hand a
large number of solutions which require careful standardization
and must be re-standardized at frequent intervals, Although
this method may be better where a large number of cases are
to be examined in a short time, yet for the ordinary observer,
who uses a method of this kind. less often and upon a small

170 PROCEEDINGS OF THE

number of cases, the one which we have used seems prefer-
able.

In both methods, Hammerschlag and Jones have found that
there is no appreciable difference due to variations of temperature
in the room. :

The results which we have obtained in making parallel tests
with the above described methods, may be summarized as follows :

The readings of the Fleisch] ran as a rule from ten to fifteen
per cent. lower than the percentage estimated from the specific
gravity. The readings of the Gowers ran a few per cent. lower than
the specific gravity. The Gowers’ instrument is liable to an error
of at least fifteen per cent. depending upon the intensity of the light.
The Fleischl instrument is liable to an error of about ten per
cent. In the specific-gravity method there is liability of error
from two sources. The drop of blood may adhere to the sides
of the jar, or some air may become mixed with it. These errors
in the specific-gravity method are reduced to a minimum by
careful manipulation.

The greatest error in this last method may be due to the table,
since of the cases from which Hammerschlag constructed his
table, a great number were primary anzemias and chloroses. For
these his table would probably be more accurate than for our
cases, as all the anemias which we examined, were secondary.
Our cases were taken as ordinarily found in hospital wards, both
medical and surgical, and covered a wide range of diseases.

We are convinced from the experience of others and from our
own observations that all of these methods are liable to con-
siderable error. Osler says that the error in the Fleischl instru-
ment may not be more than two per cent. in blood, which is
nearly normal, but cites Neubert and Letzius as having shown that
in a much impoverished blood the error may be as much as
twenty per cent.

The specific-gravity method has the advantage of cheapness
and convenience ; of taking but little blood, and of not being a
color test. This last is of the most importance since the accuracy
of the test does not depend so much upon the judgment of the

AMERICAN MICROSCOPICAL SOCIETY. si

individual, and makes it practical for observers who lack sufficient
appreciation of colors and shades.

In following up a case with a color test, an error of five per
cent. too low might be made at the first reading, and one of five
per cent. too high at the second and the patient be supposed to
have improved to that extent, when, in reality, his condition had
remained unaltered. With the specific-gravity method this error
is less likely to occur.

It has been found that while the specific gravity may vary at
different times of the day, being influenced by sleep, food, drink,
exercise, etc., the hamoglobin, under similar conditions, varies
also.

From the Laboratory of Pathology, University of Buffalo,
August 21st, 1895.

THE HISTORY OF THE SEX=-CELLS FROM THE TIME OF SEGRE-
GATiION TO SEXUAL DIFFERENTIATION IN
CYMATOGASTER.

Proressor C. H. EIGENMANN, Bloomington, Ind.

ABSTRACT.

Cymatogaster is a small fish, abundant along the coast of Cali-
fornia and extending as far north as Alaska. It inhabits the
shallow water of bays. It is a member of the viviparous family
Embiotocide, and in it viviparity has produced changes greater
than in any other member of the family with the possible excep-
tion of Adcona minima. I have described the general develop-
ment elsewhere and wish to give an account of the reproductive
cells from their first appearance till the sexes have become dis-
tinct. An account of the early history of these cells I have pub-
lished in the Journal of Morphology for 1893. I there traced
these cells from their early segregation till they became located
in the mesentery of the hind gut in larvae 2.5 mm. long. Dur-
ing this time they did not divide. These same cells that origin-
ally became segregated as sex cells migrated with the develop-
ment till they became located as noted above. In larvae 5 and
7 mm. long the cells still retain their individuality, but have
undergone a measurable change. Soon after the 7 mm. stage is
passed the sex cells begin to divide. In the meanwhile they
have migrated laterad and lie for the most part in a longitudinal
fold of the peritoneum where they are mixed with a few cells of
peritoneal origin which later give rise to the entire stroma of the
sex glands. In one case such a sex ridge was formed much
further forward than usual, in connection with a few sex cells
which were accidentally belated in their migration. Behind, the
sex ridges of the two sides are united into a single ridge. The
descendants of one of the original sex cells divide rythmically so
that in later stages little nests of sex cells in the same stage of

AMERICAN MICROSCOPICAL SOCIETY. r73

division are frequently found. There is considerable variation
in the rate of segmentation in larvae of the same size, but the
following will give an idea of the segmentations and the number
of sex cells in differerent stages :

Size of Larva. No. of Sex Cells. ne ee wee
.45— 5 mm. 9— 15 5
8 mm. 22 6
10 mm. 28-— 183 6— 9
12 mm. 39— 143 7— 9
15—17 mm. 638—2, 280 11—138 Sexes distinct.
16—25 2,200—8, 000 135—15

The sexes can first be distinguished after about eleven seg-
mentations from fertilization. The differences are first apparent
in the tunic of peritoneal cells which has become much thickened
on the median side of the sex ridge. A small groove on the
outer ventral part of the sex ridge is the first indication of the
ovarian cavity and is the surest criterion of the female. In the
male the sex gland remains much more circular in cross section
and no groove is developed. Much later, histological differences
in the sex cells can be made out. These differences consist in
the long, slender chromatin threads of the female cell just before
division, being represented in the male of the same stage by short
thick bars. The paper was illustrated by black board sketches
and about three hundred figures. In summing up it was con-
cluded that the peculiarities of the sex cells are due to histo- .
genesis, and their function to the division of labor and not to the
transmission from generation to generation of unchanged germ-
plasm. Many of the causes assigned as determining sex are not
applicable to the present case. While the when, the where and
the how, the sexes become distinct is determined for Cymatogaster,
the why is still left in doubt.

INTERCELLULAR SPACES IN THE EMBRYOS OF ERECHTHITES
HIERACIFOLIA AND BIDENS CERNUA.

Kart M. Wriecanp, Ithaca, N. Y.

Some months ago the writer had occasion to investigate the
structure of the achenes of many of our native Composites.
Among other interesting phenomena observed, were found well
organized intercellular spaces in the embryos of 4rechthites
hieracifolia and Bidens cernua. Since the occurrence of inter-
cellular spaces in embryos is comparatively rare, it was natural
that surprise should be occasioned by finding them developed to
such a degree in these two species. This was more noticeable
since they were not found in any other Composites examined.
Deeming it important that some note of this should be made, the
following descriptions have been prepared :

Erechthites hieracifolia (L) Raf—The embryo is oblong with
the cotyledons somewhat wider than the hypocotyl, but each not
over one-half as thick. The cotyledons about equal the hypo-
cotyl in length, and there is an indication of the growing point in
the axil between them. Some of the cells of the embryo have
already begun to differentiate into a portion destined to become
the vascular portion of the plant. This now consists of a central-
cylinder in the hypocotyl and three branches in each cotyledon,
one of which lies in the center and one near each edge. Immedi-
ately adjoining each of these strands and toward the circum-
ference of the seed is a cylindrical cavity extending the whole
length of the cotyledons. Each is about 25 microns in diameter
and in cross section appears to be surrounded by four cells of such
a shape that together they form a ring. These cells, like other
cells of the embryo, are approximately isodiametric and have
similar cell-contents.

Tracing these spaces down toward the hypocotyl, the two
lateral ones are found to disappear in the vicinity of the plumule,
while the other two continue on either side of the central cylin-
der for about one-third its length, when they too, disappear.

AMERICAN MICROSCOPICAL SOCIETY. 175

Meanwhile two larger spaces have arisen in the axil of the coty-
ledons and continue down the hypocotyl close to the central
cylinder until they reach a point not far from the root cap, where
they each divide into two branches. The cellular sheath of all
of these spaces in the hypocotyl is similar to that described for
the spaces in the cotyledons.

Lidens cernua L.—The embryo of this plant is similar to that
of Erechthites except in the following features :—1. It is a great
deal larger and longer; 2. The cotyledons are much broader
than the hypocotyl; 3. The general form of the hypocotyl is
elongated conical rather than oblong. It presents the same
traces of the future fibro-vascular bundles in the cotyledons, and
central cylinder in the hypocotyl. The area of the latter is, how-
ever, larger and less distinctly marked off.

The spaces in this plant are wholly confined to the hypocotyl,
and are much more numerous than in Eyvechthites. They consist
of cavities between the adjacent cells in the tissue between the
central cylinder and the epidermis. Since in general the cells
are arranged in vertical rows, the spaces are much elongated and
extend for a considerable distance in a longitudinal direction in
the hypocotyl. In case of the larger ones they often extend
through its entire length. Viewing them in cross section the
tendency seems to be for the cells about the spaces to arrange
themselves in the form of a circle enclosing the space within.
This is similar to the structure in Lvrechthites and somewhat
analogous to the peculiar arrangement about resin ducts in
coniferous plants. The exact structure in both embryos will be
more apparent on reference to the drawings of cross and longi-
tudinal sections of each.

To determine the effect of water and growth on these spaces
some seeds were soaked for forty-eight hours and others were
germinated. In both cases the specimens soaked showed a
decided increase in the diameter of the spaces. Investigation of
the seedling, on the other hand, gave rather astonishing results.
It was found that in Erechthites they disappeared almost immedi-
ately. They could not be distinguished in the hypocotyl at all

176 PROCEEDINGS OF THE

after germination but were present in the cotyledons for a short
time. In Bidens they seemed to increase slightly in size, but how
long this increase continued was not determined owing to the
death of the seedlings.

To determine the nature of the contents of these spaces some
specimens were treated with strong sulphuric acid. In the case
of Bidens an embryo was taken that had been soaked in water
for some time and was presumably nearly ready to germinate.
When cleared by the acid, the spaces were seen to be filled with
some gas, which became very conspicuous in the hypocotyl, but,
as would be expected, was not present in the cotyledons. A
young germinating hypocotyl treated in the same manner showed
the presence of the gas ina still more marked degree. vech-
thites, on the other hand, showed no gas at all; but when the
cotyledons of a young seedling were treated, a series of small
globules of some fluid matter was left along the path formerly
occupied by the spaces. A fresh cotyledon was treated with
Fehling’s solution asa test for sugar, but no reaction was
obtained. A test was then made for oil with alcanna. The
result here was more favorable. Bright red drops were plainly
visible not only in the general tissue of the cotyledons, but also
arranged in a row in each of the spaces.

From the above experiments alone it is not possible to under-
stand the exact uses of these structures to the plant. Perhaps in
Lidens they in some way have to do with the transmission
of gases due to growth at the growing point. In &rechthites
where oil is found in the tubés, the function may be one of food
transmission.

Many investigations have been carried on with reference to
intercellular spaces which contain gases. The papers of Martins,
Mangin, Schenk, Schrenk, Devax and Rowlee are some of the
more important. A list of papers relating to the subject is given
in Professor Rowlee’s paper published in the Proc. Amer. Micro.
Soc., p. 143 (1894.) The work has been almost entirely confined
to organs of mature plants, and as far as the writer is aware such
spaces have not been observed in mature seeds.

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178 PROCEEDINGS.

EXPLANATION OF THE PUAiE:

1. A cross-section of the hypocotyl of Hrechthites hieracifolia, before
germination, showing two large intercellular spaces (s.)

2. A vertical section of the hypocotyl of the same showing one
space (s.)

3. A cross section of two cotyledons of the same showing the three
spaces (s) in each.

4. A cross-section of the hypocotyl of Bidens cernua before germination,
showing the spaces (s) scattered throughout the tissue between the central-
cylinder and the epidermis.

PLATE

2

THE ACTION OF STRONG CURRENTS OF ELECTRICITY UPON
NERVE CELLS. .

PrerRE A. Fisu, D. Sc., Washington, D. C.

(PRELIMINARY NOTE.)

The researches of Hodge, Mann and Vejas have demon-
strated that weak electric currents sufficiently prolonged, have
been able to produce unmistakable changes in the Structure of
the nerve cell, as an evidence of fatigue. Such changes have
been found in the cells of the cerebrum as well as in the spinal
and sympathetic ganglia.

If instead of a weak and prolonged current a much stronger
and shorter current be applied, ought more emphasized symptoms
of fatigue to be expected ?

If fatigue be continuous, with no opportunity for recuperation,
death would ultimately ensue. Since a Weak current applied
directly to the nerves causes exhaustion, it would seem reason -
able to infer that a very strong current applied to the skin at the
proper places would tire a person to death very quickly, and, at
least, leave as marked changes in the nerve cells as the weak
current does.

This question was of paramount interest, when, in April, 1894,
there came into my possession while at Cornel] University, —
Ithaca, N. Y., a portion of the cervical] myel of L. R. W., a vic-
tim of an electrocution.

Such statements as the following might lead one to expect
some very radical change in the appearance of the cells: « 1,740
volts were sent coursing through the body, pounding at his nerve
centers with all the force of so many trip-hammers,” and again,
“the current shattered his nerve cells.”

In the case of L. R. W. portions of the cervical myel were
cut in two planes, transverse and sagittal. The cells in the
ventral horns were examined particularly and the conditions

180 PROCEEDINGS OF THE

found in Figs. 3 and 4 were noted. Vacuoles varying in size
and number were located throughout the cell-body, intruding
more or less upon the area of the nucleus. This intrusion
appeared to me not as a direct invasion within the nuclear area,
but as an overlapping of the nucleus by the vacuoles. In many
cases the margin of the vacuole abutted against that of the
nucleus, and occasionally there appeared to be a slight indenta-
tion in the latter at this point The nucleolus was well marked.

If judgment were to be passed after the examination. of the
material from this individual only, it would perhaps be most
natural to conclude that the vacuolation of the nerve cells was
due to the action of the electricity.

It was not generally believed that the murderer was insane,
nor that he was an excessively heavy drinker of alcoholic spirits,
either of which, as well as other diseases, are said to cause
vacuolation in the nerve cell.

In April, 1895, I was enabled, personally, to procure some
more material. The tissues were in the fixing reagents within
four hours after the electrocution. They were selected from the
same regions as in the former case and some of them hardened
and examined by the same methods for exact comparison.

In this individual the nerve cells showed the normal conditions
so far as the microscope could revealthem. Very rarely, indeed,
there could be detected a slight suggestion of a vacuole ina cell.

The murder committed by this man was of the most wanton
and brutal character, and it was believed by some that he was
insane. He brooded over the conditions of his birth, (he was an
illegitimate child) his anxiety to know of his parents, coupled
with the shame they had bestowed upon him, worried him greatly
and may have affected his mental balance.

The appearance of the nerve cells, however, did not indicate
this so much as in the first case, where no special claim of insanity
was made. The gross aspect of the brain presented nothing
uncommon, except that its weight was a little more than the
average.

Age conditions neea not enter, for both were young men, the

! ="

er a

AMERICAN MICROSCOPICAL SOCIETY. 18t

former, L. R. W., being about thirty-five and the latter, W. L.,
about twenty-four years old.

The evidence based upon these two cases is conflicting and
unsatisfactory ; but when compared with the results of others
from similar material, the condition found m the second case
(W. L.) seem to correspond, namely : that no apparent abnormal
phenomena are shown. This also holds true for other: than
human tissue, for in the brain of a calf experimentally electro-
cuted and examined histologically, Dr. Wm. C. Krauss, of
Buffalo, N. Y., in 1890, found ‘the result of the microscopic
examination negative as far as the physical condition of the
separate brain elements are concerned.”

The questionas to the instantaneity of death 1s still a matter of con-
troversy and does not properly come within the scope of this paper.

The rapidity of the electric current depends upon conductivity,
amount of potential, as well as other things.

It is certain that a nerve is not nearly as good a conductor
as a copper wire, nor is it said to be as good as the blood.

Even if the nerve be a relatively poor conductor of electricity,
the estimated rate of passage of the current, as compared with
that of a nerve impulse, would leave the balance considerably in
favor of the electricity, making it probable that the current
arrives at,and paralyzes or kills the nerve cells before the sensa-
tion can be conveyed there.

Experiments performed upon dogs, by those interested in the
subject, show that the heart is the first of the vital organs to cease
its visible action. This has been observed by first anzesthetizing
the animal, removing a sufficient portion of the parietes, keeping
up respiration by artificial means and then applying the current ;
instantly the heart was seen to stop its beating. The effect was
just as marked when the experiment had been carried so far as to
cut the vagus nerves. Inthe majority of cases the heart was still
before the respiration had stopped.

Of twenty-four dogs tested in this respect there were only three
in which this was a matter of doubt, and in these three ‘‘ no
priority could be assigned to the failure of either function.”

182 PROCEEDINGS OF THE

Dr. A. M. Bleile (Electrical World, N. Y., July 6, 1895) be-
lieves that death from electric shock is due to the contraction
of the arteries, caused by the action of the current on the vaso-
motor center, and that this constriction of the arteries offers a
mechanical impediment to the flow of the blood which the heart
is not able to overcome. When nitro-glycerine or nitrite of amyl
was given to counteract this effect, greater currents of electricity
could be borne.

The question arises, if the use of these reagents, after the
electrocution, would not relieve this constricted condition, or, as
has been suggested by others, that an application of a medical
electric current might bring about the same result and promote
resuscitation.

Amperage must be taken into account as well as the voltage.
A continuous current of very high voltage may be received with-
out fatal results. A very rapid, alternating current, in which
the alternation is so rapid as to be practically continuous, like-
wise is not necessarily fatal.

The current usually employed at electrocutions, 1,700 volts
and eight amperes, is said to be equal to about twenty horse-
power. Such a force turned loose into a human body must
effect wonderful changes in the living tissue, although for the
most part subtle enough to evade detection by the microscope.
The constituents of living protoplasm are too little known to en-
able us to understand what changes are effected or just how they
are brought about. It is not probable that one tissue or system
of tissues is selected by the electricity as it traverses the body,
although there may be different degrees of susceptibility. Death
may be brought about by the killing of the cells in the nerve
centers. The electricity in this case acting asa fixing agent, for
as in histology, when certain reagents or in some cases simply
their vapors are allowed to act on living nervous tissue, it not only
kills, but fixes or retains the elements of the tissue in the position
they held at the time of the action of the reagent.

This reaction is of a chemical nature ; it probably is in the case
of electricity. When properly employed the reagents cause no

' AMERICAN MICROSCOPICAL SOCIETY. 183

visible change in the form of the tissue elements, neither ap-—
parently does the electric current.

Whether this hypothesis of electric fixation is applicable to all
tissues of the body, it is now too early to say, thus far it seems
to be a rational one.

Such lesions as have been noted seem to be of a secondary
character, such as minute effusions in the heart caused by capil-
lary rupture due to too violent contraction ; the crenated appear-
ance of the red blood corpuscles taken from the body in the
region of contact of the electrodes some seven minutes after the
execution, as described by Dr. Fell.* While corpuscles taken
one-half hour after the execution from a region farther removed
from the electrodes presented no abnormal appearance as to size
or uniformity of outline.

* Proceed. Amer. Soc. Microscopists, Vol. XII., pp. 1-34, 1891.

184 PROCEEDINGS.

EXPLANATION OR. PALE

d dendrites.

n ?—=neurite.

nl nucleolus.

nr—neuroglia or spider cells.

ns —=Nissl’s spindles.

nu—nucleus.

v =vacuole.

Figs. 1 and 2 From cervical myel of case No. 2(W. L.) Sagittal plane.
Figs. 3 and 4. From cervical mvel of case No. 1 (L. R. W.) Sagittal plane.

The methods used in the preparations from which Figs. 1-4 were taken,
were not adapted to bringing out the appendages of the cell to any great
extent. Such parts of the cell processes as are shown are undoubtedly
dendritic, the neurite not appearing.

Fig. 5. Pyramidal cell from the cortex of the precentral gyre. Case
No. 2(W. L.) Formalin-bichromate-silver preparation,

vs

POA LE.

3 yj P. A, FISH, del,

~ COMPARATIVE MORPHOLOGY OF THE BRAIN OF THE SOFT-
SHELLED TURTLE (AMYDA MUTICA) AND THE ENGLISH
SPARROW (PASSER DOMESTICA).

SUSANNA PHELPS GAGE, Pu. B., Ithaca, N. Y.

The papers presented before this society have so wide a range
that this subject may be admitted with the rest, though its one
claim of unity with other papers of the meeting is that the
methods and apparatus used are those which it is the primary
object of this society to consider and perfect.

The brains studied are too small for thorough observation by
gross methods, hence serial sections through the head after
decalcification and serial sections stained by the Weigert method
have been made, drawn by the camera lucida, and carefully
reconstructed into pictures. With these methods all are familiar,
as they have been discussed before this society.*

It may be asked, why should such widely different forms of
animal life as a soft-shelled turtle and a sparrow be brought into
comparison, the one from the lakes and streams of the middle
United States passing a self-controlled, watchful, quiet existence,
alternating with rapid and vigorous action, the other, the alert,
restless, quarrelsome creature brought from Europe, and become
the pest of our door yards.

To the older anatomist, who worked with such remarkable

* Material: Amyda mutica, gross preparations of the brain of the adult ;
serial, sagittal sections through the decalcified head (Gage, S. H. ’92) of a
specimen 6 cm. across the carapace ; serial, sagittal, frontal and transec-
tions of the head of four specimens 13 cm. across the carapace, stained in
hematoxylin and eosin ; frontal and transections of the brain of two such
specimens, stained by Weigert’s hematoxylin method.

Passer domestica, adult, gross preparations of the brain and sections in
the three planes through the decalcified head; embryos of early and middle
stages of development sectioned through the entire head and compared
with the entire medisected head of specimens of the same age.

The sectioning was done by the collodion method, as modified by Fish
(93) and Gage (’95),

13

186 PROCEEDINGS OF THE

manual skill and accuracy, without the newer appliances, such a
comparison would not be surprising, for to him the differences to
be seen with the eye alone and the correlation, as far as possible,
with the difference in physiological function, were the aims to be
sought. Nowhere can a statement showing clearer insight into
the great problems now confronting the comparative neurologist
be found than that given in the introduction to the great work
on the brain by Tiedemann in 1816 ('16). With the rise of Dar-
winism, and the universal interest in evolution as a process of
creation, and with the new methods which (by means of micros-
copes and microtomes) brought difficult research within the reach
of students of average skill, another bent was given to anatomical
study. The comparative morphologist has been searching for,
and working upon, generalized types, in order to trace the evolu-
tion of animal forms from the simple to the complex.

Perhaps now the time is ripe for the pendulum to again swing
back, and, with the new thought and the new methods, to make
more careful comparisons of widely different forms than before
has been possible, in order to obtain from exaggerated structure
and highly specialized function, added light upon the problems in
which we are all interested.

The work here presented is part of a larger plan in which it is
proposed to bring into comparison certain features of the brain of
one or more species from each of the great groups of vertebrates,
from the period when the brain begins to differentiate from the
initial common plan toward the adult condition. This means
that many hundreds of successful series must be made and
studied. Preliminary work has been done upon certain species
of all the great vertebrate groups, and that upon the amphibian
brain has been published (Gage, S. P., ’93).* The study of two
forms, the turtle and the sparrow, has been carried far enough to
permit a preliminary report, leaving details to be filled in as
material accumulates.

The problem before us must be attacked in two ways, first, by

*As the present paper is a continuation of the work done upon Diemye-
tylus, Amia and lamprey, no further reference will be given to that article.

AMERICAN MICROSCOPICAL SOCIETY. 187

determining the exact morphological equivalent of each part in
the animals compared by means of their development, and by
their connections in the adult; second, by close observation of

. the animals, to determine their normal physiological activities and

the correlation of these with the parts of the brain which control
them. With the plan in mind a beginning can be made, and
below are given some of the points that have been studied.

This work is more or less connected with that done in mor-
phology at. Cornell University by Professors Wilder and Gage,
and represented in special papers by them and Drs. Fish, Hum-
phrey, Kingsbury and Stroud. The articles which are most
closely related to the present are by His, Studnicka, Burckhardt,
Herrick, Humphrey and Turner. Herrick (’91) figures the
dorsal, ventral and lateral aspect, and six sagittal sections of a
young Aspidonectes spinifer, another genus of soft-shelled turtles,
closely related to Amyda. In four pages of text he makes some
general comparisons with the brain of other reptiles, and discusses
the histology. In general features the brain of Aspidonectes
agrees with that of Amyda, for example, the coalescence of the
olfactory lobes. Some points discussed are aside from the main
purposes of this paper, others will be further mentioned. Hum-
phrey (’94) shows the external form of the brain of the snapping
turtle (Chelydra serpentina) both adult and embryo, figures the
mesal aspect of this and of the green turtle (Chelone midas) with
diagrams of fiber tracts. Turner (’91) figures and describes a
few points in the anatomy of the brain of a sparrow, as the union
of the olfactory lobes, and from the form of the brain in different
birds draws conclusions as to their relative rank, placing the
sparrow among the more highly specialized. Other articles upon
general or special problems touched upon will be referred to in
phe text.

Among the turtles, the soft-shells are far from the type, not
only in external form, but in habits. The body is depressed.
The leathery carapace and the combined zrial and aquatic respira-
tion suggest that they may be lonely representatives of a primal
stock from which the other turtles are an offshoot, but the union

188 PROCEEDINGS OF THE

of the olfactory lobes indicates specialization rather than a prim-
itive condition. As compared with a snapping turtle (Agassiz,
’57) it is seen that the small head of the soft-shell does not indi-
cate a relatively small brain, but a more compact arrangement in
a cavity which is none too large, while the great jaws and power-
ful muscles of the adult snapping turtle require a large skull, and
leave a spacious cranial cavity in which the comparatively small
_brain stretches itself out freely upon the floor in such a way as
to show all the segments in any view, and thus to make ita
brain easy to study (Humphrey, ’94).

Among the birds, the group to which the sparrow belongs
shows a tendency similar to Amyda toward a thinning of the skull,
and a crowding and overlapping of the segments of the brain. In
the sparrow these are indeed carried to an extreme.. Among the
more general forms of birds, as the turkey (Turner, ’91, Carss,
’95) the optic lobes or gemina appear on the dorsal side, but in
the sparrow they are visible from the ventral aspect. They are
covered by the large cerebrum and cerebellum, which together
give an approximately spherical outline to the brain. A com-
parison of the dorsums of the sparrow and turtle brains (Figs. 19,
28) shows the great difference of form. The turtle’s brain shows
a portion of all the important parts; the sparrow’s brain is so
warped from the simple type that the olfactory lobes, the gemina
and the oblongata are entirely hidden by the great cerebrum and
cerebellum, thus resembling higher mammals. As seen from the
meson (Fig. 29), this turtle’s brain is one of the most evenly
developed and symmetrical brains that it would be possible to
find ; no part unduly crowds or overtops the rest; each has place
sach part is equal to the others. In the sparrow (Fig. 20) these
conditions are all changed; crowding and disproportionate growth
are the rule. In the younger turtle greater crowding of parts
exists, and in the embryo sparrow the form more nearly ap-
proaches that of the turtle (Figs. 11, 29), while, could we trace
still farther back, it would be found that the two brains are quite
similar in form, the two in course of development taking different
paths, the sparrow’s tending to extreme condensation and to ex-

.

AMERICAN MICROSCOPICAL SOCIETY. 189

cessive growth of certain parts, the turtle’s to uniform growth
and comparative elongation.

It is desirable to see what relations of the brain to the body
and the habits can be established. The brain of the sparrow has
at least twice the size of that of the turtle, while the body
weight of the sparrow is only a fraction of that of the turtle.
The greater absolute size of the brain is accounted for by the
relatively enormous development of cerebrum, cerebellum, gemi-
nums and the optic tracts, while the myel and oblongata do not
attain to as great size as in the turtle. In the turtle a large part
of the activity of the muscles of the body is under the control
of the myel and local nerve centers, as can be inferred by the
long time that the muscles respond to the stimuli after severing
the head, and the long time which it takes to kill the animal by
chloroform. In the sparrow death soon occurs by either
method. The brain of the turtle is then very largely devoted to
the functions which pertain to the head and throat or are under
the control of cranial nerves. This turtle can fill its lungs with
air and then dive under the water and remain for hours together,
the rial respiration being supplemented by an aquatic (Gage,
S.H.,’92). In the brain a modified respiratory center corre-
sponding to this habit is to be expected, but this has not yet been
actually demonstrated, as too many simpler problems had first to
be solved. In the sparrow, in addition to the functions of sensa-
tion, etc., there is much more intimate control of the body
muscles than with the turtle. The mantle of the brain, the part
so greatly developed in man, and forming the highest center for
the control of muscular activity, is little developed in both turtle
and sparrow. In the sparrow the striatums are the most bulky
part of the cerebrum. In them are gathered and from them pro-
ceed the fibers, which are comparable to the motor tracts of
mammals, which connect the higher with the lower nerve centers.
In the turtle, though the striatums occupy a large part of the
cerebrum, they and their motor tracts are much smaller than in
the sparrow, Figs. 29, 36, 30, 31, 22,23. From experiments of
different kinds it is believed that this part of the brain in mam-

190 PROCEEDINGS OF THE

mals has largely to do with the co-ordination of muscular move-
ments. The ceaseless activity of the sparrow, its movements
implying rapid and accurate co-ordination certainly point to a large
controlling center. The turtle which basks for hours in the sun,
or for hours lies at the bottom of the stream, watching for prey
or hiding from enemies, only occasionally making a rapid move-
ment and then of a comparatively simple kind, apparently does
not need so complex or large a center for muscular co-ordina-
tion. The removal of the cerebrums from a frog with subsequent
regaining of the power to balance himself and to perform a few
acts requiring co-ordinated movements, seems to show that
higher centers in the cerebrum are not the only centers, but on
the other hand the striatums in the frog are insignificant in size
and are simple in structure. Other questions which have arisen
as to the relation of function to form can be better treated under
special headings.

The differences of form are obvious, but if the problem is
viewed in a more fundamental way the tendency is to swing to an
extreme and see only the unity of plan in construction and for-
get the differences. Long ago Wilder* urged the importance, in
brain comparison, of the parts which lie exactly upon the middle
plane or meson, and now (Burckhardt ’93) this is coming to be
accepted as the region in which to seek for those structures which
most clearly indicate a common inheritance. Looking closely at
the mesal view of these brains (Figs. 20, 29) great landmarks at
once appear ; the precommissure which unites the cerebrums at
the base, the postcommissure at the dividing line between the
mesencephal and the diencephal, the crossing of the fourth
nerves in the valvula, the cerebellum, the oblongata, the infundi-
bulum and the hypophysis, the chiasma, the epiphysis and the
membranous roof of the diencephal. These are different in size

* The special course on Comparative Neurology given in the spring of
1876 by Professor Wilder, was illustrated by serial sections and by pre-
parations and drawings of the mesal aspect of many vertebrate brains made
by him during the previous summer and earlier. Since that time his
published papers have borne constant evidence of his appreciation of the
morphological significance of structures occurring on the meson.

=

AMERICAN MICROSCOPICAL SOCIETY. IgI

or in location, but from essential relations are found to ‘be
homologous structures. A less fundamental similarity consists
in the union of the olfactory lobes, while striking differences are
found in the condition of the mesencephal, and the presence in the
turtle of a very large medicommissure which is entirely lacking
in the sparrow.

:

MEMBRANES.

The membranes have been only casually studied. In the
adult sparrow they are very thin, crowded together between the
brain and the skull. In the embryo sparrow they are not differ-
entiated into distinct layers (see plate I and II), but the spaces
between the segments are filled by an arachnoid-like tissue in
which are scattered blood vessels. The condition is much like
that found in lamprey, Amia, etc. The membranes of the
turtle’s brain are much like those of the snapping turtle (Hum-
phrey, 94) ; a thick dura, peeling easily from the skull, especially
on the dorsum; an arachnoid especially noticeable about the
mesencephal; a pia lying close to the surface and in many
places, especially over the gemina and cerebellum and between
the cerebrums, clearly seen to be attached by filaments (Figs. 32,
34), as occurs in Demyctylus. Blood vessels of considerable
size, especially in the sparrow, enter the brain substance and
divide into a capillary net work (Fig. 31), as in Avzza and mam-
mals, instead of forming loops, as in Deemyctylus.

MEMBRANOUS PARIETES AND PLEXUSES.

The original roof of the brain-tube is a membrane or tela, and
in the forms under consideration many remains of this primitive
condition persist.

The metatela (mt.), protruding as a sac over the myel in the
early embryo sparrow (Figs. 1, 7), isa prominent feature, but
with development of other parts becomes less so (Figs. 11, 17,
18). It would be in this region that a metapore would be looked
for, but although the membrane is extremely thin it seems to be
continuous, and up to the present no distinct metapore has been

192 PROCEEDINGS OF THE

found. In the turtle this sac is present, though not as large as
in the snapping turtle (Humphrey, ’94). It has a considerable
lateral extent and at the sides the cells become attenuated, but
in all specimens examined this part was so obscured by a plug
of material containing granules that it is impossible to state
whether a metapore exists or not.

In the embryo sparrow, between this sac and the cerebellum
are a few folds in the membrane, the simple beginning of the
metaplexus (mtp., Fig. 1). In the older embryos (Figs. 11, 17),
this plexus is seen to be greatly developed, and to arise in part
of its extent between solid walls, both of which belong to the
cerebellum, thus forming a true efzplerus. In the adult (Pig.
20), with the growth of the cerebellum it becomes an insignif-
cant feature, both on the meson and as it continues along the
caudal border of the cerebellum to the flocculus. In the turtle
(Fig. 29), the metaplexus occupies an extensive portion of the
metaccele. In its caudal part it crosses the meson as a deep in-
folded membrane (Fig. 30), but cephalad it is formed by intru-
sions of pia and covering endyma through the slit (Fig. 36).
The elongated folds interlace across the meson. In the green
turtle (Humphrey, ’94) the plexus is a nearly drum-head-like
membrane with a few slight folds occupying a V-shaped opening
between the short cerebellum and the oblongata. A growth
caudad of the cerebellum narrowing the V-shape to a slit would
produce the result here attained, pushing the lateral part of the
plexus into the cavity and crumpling the*mesal union into a com-
pact mass at the caudal extremity.

In an early embryo sparrow the roof of the epicoele is a narrow
membrane or cfitela ( Figs. 1,6), which is replaced later by a
median lophius, while a raphé-like appearance exists between the.
two halves. In the turtle such a lophius is a marked feature of
the cerebellum and will be discussed under sz/cz.

The membranous roof of the mesencephal, mesotela (mst.), is
found in all stages of the sparrow’s growth, becoming exagger-
ated with age (Figs. 3, 23), while in the turtle it is obscured com-
pletely. (See below under mesencephat ).

AMERICAN MICROSCOPICAL SOCIETY. 193

The roof of the diaccele always remains membranous. Its
original condition, an unfolded tela, is seen in the young embryo
sparrow (d7¢., Figs. 1,2). The folding which occurs later is in the
form of a mesal plate with secondary foldings (dp. Figs. 15, 16),
a condition remaining unchanged in the adult (Fig. 20). In the
turtle the exaggerated tubular condition of the roof of the dien-
cephal described by Humphrey in the snapping and green turtles,
and which was originally mistaken for an epiphysis, does not
occur. But on the other hand, though ventrad of the epiphysis
the plexus takes a mesal position ( Fig. 42), most of the plexi-
form folds are from the lateral walls and are so continuous with
the auliplexus as to make a boundary line between the two im-
possible. The condition in the turtle seems to be one which
could be transformed more readily into the mammalian type
(Wilder, ’89), with two parallel plexuses from the roof of the
diencephal, than the condition found either in the sparrow or in
Amphibia (Diemyctylus and Desmognothus, Fish, ’95) where there
is a single mesa] plexus.

The roof of the prosoccele involves questions of morphology
which will be reserved for the latter part of this paper. There
is in both the sparrow and the turtle, a mesal portion of the
plexus which will here be called audplexus (Figs. 39, 14, 15),
though sometimes called the velum. It gives off the para-
plexus on either side. The faraplexus in the turtle, outlined
faintly in Figs. 29, 36, is of somewhat remarkable form. At the
porta it divides into two portions, a dorsal and a ventral; the ven-
tral passes obliquely cephalo-ventrad, to the angle fc’ in the
paraccele, the dorsal sends a branch cephalad even into the
rhinoccele, and another branch caudad along the dorso-caudal
angle of the paraccele, pc’, to near the tip of the medi-
cornu, fc., Figs. 40-42, 29-31.* This last-named part has no
connection with the paratela. In the sparrow the paraplexus
consists of a single portion outlined faintly in Figs. 1, 11, 20.

*In the article by Herrick (90) on the alligator’s brain, no plexus was
shown, but it really exists in the young, at least, and is very similar to that
of Amyda.

194 PROCEEDINGS OF THE

In the youngest specimen here figured it is merely a slightly
corrugated membrane lying close against the great mesal mass
of undifferentiated membrane (Fig. 2). It is noticeable in the
older embryo that the union of the auliplexus with the para-
plexuses lies dorsad of the porta (Fig. 14).

The faratela in both forms is a membrane extending from the
porta to near the tip of the medicornu. In the turtle it forms a
large thin membrane (Figs. 31, 40-42, vm.) of endyma and pia
stretched over the thalamus, and could we imagine the object
large enough the glistening white of the underlying optic tracts
would shine through it. It is bounded dorsad by the fimbria,
ventrad by a similar edge or ripa, the tenia, lying next the
striatum. (See sa/cz below.) In the adult sparrow it is doubt-
ful if the extensive thin portion of the cerebral wall occupying
a similar position can all be called paratela, because it is traversed
by bands of fibers which converge toward the base of the cere-
brum (Fig. 11), but in the half-grown embryo there is a
definite paratela (Figs. 14-16, 7z.). In the earlier embryo (Figs.
2, 3) the part so designated has a barely appreciable amount of
wall which has the general appearance of nervous substance.

MEMBRANOUS OUTGROWTHS.

Small pockets, or offshoots of the endymal lining of the cavi-
ties, have been observed in both forms, and cannot be omitted,
because of the significance which must attach to any structure
which exists in these parts, where there is preserved most accur-
ately the original pattern. The embryo sparrow has such a
pocket (Fig. 1, R) extending around the dorsal thickened portion
of the terma,in which later appears the precommissure. It seems
like the Lodus olfactorius impar of Kupffer, but no trace of
it is found in the later stages. The adult turtle has two pockets,
nearly meeting about the common projection, formed by the
precommissure and the callosum (Fig. 29). The ventral one
is quite deep (Fig. 35, 4), and is homologous with the preoptic
recess of Amphibia. A pocket of endyma caudad of the post-
commissure in the embryo sparrow (Fig. 11, S), is a strong

AMERICAN MICROSCOPICAL SOCIETY. 195.

reminder of the pocket of endyma, which in the lamprey extends
cephalad of the postcommissure, and in the frog (Ecker ’89) and
shark occupies a similar position. Their homology has not yet
been determined.

The epiphysis of the turtle curves cephalad from the roof of
the diencephal. Between the postcommissure and supracommis-
sure its occluded tube can be traced to join the endyma of the
cavities'(Fig. 29). The condition is not as described by Herrick
(’91) in Asfidonectes, a tube opening “into the canal connecting
the optic ventricles with the dorsal part of the third ventricle.”’
It seems as though this must be a misinterpretation, which would
not have occurred if transections as well as sagittal sections had
been studied. The epiphysis of the sparrow has an elongated
stalk continuing to the skull, along with the great growth of the
cerebrum (Fig. 20). In the embryo it is nearly sessile, and the
tube is open (Fig. 11). The end of the epiphysis in the earlier
embryo (Fig. 1) has not yet formed the complex foldings found
in the adult. It does not appear from the late formation and
slight development of the supracommissure in the sparrow that
this is an essential landmark of the entrance of the epiphysis,
while in the adult the long stretch of membrane between this
entrance and the postcommissure (Figs. 20, 25) shows that varia-
bility in the details of arrangement in this region may be ex-
pected. In neither of the forms is there any appearance of an
eye-like structure in the epiphysis.

It is now known that @ paraphysis exists in a number of verte-
brate groups including the human fcetus (Francotte, 88, ’94). It
is found in the adult Amphzdza and the snapping turtle. It also
exists in this soft-shelled turtle (Figs. 29, 40). As with the
snapping turtle (Humphrey, ’94) the character of the endyma
lining the epiphysis is different in appearance from that covering
the plexuses. It was by this difference alone that it could be
distinguished in many sections from the plexuses, a difference
very marked in Weigert preparations where the nuclei of the
paraphysis stain very deeply. Its function is unknown, but it
furnishes another landmark by which may be determined more

196 PROCEEDINGS OF THE

exact homologies of parts. In the adult sparrow no trace of
this structure was found, but in the younger embryo (Fig. 2) its
presence as a minute pocket was clearly seen. The paraphysis
in Amyda, as in Amp/ibia, lies between the auliplexus with its
branches and the diaplexus, but its ramifications, extending both
cephalad and caudad make it difficult to state with any such defi-
niteness as with the Amzp/fidia the relation of the opening to the
cavities. In the young sparrow (Fig. 2) the paraphysis occurs in
the midst of a mass which gives off the paraplexuses, and it opens
directly dorsad of the portas, z. ¢., into the aula.

RHINENCEPHAL.

This term is used here for convenience alone, since evidence of
segmental value is in these forms purely negative. The olfactory
lobes of these two forms have a feature in common, that is, they
are united across the meson. In the sparrow the union is very
close, the cinerea forming a core containing no cavity and show-
ing no indication of division into two halves (Figs. 20, 21, 27),
while in the turtle the concentric layers about the cavity of each
side are complete, not fusing across the meson (Figs. 30, 31, 37).
In the turtle myelinic fibers lie parallel to the meson, but do not
cross from one lobe to the other. The lobes are, however, united
by aclose meshwork, in which a few blood vessels indicate that the
condition is secondary. A few cells lie exactly upon the meson, but
from the appearance in Golgi preparations they seem to belong to
the neuroglia. The glomerular layer of each lobe is separated from
that of the other by pia. In the sparrow this union was not found
to exist in the younger embryo (Fig. 1), but was fully established in
the older(Fig.11). Thus it appears that this peculiarity of the soft-
shelled turtle (Herrick, ’91), the higher birds (Turner, 91), and the
frog (Ecker, 89), is not one that indicates relationship, but is a
condition incident to other specializations, established compara-
tively late in embryonic development. Without doubt it indicates
the less relative importance of olfaction to these highly specialized
representatives of different vertebrate groups. The process of
degeneration has been carried in the sparrow to an extreme, and

AMERICAN MICROSCOPICAL SOCIETY. 197

its olfactory lobes, from their intimate union and small size, indi-
cate very little functional activity. In the turtle there are two
olfactory nerve roots on each side, a large one from the ends of
the lobe, anda smaller one from the caudo-mesal angle (Figs.
30, 37). Each root has its independent glomerular layer, as
described by Herrick (‘91). The two roots soon unite, and the
course of the olfactory nerves so formed continues independently
to their distribution in the nose. In the sparrow there is one
nerve on each side, and that a small one. The cavity of the
cerebrum does not reach the olfactory lobe in the adult sparrow.
In the turtle the rhinoccele is large, with a lateral extension
(Fig. 31), and itis distinctly marked off from the paraccele by both
dorsal and ventral constrictions, while externally the lobes are
demarcated from the cerebrums by a slight furrow laterad (Fig.
28) and a deeper one mesad, in which are blood vessels (Figs.
20,30).

PROSENCEPHAL.

In any section of the cerebrum of either form under discussion,
it is seen that the mesal and caudal walls are thin. In the adult
sparrow, in many parts, they approximate membranes. In either
case the greater part of the mass is in the body called here the
striatum, in accordance with older usage, though some recent
writers (Herrick, ’91), propose to call it the axial lobe. Spitzka,
(81) pointed out that in birds the striatum in its growth crowded
against the adjacent parts and fused with them. The appearance
in the soft-shelled turtle and in a young alligator, and a compart-
son of sections of the different stages of development of the spar-
row, leads to a similar conclusion. While, therefore, the main
part of the mass is homologous with the striatum of mammals,
the comparatively thin lateral portion fused with it should be
excluded. Curving around the endymal surface of the striatum
in the turtle, and forming the larger part of the protrusion into
the cavities (Plate IV, V), is a portion which seems comparable
with the caudatum of mammals, but in the caudatum of the turtle
the cephalic end is narrow and the caudal wide, thus reversing

198 PROCEEDINGS OF THE

the conditions in mammals (Figs. 38-42); and its caudal tip pro-
jects freely into the cavity. In the embryo sparrow a similar
elevation can be seen (Figs. 1-4, 11). In the older embryo (Fig.
16), the rest of the striatum has grown so much that the cauda-
tum is less clearly seen, and in the adult it is very slightly raised
above the general level.

Following the transections of the turtle’s right cerebrum from
the caudal tip it is seen that the wall has the form of a Greek
delta.

STRIATAL.

The ventral limb -epresents the striatum, the right limb the
pallium, the left represents the combined hippocampal and tenial
borders. The ventral limb is soon increased by the addition of
the caudatum (s¢. Fig. 42), which in a few sections more is fused
with a part of the pallium (Fig. 41), and can be traced to the
olfactory region, where it is gradually thrust away from the endy-
mal surface by the proper olfactory structures. The pallial limb
of the delta occupies at first the lateral portion (Fig. 42), but
cephalad a free portion is confined to the dorsal aspect (Fig. 38),
and is gradually lost in the olfactory region. The mesal limb
soon divides into a dorsal, the hippocampal and a ventral, a
scarcely appreciable ripa, the tenia, seen at the edge of the stria-
tum (Fig. 42). As the porta is approached, the tenia joins with
the wall of the thalamus (Fig. 40) and an outgrowth from this
united portion (Fig. 39), apparently a thickened portion of the
original terma, hence called termatic, joins with the hippocampal
limb (Fig. 38 ) to form a single mesal limb, thus re-establishing
cephalad of the porta, the delta-form. This mesal limb is also lost
in the olfactory region. In the Déemyctylus, sections show this
delta-form of the cavity still more clearly, for the striatal limb is
not obscured by the growth of a caudatum. The portion of the
mesal wall in Dzemyctylus was called the callosal eminence, but

AMERICAN MICROSCOPICAL SOCIETY. 199

here is called the hippocamp. The small hippocampal region of.
the sparrow is, as in mammals, related to the reduced olfactory
lobes, and this segment and the pallial are not well separated in
the adult, although in the embryo (Fig. 2) they are distingutsh-
able.

In the mesal views a projection of the outline of the cavity is
. indicated by interrupted lines. In Fig. 29 an attempt is made to
indicate the two wings of the paraccele of the turtle which are
due to the position of the striatum (Fig. 42). The lateral wing
extends quite far cephalad (Fig. 40 pc’’’.) while the mesal wing
(fe.) in the neighborhood of the fimbria and paratela can be
strictly compared with the medicornu of mammals. The caudo-
dorsal angle (fc’’.) demarcating the pallium from the hippo-
campal region (Figs. 30, 41-42) becomes rounded cephalad
(Fig. 38), and caudad it continues to the tip of the paraccele. I
will refrain from using the term post-cornu for this part because
it suggests a close homology with the primate brain, implying
the presence of a calcar (Wilder, 89). However, the calcar is
due to a total fold of the wall of the cerebrum which does not
occur outside of the primates, but the angle which the calcar
projects into, and which forms one essential part of the post-
cornu, probably existed prior to the intrusion of the calcar. The
precornu (/c’.) dips ventrad so that the cephalic continuation of
the cavity into the rhinoccele (vc.) is from the dorsal partion of
the narrowed cavity. In the sparrow a true rhinoccele does not
exist in the stages examined, but the projection of the cavity
toward the olfactory region is dorsad of a portion of the cavity
(Figs. 1, 11, 20, fc’.) which is homologized with the precornu
of the turtle. The medicornu and the lateral wing around the
striatum are clear in the embryo sparrow (Tig. 2, fc.), and the
latter becomes much exaggerated in the adult, but in no stage is
the caudo-dorsal angle so evident as in the turtle.

DIENCEPHAL.

In the turtle the most marked feature of the diencephal is the
great mesal union of the two sides by means of the medicommis-

200 PROCEEDINGS OF THE

sure. Such a connection is present in mammals and reptiles,
but in both it is due to a secondary thickening of parts and arises
late in embryonic development, or in man it is sometimes absent.
( Wilder, ’89). In birds it is not present and hence an
unobscured picture of the more fundamental conditions may be
looked for in them (see sulci). In the turtle a few very delicate
myelinic fibers cross the meson in this commissure (Figs. 42,
30), a condition said to be present in man (Quain, ’92). The
other,commissures, nidi and tracts, which make up the main body ~
of the thalamus, will be discussed later.

The part of the ¢evma between the chiasma and precommis-
sure is a thin narrow membrane (Figs. 1, 5, 11, 13, 20, 29). It
is much elongated in the sparrow, while the corresponding
part in Demyctylus and lamprey is shorter, in Ama it is very
much reduced. In the last three it forms a part of the ventral
wall. At the ventral end of the terma occur the optic recesses
which, in both sparrow and turtle, form pouches with thin cephalic
walls hanging at either side of the chiasma. At the dorsal end
of the terma in the turtle occurs the preoptic recess (igs. 29, 35
6). This was not found in the sparrow.

In fishes the zxfundibulum reaches its maximum development.
It was found in Awa that there are from this region two unpaired
caudal extensions; a ventral, the saccus vasculosus, a dorsal,
called by Herrick (’93) the “ mammillary body ;” and dorsad of
them a pair of projections, the ypfoaria; and a pair extending
cephalad and surrounded by the hypophysis. A sulcus con-
tinues from these last to the saccus. The Amyda has from the
caudal part of the infundibulum three mesal projections (Figs.
29, 30, 42). The ventral has a peculiar wall composed of clear,
columnar cells. It is somewhat ramified, and as it is surrounded
by the hypophysis it is not strictly comparable with the saccus
of Ama except in position. The hypophysis* is separated from

the infundibulum by pia and is composed of two portions, an ectal

*The term hypophysis, as here used, agrees with the usage in immam-
malia. In mammalia, a part of the brain wall is frequently included in
the term or the two parts are distinguished as pre- and post-hypophysis.

AMERICAN MICROSCOPICAL SOCIETY. 201

and an ental of somewhat different microscopic appearance. The
next dorsal of the two mesal pits mentioned is much the wider
of the two. Continuing cephalad from the ventral of the three
is a sulcus on either side which extends to a point ventrad of the
infracommissure, z. ¢., to a point quite similar in position to the
cephalic pair found in Ama. In the bird the infundibulum is
comparatively much simpler. In the younger embryo (Fig. 1)
the hypophysis is composed of tubules, and a duct connects it
with the enteron. It lies cephalad of the ventral prolongation
of the infundibulum. In the adult (Fig. 20) it is applied along a
greater extent of the infundibulum, and the latter has a caudal
and a slight cephalic projection. No others were discovered,
unless the wavy outline of the caudal boundary of the infundi-
bulum (Fig. 1) indicates such in the embryo. In both animals
the infundibulum is far from the fish-like form and particular parts
could only be homologized after more thorough study.

MESENCEPHAL.

In the turtle the roof of the mesencephal is a solid structure
with great commissural systems uniting the gemina (Fig. 29,
gm.cm.). Inthe adult sparrow, at the part which lies next the
postcommissure there is a slight union of the two gemina across
the meson by fibers (Fig. 20 gm. cm.), and all the rest of the roof
is a membrane which is stretched (Fig. 23) between the widely
separated gemina. In the embryo the solid parts of the roof are
close together (Figs 1-5, 11) the membrane being a mere
narrow strip. The adult condition is a strong reminder of that
in the young lamprey, where it was found that not only was the
roof a membrane, but it forms a plexus. In the mouse a
similar thin membrane was found in this situation by Professor
H. E. Summers (Unpublished work done at Cornell University,
1886-8). In the turtle as in the Diemyctylus, a trace of this
membranous condition can be found in the adult in a small
mesal lophius and in cells which extend along the meson far
toward the pial surface. Here a curious problem arises. The

sparrow, like other higher birds, seems to be pre-eminently a
14

202 PROCEEDINGS OF THE

seeing creature, and the parts connected with vision are all large
and well developed except this. In the turtle in which, from
general development of parts, vision apparently is far less im-
portant, a union occurs along the whole meson, in which great
commissural tracts cross from side to side, and mingle with fibers
from the optic nerves. Mere stretching or crowding by other parts
cannot account for the difference. More fibers could have taken as
long a course as between these divaricated gemina had there been
continued in the birds the need of so intimate connection of the
parts as existed in their reptilian relatives. The significance of
this condition, its connection with vision, the question as to
whether it arises from atrophy producing reversion to an original
type, or whether it is a direct inheritance froma form in which
~such union never took place, all remain to be studied. In the
early embryo sparrow the gemina are prominent upon the dorsal
surface (Fig. 1) and have a position and relative size comparable
with the adult turtle. In the next embryo (Fig. 11) the relative
shape and position have become markedly changed. The cere-
brum and cerebellum are not large enough to produce any such
crowding as to account for this change, hence the conclusion is
reached that it is the inherent growth and development of the
gemina themselves that has led to the displacement from their
typical position. Compared with the cellular portion of the brain
the myelinic fiber tracts are fixed. The optic tracts in the second
embryo are well developed with two strands on either side (Fig.
12), one arising near the everted tip of the geminum and crossing
entad of the other tract. This ental tract is, in the younger
embryo, little developed, and as it has already been seen, the tip
of the geminum is also little developed. With a large develop-
ment in the region of the tip, and with fiber tracts early becom-
ing myelinic, the tendency must be, with the growth of other
parts, to hold the tips in a relatively fixed position, thus stretch-
ing the thin membranous roof, and leaving the gemina at the
level of the base of the brain. The cerebrum and cerebellum in
their growth cover the gemina, but do not push them aside. In
the turtle two similar optic tracts (see description of Fig. 36) are

AMERICAN MICROSCOPICAL SOCIETY. 203

found, but neither the ental tract nor the tip of the geminum
takes on such marked development. It is observable that the
gemina in the second embryo sparrow are really nearly as great
in length as in the adult (see description of Fig. 20), that is, be-
fore the cerebrum and cerebellum have made much progress
toward developing the geminum has approached its maximum
size, and this may be adduced as another argument against the
theory of crowding.

In the younger embryo (Figs. 2-4) the wall of the geminum
is comparatively thin, and the opening from the mesoccele into
its lateral recess is large (Fig. 1), in the second embryo the
walls are rapidly thickening (Fig. 13) and the entrance into
the lateral recess is diminishing (Fig. 11). These processes
continue until, in the adult, the recess is constricted and the
opening a mere point. One feature of the wall is remark-
able. On the mesal surface of the geminum (Fig. 1) is seen a
furrow (G); corresponding to this on the endymal surface is a
ridge (Fig. 4), which as it passes ventrad becomes wider and
divides the recess into two parts (Fig. 5). In the second
embryo in the corresponding region of the geminum (Fig. 13, N),
there is only one pocket; the other has become consolidated
and is represented by a cell-mass. From a comparison of the
position of this cell-mass and of the connections of a fiber tract
in this region it seems probable that the nidus (q, Fig. 36) of the
turtle corresponds to this cell-mass ; also that there may be rep-
resented the post-geminum of higher forms, and that in all forms
at some time a pocket of endyma may close and give origin to
the cell-mass of the post-geminum.

In one specimen over an extensive area lying between the
letters g. and m. fp. (Fig. 29) the endyma of the two sides has
coalesced and degenerated, leaving a narrow tube ventrad, and a
somewhat wider one dorsad of it to connect the cephalic and the
caudal parts of the brain cavity. It seems like an incipient nar-
rowing to produce an iter like that of the mammalian brain or
the narrow mesoccele of the adult sparrow. A pit at the cephalic
tip of the oblongata has been identified in the turtle and in embryo

204 PROCEEDINGS OF THE

sparrows as the mesencephalic pit (Figs. 1, I1, 29, m. p.), but it
could not be found in the adult sparrow.

The direct short course taken by the optic tracts in the spar-
row has already been mentioned. They retain in the chiasma
their relative position (Fig. 12), and in addition there is a distinct
tract arising in the thalamus and crossing in the chiasma dorsad
of the others. The long course of these tracts in the turtle is as
described under Fig. 36. The optic nerves of the turtle are
pressed closely together for some little distance, when they turn
abruptly toward the eyes. Each is deeply folded upon itself, the
pia dipping into the fold. The chiasma and optic nerve of the
sparrow are extremely large.

OBLONGATA.

For the purposes of this article the limits of the ventral
portion of the metencephalic and epencephalic segments will not
be considered since no new light has.been thrown upon the sub-
ject; hence the whole floor of the caudal part of the brain will
be considered under the heading od/ongata, and only a few facts
noted with regard to the cranial nerves will be mentioned.

In the turtle all the nerves controlling the muscles of the eye,
the third, fourth and sixth, are relatively large, and it is interest-
ing to note that, while every other part of the body, except the
swelling throat, may be kept in a condition of apparently abso-
lute quiet, the eye is tirelessly turning as indicated by the hori-
zontal bar across it, with every slight movement of the observer.
This turtle seems to have adopted the motto of ‘ eternal vigil-

’

ance ’’ in place of an armored.defence. In the sparrow the third
nerve is very large, but the fourth and sixth are relatively small.

In the turtle, as in the mouse, a very large branch of the fifth
nerve passes to the tip of the snout. The vibrissae of the mouse
and the long, pointed, comparatively thin skinned tip of the
turtle’s snout evidently have comparable functions. A large and
valuable part of the information carried to the brain must be
through this channel. Much of the time the turtle is completely
submerged in the water, except the tip of the nose, or if buried

—

- =

AMERICAN MICROSCOPICAL SOCIETY. 205

in the sand, the tip of the nose, through which it breathes, is just
exposed. In other turtles and in the birds the horny beak cannot
be as delicate an organ of touch, and in the sparrow the corre-
sponding branch of the fifth nerve is only of moderate size. In
the turtle the fifth nerve has a large Gasserian ganglion. In the
sparrow this ganglion is not so large. In the turtle the fifth
nerve arises upon the cephalic side of an enlargement of the brain
cavity in that position, the seventh and eighth upon the caudal
side of the same enlargement, while the tenth is at the widest
point of the next succeeding enlargement of the cavity. In the
young embryo sparrow similar relations were observed, but in
the adult the thickening of the oblongata obscured the appear-
ance. The ninth nerve in the turtle has a ganglion independent
of the ganglion of the tenth, while the numerous nerve roots of
the ninth unite and then pass into the ganglion of the tenth. The
seventh and eighth nerves have much less intimate connec-
tion than in the sparrow or in the snapping turtle (Humphrey,
94). The seventh has three branches, the first dividing into two,
each of which has an independent ganglion, as does the second
branch; the third branch joins the ganglion of the eighth.
These ganglia indicate sensory functions, and two of the above-
named branches pass into the cephalic parts of the ear capsule,
and may really be part of the eighth. In the sparrow the
seventh and eighth unite more completely in a ganglion common
to both, and the eighth has a very large band of conspicuous
fibers, which pass mesad and immediately ventrad of the endyma
cross the meson. This is one of the two instances where a com-
missural or decussational connection across the meson is noticeably
greater than in the turtle. The sense of hearing certainly is
keener in the sparrow than in the turtle. The turtle will move
when a distinct jar is given the vessel containing it, but even
quite a loud noise does not appear to give it any uneasiness,
while the sparrow is startled by any slight sound.

CEREBELLUM.

The function of the cerebellum is still a matter of great doubt,
but in the two forms considered the great peduncular tract (32),

206 PROCEEDINGS OF THE

(Figs. 11, 13, 36, 30), coming from the alba of the cerebellum and
decussating across the meson bends over among the fibers of
the eighth, becomes diffuse at this point in such a way as to lead
to the inference that certain functions of the ear must in these
forms be largely regulated by the cerebellum. The intimate con-
nection of the acoustic eminence and the cerebellum in the
Sauropsida was noticed by Spitzka (81). The large, complex
cerebellum of the bird and the simple one of the turtle would
appear to harmonize with the facts concerning the eighth nerve
and the sense of hearing already mentioned, and perhaps also
points to a connection with the still only partially understood
sense located in the semi-circular canals of the ear.

In the development of the cerebellum of the bird the roof is
at first a mere membrane (Fig. 1), the thickened portions not
having passed across the meson, a condition which corroborates
the position taken by Stroud (’95) and Schaper (’94), that the
cerebellum is originally a paired lateral outgrowth. In the next
stage the union has taken place across the meson (Fig. 11), and
with the exception of a few folia upon the surface, it has a marked
resemblance to the turtle’s cerebellum, and an even more marked
resemblance to that of the alligator, for in that the caudal part
bends over more than is the case in the turtle. In the second
embryo of the sparrow the pit in the skull (Figs. 13, 17-18)
which later will be occupied by the flocculus is filled by arachnoid
tissue, and the flocculus is a mere projection pointing toward the
pit, z. e., the bony wall undergoes the modification necessary
for the reception of the flocculus before that comes in contact
with it. In the adult sparrow the cavity of the cerebellum is
small and at the middle is actually closed by the crowding of
parts together, the endyma having become obliterated (Fig. 20).
There are thirteen folia seen at the meson. ‘Traced laterad in
serial sagittal sections it is seen that the central folia, the seventh
and eighth pass slightly beyond their neighbors giving the
appearance of a lateral lobe, while the caudal ones, the eleventh
to the thirteenth, forming the caudal rim of the cerebellum, fuse
laterad and at the tip of the lateral recess of the cavity form the

AMERICAN MICROSCOPICAL SOCIETY. 207

flocculus by uniting with the lateral extension of the cephalic part
of the cerebellum. The /locculus was originally given the name
because of its resemblance to that organ in mammals, but it has
been questioned whether the homology was correct. Comparison
has been made with man and the cat, and from the evidence at
hand it would appear that the homology originally given was
EOrrect.

In Quain (’92), the essential relations of the flocculus of man
are as here described, and Stroud’s paper upon the cerebel-
lum (’95) shows the same relations in its development. Dr.
Stroud considers the mammalian flocculus to be a complex organ,
a portion of which, the pugnus, in rodents, some carnivora, etc.,
lodged in a cavity of the petrous bone, is developed from a part a
little removed from the caudal margin of the cerebellum. The
question arises whether these facts can be brought into harmony,
and the term /locculus be applied to the organ occupying a
depression in the skull from whichever of the folia it arises, or
whether there are really two different organs that have been
called by the same name.

The simple flocculus of the sparrow is seen to be an _ offset
from a primitive part of the cerebellum which lies next the edge
to which the plexus is attached, with a special prolongation at
the tip of the lateral recess. Fig. 24 shows with diagrammatic
clearness the relations of this body to the mass of the cerebellum.
The sheet of cinerea which in general is far from the cavity passes
into the floccu/us and with the last folium comes in contact with
the caudal part of the cavity, In the embryo (Figs. 17-18) the
condition is simpler, but still recognizable, though the caudal wall
of the cerebellum is not so greatly developed. In the younger
embryo the part could not be distinguished with certainty. In
the Amyda a distinct flocculus was not found, but a ripa between
the edge of the cerebellum and the plexus (Fig. 30) may be
considered as the proton of the floccudus. In the alligator a still
more marked rudiment of the part exists and the appearance
in frontal section is quite similar to the embryo sparrow

(Fig. 17).

208 PROCEEDINGS OF THE

FIBER TRACTS AND COMMISSURES.

In the Amyda were traced over thirty fiber tracts, most of
them being myelinic. These are, with their commissures, de-
scribed under figure thirty-six ( Fig. 36), where a numerical sys-
tem of designation is adopted, in order to avoid too exact homol-
ogizing. In the turtle one is struck with the fact that few tracts
are long. Tract 24, the posterior longitudinal fasciculus, is a
marked feature of the Amyda’s oblongata; it extends into the
myel and is not as prominent as in the snapping turtle (Hum-
phrey, 94). In the sparrow this tract is comparatively small.
The optic tracts of the turtle are long, but as shown above, they
are comparatively much shorter in the sparrow. In the base of
the cerebrum of the turtle, and extending into the base of the
oblongata, is an amyelinic system of fibers (2, 2, 2) which
at first appears continuous. On closer scrutiny it seems to be
interrupted by a cell nidus (k), which is so attenuated at its
middle as almost to form two nidi. The remaining tracts are
shorter, and in most cases it cannot be said that one, even with
the intervention of a nidus, is a direct continuation of another.
One tract fades out, another gradually increases, and from this
fact one would suppose that the turtle’s mental processes must
be by slow and indirect methods.

In accordance with the greatly developed striatum in the spar-
row, the union of the thalamus and striatum by tracts is corre-
spondingly large. The tracts at the base of the striatum, extend-
ing cephalo-caudad through a great part of its length are also
large. The mesal wall of the sparrow's cerebrum, as with some
other birds (Bumm, ’83, and Carss, ’95) shows a great fan-like
spreading of fibers, the handle of the fan passing into the thalamus
mesad of the optic tract and disappearing in the caudal region of
the infundibulum (Iig.11). This seems to represent one portion
of the complex system of fibers which in mammals is comprised
under the name fornix. It is gathered from the entire hippo-
campal segment, both cephalad and caudad of the porta. In the
Amyda only a few fibers could be in any way homologized with
this tract.

AMERICAN MICROSCOPICAL SOCIETY. 209

The precommisure in both animals has a well developed strand
passing caudad into the edge of the striatum. In the sparrow
only this one portion could be distinguished. In the turtle it
was seen to be composed of two parts, a myelinic and an amyel-
inic, while another part consisting of amyelinic fibers turns ceph-
alad, but could be traced for only a short distance.

This brings us to the vexed question of the presence or
absence of a callosum in these forms, in which, highly specialized
as they are, a callosum might be expected to appear if it occurs
in any forms below the mammals. In both cases a small com-
missure does exist dorsad of the precommissure in a_ position
assigned by Osborn (86-87) to the callosum. Smith (’94) finds no
evidence of a true callosum in the lowest mammals, and various
investigators, both before and after Osborn are inclined to believe
that in forms below mammals the commissure mentioned is a
hippocampal or forni-commissure. Bellonci, ’87—’88; Meyer,
’95; Kingsbury, ’95, are inclined to consider it is not a callosum.
The name callosum is retained in this article for convenience
merely. In the soft-shell as in the snapping turtle there are two
amyelinic bundles, one passing dorso-caudad into the hippo-
campal segment as a kind of fornix, the other dorso-cephalad into
the termatic eminence and thence into the hippocampal segment.
Perhaps these, as Herrick, (’93), supposes, represent two distinct
commissures, the forni-commissure and the callosum. As yet a
complete homology of either of these parts with the so-called cal-
losum of Amphibia does not seem to have been established. In
the sparrow this commissure is so small as almost to escape obser-
vation ; still, as in the turtle, it is distinct from the precommis-
sure. In the latter quite a notch of endyma intervenes between
the two (Fig. 29). In the younger embryo sparrow (Fig. 1)
neither of these commissures had appeared in the thickened terma
although the postcommissure is well developed.

The supracommissure of the Asmyda is strongly developed ;
one portion lies in close proximity to the tube of the epiphysis,
but no fibers could be traced to the epiphysis, as Herrick (’91)
found to be the case in Aspfidonectes. In correspondence with

210 PROCEEDINGS OF. THE

the small habenz the supracommissure of the sparrow is very
slight. Even in the adult it was recognized with great difficulty
and it is late in development, not a trace of it being found in
the second embryo. This difference would be accounted for if
the theory is correct which holds that the supracommissure is
correlated in part with the epiphysis, especially in those forms in
which the latter has an eye-like structure. The turtle is much
more nearly related than the sparrow to forms in which a dis-
tinct eye-like structure is found.

The geminal (Sylvian) commissure is in the turtle well developed,
in fact there are extensive commissural connections between two
distinct sets of fibers, that lying ventrad of the great balloon cells
being composed of large fibers, that lying next the endyma of
fine fibers. In comparison with the large gemina only a small
geminal commissure is present in the sparrow, the fibers taking
a long course between divaricated gemina.

In the cerebellum of the turtle very close to the decussation
of the fourth nerve in the valvula are two decussational tracts,
the one from fibers of the cerebellum itself (tract 30, Fig. 36),
the other more cephalic in position composed of fibers from the
lateral surface of the oblongata cephalad of the fifth nerve (tract
29, Fig. 36). The latter is reinforced by fibers coming from a
poiat caudad of the gemina and thus forms a great mass of fibers,
which from its situation reminds one of the pons of mammals,
but though fibers from it pass far toward the ventrimeson, none
were found actually to cross. Arcuate fibers independent of this
tract do cross the meson. Any conclusion with regard to a
rudimentary pons in reptiles must depend upon a more thorough
study of the origin of the pons in mammals. The commissure
of the cerebellum in the sparrow (Figs. 11, 20, 24) receives fibers
from the lateral parts of the cerebellum and even the flocculus, in
this respect reminding one of the more caudal of the decussations
in the turtle, and connects the great columnar peduncles which
pass over into the acoustic eminence. There is no appreciable
constriction separating the lateral part of the cerebellum from the
acoustic eminence. This commissure of the cerebellum is the

AMERICAN MICROSCOPICAL SOCIETY. 211

second instance of a more complete union between the two sides
in the sparrow than in the turtle.

CINEREA, NIDI.

In the adult Avzphzbia the cinerea is collected closely around
the endyma, except in a few places where an incipient ecto-cinerea
is found. It seems to be typical that cinerea should lie next the
endyma, but it may be displaced from that position by growing
fiber tracts, or fibers may so increase among the cells as to sep-
arate them. In both the turtle and the three stages of the spar-
row, there are in many parts cells somewhat evenly distributed
throughout the nervous tissue. In the youngest sparrow (Plate
I) a concentration of cells is seen in most parts around the
endyma, but already indications of layers and nidi of cells appear.
In the adult the formation of layers and nidi has progressed
much farther than in the turtle. In the latter there are around
the rhinoccele three layers, more or less complete, separated from
the endyma and from each other by fibers and surrounded by
the glomerular layer (Fig. 37). In the sparrow only one group
of cells could be distinguished, and that formed a central core for
the coalesced olfactory lobes (Fig. 27).

In the turtle an incomplete layer of ectocinerea (Figs. 36, 31,
38-42) is especially noticeable in the pallial and hippocampal
segments. In the sparrow a similar layer can be found only in
the hippocampal region. The striatum in both animals is a mass
of cells marked off by prominent bands of alba (Figs. 22-23).

In the habenz the cells are arranged in hollow spheres, as
shown by the rings of cells appearing in sections made in differ-
ent planes ( Fig. 25).

The geminum of the young embryo sparrow already shows
indistinct layers (Figs. 2-5). Three such layers of greater con-
densation are seen, which become more pronounced in the next
stage (Fig. 12). In this stage the number of distinct layers is
the same as with the turtle (Fig. 31). In the adult sparrow the
cinerea is divided into eight layers of cells, exclusive of the
endyma, by fiber tracts.

212 PROCEEDINGS OF THE

The cinerea of the cerebellum of the turtle has an amphibian
character in that the cells are near the endyma. They are inter-
spersed by myelinic fibers and bordered by a layer of Purkinje
cells. In addition to this, throughout the ectal alba, radiate rows
of cells, and over the entire surface is a layer of small cells (Fig.
31). In the younger embryo sparrow (Fig. 6) a layer of cells
preserving continuity with the endyma at the tela, is the only
differentiation. It appears to be the same as the cellular layer
which covers the surface in the next stage (Figs. 17—18), but in
addition are found distiact nidi of cells. This ectal layer is con-
firmatory of the theory of Herrick (g1) that the cinerea of the
cerebellum arises from the union of the solid parietes with a tela.
In the adult the cinerea has assumed the seggregated character
found in mammalia, but retains its position next the endyma
only at the meson, in the first and last folia (Fig. 36).

In structure as well as form the geminum and the cerebellum
of the turtle have advanced about as far as the second embryo
sparrow, while all the parts, including the cerebrum, have made
about equal progress toward complete evolution, again showing
the well-balanced condition indicated by the form. In the adult
sparrow the cinerea of the cerebellum and gemina is highly
developed, but in the cerebrum the arrangement of cells indicates,
like the form of the parts, a high specialization aside from the
more usual type of the mammalia.

As shown in the turtle (description of Fig. 36), there are besides
these layers of cinerea at least twenty-six more or less distinct
nidi of cells.

SULCI.

Perhaps one of the greatest contributions of recent years to
the morphology of the brain is the discovery by His (90) of the
building up of the oblongata by a series of unions of the mem-
branous portion with the edge of the solid parietes. Each so
formed consolidation is a center for the proliferation of cells.
This work is supplemented. by that of Herrick (’91) on the cere-
bellum in which he found a similar process taking place. In
1893 in the article on Diemyctylus the writer recognized that in

AMERICAN MICROSCOPICAL SOCIETY. 213

other regions, as in the diencephal, definite furrows called sa/ez
occur which have a morphological value in determining homo-
logies, and which, at their deepest part, give origin to special cell
masses. Humphrey (’94) in his study of the snapping turtle
came to the conclusion that these sulci are of little morphological
value and are determined mainly by fiber tracts or are incident
to foldings of the wall. Since that time some progress has been
made in studying these sulci. Those found in the turt'e are
enumerated under the description of Fig. 29. There appear to
be three kinds of sulci; Ist, those just mentioned as discovered
by His; 2d, those formed as the result of growing together of two
symmetrical parts united by a membrane at the meson. The
membrane apparently forms a U-bend and the original line of
union on either side with the solid parietes forms the sulci, a
lophius or ridge forming between them. What seems to be a
striking example of this is seen in the roof of the cerebellum.
Originally a membrane forms a roof-like connection between the
two lateral halves (Fig. 6). In later embryos of both sparrow
(Fig. 11, ~) and the cat a small lophius is found extending a con-
siderable distance along the meson. No trace of this was found
in the adult sparrow, but in the turtle and the alligator a promi-
nent feature of the cerebellum is a median lophius (Figs. 29-31,
- £) occupying a perfectly comparable position. The roof of the
mesencephal in the turtle shows a similar. sulcus and lophius
OPIS) 26; 5).

At the crista in these lower forms the solid parietes approach
each other, but do not unite. There is a distinct U-bend with a
sulcus on either side. The endymal cells forming the U are
generally elongated and of a peculiar appearance. Figs. 8, 16,
26, 34, show that it occurs in the turtle and in different stages
of growth in the sparrow. It always contains a blood vessel and
sometimes more than one. Studnicka (’95) figures a similar
appearance in Amphibia and shows its relations. He calls it the
terma, but here is preferred the word crzs¢a, introduced by Wilder
(80), to indicate a small outgrowth upon the cephalic wall of the
aula. . This crista of the higher forms was identified by the writer

214 PROCEEDINGS OF THE

with an intrusion of endyma into the aula of Dzemyctylus and
Amia and has since been found in specimens of all the groups
examined, except lamprey and shark, and with further study it
is hoped to identify itinthem. Inthe adult mammalit is obscured
by the growth and crossing of hippocampal or fornix fibers
cephalad of it, thus cutting it off from the pia and leaving it as a
kind of record of development. It forms a valuable point of de-
parture in determining the homologies of the region.

In the snapping turtle Humphrey (’94) figures a small mesal
lophius or ridge in the floor of the metaccele but considers it
without significance. A similar fold is found in the soft-shelled
turtle (Figs. 29, 43, 2), where it has only a short cephalo-caudal
extent. It was sought in the sparrow, and in the embryo a
mesal lophius was found, extending the length of the oblongata.
Fig. 9 shows that from the sulci at either side extend layers of
cells into the raphé, and that the slightly differentiated endyma
at the side of those sulci corresponds with the limits of the
raphé. The suggestion is made that these sulci give rise to the
cells of the raphé, including the nidus x (Fig. 36). A mesal
lophius occurs in other parts of the floor of these brains, but
special attention has not been given them.

Just ventrad of the postcommissure in both forms is the sulcus
(0). On closer examination this is found to be really a paired -
sulcus (Figs. 24 A, 32, 33), and hence can be homologized with
the sulci composed of similar long, clear cells found widely sep-
arated veatrad of the postcommissure in the lamprey. All forms
of vertebrates examined have such sulci. In the article on
Diemyctylus they were homologized, following Rabl-Riickhard
(83) with the torus of fishes. There is now thought to be no
such homology. The work of Locy (’93) upon the existence of
a third pair of rudimentary eyes in sharks and Amphibia, is looked
to with interest as throwing possible light on these structures,
for they occur caudad of the entrance of the epiphysis, which he
considers to result from the fusion of the second pair.

Sulci of a third class are those which occur upon the endymal
surface of solid parieties, and give rise to nidi by complete union

AMERICAN MICROSCOPICAL SOCIETY. 215

of the walls of the sulci or by proliferation from the deeper layers ;
the sulcus in the latter case continues as a feature of the endymal
surface. In the brain of the young embryo sparrow are excel-
lent examples of this class (Fig. 1, 2, 7, p.). Sulcus (%) is a
deep furrow, forming the caudal boundary of a lophius, which in
frontal section (Fig. 3) is a noticeable feature just caudad of the
porta. Caudad of the sulcus is the end of a fiber tract (5, in
Fig. 1), which curves ventrad of it, then turns apruptly laterad
into the striatum. In the second embryo the lophius and sulcus
have almost disappeared (Figs. 11, 16, 2), but the tract (5) re-
tains its ventral curve, anda mass of cells shows the original
extent of the sulcus. This mass of cells is recognizable in the
adult. In the turtle the dorsal limb of the tract marked (5) in
Fig. 36, takes a similar course to the above and ends at a great
nidus (d), which seems to be homologous in position with the
above mentioned mass of cells, although its relations to the
endyma are obscured by the union of the endyma across the
meson in the medicommissure. Sulcus (¢, Fig. 29) just cephalad
of the medicommissure may represent sulcus (/) of the sparrow.
It is hoped from material now ia hand to settle the matter of
development of this and similar parts in the embryo soft-shelled
turtle.

The sulci z and fare found in the embryo sparrow and in the
turtle (Figs. 1, 4, 29, 32, 33). The first is comparable in posi-
tion, according to Quain (’92), to a slit which “leads into the still
hollow geniculate body.’ In the sparrow this sulcus disappears,
but a cell nidus takes origin from it. The sulcus (f) does not dis-
appear in either form, but in the early stage of the sparrow it is
connected with a cell-mass which later separates from it. These
two sulci give off nidi, which if compared with those of the
turtle, seem to be homologous in position with nidi (i, j Fig. 36).
One of these apparently represents the pregeniculum or “‘ exter-
nal geniculate body ” of man.

On the mesal aspect of the caudal part of the geminum of the
embryo sparrow isa furrow (Fig. 1, G). This forms a total fold
of the wall and corresponds with an endymal ridge projecting

216 PROCEEDINGS OF THE

into the cavity (Fig. 4). This, soon uniting with the adjoining
wall, divides the cavity into two parts, a mesal and a lateral (Fig. 5,
vr. mc.). In the next stage the mesal cavity is not present, but a
nidus (Fig. 13, V) staining more deeply than the others, is seen
to occupy (with reference to the cavities) a position which, when
change of shape is taken into consideration, is found to be ex-
actly comparable with the mésal part of the cavity above de-
scribed. Nidus (q) (Fig. 36) of the turtle holds the same rela-
tive position, and from the fiber tracts of the region may be con-
sidered as the representative of a postgeminum, as. suggested by
Humphrey ('94).

Other sulci have masses of cells from their deeper parts which
have not yet been identified. . Some of these have morphological
significance, however, independent of the nidi, as for instance
those which enter the porta. In the turtle (Fig. 29) there are
four sulci entering the porta which may materially aid in deter-
mining the real relations of that puzzling region. First, a small
sulcus (@) is seen on the mesal wall of the projection of the aula
cephalad of the porta. This enters the porta and turns cephalad
upon the mesal wall of the paraccele and continues to the precornu
(fc’.). It indicates the line of junction between the hippocampal
and termatic segments of the mesal wall of the cerebrum men-
tioned above. Second, a sulcus arising in the preoptic recess (0,
Fig. 34) passes caudad of the precommissure, enters the porta
(Fig. 35) and passes cephalad into the precornu (gc’., Fig. 29).
Third, the sulcus (ad) (Figs. 29, 35, 36) enters the porta and
passes ventro-caudad to the medicornu, (fc ). Fourth, the sulcus
forming the dorsal boundary between the habena and diatela
(Fig. 31, ¢) passes into the porta upon the mesal wall of the
paracce'e and forms the boundary between the tenia and the
paratela.

The relations of the fourth sulcus show that the paratela is
continuous with the diatela or roof of the diaccele and is thus in
reality part of the roof of the prosoccele. This sulcus is morpho-
logically at the lateral edge of the rima. At the opposite edge
of the rima the sulcus there found unites with its opposite in the

AMERICAN: MICROSCOPICAL SOCIETY. 217

region of the crista. In the sparrow this relation of the paratela
is not quite as clear owing to the attenuation and distortion of
adjacent parts.

Between the third and fourth sulci of the turtle a part of the
thalamus lying cephalad of the medicommissure projects into the
paraccele as shown at the right of figure 30. In this respect it
agrees with the relations in the embryo cat as shown by Hoch-
steter (’94), a condition by no means to be confounded with the
exploded notion that the rima allows a portion of the pial surface
of the thalamus to enter the paraccele (Wilder, ’84, ’88). In the
sparrow (Fig. 16) there is no medicommissure, but the essential
relations are the same.

The mass included between the second and third sulci and
theit extensions represents the union of the striatum with the
thalamus. According to His (’92) this mass should represent
the cephalic continuation of the ‘ Grundplatte’’ or ventral of the
two segments into which he divides the lateral wall of the brain
tube. In neither of the forms could be found that connection of
either of the sulci bounding the mass with any more caudal
sulcus which would indicate agreement with the explanation of
His. The caudal sulcus in the embryo sparrow (Fig. 1, 2) be-
comes shallow, but seems to be continuous with a sulcus extend-
ing to the optic recess.

In the sparrow (Fig. 1) occurs a marked peculiarity. The
mass between the two sulci (4 and @) rises flush with the gen-
eral level of the cavity. It is an excessive growth of the cauda-
tum formiag a part of the lateral wall of the awa, and dividing
the porta into two parts (Fig. 3). This becomes less apparent
in the next embryo (Figs. 11, 16) and in the adult the growth
of surrounding parts makes this condition noticeable only on
account of its history. At first sight this seems to be quite dif-
ferent from the condition in the turtle; but the other relations of
the sulci, including the mass, are identical.

Between the first and second sulci (Fig. 29, a, 4) of the turtle
lies the termatic outgrowth, extending from the thickened part

of the terma, in which the commissures occur. It includes a part
15

218 ‘ PROCEEDINGS OF THE

of the preportal aula, and extends into the paraccele to the tip of
the precornu. In the sparrow there is no preportal aula, but the
part of the termatic outgrowth which bears the crista (Fig. 3, ¢.
é.), is included wholly within the paraccele on account of the
caudal extension of the mesal wall of the cerebrum, and the
bounding sulcus can be traced to the precornu as in the turtle.
A segment similar to this was found in Dzemyctylus and recog-
nized as termatic in its origin.

Dorsad of the first sulcus (Fig. 29, a) of the turtle is an aulic
portion of the hippocampal segment. In the sparrow this, like
the termatic segment, appears as if drawn entirely within the
paraccele.

Thus upon the aulic surface of the cavities of the turtle, and
potentially so in the sparrow, are represented all of the segments
of the cerebrum, except the pallium.

The caudal boundary of the aula seems to coincide with the
sulcus (@), or perhaps better the lophius caudad of it, extending
from the porta to the optic recess. If this interpretation is cor-
rect it would appear to be more in accord with the researches of
Studnicka than of His. Studnicka (’95) shows that in the early
embryo of the lamprey and Amphibia there is a neuromeric,
dorso-ventral, endymal furrow which bounds the ‘ massive
Anlage des Hemispherenhirns,” and that the division of the
cerebrum itself into parts is by the formation of an independent
intrusion of the cavity into the wall. This being the case, the
pallium would not necessarily have a special aulic representative.

ADDENDUM.

The foregoing paper was read at the meeting of the society in
August. While it was passing through the press the article by
Burckhardt, ““ Der Baup!an des Wirbelthiergehirns ”’ (Schwalbe’s
Morphologische Arbeiten, Vol. IV., 1895, pp. 131-149, 1 pl.),
came into my hands. The colored diagrams show with great
clearness his morphological views, and the text discusses with
admirable force and directnesss the plan of the vertebrate brain,
especially as shown by mesal structures. From a study of the

AMERICAN MICROSCOPICAL SOCIETY. 219

amphibian brain he derives a plan of th segments extending from
the caudal end of the brain tube into the cerebrums. This plan
is an extension and elaboration of the idea of His mentioned above
(p. 217). The present paper reaches conclusions from entirely
independent data ; some of them agree with and some differ from
those of Burckhardt, but more extended notice than could be
given at present would be necessary justly to estimate the facts
and bring them into that harmony which must be the ultimate
result of complete knowledge.

SUMMARY.

The points touched upon in this paper are :

1. The importance of comparing through all stages of develop-
ment widely different forms of brains in order to gain from ex-
aggerated form and specialized function more light upon the
truths of morphology and evolution.

2. The overlapping and crowding of parts of the brain in
these, which in comparison with others of the same groups, are
highly specialized forms.

3. A degenerate condition of the olfactory lobes resulting in
union due to crowding, not to a crossing of fibers from one lobe
to the other. It is a feature incident to other specializations.

4. Although the parts connected with vision in the sparrow
are highly developed the union of the gemina across the meson
by a relatively small commissure would indicate an independence
of action of the two sides in contrast with the condition in the
turtle and other forms where the connection between the two
sides is far more intimate.

5. The tip of the snout is a more important tactile organ in
the turtle than in the sparrow, as indicated by the large branch
of the fifth nerve distributed to it in the former.

6. The eighth nerve has reached a higher development in the
Sparrow than in the turtle as indicated by its intimate connection
with its opposite across the meson and its apparent connection
through the auditory eminence with the column-like peduncles
of the cerebellum, which in their turn form a large commissural

220 PROCEEDINGS OF THE

connection in the cerebellum. These complicated and extensive
structural developments and relations of these parts are probably
associated with higher and more complex functions than the
simpler conditions in the turtle.

7. The flocculus of the sparrow is probably homologous
with the organ of the same name in man, and has a proton in
the turtle and alligator. The pit in the skull for the reception
of the flocculus is formed before the flocculus has grown suffici-
ently to enter it.

8. Twenty-six nidi and more than thirty fiber tracts with their
commissural connections were found in the turtle and many ap-
parent homologues were recognized in the sparrow. Especially
in the turtle there is not the continuity of nerve tracts which one
is led to believe occurs in mammals, but there is rather a more
or less independent, overlapping series of tracts.

g. The pons is not present.

10. In the sparrow a large fiber tract from the mesal wall of
the cerebum strongly suggests the fornicolumn of mammals, but
it has more extensive relations.

11. The conclusion is adopted that the so-called callosum of
birds and reptiles is the rudiment of a fornicommissure with a
few fibers which may be truly callosal.

. A metapore was not demonstrated in - either the sparrow
or = Bai turtle, although the tela is very much attenuated in
the position usually assigned to the metapore.

13. The metaplexus is apparently formed by crowding a V-
shaped membrane between two nearly parallel edges of the cere-
bellum and the oblongata.

14. The roof of the epiccele is at first amembrane. The union
of the lateral halves of the cerebellum across the meson is sec-
ondary, the connecting membrane being replaced by a mesal
lophius.

15. The widely divaricated condition of the gemina in birds is not
due to crowding by the cerebrum and cerebellum, but to their in-
trinsic growth, nearly completed before any crowding could occur.

16. There is suggested the possible identity of the double sul-

AMERICAN MICROSCOPICAL SOCIETY. 221

cus ventrad of the postcommissure with the pair of lateral out-
growths occurring caudad of the epiphysis, discovered by Locy.

17. The diaplexus of the turtle consists, in large part, of fold-
ings of the membrane at either side of the meson. In this re-
spect it has a closer relationship with the mammalian type than
the mesal plexus of either the bird or the amphibian.

18. In both turtle and sparrow the paratela, oocupying the
rima, or interval between the fimbria and the tenial edge of the
striatum, is morphologically a part of the roof of the prosoccele.

19. Various pockets of endyma were found upon the meson,
which have great significance for morphology, but are physiol-
ogically of slight importance. Among these pockets is the
paraphysis, found in the adult Aszyda and in the embryo
sparrow. ;

20. In Amphibia, turtle and sparrow, a transection of the
hemi-cerebrum shows essentially a delta form. Caudad of the
rima the three limbs are: (1) The ventral or striatal; (2) the
lateral or pallial; (3) the mesal. The first two form segments
extending from the caudal tip to the olfactory lobes. The rima
divides the mesal segment into two parts, the dorsal or hippo-
campal, and the ventral or tenial. At the porta the tenial unites
with the thalamus. Cephalad of the porta the hippocampal
unites with an outgrowth of the terma, the termatic segment ; so
that in the cephalic part of the brain the same complete delta
form is re-established.

21. Sulci which enter the porta indicate that the hippocampal,
termatic, striatal and tenial segments of the cerebrum have a rep-
resentative in the mesal wall of the aula (cephalic part the third
ventricle).

22. In both the sparrow and the turtle the striatal limb of the
delta has a secondary thickening, which is comparable with the
caudatum of mammals.

23. The porta of the embryo sparrow is bifurcated by the
intrusion of the caudatum into the aula. In the adult this intru-
sion is crowded into insignificance by surrounding parts. The
two sulci of the aula which enter these parts of the porta can be

222 PROCEEDINGS OF THE

traced upon the wall of the paroccele, one extending cephalad
and the other caudad. On the aulic surface these sulci pass ventrad
with no appearance of turning caudad to form the aulix or sulcus
of Monro, as the theory of His would seem to demand. Compar-
able sulci entering the porta were found in the turtle, although
the caudatum does not intrude into the aula.

24. The significance of other sulci was considered. (1) Those
which indicate the boundary of a primal mesal membrane as in
the cerebellum and at the crista ; (2) those occurring at the edge
of solid parieties as in the formation of parts of the oblongata, as
shown by His, or of the cortex of the cerebellum, as shown by
Herrick ; (3) those occurring in more solid parts, and whose walls
finally coalesce to form a cell nidus.

AMERICAN MICROSCOPICAL SOCIETY. 223

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Gage, S. H.—Methods of decalcification in which the structural ele-
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Gage, S. H.—The comparative physiology of respiration. Proc.
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PROCEEDINGS OF THE

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Herrick, C. L.—Comparative anatomy of the nervous system. Ref-

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His, Wm.—Die Entwickelung des menschlichen Rautenhirns vom
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His, Wm.—Zur allgemeinen Morphologie des Gehirns. Arch. f. Anat.
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Humphrey, O. D.—On the brain of the snapping turtle (Chelydra

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plates.
Kingsbury, B. F.—The brain of Necturus maculatus. Jour. Comp.
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Kupffer, C. v.—Studien zur vergleichenden Entwicklungsgeschichte

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Locy, W. A.—The derivation of the pineal eye. Anat. Anz., Vol.
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Meyer, A.—Ueber das Vorderhirn einiger Reptilien. Zeit. f. wiss.

Zool., Vol. LV. (1893), pp. 63-133. 2 plates.

Meyer, A.—Zur Homologie der Fornixcommissur u. d. Septum luci-
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86-87. Osborn, H. F.—The origin of the corpus callosum ; a contribu-

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"So.

if.) Gs

tion upon the cerebral commissures of the Vertebrata. Morph.
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Schaper, A.—Die morphologische und histologische Entwickelung

' des Kleinhirns der Teleostier. Anat. Anz., Vol. IX. (1894), pp. 489-

501. 20 figs.

Smith, G. E.—A preliminary communication upon the cerebral com-
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Spitzka, C.—Some new features of the anatomy of the corpora gem-
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Spitzka, C.—Further notes on the brain of Sawropsida. Science,
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Stieda, L.—Ueber den Bau des centralen Nervensystems der Schild-
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’89.

$2.

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Stroud, B. B.—The mammalian cerebellum. Part I, The develop-
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Studnicka, F. K.—Zur Lisung einiger Fragen in der Morphologie des

’ Vorderhirns. Anat. Anz., Vol. IX. (1894), pp. 307-3820. 2 plates.

Studnicka, F. K.—Beitrige zur Anatomie und Entwickelungsge-
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Tiedemann, F.—Anatomie und Bildungsgeschichte des Gehirns im
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Turner, C. H.—Morphology of the avian brain. Jour. Comp. Neur.,
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Wilder, B. G.—The crista fornicis, a part of the mammalian brain
apparently unobserved hitherto. Amer. Assoc. Adv. Sci. (1880), N.
Y. Med. Record, Vol. X VIII. (1880), p. 328.

Wilder, B. G.—The foramen of Magendie in manand thecat. N.Y.

Med. Jour., Vol. XXXIX. (1884), p. 458.

Wilder, B. G.—The relation of the thalamus to the paraccele (lateral
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Wilder, B. G. and Gage, S. H.—Anatomical technology as applied to
the domestic cat. New York (1882).

AMERICAN MICROSCOPICAL SOCIETY.

227

EXPLANATION OF PLATES.

Roman numerals, I. to XII, indicate the cranial nerve roots. Arabic

numbers indicate fiber tracts.

indicate nidi.

see explanation of Figs. 29, 36.

ap.—auliplexus
au.—=aula
b.v.=blood vessel
cal.=rudimentary cal-
losum or fornicom-
missure
cbl.=cerebellum
c.e.=termatic eminence
ch.=chiasma
er.=crista
Dien.=Diencephal
dp.=diaplexus
dt.—diatela
ec. =epiccele
el.=endolymphatic sac
end.=endyma
Epen.=Epencephal
epi.=epiphysis
et.—=epitela
F.—Fissure between ol-
factory lobe and cere-
brum
fim.=fimbria
fl.—flocculus
G.—Fissure on the gemi-
num

ABBREVIATIONS.

gl.=glomeruli
gm.—=geminum
hb.=habena
hy.—=hy pophysis
inf.=infundibulum
inf. em.—=commissures
of the infundibulum.
mb.—=membranes
me.—=mesocoele
mem.—=medicommissure
Mesen.—=Mesencephal
Meten.—=Metencephal
m.p.—=mesencephalic pit
mst,—mesotela
mt.—=metatela
mtc.—=metaccele
mtp.—=metaplexus
my.—=myel
myc.—=myeloccele
N.=Nidus in the gemi-
num
ne.—=notochord
obl.=oblongata
op.7r.=optic recess

Italic letters represent sulci,
For a complete enumeration of the nidi, tracts and sulci

Roman letters

P.=Purkinje cells
par.—=parapnysis
pc.=paraccele
pcem.=precommissure
pia.=pia
pl.=pallium
pocm.=postcommissure
pp.—=paraplexus
Prosen.—=Prosencephal
pt.=porta
f.=Recess from aula
ventrad of crista
re.=rhinoccele
Rhinen.—Rhinencephal
rm.=rima
rmec.=recess of mesoccele
S.—=Endymal pocket cau-
dad of postcommissure
scm, —=supracommissure
st.=striatum
ter.—terma
th.—=thalamus
v.cm.=ventral commis-
sure

The mesal views are reconstructed from sagittal, frontal and transections.
Faint outlines indicate the hidden parts of the cerebrum, the geminum and

the paraplexus.
lines.
rupted lines.

The outlines of lateral cavities are indicated by interrupted
The position of the eye and of nerve roots is indicated by fine inter-

Numbers and reference lines indicate upon the mesal views the level at
which the sections figured were made.

228 PROCEEDINGS.

PLATE I.

Fig. 1. Mesal view of the brain of an embryo sparrow, the general state
of development of which corresponds quite closely with a chick of 10 days’
incubation as figured by Duval (’89). x 17.

Special attention is called to the following: The recess (R.) cephalad of
the thickened terma in which later the cerebral commissures develop; the
membranous roof of the diacoele (dt.); the large, open epiphysis; the ab-
sence of a supracommissure; the post and geminal commissures; the large
opening of the mesoccele into its lateral recess; the fissure (G) upon the me-
sal surface of the geminum; the membranous roof of the mesoccele (mst.),
and the epiccele (et.); the small metaplexus (mtp.); the thin walled sac ex-
tending over the myel (mt.); the deep sulci on the endymal surface; the
appearance of a portion of the striatum in the aula, dividing the porta into
two parts; the independence of the olfactory lobes.

Fig. 2-7. Frontal sections of the series from which Fig. 1 was con-
structed. x 9. The undifferentiated membrane (mb.) from which pia and
arachnoid develop, is shown at the left.

Fig 2. Shows the paraphysis in the midst of undifferentiated membrane ;
the rudiment of a paratela (rm.) and paraplexus; the union of the left re-
cess of the mesoccele with its dorso-mesal extension.

Fig. 3. Shows the intrusion of the striatum into the aula and the divi-
sion of the porta into two parts.

Fig. 4. Shows recess (R) cephalad of the terma; the aulic and paraccelic
parts of the sulci (b and d); the deepest parts of sulci (h and n); the fibers
to the postcommissure.

Fig. 5. Shows sulci (j. k.); the two parts of the recess of the mesoccele
produced by the total fold (G).

Fig. 6. Shows lateral halves of the cerebellum connected by a membrane
(et.); ectal cells (jl) continuous with the membrane.

Fig. 7. Shows lateral extent of metatela.

Fig. 8-10. Details from the same series. x 45.

Fig. 8. Shows entrance of sulcus (b) into the ventral angle of the porta;
the crista between the wide termatic segments (ce).

Fig. 9. Shows mesal lophius between sulci (z) and cells extending from
them into the raphé.

Fig. 10. Shows sulci(o and p) which pass ventrad of the postcommissure.

9 ut =>.

WU err

. ’ y
MI Uo QU

230 PROCEEDINGS. .

PLATE II,

Fig. 11. Mesal view of the brain of ahalf-grown embryosparrow. 16.5.
Compared with figure 1, the mesal extent is seen to be not much greater;
development has consisted in lateral growth, thickening of parts and change
in direction of the drain tube.

Attention is called to the fibers on the mesal aspect of the cerebrum which
unite and pass into the thalamus; the small protrusion of the striatum
into the aula; the decreased sulci of thecephalic region; the small entrance
from the mesoccele to its lateral recess; the close approach of the well de-
veloped geminum to the chiasma; the union of the olfactory lobes.

Fig. 12-13. Frontal sections from the series from which Fig. 11 was
constructed. 7.

Fig. 12. Shows short course of optic tracts (11) from chiasma to gemina.

Fig. 13. Shows nidus (NV) occupying the position of the mesal part of the
lateral recess (Fig. 5). Nidinear the raphéare of the third and fourth nerves.

Fig. 14-18. Details from the same series. X12.

Fig. 14. Shows relations of auli and diaplexuses dorsad of the porta.

Fig. 15. Shows the portas and plexuses.

Fig. 16. Shows at the left, the bifurcated porta; a the right the union
of the striatum with adjacent parts ventrad of the porta; the aulic and
paraccelic parts of the sulci (b and d); the united hippocampal and termatic
segments (c. é.), containing fornix fibers; a tract crossing the striatum from
the thalamus; a tract from the thalamus to the cerebrum; the mesal dia-
plexus.

Fig. 17. Shows, at the left, the flocculus and its relations to the depres-
sion in the skull filled with connective tissue (mb); at the right the entrance
of the epiplexus which is (Fig. 18) continuous with the metaplexus.

EAT.

PEAT

232 PROCEEDINGS.

PLATE IIL.

Fig. 19. Sketch of the dorsal view of the brain of an adult English spar-
row. x1. It shows the globular form of the brain and the overlapping of
the segments, the mesencephal and the metencephal not being visible.

Fig. 20. Mesal view of thesame. X 8.5. Compared with similar views of
the embryo (Fig. 11), it shows that the eye and the brain have become more
separated; and that, considering the relative magnification (at the same
scale as Fig. 11, Fig. 20 would be twice as large), the area of the chiasma
has greatly enlarged; in length the cerebrum has increased threefold, the
cerebellum five and the oblongata two, the union of the olfactory lobes
threefold and the geminum one-third. Attention is called to the compara-
tively smooth endymal surface; the well marked ectal cinerea of the cere-
bellum and its 13 folia; the epiccele partially occluded at the meson; the
caudal wall of the infundibulum nearly fused with the oblongata; the
greatly elongated, partially closed tube of the epiphysis; the small supra-
commissure; the long stretch of membrane between the entrance of the
epiphysis and the postcommissure ; the minute opening of the mesoccele into
its lateral recess.

Fig. 21-23. Transections of the same. X 3.3.

Fig. 21. Shows the cephalic, solid portion of the cerebrum and the united
olfactory lobes.

Fig. 22. Shows the small portas and paracceles; the layers of alba and
cinerea in the striatum; the ventral position of the gemina.

Fig. 23. Shows the opening of the mesocoele into its lateral recesses and
its wide membranous roof. :

Fig. 24. Frontal section of the same. The level at which this section
was made is indicated by the (24) at the right of Fig. 20. This figure shows
the cerebellum, its commissure, cavity and a nidus at either side; a continu-
ous sheet of ecto-cinerea extending into the flocculus and caudad of the
epiccele forming an ento-cinerea.

Fig. 24 A. An enlarged view of the double sulcus (0) on the cephalic as-
pect of the postcommissure.

Fig. 25-27. Details of the same series. 22.

Fig. 25. Shows habenas with their peculiar annular arrangement of cells;
the diaplexus and paraplexuses; ventrad of the diaccele, the membrane
connecting the tube of the epiphysis and the postcommissure.

Fig. 26. Shows the crista; the wide termatic eminence (¢. @.); the en-
trance of the sulcus (d@) into the paracoele at the cephalic angle of the porta.

Fig. 27. Shows the fusion of the cinerea and of the glomerular layer in
the olfactory lobes.

Ill

PLATE

Md UO:

usog«

‘UM?

19?

234 PROCEEDINGS OF THE

PLATE IV.

Fig. 28. Sketch of the dorsal view of the brain of an adult Amyda
mutica. xX 1. It shows a portion of all the segments.

Fig. 29. Mesal view of the brain of a young Amyda mutica (13 em,
across the carapace). xX 9. Especial attention is called to the symmetrical
development of the segments, the uniformity of commissural relations in
the different segments; the union of the olfactory lobes; the paratela,
faintly outlined, extending from the porta to the medicornu (pe.); the pro-
jection into the cavity of the callosum and precommissure; the close ap
proach of the paraphysis and epiphysis dorsad of the diaplexus.

Sulci.—(a) From preportal aula into the precornu; (b) from preoptic re-
cess to precornu; (c) laterad of crista; (d) from aula to medicornu; (e)
cephalad of medicommissure; (f) defines medicommissure; (g) dorsad of
habena joined by (i) ventrad of habena, the sulcus formed by the union of
these passes through the porta to form the boundary between the paratela
and tenia; (7) in the roof of the diaccele at either side of the paraphysis
and epiphysis; (j. k.) in the optic recess; (J) the deep infundibular sulcus
with its branches to the different recesses of the infundibulum; (m) sulcus
between the ventral and infundibular commissures; (”) cephalad of the
postcommissure; (0. p.) following the outline of the postcommissure; (q}
extending into the cephalic end of the slit-like passage from the mesoccele
to its lateral recess; (7. s.) entering caudal end of the same; (p) defining
the mesal lophius of the cerebellum; (7) bounding the entrance of the
metaplexus (compare Fig. 36); (v. w.) ventrad of (u); (7) laterad of the
posterior longitudinal fasciculus; (y) in the metatela; (2) defining the mesal
lophius of the oblongata.

Fig. 30-31. Frontal sections of the same. xX 4.2. The union of the ol-
factory lobes is shown, the cavities and plexuses of the cerebrum; the nar-
row cerebellum with its mesal lophius (f), entocinerea and Purkinje cells.
For tracts and nidi, see Fig. 36.

Fig. 31 shows meso-dorsal part of mesoccele and its lateral recesses.

Fig. 32-35. Details fromsame series. < 19.

Fig. 32-33. Show relations of sulci in the region of the postcommissure.
The double sulcus (0) extends upon a mesal fold of tissue caudad of the
postcommissure.

Fig. 34. Shows the crista and the entrance of the sulci fon the aula to
the paracoele at the ventral angle of the porta.

Fig. 35. Shows the preoptic recess cephalad of the precommissure and
the origin in it of sulcus (b); the relation of other sulci to the medicom-
missure,

—UISOAT

‘ejay yy QC

236 PROCEEDINGS.

PLATE V.

Fig. 36. The outlines are the same as those of Fig. 29. Shows nidi (sur-
rounded by interrupted lines); fiber tracts; commissural fibers (indicated by
dots); cells upon the meson (indicated by small circles); the dorsal limit of
the diaccele (interrupted line 7) after removal of mesal structures; the space
(w) at which the metaplexus enters.

Nidi: (a) in the termatic segment; (Db) near the porta; (@) in the base of
the mesal wall of the cerebrum and extending into the thalamus ventrad of
the precommissure; (d) near the meson next the medicom missure, the cen-
tralis of Humphrey, (Fig. -40—-42); (e) a less defined nidus caudad of (d); (f)
in the habena; (g) mesad of the optic tract (Fig. 42); (I) cephalad of the
postcommissure; (i) near the meson in the cephalic part of the geminum;
(j) laterad of the postcommissure; (Ik) mesad of the optic tract in the path
of tract 2; (I) the nidus of the third nerve; (m) the nidus of the fourth
nerve; (m) dorsad of (1 and m); (0) at the cephalic end of the oblongata (in-
tercrural); (p) in the caudal part of the geminum (interoptic of Spitzka);
(q) in the postgeminum; (1°) ventrad of the cerebellum; (S) among the roots
of the fifth nerve; (t) among the roots of the seventh nerve; (u) among the
roots of the ninth nerve; (vw) among the roots of the 10th and 11th nerves;
(x) in the raphé; (y) near caudal root of the fifth nerve; (z) the large cells
in the roof of the mesencephal; (P) Purkinje cells of the cerebellum.

Tracts: (1, 1) myelinic fibers extending from the olfactory region
through the base of the cerebrum (Fig. 38-42); (2, 2, 2) a system of am-
yelinic fibers from the base of the cerebrum connecting with a curving
branch in the region of the terma, extending through the thalamus near the
optic tract (Fig. 38-42) to cell nidi (k) where apparently new fibers arise and
continue into the base of the oblongata where they gradually disappear;
(3) myelinic fibers in the hippocampal segment (Fig. 38) passing to nidus
(a); (4, 4) amyelinic fibers from the fornix passing cephalad and crossing
in the fornicommissure, also fibers from the same commissure passing ceph-
alad into the termatic segment and hence possibly to the hippocampal
(Fig. 88-89); three sets of fibers crossing in the Precommissure, (a) amyelinic
turning cephalad and traced only a short distance, (b) myelinic and (c) am-
yelinic together forming the chief part of the commissure and passing
caudo-dorsad of tract 5 to the cerebrum; (6, 5, 5), fibers from the cephalic
part of the striatum (Figs. 88-40) passing in a compact bundle into the thal-
amus (Fig. 40), turning caudad and dividing into three strands, one passing
into nidus (d), (Fig. 41), another to nidus (e), a third is lost in the region of
the infundibular commissures (16, 17); the third is joined by a more caudal
tract from the striatum; (6) a tract arising between the caudal ends of
tract 5, passing in a curved course to mingle with the fibers of the post-
commissure; (7) Meynert’s bundle from nidus (f) of the habena passing

PLAGE. V..

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.

AMERICAN MICROSCOPICAL SOCIETY. 237

obliquely cauded; ($) the swpracommissure having two parts, the dorsal
closely connected with the tube of the epiphysis, its fibers passing on either
side to the nidus (f) of the habena; the ventral and larger portion of the
fibers passing to the surface of the thalamus, dividing into two portions,
one extending into the basal part of the cerebrum (Fig. 41), the other con-
tinuing toward the nidus (b); (9) a tract between the eighth and eleventh
tracts passing through nidus (i) to nidus (g); (10), fibers entad of the optic
tract crossing in the infracommissure (Fig. 39); (11), the optic tract, be-
ginning with the chiasma, forms a compact bundle, becoming ribbon-like
in section as it passes caudad over the surface of the thalamus (Figs. 30,
28-42); it divides into three parts; one as a distinct bundle passing dorsad
on the cephalic aspect of the geminum (Figs. 30-31), then spreading out to
form the ectal layer of that part; the other spreads out in fan shape upon
the latero-caudal aspect of the geminum; a third part consists of fibers
which pass into nidus (g); (12), a tract separated from the optic tract 11 of
the geminum by a layer of cells, its fibers passing, in general, parallel with
the optic (Fig. 31), a part cephalad passes through and around nidus (i), a
part passes ventrad with the postcommissure, a part joins with the tract
13, and a part passes cephalo-ventrad into the tegmentum and nidus (g); a
part caudad crossing the meson in the postgeminum, or passing into the
nidus (q); (18) a tract partially separated from tract 12 by cells (Fig. 18),
its fibers at right angles to tracts 11 and 12; these cross the meson as the
geminal commissure, laterad pass into the tegmentum, and caudad have
an apparent connection with nidus (p); (14) a tract, separated from tract
13 by a layer of cells, composed of a few very fine fibres (Fig. 31), part of
which cross the meson ventrad of the great cells of nidus (z) and a part in
the caudal part of the geminum pass ventrad into the tegmentum; (15) a
tract passing from the region of the postgeminum caudad; three tracts
from the postcommissure; the fibers of the cephalic part pass as a thin
layer (Fig. 32) dorsad of the sulcus (0) into an indistinct nidus (h) of the
diencephal, the middle part curves cephalad as tract 6, the caudal part
sends fibers into the tegmentum, but is reinforced by fibers from tract 12 in
such a way as to make this tract appear very large. (Fig. 33); (16, 17)
fibers from the infundibutar region crossing at the meson; (18) a large
number of fine fibers lying next the endyma (Figs. 38-42) with a few cross-
ing the meson; (19) a few fine fibers crossing the meson at the middle
of the medicommissure; (20) fibers forming a ventral commissure
just cephalad of the mesencephalic pit; (21, 22, 23) arched fibers cross-
ing the meson ventrad of the endyma, at the middle and ventral part of the
oblongata, throughout its entire extent or turning abruptly into the raphé;
fibers of 22 are especially concentrated in the region of the fifth nerve; (24)
the posterior longitudinal fasciculus, beginning near the nidi of the third
and fourth nerves, continues caudad forming the lophius bounded by sulcus
(x) (Figs. 29, 43); it is perforated and probably increased by arched fibers and_.
is the most marked and direct connection between the brain and the myel;
(25) the solitary bundle following the roots of the 11th, 10th and ninth
nerves to the seventh, where it disappears; (26) caudal root of the fifth

238 PROCEEDINGS.

nerve extending along the nidus (y) taking an oblique course into the myel;
(27) the tegmentum, a somewhat diffuse mass of cells and fibers with a
general cephalo-caudal direction, occupying a large portion of the oblongata
and perforated by nerve roots and arched fibers; (28) the decussation of
the fourth nerve in the valvula with its continuation to nidus (m); (29) fi-
bers lying between the roots of the fifth and eighth nerves at the surface of
the oblongata, passing through the decussational area at the base of the cere-
bellum; (30) fibers passing in a general cephalo-caudal direction among the
cells of the cerebellum to a decussational area just caudad of the preceding ;
(31) a few fibers next the endyma in the cerebellum passing ventrad to join
the tegmentum; (32) a great tract gathering near the surface of the cere-
bellum (Fig. 30), passing ventrad among the fibers of the eighth nerve, then
caudad and disappearing, as a compact mass, among the fibers of the ninth
nerve.

Figs. 37-42. Transections from one of the series from which Figure 29
was constructed. 95.5.

Fig. 37. Shows thesecond olfactory nerve root (I') ; the glomerular layer of
the first root; the complete encircling of the cavity of the olfactory lobe
by one layer of cells, the other layers being incomplete; traces of pia be-
tween the lobes.

Fig. 88. Shows the precommissure and callosum; the preportal aula and
the sulcus (a) at the junction of the hippocampal and termatic segments;
the sulcus (b) on both aulic and paraccelic wall.

Fig. 39. Shows the notch of endyma (b), between the callosum and pre-
commissure, and its continuation as sulcus (b); the relation of the auliplexus
to the paraplexus.

Fig. 40. Shows the medicommissure; paraphysis; diaplexuses; the para-
tela (rm); the passage of tract 5 from the striatum to the thalamus.

Fig. 41. Shows the passage of tract 8 frora the supracommissure to the
base of the cerebrum; nidus (d) at either side of the medicommissure.

Fig. 42. Shows the three caudal pockets (1) from the infundibulum: the
epiphysis; the optic tracts extending toward the gemina.

Fig. 43. Detail of the same series, to show the small lophius occurring
at the meson. c

ON THE FLAGELLA OF MOTILE BACTERIA.

VerRANuS A. Moore, Washington, D. C.

(ABSTRACT.)

During the past three years several new methods for demon-
strating flagella have been announced. Up to the present, how-
ever, a perfectly satisfactory process has not been devised and
the results obtained by different workers have been in many in-
stances quite contradictory. The efforts to fix upon the flagella
specific characters have also failed, although much advance has
been made in that direction.

THE NATURE OF THE FLAGELLA.

Notwithstanding the somewhat definite results which have been
obtained in reference to the structure of the flagella, it appears to
be of the highest importance that their nature should be more
fully determined before they are accepted as constant and integral
parts in the morphology of individual bacteria. The examina-
tion of a large number of preparations stained by the same
method, and frequently a single specimen, will reveal quite dif-
ferent appearances. In some instances, and in my experience on
a large majority of the bacilli, the flagella appear as appendages
radiating from the body (nucleus according to Biutschli) of the
organism. I have occasionally observed a narrow unstained or
more feebly-tinted band separating the body of the organism from
a deeply-stained ring of which the flaggella appeared to be pro-
jections. This capsule-like appearance has been illustrated by
several observers. Biitschli, Zettnow and others, hold that the part
of the bacillus which is easily brought out by the ordinary
staining methods is the nucleus only, and that ‘the additional
portion of the organism demonstrated by Leeffler’s method is
plasma which surrounds the nucleus. Heckle, on the other
hand, states that they have no nuclei. For this and other rea-

240 PROCEEDINGS OF THE

sons he refers bacteria to the animal kingdom, placing them in
the first class of Archezoa.

Farrier has recently published a series of interesting experi-
ences in which he shows that flagella on a single species of bac-
teria
to variations according to the conditions under which the organ-

as determined by the study of several forms—are subject

ism is cultivated. Thus he found that Bacillus coli communis,
cultivated at the temperature of the body, possessed several flagella,
but when grown at a much higher temperature (46°C. maximum
temperature for this bacillus) flagella could not be detected.
If grown at 44°C. a few of the individual bacteria possessed these
appendages. The age of the culture and the presence of a non-
fatal quantity of an antiseptic in the culture media were likewise
found to have appreciable effects. He states that this pleo-
morphism is due to their protoplasmic nature; the hypothesis
assumed being that when the bacteria are subjected to degenera-
tive agencies, such as high temperatures or antiseptics, the plasma
contracts in a ball-shaped mass (presumably about the organism),
but when the bacillus is again brought under favorable conditions
the plasma resumes its motile form.

Accepting this explanation, it is difficult to understand why
the motile bacteria possessed of capsules such as Micrococcus lan-
ceolatus are not, under certain conditions motile, or why the
methods employed satisfactorily in staining the capsule will not
act as well in bringing out the flagella. I have tried repeatedly
to stain the flagella after these methods, more particularly the
one used by Prof. Welch in staining the capsule on AZicrococcus
lanceolatus, but invariably the results have been negative. Why
there should be such a marked difference between the motile and
non-motile forms in the reaction of the “ capsular” plasma to
staining fluid has not yet been explained.

[ have sought for an explanation of the structure of the flagella
producing substance in the cilia or flagella of the zodspores found
in certain of the fungi, but thus far my efforts have not been re-
warded, although much assistance may be obtained from a study
of those forms. It is quite probable that certain observed

AMERICAN MICROSCOPICAL SOCIETY. 241

phenomena, especially in reference to the free flagella and the
formation of the rings and hooks frequently observed both on
the distal ends of the flagella, and separated from them, may be
explained by the same theories as those of zoospores. There
are two views as to the disposition of the flagella of swarm spores.
One is, that they are cast off, and the other, that they are absorbed
into the body of the spore. Rothert shows, in a recent article,
that both views are correct. ‘In the second swarm stage of
saprolegnia and in the peronosporee, the flagella are either cast
off as soon as the spores come to rest, or soon after, or else they
remain attached to the spore indefinitely even after germination.
In the first swarm stage of saprolegnia, however, he found, to
his surprise, that they are uniformly drawn back into the body
of the protoplasm, the withdrawal being slow at first, and then
quite rapid. The loops are formed either while the flagella are '
attached to the spores, or after they are cast off.” He suggests
the possibility that the flagella are formed out of special cyto-
plasm existing only in small quantities. It is highly probable
from certain opinions and results herein cited, that there is a close
resemblance between the flagella of bacteria and those of the
swarm spores.

The observations of Stocklin and Bunge that several bacilli
are sometimes included within the same capsule from the per-
iphery of which flagella radiate is exceedingly interesting. This
phenomenon is explained in two ways, one that the surrounding
plasma of two or more bacilli runs together, thus enclosing the
bacilli in a common capsule, and the other is that the variable
number of bacilli included within the same capsule is due to the
multiplication of the organism within the capsule. These obser-
vations strengthen the hypothesis that bacteria have nuclei and
surrounding plasma.

THE FOOD SUPPLY OF THE GREAT LAKES; AND SOME EX-
PERIMENTS ON ITS AMOUNT AND DISTRIBUTION.

Henry B. Warp, Pu. D., Lincoln, Neb.

The subject of agriculture has received for many years the
closest attention of scientific workers. Not only the character
of the different products, their food value for different uses and in
connection with the raising of different kinds of stock; but also
the value of the soil, the use of each element in it, the exact
relation of each individual particle in the entire chain of biological
relations from the unorganized matter to the saleable beef or
pork, has received from the experiment stations of the country
the most careful study. Every one of the pests, of the enemies
which threaten any agricultural product, be it plant or animal,
has been the object of similar active inquiry.

There is, however, one subject of economic importance which
has never received the same treatment in scientific hands.
Despite the painstaking investigations of a few scientific workers
and the encouragement of some official boards with limited means,
aquaculture has been almost as much neglected as agriculture
has been advanced, and all because continued and systematic
studies of its problems have not been supported and encouraged.
The incentive given by the early work of Hoy, Milner and
Forbes on the Great Lakes a quarter of a century ago has not
been followed up; chance has been relied upon to control the
conditions in these vast inland seas, and the fundamental features
of the problems are as little understood to-day as when there
was no drain upon the life in these waters. No farmer is so
ignorant as to suppose he could scatter the seeds of a grain whose
development was entirely unknown over land of which he was
equally ignorant, and leaving the land could hope on his return
in the fall to reap a bountiful harvest. And yet this is just what
has been looked for in the case of the white-fish, To use

AMERICAN MICROSCOPICAL SOCIETY. 243

the expression of Professor Reighard, published in his report as
scientific expert of the Michigan Fish Commission, a year ago, .
the fish culturists in the Great Lakes have been endeavoring to
‘take the young white-fish just hatched, an absolutely unknown
quantity, and put it into the water of the lakes, another
equally unknown quantity, and hoped from the union of two
unknowns to secure marketable white-fish. Putting the question
in that way it is evident that only the good fortune of a blunder
of some sort could result in practical benefit. Asa matter of
fact, when we examine the figures showing the results of white-
fish culture, we find that while there have been poured into the
waters of the lakes at different points some hundreds of millions
of young fry every year, the number of white-fish caught, the
supply present in the lakes, has decreased steadily year by year.

The question must be attacked from the other direction. We
must find at the first something of the biological conditions in
the life of the white-fish ; we must be able to say something of
where it goes, what it ought to do, with what enemies it has to
contend, what food it needs, what temperature, what general
conditions for development, before we can hope that the result
of white-fish culture will be more satisfactory. The conditions
of domestic economy on the land are so evident to our eyes that
we hardly stop to think of them. We see the large plants, the
grasses particularly, derive their nourishment directly from the
soil, the water and the atmosphere, turning inorganic matter into
organic ; we see this organic material in the domestic economy
of the cattle turned into beef-steak, and so the transition is direct
and immediate. It goes on, as we think, before our eyes, and
while the question is, naturally, more complicated than can be
outlined in a sentence or two, we think that we understand it.
The conditions in the Great Lakes are so utterly different that
when we come to speak of them we can afford no explanation
of what is the series of changes from the inorganic matter to the
organic, and from that to the white-fish in this particular
instance.

Without stopping to outline extensively the characteristic feat-

244 PROCEEDINGS OF THE

ures of the lakes themselves, I will simply enumerate two or

three characters which willappeal to you as peculiar. The absence

of large plants, 7. ¢., the absence of vegetation in the more

popular sense of the word, is important. - Shore plants are almost |
entirely lacking; the storms and the shifting character of the

shore itself, as well as the very small area of shallow water,

tend to reduce the possibilities for existence for the higher plants,

such as Phragmites, Scirpus, Potamogeton and others, that grow

in the littoral zone. The Characez, water plants that are found

in most or all of our shallow lakes in enormous numbers, so that

for instance in Lake St. Clair the bottom may be said to be

carpeted with them as thickly as a thrifty field of clover—these

are found in the Great Lakes only in comparatively isolated

patches.* While it is true that so far as I can find, with the

exception of a few rather scarce species of Gastropods, there is

no water form that feeds directly upon these plants, still they

vive shelter to an enormous number of minute forms, forms which

depend upon them for protection and find on them the growth of
smaller plants on which they live. The absence of these plants

in the Great Lakes reduces at once one element which is
important in all other fish regions: there is ordinarily only a
scanty bottom flora and fauna present.

Another source of food supply might be supposed to be the
different streams, which would bring to the lake nourishment in
one form or another. But if you examine the maps you will see
at once that the inflow into the Great Lakes is very small in com-
parison with the total volume of the lakes themselves. The con-
ditions there are practically stable. The inflow and outflow alter
the volume only by a very small per cent.** This is due, of
course, to the enormous area and depth of the lakes, and Lake
Michigan alone is said to contain more than one-tenth of all
the fresh water on the face of the globe.

In the next place from the atmosphere comes a little inorganic

* Tam speaking here of the deeper northern lakes. Of the conditions in
Lake Erie I can not speak from personal experience, and of Lake Ontario
I am entirely ignorant.

** Probably about one per cent yearly.

AMERICAN MICROSCOPICAL SOCIETY. 245

material, and also a very small percentage in the shape of insects
carried into the water by the wind. Together these constitute
but a small fraction of the whole, not enough to make them
more than secondarily important. The source of food must be
sought, then, within the waters themselves. If you take a net of
fine gauze, so fine that the meshes measure from .O1 to .oo! of a
square inch in opening, and draw this through the water, you will
collect by it a mass of minute microscopic forms, forms that are
known to most of you ; though very varied, they are all included
under the recently-coined scientific term of “plankton.” The
plankton includes all those minute forms of life which, floating free
in the water, are unable by their own efforts to effect their distribu-
tion. Inthe case of the animals, their individual motion will change
their position somewhat, but only by a distance, which is nothing in
comparison with the size of the lake and the conflicting agencies
of current and wave. Their distribution, then, is effected by
winds, waves, currents or storms, z. ¢., movements of the water
or of the atmosphere. They are passively distributed. They
differ, hence, from the larger forms, such as the fishes, or the
larger Crustacea, in that they are entirely dependent on their en-
vironment ; they can not alter it at will.

Within this group of forms are found in the first place as the
fundamentally important food element, the unicellular Alga, the
desmids and diatoms largely, forms which are present in an
enormous number of individuals, but, as our work in the lake
shows, in a comparatively limited number of species and genera.
Then there are the unicellular animals, the protozoons, present
also in limited number of genera and species, but also in enorm-
ous numbers of individuals. Beyond these the plankton of
the northern lakes shows only two forms, or groups of forms
rather, the rotifers, somewhat numerous, insignificant in size, and
for plankton purposes, I believe, unimportant ; and finally the
microscopic Crustacea, including such forms as the different genera
of Daphnids, the species of Cyclopida and Calanidze and other re-
lated forms in the Entomostraca. These together constitute the
elementary food supply of the water.

246 PROCEEDINGS OF THE

The unicellular plants float as it were in a nutritive fluid, the
lake water containing in solution inorganic matter derived from
the air, the bottom, the shore and brought into the lake by the
rains or in the inflowing streams. In the presence of light these
plants are capable of transforming this inorganic substance into
living matter.

It is a well-known fact that chemical action proceeds most
rapidly where the proportion of surface to volume is the
greatest. Leuckart brought this forward prominently in connec-
tion with the biological conditions of existence among the minute
organisms, to explain the extremely rapid reproduction and
multiplication of these forms. Microscopic, as they are, the pro-
portion of surface to volume is exceedingly large, beyond com-
parison greater than that of larger forms. They, of all organisms
are, then, capable of producing the most active chemical proc-
esses; in other words, they are capable of most rapid growth,
and since with them reproduction is clearly growth beyond the
limits of the individual and division into two, reproduction or
multiplication goes on with extraordinary rapidity, the number
that can be produced, and consequently the number that can be
consumed, in an hour, being almost if not entirely beyond ex-
pression in figures.

- The Protozoa live upon the unicellular Algz, the rotifers upon
both Protozoa and Algz ; the Crustacea are omnivorous, subsist-
ing upon all of the other forms, either dead or living, eating
everything which falls in their way; and it is this group, the
Crustacea, which forms the immediate source of fish food. The
smaller forms are eaten by the fish fry directly, and are some-
times the immediate food of the larger fishes also. One in-
stance that may be cited is the case of the lake herring, which
feeds directly upon these smaller crustaceans. With one inter-
mediate stage these forms constitute the food of the white-fish*
and of the larger fish of the lake. The immediate food of the

*Under the term ‘‘ white-fish” I wish to include not merely the particu-
lar species known properly under that name, but also the forms called
‘“long- jaw," ‘‘ black-fin,” ‘‘ tullibee,” etc., which are closely related species.

AMERICAN MICROSCOPICAL SOCIETY. 247

white-fish, as was shown by the investigations of the Michigan
Fish Commission at Charlevoix, last year, consists of JJysis,
Pontoporeta, severai different forms of gasteropods, small
Spheria, Pisidia, etc., among the lamellibranchs, and a single
form of insect life, two species of Chironomus \arvee, which are
exceedingly abundant at the bottom of the lake.

This is discussed very briefly, simply to show in outline the
connection between the plankton and the white-fish. The plank-
ton bears exactly the same relation to the lake herring that the
erass does to beef steak. It bears the same relation to the white-
fish, only that the relation is one step further removed. As I
might say, if cats were a source of revenue, the raising of cats
would depend upon the grass through the medium of the beef
eaten by the cat; so the store of the white-fish depends upon the
plankton through the medium of the larger forms, Myszs, Ponto-
poreia, etc., which feed directly upon the plankton. .

If the plankton is, then, the productive source of food supply
for the lake fishes, it becomes important to know something with
reference to its distribution, horizontally in area, and vertically in
strata, as well as its distribution in the different seasons
of the year and under different conditions of weather.
The questions of distribution in different seasons, and dif-
ferent conditions of weather, are so great that not one year
but many years of closest observation are necessary to give
even an approximate idea of them. However, the distribution
vertically, and to some extent horizontally, can be ascertained by
more restricted investigations. The Michigan Fish. Commission
for two years has maintained an investigating party in the field ;
in 1893 it was in charge of Prof. J. E. Reighard, and in 1894,
during his absence in Europe, I had the privilege of directing it.
During the first year the work was on Lake St. Clair; the sec-
ond year it was carried on at Charlevoix, Mich., which is in the
Traverse Bay region, in which the white-fish is caught during the
entire year.

It is possible here only to outline briefly the chief results, with-
out going into details with reference to the methods used, which

248 PROCEEDINGS OF THE

are given in full in the report of Prof. Reighard and in my own.*

The amount of plankton obtained has been measured and com-
pared for different places. The first chart (Plate I.) shows the most
important results, tabulated in a way to admit of ready comparison.
It was customary to select points on the lake at certain distances
from each other, with certain relations to shore, shallow water
and fishing grounds ; to make hauls with a vertical net at each of
these places. The material obtained was carefully preserved, and
later measured, in order to obtain the amount of plankton in the
water strained by the nets.. By calculating the amount of water
filtered, and the area of the net, the amount of plankton under
one square meter of surface is found, and this is used in the plate.
The dotted line (D) shows the depth of water at the different
stations. The total amount of plankton under one square meter
of surface, as measured in cubic centimeters, is shown by the solid
line (T), each station being represented by a vertical line with
Romannumeral. By dividing the total amount of plankton obtained
by the depth of the water, it is evident that the amount per cubic
meter of water is found, and this is expressed by the line (R) of
relative volumes.

Two or three points are very evident on the examination of
this chart. In the first place it will be seen, by comparing the
line of depth with that of total volumes, that, while the latter
suffers some fluctuations, it increases more rapidly than the depth,
up toacertain point.f For some distance then the line indicates no
decided change, but finally, with the sharp downward turn of the
line of depth at X, it makes a prominent though less decided
bend than that in the line of depth. Up to a depth of about 30°
meters, then, the total amount of plankton increases more rapidly
than the depth, but beyond 50 meters less rapidly.

The amount per cubic meter, as represented by the line of

*Bulletin of the Michigan Fish Commission, Nos. 4 and 7.

tThe extreme fluctuation at XIX is due to the presence of foreign matter
(sand) in the volume measured, and the dotted line from XVIIT to XX
represents more nearly the true course of the line of total volumes (T) after
eliminating this error.

AMERICAN MICROSCOPICAL SOCIETY. 249

relative volumes (R) is seen to decrease somewhat irregularly
with increase in depth; still it follows closely the average, as
indicated by the dotted line. In other words, the total amount
of plankton in the water increases with the depth, and the
average amount per cubic meter as clearly decreases at the same
time.

The explanation of this appears when we come to consider the
amount of plankton in the different layers or strata of the water.
By hauling the net from different depths, first from two meters,
then from five, twenty-five, fifty, one hundred meters and the bottom,
or wherever the bottom haul might be, we are able to distinguish
different artificial strata. We can then measure the amount of
plankton collected from the different depths, and by subtraction
are enabled to ascertain the amount that will be found in differ-
ent strata. It may be said that the amount in the first two
meters, the surface stratum, far exceeds in amount per cubic meter
that in all the rest of the water. In other words, proportionally
much the largest part of the plankton is accumulated in this sur-
face stratum. This is shown in the accompanying chart (Plate
If). The upper line (S) which crosses the chart variably shows
the amount of plankton per cubic meter in the surface stratum.
This amount appears to be independent of the depth, and similar
charts show no apparent relation between its fluctuations and
those of the line of temperature or of the time of day.

The amount of plankton per cubic meter of water contained in
the three intermediate strata (2-5 m., 5-10 m., and 10-25 m.,)
is not far from half that in the surface stratum, and is nearly
equal for the three strata. Curiously the amount in the middle
stratum is a little greater than that in the upper, while the latter
has even less than the lower of the three strata in some cases.
They are all as distinct from the deeper strata as from the sur-
face stratum. In fact the deeper strata (25-50 m., and 50 m.—
bottom) possess extremely little plankton, so little that it appears
as a negative quantity in some instances. This will occur in
applying the method of subtractions when the total amount pres-

ent in any stratum is less than the errors of the method or than
17

250 PROCEEDINGS OF THE

the normal fluctuations in the amount of plankton found in the
superjacent water.

With reference to the areal distribution of the plankton, it may
be said that so far as can be ascertained the amount in one part
of the lake is the same as the amount of plankton in any other
part of the lake; not only that, but there is no great difference
in the amount of plankton in the upper end of Lake Michigan
and the amount of plankton taken from the western end of Lake
Erie in some hauls made by Professor Reighard two years ago.
In other words, the conditions of existence are so uniform
throughout the lakes, and if you consider the temperatures, the
light and the other conditions, you will see that this might have
been expected to be the case—the conditions of life are so uni-
form that in the Great Lakes we find corresponding amounts of
plankton in widely separated regions. The conditions of existence
in the surface two meters are so similar that we find approximately
the same amount in the surface two meters everywhere. But
the conditions below that alter so rapidly with the lowering of
the temperature, the withdrawal of the light, and the change in
pressure, which are undoubtedly the three great conditions influ-
encing them, that below the two-meter line the amount of plankton
is very much less than in the upper two meters. It must not be
thought, however, that this is a fixed limit; the strata are purely
artificial, and the mass of life in the upper two meters may, so far
as our present knowledge is concerned, be accumulated in any
part of the stratum. In Lake St. Clair Reighard found that the
upper one and one-half meters held the bulk of the plankton.
The more exact limits of the water within which the plankton is
most crowded are yet to be determined.

Finally, the distribution of the plankton is merely the sum of the
distributions of its component species, and on the investigation of
the latter must wait the solution of the various peculiarities in the
former. I should like to call your attention here to the splendid
work recently done by Birge in Lake Mendota, Wis., and Marsh
in Green Lake, Wis. By such work is accumulated most valu-
able evidence on the biological conditions in smaller bodies of

AMERICAN MICROSCOPICAL SOCIETY. 251

water. But fish culture will never attain its proper results until
it receives from the hands of the people the same favors that have
been extended to agriculture, the establishment of permanent and
well-equipped experimental stations, where trained workers shall
devote all their time and energy to the solution of its problems.
The Great Lakes furnish a cheap and valuable food supply to one
third of our entire population ; this food supply is rapidly becom-
ing depleted ; how long must such important interests await their
just recognition and adequate protection ? And if properly devel-
oped who can limit the possibility of these inland seas in supply-
ing the nation with food ?

eRe PROCEEDINGS.

EXPLANATION OF PEAW Eas:

.
Vertical lines indicate stations, each of which is designated by a Roman
numeral.
Horizontal lines denote amount or distance according to the figures on
the margin.
A, hauls in Lake Michigan.
B, hauls in Round and Pine Lakes.

PLATE I.
D---—D, indicates depth of various stations.
T——T, indicates total volume of plankton in bottom hauls with vertical
net.
R—R, indicates volume of plankton per cubic meter of water insame

hauls.

PLAGE. I.

MY deo Wy ett

souludeg Wort

‘wl 24°/d

mpbry 31790 aWor)

"S@yary aig We PUNO
‘eT 94%Fd

\

ay
umanae
ener
Tl)

Miike

hee aes

mi!

~ed

254 PROCEEDINGS.

PLATE IL

D-—-D, as before.
Ss S, amount of plankton in hauls from 2 m. to surface.

2— 5m., amount of plankton in hauls from 5m. to 2m.

5-10 m., amount of plankton in hauls from 10 m. to 5m.
10-25 m., amount of plankton in hauls from 25 m. to 10 m.
25-50 m., amount of plankton in hauls from 50 m. to 25 m.

50 m. — bottom, amount of plankton in hauls from bottom to 50 m.
For further details see text.

PLATE. i:

A NEW JIETHOD FOR THE QUANTITATIVE DETERIIUNATION OF
PLANKTON HAULS.

Henry B. Warp, Pu. D., Lincoln, Neb.

Only two methods of estimating the quantity of plankton
obtained in a haul of the vertical net are known to me, These
two may be called the volumetric and the gravimetric. Both
have been used by observers in various parts of the world, but so
far as I know have never been comparatively tested in order to
ascertain the relative value of results obtained by the two. The
experience of the past year has shown some disadvantages in the
first method, while the second has certain evident objections
which rendered its employment in this case out of the question.
In the course of the work outlined in another paper (Ward 95)
I worked out a modification of the volumetric method, or
rather, perhaps, a way of combining the two methods so as to
control and correct each other; this combined method is briefly
outlined in the following, but for its better understanding a short
account of the two previously existing methods is prefixed.

The volumetric method of measuring the plankton is as follows :
The material obtained in a single haul of the vertical net is pre-
served zz toto under precautions which prevent any appreciable
loss* ; this mass is brought with care into an accurately graduated
glass tube and allowed to settle twenty-four hours ; at the end of
that time the exact volume in cubic centimetres is read from the
scale, or estimated if the upper surface is not perfectly horizontal.
This approximation can be made sufficiently accurately, but I have
found by a series of experiments, that the amount obtained by
these measurements is variable and depends on several condi-
tions. In the first place it fluctuates with the size of the tube.
In the larger tube the plankton settles more thoroughly and gives

*For a more detailed account compare Reighard 94, pp. 26-28.

256 PROCEEDINGS OF THE

a smaller volume; in the smaller tube it stays up and yields a
-larger volume. Again the result varies with the character of the
material. A diatomaceous plankton settles much more com-
pactly than one consisting largely or entirely of Crustacea. If
there is any considerable amount of filamentous material, like
Lyngbya, or some of those forms, the plankton will not settle at
all thoroughly, and the volume reached then will be much
greater than is really represented. In the third place the amount
obtained by the measurement depends upon the time which the
plankton is allowed to settle, since I have had tubes standing up
to two or three months and the plankton settled all the time, not
regularly, but somewhat even to the end of the period. Finally
the volume depends upon the conditions as regards movement—
whether the tube be in absolute quiet or in a room where there is
constant vibration. The amount of variation possible, z. ¢., the
limit of accuracy in this method, will be discussed more fully in
another paper.

The second or gravimetric method of determining the volume
of plankton has been recently advocated by Zacharias (95). The
material obtained in a single haul of the vertical net is brought
with care on to a filter paper and thoroughly drained ; while yet
fresh and moist it is weighed. The amounts thus obtained, ex-
pressed in milligrams, afford the basis for comparison of differ-
ent hauls. One great objection to this method is advanced by
Zacharias himself. A complete drying out of the mass is im-
practicable and the amount of moisture which remains is undoubt-
edly variable, being evidently greater in hauls of larger volume
than in those which are scanty. Zacharias estimates the quantity
of moisture held in the mass of plankton as one-fourth of the
entire weight. No evidence is given to support this statement,
neither is it discussed at all, and I cannot see that it has more
weight than a purely personal opinion. Nor is it evident that
greater accuracy is reached in this way than by the volumetric
method. The second objection to this proceedure, however, is
more weighty and absolutely prohibits its employment in many
cases. The material obtained in the haul is entirely destroyed

AMERICAN MICROSCOPICAL SOCIETY. 257

and cannot be used for subsequent qualitative examination of any.
sort, whereas the measurement of a haul by the volumetric
method does not in the slightest injure the material or in any way
interfere with future operations involving its use.

But beyond all these variations in the amount as determined by
both methods, there is at least one source of error in the relation
expressed by the actual figures. It is no infrequent matter to
find in those hauls made from the bottom a certain amount of
foreign material, sand, etc.; this is either stirred up when the
bucket strikes the bottom, or, being carried along by currents, is
swept into the net. It is, of course, preserved and measured with
the true plankton material and falsifies the actual amount of the
haul according to the value which may be included. In one case
at least (Ward, 95, p. 248,) the amount of sand swept into a bottom
haul by a current was sufficient to raise the total mass not less
than one-third. The modification here suggested was evolved to
meet this difficulty and to correct these errors.

The entire mass of plankton in a single haul is measured in a
graduated tube, as in the volumetric method, and the volume
recorded. A certain amount is then poured into an evaporating
dish, care being taken that the entire mass of the haul has been
thoroughly shaken up so that the different constituents are
equally distributed through the volume. Ordinarily one-fifth
to one-half of the original mass will be poured out, and the re-
mainder is then carefully measured as before by the volumetric
method and the volume noted. The portion of the plankton re-
moved is next air-dried, weighed, then dried to a constant weight
at 100°C., and the amount recorded. The ash is next obtained
and weighed, and finally by digestion with concentrated HCI, wash-
ing, decanting and drying, the weight of the sand present is found.*
To illustrate the method three hauls made in Lake Michigan
are taken: XIII* and XIX‘ are partial hauls, free from impuri-
ties; and XIX® is a bottom haul which was very much polluted

*The processes were very kindly carried out for me in the Chemical
Laboratory of the University of Nebraska, by Mr. E. E. Nicholson, to whom

my sincere thanks are due for his kindness.
18

258 PROCEEDINGS OF THE

by sand. The results obtained are expressed in the following
table :

‘Depth in Metres, Volume in ce. of Pereentage| Weight in Milligrams.
Number of | Haul. of Original te a mt oe
of Haul oe a Volume a Constant Ash | Sand
. ier. _| Origi- After aken as a) Driec eightof, in in
Haul. | W ae) nally. removal.. Sample. |Sample| Sample. Sample| Sample
= —- Bs ————— ——| —— -~- = =|

XIII* 25 30 | 9.385) 488 47.8 53 50 “6 2.3
XIX# 25 "| 36 | 6.55} 1.00 | 84.8 61 57 8.0 mal
xX1X5 30 if) 0.60) oil lt90 se OU Gian amore 207 206 {60 0 | 112.0

From this table data with reference to the composition of the
entire haul may be obtained by simple calculation. In this way
we arrive at the results given in the next table.

Weight in Milligrams for Percentage in Air-dried Sample | Welt
Number Entire Haul of of 1sq. m. of surface.
Ce neneical Organic | Entire |Ash,not Air-dried

- Cc { 7 | a

Plankton. Ash. | Sand. Material.) Ash. | Sand. Sand. Material. Ash.

= = | | = | —— — =
XIIi4 110 16 4.8| 85.7 | 143) 10.7 | 3.6 | 1.9982 | 0.2860
xXIx4 72 9 2.5 | 86.8 | 13.2 9.7 |} 3.41 1.3050 | O:2708

0 22.6 | 77.4 |. 28.2 | 54.2

XIX*® | 1034 | 800 | 560.

18.7064 |14.4710
From the average of XIII* and XIX‘ it appears that in an
ordinary haul, comparatively free from foreign material, there is
about 3.5 per cent. of silicious matter, 10 per cent. of other
earthy substances, chiefly calcareous, and 86 per cent. of organic
material. The plankton is thus very nearly pure food matter.

If this be compared with the results given for XIX‘, it will be
seen that the latter haul contains an excess of sand equal to more
than 50 per cent. of the total weight and an excess of total inor-
ganic matter equal to 64 per cent. This is in itself sufficient
proof of the contamination of the haul, a fact equally clearly
established by microscopical investigation of the material. By
this method, however, one can compute with considerable ac-
curacy the amount of foreign matter present and thus obtain a
correction by which the recorded volume of any haul may be
reduced to the true volume.

The method is further applicable to those cases when the

AMERICAN MICROSCOPICAL SOCIETY. 259

plankton does not settle readily or fully. Here a correction is
found for the excess of volume over the true amount.

In other words the results obtained by the volumetric method
may thus be controlled and corrected through the detection of
errors due to false measurements, and of those due to the
presence of foreign inorganic substances in the haul. The latter
is not shown in any way by the simple volumetric or gravimetric
method. At the same time a much more accurate idea of the
true weight of the plankton is offered than by the gravimetric
method, since there is no variable and uncertain quantum of moist-
ure to be estimated. The method is further applicable in that
number of cases when the plankton is also to be used for the
qualitative examinations, since the part not weighed is entirely
uninjured, and may be treated in any desired manner.

I have used this method further in determining the actual
weight of the plankton in Lake Michigan, in order to ascertain
in this way the actual amount of food material present in the
water. Further details on the application of the method may be
found in my forthcoming report on the plankton of Lake Michi-
gan, to be published as a Bulletin of the Michigan Fish Com-
mission.

260

PROCEEDINGS.

PAPERS {Gite

94. Reighard, J. E.—A Biological Examination of Lake St. Clair. Bul-

95.

95.

letin of the Michigan Fish Commission, No. 4 ; 60 pp., 2 plates and
map.

Ward, Henry B.—The Food Supply of the Great Lakes ; and some
Experiments on its Amount and Distribution. Proc. Am. Mic. Soe.
Vol. XVII., pp. 242 to 258.

Zacharias, O.—Ueber die wechselnde Quantitit des Plankton im

Grossen Pléner See. Forschungsberichte aus der Biologischen Sta-
tion zu Plén, Theil III., Seite 97-117.

THE SPERMATHECA AND IIETHODS OF FERTILIZATION IN
SOME AMERICAN NEWTS AND SALAMANDERS.

B. F. Kinespury, Pu. BD. ,~ Defiance, O.

In the proceedings of the American Microscopical Society for
1894,in a paper upon the “ Histological Structure of the Enteron
of Necturus maculatus,’ the writer alluded to the presence of
“ Receptacula semints,”
female in this animal. Certain inconsistencies between the con-

in the dorsal wall of the cloaca of the

ditions in this one form, and statements made by investigators
who had worked upon European salamanders alone, showed the
desirability of a knowledge of the relations existing in a wider
range of forms, and a study of the cloaca in the female of six
species of American urodeles was undertaken.* These forms were
Necturus, a perennibranchiate purely aquatic form ; Déemyctylus
viridescens, aquatic in its larval andadult state, but passing through
a land stage; Amzblystoma punctatum, a terrestrial salamander in
the adult, except at the breeding season ; Desmognathus fusca and
Spelerpes bilineatus, forms which seem to adapt themselves readily
to either a land or water existence; and Plethodon erythronotus
and g/utinosus, forms which are said to (Cope) pass no period, even as
larve, of their existence in the water. These are representatives
of five families and two orders of Amphibia, and present in varia-
tion of habit of life a good series from a purely aquatic to as
purely a terrestrial existence.

The general result has been the recognition in all of structures
in the cloacal wall of the female which serve as reservoirs in
which the zoosperms of the male are received, functionally com-
parable, therefore, to the Receptaculum seminis of certain insects
and other Arthropods. This term, however, by which they
have been designated hitherto, is not strictly applicable to the

*This paper was prepar at in the anatomical laboratory of Cornell Uni-
versity. I desire to express my appreciation of the abundant material and
facilities placed at my disposal. At the suggestion of Professor Gage this
investigation was made, and to his interest and advice is largely due what
of value is herein contained.

262 PROCEEDINGS OF THE

organ as a whole, since in certain urodeles, Vecturus for example,
there is no unity of structure, there being many pouches or recep-
tacula. Instead of receptaculum seminis, spermatheca is preferred
as a euphonious mononym, and when there are many discon-
nected tubules which function as reservoirs for the zoosperms,
each will be called a spermatheca. Therefore, in such forms
many spermathecas would be recognized.

Strictly the ascertainment of the existence, state of development
and structure of the spermathecas in the female would belong to
an investigation of the development and life history of each of
the forms here studied, and a discussion apart from a considera-
tion or knowledge of the mode of mating, fertilization and ovula-
tion is,in some respects, disadvantageous. Since, however, these.
related facts have been treated of somewhat monographically in
the case of two of the species here studied, and either the presence
of spermathecas has not been considered at all, or assumed to
be the same as in the European forms in which it has been
studied, the present research seems fully justified, especially as it
is hoped that in the case of Plethodon and Desmognathus it may
be but preliminary to the ascertainment of the mode of mating in
these peculiarly interesting forms.

Although generalizations and distinctions. of wide application
should be made with caution, it appears to be a constant differ-
ence between the tailed and tailless forms of Amphibia, that in the
former fertilization of- the ovum is internal, in the latter external.
Indeed, it is probable that in all the Uvodela not only is fertiliza-
tion internal, but it is accomplished by the same mode of mating.
In all the forms so far studied it consists in a more or less com-
plicated ‘‘ courtship,” which culminates in the deposition by the
male of one or more spermatophores, consisting each ofa gelatinous
body bearing on its summit a mass of zodsperms. Over these the
female passes, and the zodsperms are either actively grasped by the
distended lips of the cloaca of the female, or cling to the outside
and enter apparently of their own activity, independent of any
efforts on the part of the female; there seems to be a difference
in different forms in this respect. Within the cloaca of the female

AMERICAN MICROSCOPICAL SOCIETY. 263

they find their way into the spermathecas, and there remain until
the time of ovulation. (See under Salamandra atra in Conclud-
ing Remarks.)

As in so many other matters of habit or anatomical detail in
the Amphibia, it was in European forms that the mode of fertiliza-
tion was first carefully observed, though a long time was required
before the matter was at all understood. Spallanzani, in 1785 |
first showed that fertilization in the Triton was internal. He
considered, however, that the zoosperms became diffused in the
water and in that way entered the cloaca of the female, a view
which later observations have shown to be incorrect. Rusconi,
likewise, affirmed the internal fertilization of the ovum in the
Triton, and that the male deposited the zoosperms externally, no
true copulation taking place. With Spallanzani he believed that
internal fertilization was accomplished by the diffusion of the
zoosperms in the water, and that they thus obtained entrance to
the cloaca of the female. He still further made the mistake of
believing that an external fertilization took place in addition to the
internal. Other writers also recognized the fact of internal fertiliza-
tion in these forms, and the views of Spallanzani and Rusconi were
commonly accepted. Tomy knowledge but one observer (Finger)
affirmed that a true act of copulation took place in the Triton.

In 1858 appeared Siebold’s significant discovery of the presence
in Salamandra atra and maculosa and Triton igneus,* cristatus

* The species are here referred to under the names used by the various
writers. Where different, the following sets forth their identity with the
genera and species recognized by Cope:

GENERAL. COPE.
SALAMANDRID.©.
Triton cristatus. Hemisalamandra cristata.
Triton alpestris. Triturus alpestris.
Triton igneus. Triturus alpestris.
Triton tzeniatus. Triturus vulgaris.
Triton abdominalis. Triturus vulgaris.
Triton punctatus. - Triturus vulgaris.
Megapterna montana. Triturus montanus.
PLEURODELID 2.
Pleurodeles waltlii Pleurodeles waltli.
Triton palmatus. Diemyctylus palmatus.
Triton helveticus. Diemyctylus palmatus.

Euproctus pyrenzeus. Diemyctylus asperus,

264 PROCEEDINGS OF THE

and ¢aniatus of groups of blind pouches in the dorsal wall of the
cloaca in the female, filled with living zoosperms.. This proved
conclusively, if indeed further proof were necessary, that in these
forms at least fertilization was internal, He was led by his dis-
covery, however, to the erroneous belief that a direct sexual com-
munication was therefore necessary.* Robin, ’74, it appears,
was the first to recognize the true mode of fertilization in the
tailed Amphibia (Axolotl and 7rztonu alpestris, palmatus, cristatus
and abdominalis, or punctatus). By him the spermatophores were
recognized and described as such. Gasco later,in 1880, also
described the true mode of fertilization in the Triton, and the
following year he was enabled to report the same mode of fertili-
zation in the axolotl. The deposition of the spermatophores was
described, and he stated that the zoosperms were actively taken
up by the cloacal lips of the female in both forms. Zeller, ’91,
later states that this is not the case in Triton; Fick, however,
confirms it for the axolotl. The mating habits of two European
genera of the //lewrodelide and one genus of the Salamandride**
were described by Bedriaga, and though his observations were
fragmentary, were found to resemble quite closely that described
by Gasco for the Triton. It remained for Zeller, ’90, to gather
together these more isolated observations, and supplement them
in an interesting paper by observations of his own upon the
mating habits of the genera 7riton, Salamandra, Axolotl, Pleuro-
deles and Diemyctylus, in all of which occurs the same fundamen-
tal plan of fertilization, the preliminary courtship, the deposition
of spermatophores by the male and the reception of the zoo-
sperms by the cloaca of the female.

To the writer’s knowledge only two American forms have been
observed at all, and but one carefully, Déemyctylus. The mating

*By both Zeller and Jordan, ’91, it has been noted how persistent was the
influence of this opinion of Siebold’s, so that in many standard text-books
of zoélogy the statement persists that a copulation takes place. Cope fur-
ther makes the unqualified statement that in tailed Amphibia a copulation
occurs.

** Pleurodeles Waltlii, Megapterna montana Savi, Glossolega Hagenmul-
leri, and Euproctus (Hemitriton) pyrenceus.

AMERICAN MICROSCOPICAL SOCIETY. 265

of this newt was first observed by Zeller who speaks of it as
Triton viridescens, later by Gage and by Jordan, ’91, and the
preliminary courtship, deposition of spermatophores and the re-
ception of the zoosperms by the female are minutely described.

Observations upon the second American form, Amzblystoma
punctatum, have been very fragmentary, and Clark, by whom
they were made, drew the conclusion that in this form fertilization
was external, and is quoted to that effect by Balfour. There
is little doubt but that what he observed was the deposition of a
spermatophore* and that Awmzdlystoma agrees with the other
urodeles in its breeding habits; indeed, the thrice-repeated
observations on the axolotl would leave nothing else to be ex-
pected. Were demonstration of internal fertilization necessary,
it has twice been furnished in the anatomical laboratory of Cor-
nell by the development of eggs laid by female Ammdlystomas
with no male present. A study of the spermathecas also demon-
strates the fact completely.**

Thus we see in all the forms so far observed*** a constant
mode of internal fertilization occurs, and as stated by Zeller, ’go,
similar mating habits will probably be found in the remaining
urodeles. On the spermathecas in which the zodsperms are
stored very little has been said. Zeller refers to them merely
without discussing their structure. Their presence has been

*Clark says, p. 106, ‘‘ the males showed no inclination to clasp the females,
but quietly deposited quite large masses of an apparently rather thick
liquid, opaque white, on the bottom of the dish in which they were kept.
Upon examination this liquid was found to consist of spermatozoa moving
rapidly in a liquid. The eggs were found to have adhering to their outer
shells, shortly after, a considerable number of these male elements, but |
could not succeed after trying a great many times in finding any sperm-
atozoa within even the outer shell. Most of the eggs were laid during
the night, and by nine o’clock the next morning the first segmentation furrow
had usually made its appearance.”

**Kycleshymer, who has worked up the development of Amblystoma,
evidently considered it unnecessary even to allude to the internal fertiliza-
tion, despite the fact that published statements had been made to the effect
that fertilization in this form was external.

***In Proteus the mere fact that fertilization is internal has been
demonstrated by Chauvin, ’83.

266 PROCEEDINGS OF THE

alluded to in Salamandra, Triton, Diemyctylus, Geotriton and the
axolotl, but in Zy7#on alone has their structure been considered
in any detail.

THE CLOACA OF THE MALE AND ITS GLANDS.

It is not the purpose to discuss the structure of the cloaca and
the glands belonging to it in the male, as it is but incidentally
connected with this investigation. The attention I have given
them is entirely superficial ; indeed the complexity of the structure
and the opportunities afforded in the glands to study the changes
undergone by the secreting cells from a resting state to one of
exhaustion, would render a special study productive of valuable
results; such a study has been made by Heidenhain in 77iton
cristatus, alpestris, teniatus and helveticus, especial attention be-
ing paid by him to one of the groups, the pelvic gland. In view
of comparisons that have been made and will be made here
between the cloacal glands of male and female, a few words must
be devoted to the relations in the male.

Our knowledge of the cloaca is, as it is hardly necessary to
say, based on the conditions in European forms, especially the
Tritons, and chiefly through the writings of Rathke, Finger,
Duvernoy, Leydig, Blanchard, and especially Heidenhain, who
has devoted a most careful monograph to the structure of the
cloaca and its glands in the Triton. As his description is the
most complete it may be made the basis of the following state-
ments on the conditions in that genus: Two portions of the
cloaca are recognized by him, an ectal, caudal, ventral chamber,
called by him the cloacal chamber, (Kloakenkammer) and an
ental, more dorsal and cephalic tubular portion, the cloacal tube
(Kloakenrohr). In the outer portion of the cloacal walls there
is on each side a furrow which runs caudad and ectad (ventrad),
and appearing upon the lips of the cloaca divides them into two
limbs, an inner and outer; both the first unite to limit the vent
caudad, while the latter do not unite, but end freely. In the de-
pression occur (in Triton) about twenty thread-like papilla at the
summits of which are the orifices of the tubules which constitute
the abdominal gland (Bauchdriise).

AMERICAN MICROSCOPICAL SOCIETY. 267

The walls of the cloacal chamber are raised into ridges which
run caudad and ventrad toward the lips of the cloaca. The
cloacal cavity does not terminate roundly in its caudal end, but
projects slightly caudad each side of the meson, constituting the
posterior recesses. On the dorsal side of the cloacal chamber is
a depression (Dorsalrinne) which proceeding cephalad becomes
T-shaped and is the direct continuation of the tubular portion of
the cloaca. The shape and indeed the depression itself is doubt-
less entirely obliterated when the cloaca is filled with feces. Vent-
rad of the depression and therefore truly from the ventral wall of
the cloaca, though in appearance from the dorsal side, extends a
tongue-like elevation whose shape and size appears to be altered
by contraction.*

Dorsal and ventral ciliated areas are recognized, the former
extending from the level of the caudal end of the kidneys
(slightly caudad of the uro-genital papilla) to the caudal end of
the dorsal depression whose epithelium is ciliated throughout.
The ventral area, according to Heidenhain, is more limited, ex-
tending from about the level of the cephalic end of the dorsal
area to the caudal limit of the cloacal tube, z. e. slightly cephalad
of the cephalic end of the cloacal opening. The remaining
epithelium of the cloaca he found to be composed of a single
layer of mucous cells which continued up to the edge of the
cloacal lips where transition to the epidermis is rather rapid,
although the cloacal epithelium adjacent to the epidermis is two
layered. Therefore Heidenhain concludes that the epithelium
of the cloaca is entodermal and considers the pelvic and cloacal
glands as entodermal in origin, while the abdominal gland tubules
opening upon papilla which are covered with the stratified
epithelium of the skin are ectodermal.

*To this had been applied by earlier writers the term penis. This is un-
fortunate ; it has long been known that this is no copulatory organ. The
name of cloacal papilla employed by Blanchard is also unsatisfactory and
apt to introduce confusion. Heidenhain has not thought fit to introduce a
new term, and since I only refer it to incidentally, it will not be necessary
to do so here.

268 PROCEEDINGS OF THE

Before Heidenhain* but two giands were recognized as belong-
ing to the cloaca of the male, the cloacal gland (Kloakendrise,
Leydig ; Afterdriise, Rathke) and the pelvic gland. Heidenhain
has recognized another group of tubules included by former in-
vestigators (except Duvernoy) with the pelvic gland, but which he
separates as the abdominal gland. The cloacal gland entirely
surrounds the cloacal chamber and forms the basis of the cloacal
wall, except on the dorsal side where the continuity of its mass
is interrupted by the dorsal depression. Its tubules are quite
straight, unbranched and open in most part into the cloacal
chamber upon the summits of the longitudinal folds mentioned
above. The pelvic gland occupies a position on the dorsal side
of the cloaca. Its tubules open upon the dorsal ciliated area.
When the dorsal depression has assumed the T-shape the open-
ings are confined to the lateral walls of the depression, that is, to the
stalk of the T; farther cephalad a few tubules open in the non-
ciliated zone separating the dorsal and ventral ciliated areas. No
openings occur in the ventral ciliated area. The tubules of the
third gland newly recognized by him, the abdominal gland, open
as stated above, upon the summit of the papillae which are situ-
ated in the depression between the two limbs of the cloacal lip.
From their openings the tubules run cephalad on each side, and
form the larger part of the gland mass which is situated in the
abdominal cavity between the abdominal muscles and the peri-
toneum. The tubules are in part forked atthe end. Froma his-
tological consideration five forms of tubules were recognized, one
which Heidenhain was inclined to regard as a special kind of
gland. He was unable to determine in how far the others should
have like recognition, or merely represented different phases of
secretive activity of the same kind of gland.

Without discussing the value of the distinction into cloacal
tube and cloacal chamber, or the entodermal or ectodermal nature
of the epithelium and glands, the following brief observations
and comparisons may be hazarded on the relations exist-

*Duvernoy, however, recognized two portions of the pelvic gland which
he named prostate abdominale and prostate pelvienne.

AMERICAN MICROSCOPICAL SOCIETY. 269

ing in the males of the five genera studied in this investi-
gation. Dzemyctylus may be conveniently considered first, since
it is the most nearly related to the Tritons and the relations
at the cloaca approximate those in that form most closely,
The three glands were readily recognized and their relations
agreed closely with those in Z77zton. The dorsal depression was
well developed and assumed the T-shape described for that form.
The dorsal ciliated area was large and lined the dorsal depres-
sion throughout. It began as a mesal area and spread laterad as
it progressed caudad until not only was the epithelium of the T-
shaped depression ciliated, but also that adjacent to it in the dorsal
wall of the cloaca. From the ventral wall an elevation projected
slightly caudad of the uro-genital papilla, which caudad became
broken up into the ridges on which the cloacal gland opened. It
was covered with ciliated epithelium which extended caudad upon
. the ridges spoken of above, almost to the cloacal lips. In the
other four genera the ciliated areas were much as in Diemyctylus.
In all, the ventral tract extended caudad upon the high ridges
which bore the mouths of the cloacal gland tubules, being there-
fore much more extensive than in the 77zton. The presence of
ciliated areas in the male cloacas of purely terrestrial as well as
purely aquatic urodeles, clearly disproves the view advanced by
Leydig and quoted with favor by Hoffmann that a ciliated con-
dition of the cloacal epithelium was to be associated with an
aquatic life. It is a peculiar fact that no ciliz were found in the
cloaca of the female in any species examined save Amblystoma
and Plethodon glutinosus, which will be spoken of subsequently.

The cloacal glands in Dzemycty/us seemed to be much as in
Triton. The pelvic gland tubules opened into the dorsal depres-
sion in four groups, viz., proceeding caudad, (a) a cephalic mesal
group of short tubules whose mouths opened at the summits of
low papillz ; (b) farther caudad lateral groups opening upon the
stalk of the T; (c) a few tubules opened in the non-ciliated area
ventrad of the dorsal ciliated tract ; and (d) a mesal group open-
ing on a small elevation in the caudal portion of the dorsal
depression, The mouths of the tubules composing the cloacal

270 PROCEEDINGS OF THE

gland were situated as stated above in the summits of the ridges
which radiated toward the cloacal lips where they were succeeded
by large villi on which the tubules opened two and three together,
around the cephalic end of the cloacal opening. The cloacal
gland in all the genera presented much the same appearance, and
the relation of the mouths of the tubules to the longitudinal ridges
on the cloacal wall was constant in all. In Mecturus and Pletho-
don these ridges were especially high and thin. In Wecturus,
Desmognathus and Plethodon the ridges were succeeded at the
edge of the cloacal lips by papilla which bore mouths of the
tubules, though in the last two genera they were short. The
tubules in the specimens examined were filled with a stringy se-
cretion which took the hamatoxylin stain. . The cells appeared
as figured by Heidenhain, reticulated and staining blue; un-
doubtedly, as stated by him, this gland is mucin-secreting.

The abdominal gland in Dzemyctylus may be easily recognized .
and its relations are much as in 77riton. As in that genus, its
tubules open upon slender papilla near the caudal end of the
vent. From their ducts the tubules pass cephalad, laterad to the
mass of the pelvic gland, to form with the pelvic gland a mass
between the peritoneum and body muscles in the caudal end of
the abdomen. In Diemyctylus the mass. of its tubules extended
ventrad to lie beneath the peritoneum dorsad of the pelvic arch.
The free ends of the cells were filled with small globules of secre-
tion which stained but lightly. In some cases almost the entire
cell was filled with these globules, the nucleus surrounded by a
scanty mass of protoplasm was cramped in the basal end of the
cell. The papilla on which the tubules opened were covered
with a stratified epithelium which resembled the epidermis.

Necturus, in the configuration of the cloaca, resembled Die-
myctylus closely. The dorsal depression, however, did not
assume the T-shape as in that form. Both the cloacal and pelvic
glands were greatly developed ; a sufficiently careful study of the
glands was not made to enable me to determine satisfactorily
whether or not the abdominal gland was present. I consider it
present though its tubules do not open from papilla, nor do they

AMERICAN MICROSCOPICAL SOCIETY. PY

extend cephalad as in Deemyctylus, but are related more as in
Desmognathus.

The cloacas of Plethodon and Desmognathus were much alike.
Slightly caudad of the openings of the ureters there is a well
marked elevation on the dorsal wall covered by ciliated epithelium
which farther caudad breaks up into small papillae bearing the
mouths of pelvic gland tubules. Just cephalad of this are lateral
grouped gland tubules which appear in Plethodon, Desmognathus
and also Ambdlystoma to be of quite a different character from the
other pelvic gland tubules, so it appears as if in these forms the
pelvic gland were composed of two distinct kinds of tubules.
The homolog of the abdominal gland could be recognized, but in
both genera its tubules were short, being more developed in
Desmognathus than in Plethodon. In the latter its tubules opened
at the caudal end of the slit-like vent upon the epidermis cover-
ing the edges of the cloacal lips, but not upon papilla. In the
former the tubules open on the dorsal side of the cloaca near its
caudal limit, upon papillz, but clearly within the cloacal epithe-
lium. These variations are of importance in considering the
ectodermal or entodermal origin of the gland.

Of Amblystoma 1 need only mention in addition to what has
already been said, that all three glands may be recognized ; the
cloacal lips are simple and do not possess the fringe of papilla
present in Mecturus and Diemyctylus.

Formerly when attention was first called to these glands
as accessory to the genital organs, it was attempted to homologize
them with glands found in higher forms, and the pelvic was re-
garded as a prostate gland (hence the name applied by Duvernoy,
and others.) Wiedersheim stated unqualifiedly that there can be no
doubt but that these glands represent the prostate and gland of
Cowper of higher forms.* There is little doubt now but that,
as stated by Heidenhain and Zeller, these glands, opening upon
the cloaca discharge a secretion which constitutes the body of .
the spermatophore, forming thus a base to give support on its

*“Tass diese Bildung der Prostata und den Cowper’schen Driisen der
héheren Wirbelthiere entspricht, kann wohl keinem Zweifel unterliegen,”

272 PROCEEDINGS OF THE

summit to the zodsperms. To find, then, in all these genera that
the same glands are present at the cloaca and well developed, is
pretty strong circumstantial evidence that the same mode of
fertilization occurs in all, though the deposition of spermatophores
has been observed only in one and probably so ina second. I
confidently believe that in the proper season of the year the ter-
restrial Plethodon will be found to go to the water for the purpose
of mating just as does Salamandra of Europe, unless the sper-
matophores may be deposited with equal efficiency on land.

THE CLOACA OF THE FEMALE AND THE SPERMATHECA-

Less attention has been paid to the cloaca of the female
than the male. This is perhaps because in the European
Salamanders its structure is so much simpler. The following
brief historical review will indicate how fragementary is the atten-
tion that has been bestowed.

Rathke (’20) and Leydig ('53) both recognized the presence
of glands, doubtless the spermathecas, in the cloaca of the female
Salamandra, though neither seems to have investigated the
Triton. ‘The latter described them as formed of cylindrical tubules
of a caliber enlarging toward their end.

Siebold (’58) was the first to detect the presence of living z06-
sperms in these tubules, describing them in Salamandra atra
and maculosa and Triton igneus, cristatus and teniatus.

Wiedersheim (’75) merely alludes to the question of the pres-
ence of spermathecas in the genera Salamandrinaand Geotriton,
whose anatomy he made the subject of a monograph.*

Blanchard (’81) next mentions these in 77iton (species not
named) in connection with the pelvic gland of the male with which
he re garded them as homologous though atrophied and function-

*Of Salamandrina he says ‘‘ Die von Siebold entdeckten soblanahbeie
migen Receptacula seminis sind auch hier in zwei Gruppen vorhanden;
jedoch gelang es mir nicht, in ihnen Zoospermien zu entdecken;” of Geotri-
ton, ‘‘auch finde ich beim Weibchen keine Spur der Receptacula seminis,
wohl aber frei in der Cloakenhéhle liegende Zoospermien, wie bei Salaman-
drina.” He is therefore misquoted by Hoffman, ’78, who states that he
found in neither form a trace of spermathecas,

AMERICAN MICROSCOPICAL SOCIETY. 273

less as glands. He, however, did not agree with Siebold as to
their functioning as spermathecas.

Heidenhain (’90) who so thoroughly investigated the cloaca of
the male Z7yiton discusses the female merely in connection with
the discovery of the rudimentary tubuli which he regarded as
representing the abdominal gland of the male.

Jordan, ’91, speaks of finding in Dzemyetylus zoosperms “in
the ducts of two groups of gland-like structures situated in the
cloacal wall just below the entrance of the oviducts.”’ He
discusses the problem of how the zoosperms become ensconced in
their ‘‘ snug resting places,” and his views will be referred to sub-
sequently. .

Stieda, ’91, furnishes us with a minute account of the cloaca
and the spermathecas in the female 77zfox. His discussion is the
most detailed of all, though he does not mention the relation of
the spermathecas to fertilization, and he does not appear to have
been fully familiar with the literature.

Fisher, ’91, in a more general article upon the anatomy of
Geotriton fuscus finds the ‘“‘ receptaculum ”’ present as a single un-
paired organ which he considers homologous with the pelvic
gland of the male. No zoosperms were found in the gland ex-
amined by him. Wiedersheim’s statement that there is no
spermatheca in Geodriton is thus disproved.

Of the above writers, the accounts of Siebold and Stieda are
most circumstantial. The salamandride have been by far the
most carefully studied ; while in the representatives of two other
families (Diemyctylus, Salamandrina and Geotriton) little more
than the mere presence of the spermathecas has been reported.

Before proceeding to a discussion of the genera examined by
me it will be well to review briefly the structure and relations of
the cloacal glands and spermathecas of the female as they have
been ascertained to be in the family of the Salamandride. As
compared with the cloaca of the male that of the female is quite
simple ; there are no representatives of the glands so greatly de-
veloped in the male except the comparatively simple spermathe-
cas (if they are representatives) and in Zrzton the rudimentary

274 PROCEEDINGS OF THE

tubuli discovered by Heidenhain. The cloacal walls are equally
simple and the configuration not so varied ; well-marked dorsal
and ventral folds exist with smaller lateral ones. The sperma-
thecas are simple blind sacks lined with a single layer of cubical
cells which seemed to Stieda, to have no secretory function; a
circular layer of plain muscle cells encloses each tubule which
is in form flask-shaped, 7. ¢., enlarged toward the blind end, with
a constricted neck. These tubules open in two lateral groups in
ths dorsal side of the cloaca immediately caudad of the openings
of the oviducts ; in 7riton the number of tubules in each group
varied from eight to fifteen; in Se/amandra they were more nu-
merous, being thirty or forty on each side. Large numbers of pig-
ment cells occurred in the connective tissue surrounding the
tubules. i

The only other gland found at the cloaca of the female is the
rudimentary abdominal gland of Heidenhain, found in 77zfon.
Whether or not it is also present in Sa/amandra is unknown. As
I shall have occasion to refer to a similarly situated gland in Deze-
myctylus and certain other urodeles, it seems worth while to speak
of this in some detail. _Heidenhain describes it at some
length. There are found in the entire circumference of the cloacal
opening, upon the surface of the lips, papillze of the integument.
Microscopic examination showed that from the summits of these
papilla rudimentary gland tubules proceeded which have the
same course as the abdominal gland tubules of the male and
therefore must only be their homologs. From their origin they
assume a strongly dorso-cephalic direction, traverse the cloacal
lips, and, where they attain a considerable length, lie upon the
ventral surface of the striated muscle which is situated on each
side ventrad of the vertebre. The length of the tubules varied
considerably in the same individual. The lining cells were in-
conspicuous and presented no appreciable structure. The tubules
were also very variable in number; as many as twelve paired and
five unpaired tubules were found in some individuals, while
in other examples there was no trace of them whatever. In re-
gard to their homology with the abdominal gland tubules of the

AMERICAN MICROSCOPICAL SOCIETY. 275

male, I think it will be evident from a comparative study that the
fact of their opening upon papille in the skin at the vent is in-
sufficient to base a homology upon. Indeed, it seems futile to
attempt to pronounce a homology between structures in male and
female which possess in the two sexes clearly different functions,
and are so variable that the gland may be well developed in
the female in one genus of a family and totally wanting in another,
even though they may occupy approximately the same position.
For similar reasons, it does not seem to the writer that the
spermathecas can be homologized with the pelvic gland of the
male as has been so confidently done by Blanchard and Fischer.
As has been said by Jordan, ’93, the question is probably a barren
one.

Diemyctylus. (Fig. 7). Of the genera examined by me this
is most nearly related to the European forms, and an approxima-
tion to the relations in those genera would be expected, and is
found. The cloaca of the female is much less prominent than in
the male; the vent is situated on rather a ridge-like elevation.
Its lips are simple and no papillae are present. The internal to-
pography of the cloaca is also much simpler than in the male ;
the walls are thrown into low folds, not resembling, however, the
ridges in the caudal portion of the cloaca of the male. In the
dorsal side is a ridge which appears’ at the caudal end of
the cloaca and steadily increases in height to within a short dis-
tance of the oviducal papilla where it stops quite suddenly. As
this ridge heightens, the cloacal cavity deepens dorsally, so that
at the cephalic termination of the ridge there exists a deep dorsal
depression in which soon appear the papillae which bear the
mouths of the oviducts. The ventral portion of the cloacal cavity
simultaneously extends itself laterally and farther cephalad be-
comes the intestine. The bladder opens upon the ventral side of
the cloaca, slightly cephalad of the oviducal papille.

The epithelium lining the cloaca was stratified in the caudal
portion and columnar in the cephalic portion where it is com-
posed of mucous cells. The stratified epithelium extends cephalad
farthest on the dorsal side, even up to the area in which the

276 PROCEEDINGS OF THE

spermathecas open. These, in their structure and distribution
seem to resemble closely those in the Salamandride. They are
flask-shaped, the neck is very constricted and the diameter of the
body of the cul-de-sac varies with the amount of distension caused
by the zodsperms contained. The shape of the lining cells was
also modified by the same cause ; in the empty tubules, however,
they were cubical. Each tubule is enclosed in a layer of plain
muscular fibers which encircle it. These tubules open upon the
lateral walls of the dorsal extension of the cavity which is divided
into two parts by the mesal elevation. There were about twenty-
five such tubules opening on each side. Cephalad of the ter-
mination of the mesal ridge, which is abrupt, the lateral groups meet
at the meson and there occur five or six tubules which cannot be
regarded as belonging to either group.

But two examples of this newt were examined. by me; one
taken March 10, the other in October. In both of them the
spermathecas contained zoosperms. An examination of a more
complete series of individuals taken at other seasons of the year
would have been made had it not been already done by another.
It should be done in an exhaustive study of the glands, for which,
however, Dremyctylus is not as suitable a genus as Ambdlystoma,
Jordan, 91, has said that he found zoésperms.in the cloaca of the
female ‘‘ in nearly all the specimens examined between the first of
May and the first of July,’ and in another place he says that
zodsperms usually occur in the spermathecas of specimens taken
in the autumn.

In addition to the spermathecas other tubules occur presum-
ably homologous with the rudimentary tubules described by
Heidenhain as representing the male abdominal gland. In Dve-
myctylus, however, they are clearly not rudimentary. The open-
ings of the tubules occur upon the epidermis of the skin at the
edge of the vent in its entire circumference. From their openings
the tubules assume a dorsal, and generally, also, a cephalic direc-
tion on each side of the cloacal cavity. The more cephalic and
also most mesal tubules do not extend dorsad, but directly cepha-
lad and thus lie upon the ventral side of the cloaca. The gland

AMERICAN MICROSCOPICAL SOCIETY. 277

mass which will be spoken of as the ‘“ ventral gland,” to avoid
homologies, lies upon the ventral and lateral sides of the cloaca.
A duct and a secreting portion of each tubule might be recog-
nized; the former was constricted and its cells -flattened; the
secreting cells were columnar with their nuclei situated in the
basalend. The cell itself appeared granular. This description
applied only to the condition existing in the Diemyctylus taken in
the spring; in the fall individual the cells were much smaller
and the entire appearance was as of a gland of less functional
activity, or one recovering from a state of exhaustion. Undoubt-
edly the latter was the case as what will be said subsequently of
the seasonal variation in Amdlystoma would indicate.

To compare Diemyctylus with the Salamandridz a close ap-
proximation to relations there is seen. In the latter only the
paired spermathecas would appear to exist. In the number of
tubules, Dzemyctylus is intermediate between the genus Salaman-
dra and the Tritons. Wiedersheim mentions both the spérma-
thecas and the ventral gland in Salamandrina.* Evidently that
genus approaches Dremyctylus closely.

Necturus.** (Figs. 1-6). As in Diemyctylus the cloaca is much
less prominent in the female Wectwrus than in the male, due to
the weaker development of the glands. There are no cloacal pa-
pillee and the lips of the vent are smooth. In the configuration
of the cloaca Necturus differs somewhat from Dzemyctylus. In
the cephalic portion the oviducal papilla, which are very large,
project caudad and ventrad from the dorsal wall; caudad there is
in the dorsal side a well-marked depression which extends almost
to the caudal limit of the cloacal cavity. The epithelium of the
cephalic portion of the cloaca and the dorsal depression is formed
of mucous cells. Upon the inside of the lips of the cloaca this

*«< Diese (Cloake) ist beim Weibchen von einem Kranz kleiner, schlauch-
férmiger Driisen umgeben, welche in den die Spalte begrenzenden Lippen
gelegen sind, und erst beim Auseinanderziehen der letzteren deutlich zum
Vorschein kommen.”

**Prof. Wilder has permitted me to examine some drawings of the cloacal
cavity and papille of Nectwrus that were made in his laboratory in 1874 by
Prof. W. S. Barnard.

278 PROCEEDINGS OF THE

merges into a stratified form of epithelium, which insensibly grades
into the epidermis in which upon the cloacal lips the character-
istic clavate cells are absent.

In the depression in the dorsal side of the cloaca there may be
detected by the naked eye some forty or more orifices ; these are
the mouths of convoluted tubules which constitute the bulk of a
gland mass of considerable size situated upon the dorsal side of
the cloaca. (Fig. 2). These tubules were of large caliber and
lined with tall columnar cells which presented the characteristic
appearance of charged mucous cells. The nucleus was situated
in the base of the cell the ectal portion of which was granular.
There was no constriction of the tubule or differentiation of the
epithelium to constitute a neck, but the columnar cells passed di-
rectly into the mucous epithelium of the cloaca, with whose cells
they appeared identical in structure.

In addition to these large tubules there open upon the epithe-
lium of the dorsal side of the cloaca in or at the edge of the de-
pression other tubules of an entirely different appearance, the sper-
mathecas. We find in them a repetition of the structure in Dze-
myctylus. They are flask-shaped, lined with cubical or low col-
‘ umnar cells and open upon the surface by a very constricted neck.
Over forty of these were counted; they occur scattered among
the other tubules over an area extending from just caudad of the
mouths of the oviducts to the caudal limit of the depression.
These almost invariably contained zoosperms. In addition to
them other tubules of an entirely similar appearance open by the
characteristic constricted neck into the free ends of the large con-
voluted tubules first mentioned. The cells lining them resembled
in form and appearance those lining the tubules which I have just
spoken of as the spermathecas, and were easily distinguishable
from the taller granular cells of the convoluted tubules. Zoo-
sperms were occasionally, though only occasionally, to be observed
in these appendages to the larger tubules and they must likewise
be considered spermathecas.

Six Mecturt were examined and zodsperms were abundantly
found in all in the spermathecas which opened on the cloacal epi-

AMERICAN MICROSCOPICAL SOCIETY. 279

thelium and rarely in those which opened into the large tubules.
All were taken in the late fall or winter. It is unfortunate that
it was impossible to examine any in which it was evident mating
had occurred and the eggs were yet in the ovary. As it is, be-
cause of lack of knowledge of the habits of the animal, it is im-
possible to determine whether the zoosperms in the spermathecas
of the forms taken in the winter were acquired ina fall mating,
without ovulation, as occurs in Dzemyctylus, or were merely such
as had been left over after ovulation in the spring or fall, if one
occurs at that season of the year. There might be a marked
difference in the state of functional activity of the two sets of
spermathecas, and those which opened into the tubules might be
in reality the more functional.

The long convoluted tubules first described resembled no other
cloacal gland. Indeed the impression is strong that they may
arise as tubular proliferations of the mucous cells of the cloacal
epithelium surrounding the mouths of some of the spermathecas,
which they thus carry down with them in their development. It
is possible that an examination of young forms would throw light
on their origin and significance.

The ventral gland (Fig. 1) is present and well developed in
Necturus. Its mass lies upon the ventral side of the cloaca ceph-
alad of the vent, and its tubules open upon the stratified epithe-
lium within the edge of the cloacal opening surrounding its
cephalic portion. The tubules are paired and are some sixty or
more in number on each side. A duct and secreting portion are
well marked, the former is constricted and lined with. flattened
cells, the latter is of a larger diameter and formed of very tall
‘columnar cells which are crowded with globules and large
granules in the specimens examined. Doubtless they become
more or less exhausted at the time of ovulation and gradually re-
gain the resting state during the year as seems to be the case in
Amblystoma.

Amblystoma. (Figs. 10-15). A more complete series of
specimens of Amdlystoma punctatum were examined and a better
idea could thus be obtained of the extent and significance of sea-

280 PROCEEDINGS OF THE

sonal changes in the cloacal glands, and more satisfactory exami-
nation of the spermathecas was also possible. Those examined
comprised seven individuals: (1) two Amblystomas taken in the
spring in which the eggs were still in the ovary and no mating
had yet taken place, as ascertained by examination which showed
entire absence of zodsperms ; (2) one in which the eggs were yet
in the ovary, but mating had occurred—the spermathecas and de-
pression were full of zoosperms ; (3) one in which ovulation had
just taken place in the laboratory ; further, there were examined
two taken in july and one taken in the winter (January).

In this form the relations at the cloaca differ markedly from
those in either of the two genera just described. The ventral
gland is present and occupies much the same position as in Wec-
‘urus; the spermathecas open upon the dorsal epithelium of the
cloaca and in addition there is another group of tubules situated
in the dorsal wall of the cloaca opening upon the epithelium far-
ther caudad. The relations of the three groups is well shown by
the sagittal section, Fig. 10, from which also a general idea may
be obtained of the shape of the cloacal cavity. The oviducts open
as usual upon papillz situated in a depression in the dorsal side
of the cloaca; farther ventrad is another depression, quite nar-
row, into which the greater number of the spermathecas open
and around which they are grouped; a few open upon the
epithelium just caudad of it. The epithelium lining this depres-
sion, and also the cloacal cavity for a short distance cephalad and
caudad of it on both the dorsal and ventral sides, consists of a
single layer of columnar cells. Ciliated cells were found upon
the ventral wall opposite the depression and cephalad of it on the
dorsal side of the cloaca. In neither place was the tract exten-”
sive and the ciliated cells were often separated by non-ciliated
ones. No cilia appeared on cells lining the depression. The
arrangement of the spermatheca tubules was quite compact

‘about the depression from which they radiated on all sides with a
more or less dorsal trend. Between them strands of plain
muscle cells passed, forming a network and encircling the de-
pression more or less completely.

_ AMERICAN MICROSCOPICAL SOCIETY. 281

The ventral gland tubules some distance within the vent opened
upon the cloacal epithelium which was there composed of two or
three layers of cells ; farther ectad it thickens to grade insensibly
into the epidermis upon the lips of the cloaca. The tubules open
upon the ventral and also to a slight extent upon the lateral sides
of the cloaca, and radiate cephalad and also somewhat dorsad so
that the gland mass is situated on the ventral and to a slighter
extent the lateral sides of the cloaca. Though the mouths of the
tubules do not occur in lateral groups, as in Vecturus, the gland
mass itself is divided on the meson into two equal lateral portions.

The second group of gland tubules which I have designated as
the dorsal gland, opens on the dorsal and lateral sides of the
cloaca and the tubules extend dorsad and cephalad.

In structure these glands were of an entirely different appear-
ance. In both, the tubules were composed of the customary
duct and secreting portion. The cells of the ventral gland were
very tall columnar cells (Fig. 11). The cell body was crowded
with large granules which took a cytoplasmic stain readily. The
cells of the dorsal glands are finely granular and stain less in-
tensely, coloring a light pink where the ventral gland stains rose.
This is the appearance in the spring specimens taken before ovu-
lation when the glands are in a resting condition and _ fully
charged. In the specimen after ovulation the glands, though not
exhausted, were yet so nearly so that an idea could be obtained
of the grosser changes in secretion. Fig. 12 shows a partially
exhausted tubule; the tall columnar cells filled with granules
have shrunken, the lumen of the tubules is greatly enlarged and
filled with the mass of secretion. A more complete stage of ex-
haustion is shown in Fig. 13, where the cells have become almost
flattened. A comparison of the two summer forms in which the
Ovaries were spent shows the gland cells in an exceedingly re-
duced condition, in many tubules consisting of little besides
nucleus. Fig. 14 shows one of the more developed tubules in
which a few granules occur in the cells which are apparently just
recovering and beginning to elaborate the secretion which will
swell the cell so enormously. In fact, these glands, as do the

282 PROCEEDINGS OF THE

cells of the oviduct, seem to prepare themselves yearly for their
period of secretive activity at the time of ovulation. The secre-
tion found in the tubules and the cloacal chamber stains deeply
with eosin. Changes in the dorsal gland seem to correspond
closely with those in the ventral. I believe that these glands
secrete the substance which forms the gelatinous mass which
binds together the eggs when laid in masses.

So exactly do the spermathecas resemble in form and structure
the gland tubules that we naturally turn to see if seasonal varia-
tions may also be detected_in them. In the spring individuals in
which the eggs are yet in the ovary and mating has not yet taken
place, the conditions are most favorable for determining whether
or not they present an appearance of cells in a resting condition,
which might be expected if they elaborate any secretion attractive
to the zodsperms. However, the cells show no traces of a stored
secretion, and compared with the cells in the spermathecas of the
specimen in which the eggs had been laid, but which still contained
some zodsperms, show no apparent differences. The spermathe-
cas contained more or less granular or stringy matter as of some
coagulated secretion, as in all the other forms in which the empty
tubules were observed—JVecturus, Spelerpes, Plethodon. The
cells resembled exhausted mucous cells, appearing finely reticu-
lar and staining lightly.

Spelerpes. (Figs. 16-17). Buta single individual of Spelerpes
bilineatus was examined, which was taken October 7. The struct-
ure of the spermatheca, however, was so unique and so suggestive
of the meaning of the single mesal spermatheca in this and the
two genera to be discussed subsequently, that a description is
siven, although no zodsperms were contained in the spermatheca
of this one individual.

The structure of the cloaca in its caudal portion presents no
unusual features; proceeding cephalad from the caudal end of
the cloacal opening, the cavity deepens gradually and its walls
are thrown into small longitudinal folds. Slightly caudad of the
cephali¢ end of the vent, a narrow dorsal depression makes
its appearance which deepens gradually until it meets the mouth

AMERICAN MICROSCOPICAL SOCIETY. 283

of the common tube of the spermatheca as it may be termed,
which extends cephalad (Fig. 17). As this depression increases,
more lateral ones appear cutting out or bordering lips for the
mesal one, thus producing the appearance of a dorsal elevation cut
in two. Cephalad of the opening of the spermatheca these folds
disappear and a mesal elevation appears which is soon divided by
a deep and narrow depression.

The spermatheca first appears in a series of transections slightly
cephalad of the caudal end of the cloaca, upon its dorsal side and
at first only loosely joined to it by connective tissue. In struct-
ure it consists of a central common tube into which near its caudal
end open twelve to fourteen tubules, flask-shaped, with the body
lined with columnar cells and opening by means of a constricted
neck—a repetition of the structure of the tubules in the genera
before described. (Fig.16). The cells lining the common tube
and the tubules opening into it are columnar and identical in ap-,
pearance in the two places ; they stain but lightly, resembling the
cells of the spermathecas of Ammélystoma. The epithelium of the
spermatheca extends cephalad beyond its mouth some little dis-
tance, covering the mesal elevation before it becomes divided and
persisting on each side for a short distance after the mesal de-
pression appears.

The cloacal epithelium in its caudal portion is a stratified one,
and two or three cells thick, which extends cephalad almost to the
opening of the spermatheca. Mucous cells first appear in the
lateral depressions alluded to above and spread over the entire
surface of the cloaca except where the epithelium of the sperma-
theca extends cephalad, from which, because of the stain taken,
they are readily distinguished.

The common tube of the spermatheca was of uniform structure
throughout, somewhat larger at its blind end. It was enclosed
in a thick layer of plain muscle cells which also surrounded each
tubule opening into it. The entire organ is enclosed by a
dense layer of pigment cells which is especially concentrated about
the tubules. (Fig. 16). When the cloaca is dissected off, be-
cause of its pigmented state the spermatheca appears as a black

PROCEEDINGS OF THE

NO
oo
nS

sack. It is impossible to resist the impression that the organ in
Spelerpes, and therefore in the two following genera, is not the
equivalent ofa single one of the many tubules of Dzemyctylus which
has become enlarged and branched, but presents an exaggeration
of the condition in Ambystoma, and represents a tubular depres-
sion of the cloaca, into the end -of which the clustered tubules
open,—which in such forms as Dzemyctylus open upon the cloacal
epithelium directly,— and thus seems to constitute a connecting link
between Diemyctyvlus, Necturus and Amblystoma on the one hand,
and Prethodon and Desmognathus on the other. The tubules as
well as the common tube contained more or less matter, granu-
lar or globular which might seem to indicate, as in Amblystoma,
a secretive activity of the cells.

Opening upon the dorsal side of the cloaca, caudad of the mouth
of the spermatheca, are some twelve tubules which were in a
very undeveloped condition and were either rudimentary or in a
state of inactivity or exhaustion. In some the cells consisted of
little more than the nucleus, in others they were cubical and in
all a lumen was present. Likewise, cephalad of the opening of
the spermatheca, were six or seven short tubules of a rudimentary
appearance. Whether these represented the dorsal cloacal gland
or were spermatheca tubules which had become rudimentary from
disuse—if the spermatheca were not in the first place modified
tubules of the dorsal gland—is, of course, idle speculation. In
the absence of a knowledge of the habits of the salamander or
specimens taken at different times of the year, any conclusion is
impossible.

The ventral gland is well developed ; its tubules open upon
the cloacal epithelium of each side some distance within the vent.
They are all paired and are some twenty in number on each side.
The more caudal tubules extend dorsad, those more cephalad
lie upon the ventral side of the cloaca and there form a consider-
able gland mass. The location of the gland, therefore, is as in
Diemyctylus. The cells of the tubules are columnar and resemble
those in that genus.

Llethodon erythronotus. As in Spelerpes, the spermatheca in

AMERICAN MICROSCOPICAL SOCIETY. 285

this salamander is mesal though it differs much in structure. A
feature peculiar to Plethodon among the forms examined is the
presence of a large papilla-like elevation which projects caudad
and ventrad from the dorsal wall of the cloaca-in its caudal
portion. Strands from the circular muscular coat of the cloaca
project up into it, so that it is doubtless more or less contractile.
Dorsad of its base the cloacal cavity extends cephalad a short dis-
tance as a cephalic recess.

Cephalad of this papilla there appears a dorsal elevation on
which opens the spermatheca. As in Sfelerpes this organ is
sack-like in shape and highly pigmented and muscular. /Vetho-
don differs, however, in that the common tube has a constricted
neck, and instead of the twelve tubules opening into its blind end
there are but four diverticula which do not show the characteris-
tic structure possessed by the tubules in the previous forms,
namely, a body and neck, though they are somewhat constricted
at the opening into the common tube. The common tube and
to a less degree each of the diverticula have an encircling muscu-
lar tunic. The circular muscular coat of the cloaca proper is
well developed and sends bundles of cells up into the elevation
on which the spermatheca opens and also into the spermatheca.

Cephalad of the spermatheca the dorsal ridge becomes thinner
and finally disappears ; the cloacal cavity extends itself laterally
and dorsally as a depression into which open the oviducts from
short papilla, and the ureters. The epithelium of the cloaca in
the caudal position is stratified, as before, and this extends farthest
cephalad upon the dorsal elevation, to slightly caudad of the
spermatheca. It is succeeded by a mucous epithelium, which
lines the cephalic portion of the cloaca, first appearing caudad on
its ventral side.

Plethodon glutinosus. (Figs. 18-19). This species when
compared with evythronotus shows close resemblances with some
marked variations. The curious tongue-like papilla in the caudal
portion is well developed and the projection of the cavity beneath
its base is considerable. The spermatheca is much as in PVetho-
don erythronotus ; the diverticula are four in number and the com-

286 PROCEEDINGS OF THE

mon tube into which they empty opens upon the cloaca by a very
constricted neck at the summit of the dorsal ridge (Fig. 19) as in
erythronotus. A difference, however, consists in the fact that the
common tube is lined with a stratified epithelium of flattened cells.
The muscularity and pigmentation of the organ are also greater.
Pigment is not confined to this region, but occurs in the tongue-
like papillae, and to a slight extent in the circular muscular coat
of the cloaca. A most peculiar difference between the two species
lies in the presence of cilia in the cloaca of g/utenosus, and their
entire absence so far as I can determine in erythronotus, the tissue
being fixed in the same way in each. In Asmédlystoma a few
scattered ciliated cells were found, but in P. g/utinosus the area
was extensive. As in evythronotus the caudal portion of the
cloaca is lined with a stratified epithelium two or three cells deep,
which extends cephalad upon the dorsal elevation to a point
slightly cephalad of the mouth of the spermatheca. In the de-
pression on each side of this elevation ciliated cells first appear; they
spread laterad and ventrad until the area encircles the entire cloaca.

Immediately cephalad of the opening of the spermatheca and
cutting into the base of the elevation (Fig. 19), the cloacal cavity
extends, laterad and dorsad as in erythronotus, constituting the
depression into which open the ureters and oviducts, the latter
upon very small papillz, if any. These depressions are clothed
with large mucous cells, which spread laterad and ventrad to
clothe the entire circumference of the cavity.

Of erythronotus were examined specimens taken in the sum-
mer (2), fall (1) and spring (4), and only in three’ in the spring,
taken April 15th and May 2d, were zodsperms found. Two
individuals of P. ¢/utinosus were examined, and in neither were
zoosperms contained in the spermatheca. There is no doubt but
that were a larger number examined, zo6sperms would be found
as in erythronotus.

Geotriton, the only European genus of this family, was ex-
amined by Fischer, and a single mesal spermatheca, as in Pletho-
don, was found, though in the specimens examined by him no
zodosperms were contained.

AMERICAN MICROSCOPICAL SOCIETY. 287

In those individuals in which no zoosperms were contained,
the spermatheca held, as in Sfe/erpes, granular and reticulated
matter, as if of coagulated secretion, and in some instances the
free ends of the cells appeared flagellate or threaded.

Desmognathus (Figs. 8 and g). The relations are much as in
Plethodon. An elevation appears upon the dorsal side of the
cloaca, and upon this, as in /Vlethodon, the spermatheca opens
(Fig. 9). This is very well developed, consisting of a common
tube with five or six large diverticula, and opens by constricted
neck with flattened cells. The muscular sheath of the common
tube and its diverticula was strong. Both the diverticula and
the common tube were lined with low columnar cells, presenting
no marked structural features. The pigment surrounding the
organ was very abundant, and rendered it easily recognizable on
dissecting out the cloaca for sectioning.

The general configuration of the cloacal chamber was much
as in Plethodon, save that no tongue-like papilla existed. The
elevation in the dorsal wall, already referred to so often, was
present. The stratified epithelium of the caudal portion of the
cloaca extends cephalad upon it up to the mouth of the sperm-
atheca. Mucous cells first appear in the depression on each side
of the elevation, whence they spread ventrad to completely clothe
the cloacal wall. Cephalad of the spermatheca the elevation
disappears and the cloacal cavity assumes a tubular form. The
oviducts and ureters open relatively farther cephalad than in
Plethodon, the former upon the summits of low papillz.

Specimens taken in the summer, fall, winter and spring were
examined, and in all the spermathecas were well filled. Though
a confirmation from the meager facts can not be had, it seems
probable that such a difference in the presence of zodsperms in
Plethodon and Diemyctylus has an accompanying difference in habits
of mating which would explain it. Though doubtless merely
indicating the way in which the zodsperms entered the sperma-
theca, the fact that in the individuals examined they almost in- |
variably lay with their heads toward the cells of the spermatheca,
might indicate an attractive principle secreted by the cells.

288 PROCEEDINGS OF THE

Neither in Plethodon or Desmognathus were ventral or dorsal
cloacal glands present.

CONCLUDING REMARKS.

From the foregoing it is seen that in the six genera examined,
including urodeles of widely different habits, functional sperma-
thecas are to be found, though with a great variation in structure.
Indeed, in Diemyctylus, Amblystoma and Necturus they may
more strictly be spoken of as individual tubules which function
as spermathecas, possibly gland tubules, as they are termed by
Gegenbaur and Claus, so exactly do they compare with the
tubules which constitute the other cloacal glands ; and it is first
in Amblystoma that there appears in the arrangement of the
tubules around a depression the suggestion of the unity of struct-
ure found in the remaining three genera, Spelerpes, Plethodon
and Desmogunathus, where there is a single mesal spermatheca.

All discussion heretofore has been purely anatomical, and only
incidentally has the function and physiological action been alluded
to. The question may be fairly put, how far are the sperma-
thecas of real importance to the salamander, and have they in
every case a role to play in the process of fertilization, or are the
zoosperms contained only used in certain cases? Our ignorance
of the habits of all but one of the genera forbids an estimate,
but as far as more isolated observations afford an answer, it is
that their functional use is complete; in other words, that the
zoosperms taken up by the female are stored away in these
tubules and are expelled from them when needed for the fertili-
zation of the egg. In some forms their functional use is great.
Both Czermak and Siebold have shown that the female of the
viviparous Salamandra atra, with but one mating in the spring, ©
yet gives birth to its young several times during the year, and
Zeller considers it undoubted that the same is true for Sada-
mandra maculosa.* For these viviparous forms, therefore, the

* “ Ja, es ist dafiir der thatsichliche Beweis durch die schon zum éftern
gemachte Beobachtung erbracht, dass isolirt gehaltene Weibchen [Sala-
mandra maculosa| nach ein- und sogar nach zweijaihriger Gefangenschaft
eine kleine Anzahl von Larven geboren haben.” Zeller, 90, p. 595,

AMERICAN MICROSCOPICAL SOCIETY. 289

spermatheca is of the utmost importance. In Triton (alpestris
and ¢eniatus) Zeller regards it as exceedingly probable that in
certain cases fertile eggs may be laid in the spring without a
previous mating that spring, though this is unusual. In this
form, according to him, the zoésperms received from a mating
will fertilize one hundred or more eggs, which are laid within
eight to fourteen days, when another mating occurs. With the
axolotl the proceedure is more rapid; ovulation generally begins
the following night and within thirty-six or forty hours 300 to
600 eggs have been deposited ; occasionally the number reaches
800 to 1,000 (Zeller): in the aquarium two ovulations may
occur in the same year, but the second is preceded by a fresh
mating. The observations of Zeller, Gage and Jordan, especially
the last, make us more familiar with the phenomena of fertiliza-
tion and ovulation in Dzemycty/us than in any other form. Jordan
says: ‘It is probable that for a single individual the egg-laying
season lasts for at least seven or eight weeks. The longest time
over which I have actually observed the laying of a single in-
dividual to extend is four weeks, but in this case the ovaries still
retained large pigmented eggs, and under perfectly normal con-
ditions egg-laying would undoubtedly have continued for some
time longer.’”’ The largest number of eggs laid under observa-
tion was ninety-six, and ovulation lasted nineteen days after separa-
tion from the male. An autumnal mating in Diemyctylus was first
_ observed to occur by Gage, and confirmation of this has been
given by Jordan, ’92. It is possible, therefore, in certain cases,
that the laying of fertile eggs in the spring may proceed without
a previous spring mating.

But a slight clue of the time and manner of ovulation can be
gained from a study of the spermathecas, since it can never be de-
termined whether the zodsperms present are an accession of a
recent mating or are those remaining after a sufficient number has
been used in the fertilization of the ova. It would appear prob-
able that in Asmdlystoma there is but one mating in the spring
followed by ovulation. Of the habits of Mecturus at present no

definite conclusion can be drawn. The fact that zobsperms were
20

290 PROCEEDINGS OF THE

universally found in Desmognuathus in the spring, summer, fall
and winter, and found only in the spring in Plethodor suggests
some difference in habit, though too few individuals were ex-
amined to afford more than a suggestion. Sherwood says of
Desmognathus: ‘1 think there are two broods annually, as I
have found eggs from July to October and have seen very small
larve as late as November 30.” He found eggs of Plethodon and
Spelerpes October 25th. Three specimens of Plethodon ery-
thronotus, taken by me between the 15th of April and the 2nd of
May, all had the spermathecas well filled, and there is therefore
a suggestion of the possibility of two broods in this form. Mere
speculation, however, is idle and direct observation of the habits
of these forms must be awaited.

Jordan, from his careful study of Dzemyctylus, concludes that fer-
tilization in that genus takes place in the cloaca as the ovum passes
through to be extruded ; his words are: ‘ The fertilization of the
egg takes place just before the egg is extruded. The sperma-
tozoa, which have long been waiting in the tubes of the recepta-
culum seminis, are either attracted from their resting places by
the passing egg or forced out by contraction of the surrounding
muscles. J] have made repeated and careful search for sperma-
tozoa in the oviducts, but have never succeeded in finding one.
Neither have I ever found in sections any indication that sperma-
tozoa enter oviduct eggs, although eggs often lie for some time
in the mouth of the oviducts. Fertilization, then, would seem to
take place only after the egg has left the oviduct and passed into
the cloaca.”’ I would say that I also have never seen zo6sperms
in the oviducts of any species sectioned by me, in almost all of
which the lower portion of the oviducts was examined. How-
ever, in the viviparous Sa/amandra artra and maculosa fertiliza-
tion of the egg must occur in the oviduct since development takes
place there. Spallanzani (as quoted by Zeller) stated that eggs
removed by him from the oviducts of the female 77zton proved
fertile and developed. Robin, also, said that examination of
the female 77z/on at the time of ovulation discovered zoésperms in
the cloaca and “three or four millimeters within the oviducts,”

AMERICAN MICROSCOPICAL SOCIETY. 291

There would seem, then, to be a variation in the different forms.

The walls of the cloaca are largely formed of plain muscular
tissue which surrounds the vent and strands of muscle cells inter-
digitate with the gland and spermatheca tubules, and it is probable
that in fertilization the zoosperms are forcibly expelled by
muscular contraction.* In the forms in which there is a mesal
spermatheca, the whole organ and its common tube are encircled
by a thick layer of muscle cells, and that such is the case is
especially evident here.

‘The question as to how the spermatozaa find their way into
these snug resting-places is one of considerable interest. Why
should they enter these small ducts and there lie dormant, in
preference to passing ex masse up the oviducts, or to entering
the alimentary canal, or even to issuing from the mouth of the
cloaca? It appears to be probable that the explanation lies in
what Pfeffer has called ‘ positive chemotaxis.’ Pfeffer found, as
is, well known, that certain chemical substances, as malic acid,
attract spermatozoa (positive chemotaxis), and that others, as
chloroform, repel them (negative chemotaxis). For example,
the mucilage in the central canal of the archegonia of Pteris con-
tains a trace of malic acid, and Pfeffer has shown that this amount
is sufficient to attract spermatozoa to the mouth ofthe canal. A
similar explanation has been given by some _ bacteriologists to
account for the gathering of leucocytes at inflammatory foci. It
is supposed that the leucocytes have been drawn thither in virtue
of their chemotactic properties which were brought into play by
the metabolic bacterial poisons, or, as now seems more likely, by
the freed albuminoid constituents of the bacterial cell.

“It seems highly probable that the pelvic gland of the female

*«The explanation of the dropping of non- fertilized eggs seems to be that in
females with full oviducts, an egg is occasionally pressed into the cloaca by
the mere elasticity of the oviduct walls and without the special cognizance
of the newt. This egg then passes out like so much excreta without the -
performance of a voluntary act of oviposition. The fact that these eggs
are, for the most part, unfertilized indicates an expulsion of spermatozoa

from the receptacle during the performance of egg-laying.” Jordan, 95,
p- 309,

292 PROCEEDINGS OF THE

newt may secrete a substance—proteid or otherwise—with a
positively chemotactic effect, and thus draw the spermatozoa into
its ducts. At all events, such a supposition may serve for a pro-
visional hypothesis.”’ (Jordan, ’g1, p. 266.)

The question set forth in the above quotation is indeed
difficult to answer. At first sight one would seem compelled to
assume such attractive secretion by the cells of the spermatheca.
Indeed, examination of the tubules in Azzdlystoma or other forms
when they are free from zoosperms, shows clearly that some
secretion is given off by the cells of the tubules, though of its
nature and significance nothing can be said. It does not seem
at all impossible, however, that the entrance of the zoosperms
may be due solely to their own activity assisted by muscular con-
tractions of the cloaca and spermatheca.

The great abundance of pigment cells surrounding the sperma-
theca tubules in all forms except Necturus and Amblystoma has
been noted in the discussion of each form. Though, therefore,
it is not a constant accompaniment of the spermatheca, the great
abundance of it surrounding the organ in the forms in which it
is most highly developed cannot but attract attention and
arouse speculation. When the zoosperm enters the pigmented
ovum it has been observed to exert an attractive influence upon
the pigment particles, which cluster to meet its entrance and fol-
low it as a pigment trail as it penetrates deeper (see Jordan, ’93,
p. 317). Were pigment cells constantly found surrounding the
tubules, it might be suggested that the attraction exerted by the
zoosperms was potent here, hence the pigmentation ; or that the
attraction between pigment and the zoosperms was mutual and
the pigment enticed them into their resting places. Pigment
occurs surrounding other gland tubules, and in other regions of
the cloaca, though less plentifully, so that its presence is doubt-
less due to something other than an attraction exerted by the
zoosperms, though this might yet account for its greater concen-
tration at that point.

Blanchard regarded the tubules which have been seen to
function as reservoirs for the zoosperms as homologous with the

AMERICAN MICROSCOPICAL SOCIETY. 293

pelvic gland of the male, as likewise does Fischer, and Jordan,
though very guardedly. As before stated, I do not believe a
homology can safely be declared between any of the cloacal
glands of the male and female, which are so evidently dependent
on their function for their existence, which is again clearly differ-
ent in the two sexes. In regard to the spermathecas, difficulties
occur in Amblystoma and Necturus in the presence of other
tubules in the dorsal wall of the cloaca, which are again so differ-
ent in the two genera. With Jordan, I regard the question as a -
barren one.

Besides the spermatheca tubules, there occurs in all, except
Desmognathus and Plethodon, a group of gland tubules upon the
ventral side of the cloaca, which has been termed the ventral
cloacal gland. In Amdlystoma, Spelerpes and Necturus there is
a second group upon the dorsal side of the cloaca, which in
Amblystoma 1 have termed the dorsal gland. The tubules in
Necturus do not seem comparable or homologous. These glands
are clearly accessory genital glands, and their secretion is with-
out doubt of use in ovulation, though the function can not be
stated more definitely as yet; possibly it serves as a cement to
cause the eggs to adhere to that on which they are laid, or to
each other. The ventral gland is best developed in Mecturus
and Amblystoma, least so in Diemyctylus and Spelerpes. Tt may
_ be pointed out that the two forms in which the ventral gland is
wanting are such as lay their eggs on land.

METHODS.

Since it is customary to give an account of the methods em-
ployed, the following is added, though no attempt was made to
develop them. The examination of the cloaca was accomplished
by means of serial sections made transverse to the long axis of
the body, to which were added in Plethodon, Desmognathus and
Amblystoma, series cut sagittally. The cloaca was dissected
off and placed for twenty-four hours in Fish’s mixture. |
(Formula: 50 per cent. alcohol, 1000 cc.; mercuric chlorid, 5
grams ; picric acid, 1 gram; glacial acetic acid, 10 cc.) It was

294 PROCEEDINGS OF THE

then washed in 50 per cent. alcohol one day, and passed through
successively 70, 82 and 95 per cent. alcohols, ether-alcohol
(equal parts), remaining one day in each. It was placed in 1%
or 2 per cent. collodion for two days, and 6 per cent. collodion
for three days, and imbedded. The collodion was hardened in
chloroform and cleared in Fish’s castor-thyme-oil mixture, in
which the sections were cut. “They were arranged in serial order
on the knife, from which they were removed by tissue paper,
placed upon the slide; all oil possible was absorbed with tissue
paper and the sections secured by melting the collodion with a
few drops of ether-alcohol. A few minutes (5 to 15) in 95 per
cent. alcohol sufficed to remove all the oil when they were
treated as usual. Haematoxylin (Gage’s) with eosin, erythrosin
or picric alcohol, as a counter-stain, were employed. Vasale’s
clear (Xylene three parts, carbolic acid one part), was used.
This was supplemented by teasing fresh spermathecas upon the
slide to detect the living zoosperms.

SUMMARY.

1. In the genera Necturus, Amblystoma, Diemyctylus, Pletho-
don and Desmognathus spermathecas are found in the dorsal wall
of the cloaca of the female, containing zodsperms. Internal
fertilization is therefore proven for these forms.

A spermatheca occurs in Sfelerpes ; in the single specimen ex-
amined (taken in the fall) no zoosperms were contained.

In Necturus, Diemyctylus and Amblystoma there are several
tubules or spermathecas opening upon the cloacal epithelium,
which serve as reservoirs for the semen.

In Diemyctylus, Plethodon and Spelerpes there is a single mesal
spermatheca.

The condition in Sfelerpes would seem to indicate that the
organ in these latter genera equals the group of tubules found in
the first genera plus an exaggerated and modified depression of
the cloacal epithelium, such as occurs in Asb/ystoma,

2. No gland-like structures in addition to the spermatheca
occur in the female of /Vethodon and Desmognathus.

AMERICAN MICROSCOPICAL SOCIETY. 295

3. In all the remaining genera a ventral cloacal gland is
present.

4. In Amblystoma, Spelerpes and Necturus, in addition to the
spermatheca tubules other tubules occur on the dorsal side of the
cloaca.

5. The secretion of the cloacal glands is employed at the time
of ovulation.

6. The three glands of the male recognized in the 7riton, the
cloacal, abdominal and pelvic, occur and are well developed in
the five genera examined. This suggests that by all of these
spermatophores are deposited.

7. A résumé of the literature and the foregoing facts points to
a uniform mode of mating and fertilization in all urodeles.

8. Dorsal and ventral ciliated tracts occur in the male of all the
genera examined (five). Cilia in the cloaca of the female were
detected only in Amblystoma and Plethodon glutinosus, where
the tract was not as extensive as in the male.

296

PROCEEDINGS OF THE

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Balfour. F. M.—A treatise on comparative embryology. Vol IL.,
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Bedriaga, J. von—Beitraige zur Kenntniss des Rippenmolches. Ver-
handl. der Kais. Gesellsch. der Wiss. zu Moskau, p. 179. 1879.
Bedriaga, J. von—Ueber die Begattung bei einigen geschwianzten
Amphibien. Zoologischer Anzeiger, Vol. V., pp. 265-268, & 357-859.
Blanchard, R.—Sur les glandes cloacale et pelvienne et sur la Papille
clocale des Batraciens et des Urodéles. Zool. Anz., Vol. 1V., pp. 9 & 34.
Chauvin, Marie von—Die Art der Fortpflanzung des Proteus angui-
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Clark, S. F —The development of Amblystoma punctatum. Part
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Claus. Grundziige der Zoologie. Vol. Il. Translated by Sedge-
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Cope, E. D.—The Batrachia of North America. Bull. No. 34 U. S.
Jatl Museum. 1889. Washington, D. C.

Gzermak, J. J.—Beitrige zur Anatomie und Physiologie des schwar-
zen Salamanders. Med. Jahrb. des Oesterr. Staates, Vol. XLV., p. 8.
Eycleshymer, A. C.—The early development of Amblystoma punc-
tatum, with some observaticns on some other vertebrata. Jowr.
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Fick, R.—Ueber die Reifung und Befruchtung des Axolotleies. Zeit-
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Fischer, G.—Beitriige zur Kenntniss des Geotriton fuscus. Verhandl.
physik.-med. Gesellsch., Wurzburg, Vol. XXV. 1891.

Fish, P A.—A new clearer for collodionized objects. Proc. Amer.
Micr. Soc., Vol. XV., pp. 1-4.

Gage, S. H.—Life-history of the vermillion-spotted newt (Diemycty-
lus viridescens, Raf.). Am. Nat., Dec. 1891, pp. 1084-1110.

Gasco, F.—Gli amori del Tritone alpestre. Genova, 1880.

Gasco, F.—Les amours des Axolotls. Zool. Anz. Vol. IV., pp. 318,
334. Bull. Soc. Zool., France, 1881, pp. 151-162.

Gegenbaur, Carl —Elements of Comparative Anatomy. Transl. by
F. J. Bell, London, 1878.

Heidenbain, M.—Beitrige zur Kentniss der Topographie und Histol-
ogie der Kloaka und ibrer driisigen Adnexa bei den einheimischen
Tritonen. Arch f. mikr. Anat., Vol. XXXV., pp. 178-274.
Hoffmann, C. K —Bronn’s Klassen und Ordnungen des Thierreiches.
Vol. VI., part 2, Amphibien.

Jordan, E. O.—The Spermatophores of Diemyctylus. Journal of
Morphology, Vol V., pp. 263-270.

Jordan. E. O.—The habits and development of the newt (Diemycly-
lus viridescens). Jour. Morph., Vol. VIII, 1893, pp. 269-352.
Lataste, F.—Sur l'accouplement chez les batraciens urodéles. Revue
internat. des sci., No. 42. Paris. 1878.

153.

°20.

AMERICAN MICROSCOPICAL SOCIETY. 20%

Leydig, F.—Anatomish-histologische Untersuchungen iiber Fische
und Reptilien. 1853.

Rathke. — Beitrige zur Geschichte der Thierwelt. Erste Abthei-
lung. Neueste Schriften der naturf. Gesellsch. in Danzig, Vol. L.,
Danzig, 1520.

Robin, Ch.—Observations sur la fécondation des urodéles. Jour. de
Vanat. et de la Physiol. norm. et path. de Vhomme et des animaux.
Vol. X, pp. 376-3890. Also, Annals & Mag. Nat. Hist., Vol. XIV.,
p. 96.

Rusconi, D. M.—Amours des Salamandres aquatiques, 4to. Milan.
1821.

Rusconi, D. M.—Histoire naturelle development et Metamorphose
de la Salamandre terrostere. Pavia, 1854.

Sherwood, W. L.—The salamanders found in the vicinity of New
York City, with notes upon extra-limital or allied species. Proc.
Linnean Soc. of New York, No. 7, 1895, pp. 21-87.

Siebold, C, Th. von—Ueber das Receptaculum seminis der weiblichen
Urodelen. Zeitsch. f. wiss. Zool., Vol. 1X, p. 468, 1858.

1785. Spallanzani, L.—Experiences pour servir a ‘histoire de la généra-

tion des animaux et des plantes; avec une ebauche de Uhistoire des
étres organisés avant leur fécondation. Par. J. Senebier, Geneve,
1785.

Stieda, Alfred—Ueber die Kloaka und das Receptaculum seminis der
weiblichen Tritonen. IJnaug. Dissert. Kénigsberg, July, 1891.
Wiedersheim, R.—Salamandrina perspicillata und Geotriton fuscus.
Genua, 1875.

Zeller, Ernest—Ueber die Befruchtung bei den Urodelen. Zeitscl.
tT. wiss. Zool., Vol. XLIX., pp. 5838-602. 1890.

Zeller, Ernest—Berichtigung, betreffend der Samenaufnahme der
weiblichen Tritonen. Zeitsch. f. wiss. Zool., Vol. LI, pp. 787-741.

298 PROCEEDINGS.

EXPLANATION OF SEE PLATES:

All figures were outlined by means of the Abbe camera lucida and details
were added free hand. The approximate degree of magnification is given
in the explanation of the figures which are placed with the dorsal side up-
permost.

PLATE:

Fic. 1. Cloaca of Necturus. A transection just cephalad of the vent.
The ventral gland tubules are shown opening upon the ventral side. On
the dorsal side are cut spermathecas and convoluted tubules, some of
the former are shown opening upon the epithelium. x 8.

v. gd.—ventral gland.
c. t.—convoluted tubules.
sp.—spermatheca.
c.—cloacal cavity. ;

Fic. 2. Cloaca of Necturus. A transection farther cephalad showing
the opening of the large convoluted tubules and spermathecas into the
dorsal depression. A fragment of the ventral gland is seen. xX 8.

v. gd.—ventral gland.
ce. t.—=convoluted tubules.
sp.—spermatheca.
c.—cloacal cavity.
Fic. 3. A spermatheca opening into a convoluted tubule. x 48.
c. t.=convoluted tubule.
sp.—spermatheca.

Fig. 4. Isolated cells from a convoluted tubule. x 312.

n.—=nucleus.

Fig. 5. Isolated secreting cells from a ventral gland tubule. xX 312.

n,==nucleus.

PLATE TE L¢°

30¢ PROCEEDINGS.

PLATE Il.

Fia 6. Cloaca of Necturus. Transection yet farther cephalad. through
the oviducal papillees. The mass of tubules upon the dorsal side is shown,
some of which are spermathecas, others convoluted tubules. The neck of
the bladder is also transected. x 8.

sp.—spermatheca.
c.—cloacal cavity.
c. t.=convoluted tubules.
ov.—oviducts.

Fic. 7. Cloaca of Diemyctylus. Transection a short distance cephalad
of the vent. The spermathecas are seen cut at different levels. The bulk
of the ventral gland lies farther caudad and only a few tubules were tran-
sected. x 26.

- ep. epidermis.
c.—cloacal cavity.
sp. —=spermatheca.
v. gd.—ventral gland.
M.=striated muscle.

Fic. 8. Desmognathus fusca. A transection through the cloaca and
spermatheca. The zodsperms are shown in the diverticula of the latter. <A
prominent dorsal fold is present on which farther cephalad the spermatheca
opens. X 26.
sp.—spermatheca.

c.—Ccloacal cavity.
ep.—epidermis.
Z.—=ZoOsperms., -

Fig. 9. Desmognathus fusca. A transection through the cloaca farther
cephalad, showing the common tube of the spermatheca in the dorsal ele-
vation just caudad of its opening. X 26.

ep.—epidermis.
c.=cloacal cavity.
c. 8. t.-common spermathecal tube.
K.—Kidney.
M.-—-striated muscle.

hel Doyo tl Ca eg EUS

Zee ib

—

—

——=

——
2

y t

\

i -
4

302 PROCEEDINGS.

PLATE III.

Fic. 10. Amblystoma punctatum. A sagittal section nearly mesal,
through the cloaca indicating the relative positions of the glands and
spermathecas. x 8.

v. gd.—ventral gland.
d. gd.—dorsal gland.
sp.—=spermatheca.
c.=cloacal cavity.
K.—kidney.

Fic. 11. Amblystoma punctatum. A transection through a fully charged
ventral gland tubule in a spring individual before ovulation. \X 205.

n—nucleus.

Fic. 12. Ablystoma punctatum. Transection through a partially dis-
charged ventral gland tubule, showing the shrunken cells and the secre-

tion in the lumen. x 205.
n.—nucleus.

8.=secretion.
Fic. 13. Amblystoma punctatum. A transection through a yet more ex-
hausted tubule, spring individual, eggs laid. x 205.
n.—nucleus.
S.==secretion.
Fie. 14. Amblystoma punctatum. A transection of a ventral gland
tubule in a specimen taken in the summer. 205.
Fic. 15. Cells from the bottom of a spermatheca in a spring specimen in
which ovulation had not yet taken place. x 205.

PLATE Vit.

Sas | re.
! aetet, phate ae : Fi ipl MN
- ad See ey Lao be: OF ih ute
5 ' ‘be : . ,
a —.F, ‘ F .
j
‘
.-
au
i tl
oa i }
i) ‘ ,
y
Cie vai bd
Yu aby 7 J
\ 7 vat" i
are i o i
he
pee. ti }
Niue ‘
he I
" f " \,
. i
2
~ i
oti he \y |
y i - ;
j \ ‘,
a? d
y! nk
i y , -
Fe :
v a sr
\
'
fer:
i |
‘ a ial
iy i
bas, eee
ra ees

304 PROCEEDINGS.

Plate IV.

Fic. 16. Spelerpes bilineatus. Transection through the cloaca and
the spermatheca, showing the common spermathecal tube and the tubules,
one of which is seen opening into it; the others opened farther caudad. x 26.

c. s. t.=common spermathecal tube.
v. gd.=ventral gland.
sp. =spermatheca.
r. t.=rudimentary tubules.
t. =tubule.
ep. =epidermis.
c.=cloacal cavity.
Fic. 17. Spelerpes bilineatus. Transection farther cephalad at the open-
ing of the common spermathecal tube. x 26.
c. s. t =common spermathecal tube.
ep. =epidermis.
c.=cloacal cavity.
v. gd.=ventral gland.
Fic. 18. Plethodon glutinosus. Transection through the cloaca and
the spermatheca. x. 31.
sp. =spermatheca
c. s. t. =common spermathecal tube.
t.=tubule.
c.=cloacal cavity.
Fic. 19. Plethodon glutinosus. Transection farther cephalad near the
opening of the common spermathecal tube. x 31.
c gs, t,=common spermathecal tube.
c.=cloacal cavity.
K. kidney.

PEATE IV.

Wee

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Ke

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ia —S

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So
NSS

A PRACTICAL METHOD OF REFERRING UNITS OF LENGTH TO
THE WAVE LENGTH OF SODIUM LIGHT.

PROFESSOR Wm. A. RoceErs, Colby University, Waterville, Me.

During the summer of 1890 Professor Morley, of Adelbert
College, suggested to the writer a method of determining the
co-efficient of expansion of a bar of metal between the limits of
the freezing and the boiling points expressed in wave lengths of
light having a known refrangibility. After a discussion of the
best form of apparatus required for this purpose, it was arranged
that I should construct this apparatus, that he should come to
Waterville duiing his summer vacation and that we should
jointly undertake the experiment.

It will be sufficient to give in this paper a general outline of
the method and of the principle upon which it rests.

In the accompanying sketch A and & are bars of metal,:
placed upon supports which move at right angles to each other
between guides (not shown in the figure.) The support upon
which 4 rests is moved by a combined wedge and screw in order
to obtain a movement which is well under the control of the
observer. The index wheel is at the right of the observer and
within easy reach. J is placed at right angles to A upon a
plate which has slight longitudinal motion. The silvered plane-
surface mirrors @ and @ are supported upon three points at the
ends of the A and B. The mirrors 6 and @ are similarly
mounted at the other ends. One of the three points of support
is a rounded projection from the end of the bar itself. The
mirrors are first of all set at right angles to the axis of the bars
by means of a collimating eye piece by the aid of screws whose
counter end-thrust is controlled by springs.

DD is called the diagonal mirror. It is placed at the inter-
section of ac and ac. This mirror is half silvered, that is, the

process of silyering is continued until one-half the light which
at

306 PROCEEDINGS OF THE

strikes the mirror will be transmitted and the remaining half
reflected. ££ is a plane unsilvered mirror having parallel faces.
It is called the ‘‘ evener”’ since it permits the passage of both the
transmitted and the reflected rays through the same thickness of
glass.

M is a plane-reflecting mirror, 7z is a condensing lens having
cross wires at the center. A is a sodium flame and Wis a lamp
flame placed in the line mk.

One-half of a ray from & is transmitted through the mirror to
a. This is in turn reflected back to 6. One-half of this reflected
ray passes through the glass. The other half suffers internal
reflection and is seen by the observer at O.

The reflected half of the original ray is reflected back to 6.
At this point it divides again and the transmitted half reaches
the point O'. Now of the two rays which reach the eye at O,
or O', the first has suffered in internal reflection and the second
external reflection. They therefore differ by half a wave length.
This condition will always exist when da equals 6@ nearly. When
therefore the cross at m on the condensing lens is seen to be
coincident with its image in the telescope at O, the field will be
filled with dark lines about one ninety-thousandths of an inch
apart. These lines of course result from the absence of light
from the monochromatic yellow flame at the points at which the
rays differ by half a wave length. If now a white light be placed
behind R at W,a condition will be obtained in which one line,
or as it is commonly called one fringe, will be black and the
remaining fringes will take the colors of the rainbow. This
condition will be obtained when the distance da is exactly equal
to 6a, a result which can be secured by moving the plate upon
which A rests by means of the screw. In the same way the
sodium line in white light will be seen at O’ when the distance 46
is exactly equal to 00’.

The method of observation is as follows: <A fiducial line is
drawn in india ink upon the mirror directly over the fulcrum
about which the mirror moves. The fringes in white light are
first formed upon the rear mirror and the sodium line in white light

AMERICAN MICROSCOPICAL SOCIETY. 3°7

is set in coincidence with the fiducial line upon the mirror. The
telescope is then moved to QO. If now the sodium line in white
light is found to be coincident with the fiducial line upon the
front mirror the two bars have exactly the same length. If it
does not occupy this position the plate dA is moved along by
means of the screw until the dark line does appear in coincidence ;
the corresponding number of sodium lines being counted during
the movement from one coincidence to the other. In my
experience, it is found easy to count about go fringes per minute
for short runs. I have frequently counted 700 lines in Io
minutes. It will be found convenient to turn off the white light
during the greater part of the time occupied in counting, using
only the sodium lines for counting till the end is nearly reached
when both lights must be in the same field of view.

The feasibility of this method was proven by the observations
made by Professor Morley and myself in 1891. Both bars
were observed for half a day in melting ice, the difference in
length remaining constant meanwhile. The ice surrounding
the bar B was then removed and the bar was kept in steam for
several hours. Under this condition, also, the difference in
length between the two bars was measured. In this experiment,
however, it was found impossible to count the corresponding
numbet of fringes on account of the impossibity of obtaining
sufficient freedom from the condensation of moisture upon the
mirrors, although the fringes were seen upon both mirrors
under the extreme conditions of temperature. Hence the only
thing which could be done was to measure the distance by
means of a microscope attached to the plate upon which the bar
A is placed, the scale being disconnected from this plate.

These experiments made it clear that all observations of this
character must be made in vacuo.

A new refractometer was therefore constructed. In doing this
several improvements were introduced. The bars were enclosed
in boxes of rolled brass, as it was supposed that the air could
not enter the boxes through the pores of this metal. In this
J was disappointed and after a trial of over a year, it was found

308 PROCEEDINGS OF THE

necessary to begin de zovo. The two brass boxes and the box
for holding the diagonal mirrors were cast solid and were after-
wards covered completely on the outside with solder to the depth
of one-eight of an inch. The magnitude of this undertaking
may be inferred when it is stated that these boxes, with their
connections, are each nearly 60 inches in length and 6 x 6
inches cross sections ; the metal being half an inch in thickness.

The construction of this refractometer was completed in 1894,
and my time until the present summer was pretty fully eccupied
in testing it in every possible way. Early in June of the present
year, I wrote Professor Morley that so far as I could see every-
thing was now ready for the final trial. He therefore came to
Waterville early in July for this purpose. While the two
experiments which were made, are not entirely satisfactory they
demonstrate with certainty the entire feasibility of the method.
In the first series, oil was forced through the packing and
dropped upon one of the mirrors when a temperature of 65°C.
was reached, thus compelling the abandonment of the experi-
ment at this point. In the second experiment, after measures
made at 65°C. it was found that before reaching 100°C., sulphur
from the rubber packing tarnished the mirrors to such an extent
that, while the fringes could be easily seen near the maximum
points of visibility, it was impossible to count them at the
minimum point, beyond the second order.

In order to obtain the value of one division of the micrometer
screw of a microscope in terms of wave lengths of sodium light, _
the apparatus briefly described here may be greatly simplified.
We need only one set of mirrors for this purpose, and Professor
Snow of the University of Wisconsin has shown that the
expensive plane mirrors are not necessary ; if one is content with
curved lines instead of the perfectly straight lines always obtained
with the plane surfaces made by Brashear. Professor Snow
made use of mirrors cut from polished plate glass, care being
taken to cut all the pieces from the same piece of plate glass, so
that they might have the same thickness, He finds the process
of half silvering not at all difficult,

AMERICAN MICROSCOPICAL SOCIETY. 309

In this measurement we can dispense with the sodium line in
white light and use only the sodium lines in making the count
corresponding to a given distance as measured under the mi-
croscope.

The reduction from the observed number of fringes to metric
units will be found from the relation :

3393.8 fringes = I000u, or Iu = 3.3938 fringes.

The co-efficient of Jessop’s steel derived from these observa-
tions agree well with each other, but they are still subject to a
slight constant correction on account of the, as yet undetermined,
corrections to be applied to the readings. of the thermometers for
the higher temperatures.

The following are the separate results of the co-efficient of
expansion for the bar of Jessop’s steel whose length is 102.7 cms.

FROM SERIES I. FROM SERIES I.
From 0° C. to Co’eff. From 0° C. to Co’eff.
18.362 10.720 21.70° 10.586
21.1 10.681 63.90 10.797
24.1 10.806
28.0 10.693
28.6 10.782

Mean—10.667
Mean—10.736

Hence for 100° Co-efficient—10. 4385 /.
The detailed account of the experiment will be given in a joint

paper by Professor Morley and the writer which will appear in
the Physical Review.

COCAINE IN THE STUDY OF POND-LIFE.

H. N. Conser, Sunbury, Pa.

Hydrochlorate of cocaine as a narcotic for forms of aquatic
life has a special value in the study of bryozoans and the encased
rotifers. Quick-killing methods cannot be used where the con-
tractile organs are so well protected as in these forms, neither can
the narcotics that kill, for they often allow disorganization of cilia
and tentacles before other parts of the organism are sufficiently
benumbed.

The method I have found most satisfactory and certain with
the fresh water Bryozoa is as follows: Several colonies are
placed in a solid watch glass with 5 cc. of water, and as soon as
the animals have expanded one or two centigrams of cocaine is
dropped on the edge of the water at two or three distant points.
In fifteen minutes the narcotic influence is sufficient, as can be
tested by touching the tentacles with a needle. One per cent.
chromic acid is now poured in to fill the watch glass and left to
act for half an hour or more when it is nearly all withdrawn and
water substituted. This process is repeated in half an hour, and
alcohol to form about twenty-five per cent. added to the water,
the strength of alcohol is increased by the addition of ninety-five
per cent. until eighty per cent. is reached. By this means the
chromic acid is washed out and the hardening accomplished so
gradually that no distortions occur. For staining, borax-carmine
or alcoholic-cochineal is used. The clearing must be gradual
and is best accomplished by adding oil of lavender to the ninety-
five per cent. alcohol in which the animals are kept, and after an
hour, bringing them into oil of lavender from which, after per-
fect clearing, they are mounted in balsam.

The free swimming rotifers readily succumb to the influence
of cocaine, but the family Melicertade hold out a long time
against it. A method for these is like that for the bryozoans

AMERICAN MICROSCOPICAL SOCIETY. 311

with the exception that only sufficient water to cover the colony
well, need be used; the quantity of cocaine must be relatively
large, and when all movements cease, killing may be done with
twenty per cent. formalin, as chromic acid precipitates cocaine,
when present in any considerable quantity. An after treatment
with chromic acid in one-half per cent. seems to give better
hardening than formalin alone. When a colony of the Meli-
certade is subjected for fifteen minutes to a half per cent. cocaine
solution and then transferred to another watch glass with pond
water, the individual rotifers come out of the tubes and attach
themselves hydra-like to the bottom of the glass in perfect con-
dition for study, thus saving the trouble of freeing the animals
from the tubes with needles.

PARAFFIN AND COLLODION EMBEDDING.

H. N. ConsEer, SUNBURY, Pa.

Paraffin and collodion, in their modifications, answer now so
nearly every requirement of embedding media, that, excepting the
rarest cases, there is no further need of the soap, gum and gela-
tine mixtures.

For a large class of tissues either paraffin or collodion may be
used. Here, then, is the province of choice in methods, but aside
from this the question is not one of respective merits, but rather
a consideration of the method indicated for the work at hand.

Some years’ experience with a wide variety of material by both
methods has led to the observations of this paper. The choice
of a clearing agent in the paraffin method is important. Not all
solvents of paraffin which are miscible with alcohol are suitable
for the purpose. An extended trial of turpentine proves it of all,
the most hurtful to delicate structures. The shrinkage is so
much by its use that its excellent penetrating qualities are not
enough to recommend it. Chloroform penetrates slowly and
does very well for small pieces only. It must be gotten rid of
entirely in the bath, for any quantity left in the object will render
it soft and unfit for cutting. Toluol (Toluene) has in my experi-
ence given the best results. For embryos stained zz ¢ofo I have
never found any other clearing agent so reliable and satisfactory.

Of the paraffinitself the medium soft melting at 48—50°C is best
for most purposes, the harder is suitable for the harder tissues,
provided the higher temperature of the bath is not objectionable.
Sections can be cut thinner from hard than from soft paraffin,
yet, for many objects, it is better to use a soft paraffin and cut at
a correspondingly lower temperature.

Tender vegetable structures if fixed from the fresh state in the
chrome salts or chromic acid usually bear the dehydration and
heating very well.

For embryos and the like of which sections contain separated

AMERICAN MICROSCOPICAL SOCIETY. 323

parts, staining 77 foto and embedding in paraffin is of all methods
far the most practical ; the sections are easily affixed to the slide
with the albumen or collodion fixatives so that loss or displace-
ment of parts seldom occurs. Cut paraffin sections are not easily
kept unless affixed to the slips, but the blocks may be kept for
years if the embedding has been carefully done with good
paraffin. I have found that most of the specimens that harden
too much for cutting after several years’ keeping to be those
cleared in chloroform or those heated too long in the paraffin
bath.

For the collodion method the use.of Schering’s celloidin is
most advised. The first steps are essentially alike in the various
modifications of the method and the only trouble likely to occur
is from giving the collodion solutions insufficient time for perfect
homogeneous penetration. Two to four days in each is enough
for average objects, but any longer time can do no harm. I
have had objects remain in collodion solutions two years without
their having been in any wise injured. Embedding on cork or
wood in a box of paper of which the cork or wood block forms
the bottom is the common procedure. If the object be firm
enough to bear its own weight, it may as well be embedded on
the cork or wood without the surrounding paper, and placed
direct in the hardening fluid, only a thin stratum of collodion
surrounds the object and the formation of bubbles is obviated.
In the matter of hardening there is diversity of usage. Chloro-
form, which must be anhydrous, gives excellent results both as
to transparency and consistency of the mass, but its rapidity of
action on the exterior, while the interior of the mass is slowly
affected, renders it less serviceable than alcohol which hardens
uniformly, and after one or several days’ action yields an excel-
lent cutting consistency. or this purpose 80 to 85 per cent.
alcohol is best. Objects embedded on cork or wood may be
kept in a jar with 70-80 per cent. alcohol, but discolor and
deteriorate after prolonged keeping. It is better to section as
soon as the mass is well hardened and keep the sections in vials
of 80 per cent. alcohol until wanted. Sections preserved in this

314 PROCEEDINGS.

way for six years are yet good. Since it is often desirable to
keep valuable material for future use, this feature of the collodion
method is a strong one in its favor.

The dehydration of sections for mounting will depend upon
whether the collodion is to be removed or retained. In the first
case absolute alcohol followed by oil of cloves is good, usually it
is better to retain the collodion, and then, of course, none of its
solvents can be employed. Ninety-five per cent. alcohol followed
by oil of bergamont, oil of origanum (the cretici) or oil of cedar
wood answer the purpose well. As to the respective uses of the
two methods it may be said that for serial sections and normal
tissues stained in toto, the paraffin method is by far the better ;
it is the one especially suited for embryology and histology.

Paraffin is not so well adapted for tissues of the central ner-
vous system, parts of the eye, cartilaginous structures, organs
containing exudations or foreign elements, nor for those objects
injured by heat ; for these then collodion is preferred and almost
exclusively used by pathologists, being the better by reason of
the above-named advantages together with the readiness with
which its sections are double stained.

While the histologist and embryologist can get along with
paraffin, and the pathologist with collodion alone, the best results
can be obtained by the use of both, each chosen for the work it
is best fitted to accomplish.

FORMALIN AS A HARDENING AGENT FOR NERVE TISSUES.

Wirniam.C, Krauss, M. D., F, R.-M. &))'BatfaloN. Y.

Formol, Formalin, or Formaldehyde was discovered by A. W.
Hoffman in 1863 while passing wood spirit and air over a red-
hot platinum spiral. If the vapor is brought into water to its
point of saturation, a forty per cent. solution of formaldehyde is
obtained, which has long been known under the name of formol.
That Formalin possessed antiseptic as well as hardening powers
we owe to the investigations of Dr. F. Blum, and these facts in-
duced the elder Blum to make an extended series of investiga-
tions in the hardening of animal and vegetable tissues in the Sen-
ckenbergischen Institute at Frankfort, Germany. His prelimin-
ary report was published in the Zodlogischer Anzeiger, 1893,
No. 434, and a more detailed report in the Berichte tiber die Sen-
ckenbergische Naturforscher Gesellschaft, 1894, p. 195. Here
he details his experience, which in brief is as follows: Several
human embryos were finely preserved in formol diluted with ten
to twenty parts of water. Small embryos with amnion intact
were preserved and the amniotic fluid remained transparent, so
that the structural parts of the foetus and the umbilical cord were
recognizable. The mouse, hamster, and porpoise, were nicely
preserved, the hair firmly in place and the eyes in better con-
dition than under the use of alcohol. Reptiles, fishes and am-
phibians were nicely hardened in one to ten, one to twenty, or
one to thirty, solutions according to the size of the object. The

fishes- retained in great part their color, while the slime and
mucus covering them was rendered transparent. Of the inverte-

brates, snails, jelly fishes, insects, spiders, etc., all were well
preserved.

Of the various animal tissues, muscles and the brain were
quickly hardened, retaining the coloring matter of the blood in

316 PROCEEDINGS.

the muscles, while in the brain the differentiation between the
white and gray matter was very evident. Fruits, flowers and
vegetables of various kinds were equally successfully preserved,
the coloring matter very little if at all impaired. Blum’s con-
clusions regarding the hardening of animal tissues may be sum-—
med up as follows : ;

Animal objects are hardened with shrinking, and without los-
ing their microscopic structure or staining properties.

The natural form and color are preserved.

The eye remains much clearer than in alcohol.

The mucus of slime-producing animals is not coagulated and
remains transparent.

The coloring matter of blood in tissues apparently disappeared,
but may be quickly restored by a high per cent. alcohol.

These experiments of Blum were pathmaking and were quickly
followed by those of Born (1), Pintner (2), Kriickmann (3),
Kenyon, Sadebeck, Mayers, and others with seemingly favorable
results as regards the preserving and hardening powers of
this compound. Besides these qualities it was especially
valuable because of its being non-poisonous, non-combustible,
of a low freezing point, and, what to scientists is quite a serious
question, very cheap.

My attention was directed to this substance about one year
ago, through experiments made at the Laboratory of the Erie
County Hospital by my interne Dr. Helvie.

Various tissues, as liver, spleen, placenta, lungs, heart, muscles,
and other tissues and organs were satisfactorily hardened and
preserved. Especially gratifying were the results obtained with
the umbilical cord, aud other myxomatous tissues. Instead of
shrivelling up and becoming opaque as occurs when alcohol is
employed, the cord retained its normal size, was transparent and
hardened to such a degree that sections were Beret and perfectly

1. Med. Sect. d. Schlesisch. Gesellsch. f. Vaterl. Kultur. 1894.
2. Ver. Zool. Gesell., Wien, 1894, p. 8.

3. Centralbl. f. Bakteriol und Parasitenk., 1894, pp. 851-57.

4, American Naturalist, Jan. 1, 1894.

AMERICAN MICROSCOPICAL SOCIETY. a7

cut with the microtome. The intestines were greatly shrivelled
during hardening, but otherwise were a success.

These results induced me to try its virtues upon the brain and
spinal cord and especially to find the earliest time when a spinal
cord so hardened could be imbedded in celloidin and sections cut
for staining and mounting. A spinal cord which to all appear-
ance was normal was cut in pieces about one centimeter long and
placed, sections of the cervical, thoracic and lumbar regions in
bottles containing a five per cent. solution, ten per cent., twenty per
cent. and twenty-five per cent. of Formalin. At the end of seven
days a section of the cord was taken from each of these solutions
and imbedded in celloidin, then placed on the microtome. The
cord was evidently too imperfectly hardened as no good cuts were
obtained. At the end of fourteen days the same procedure was
followed, likewise on the twenty-first day and twenty-eighth day.
From each of these intervals excellent cuts were obtained, the
cord retained its external contour and appearance, but the dif-
ferentiation between the white and the gray matter was not as
well marked as when alcohol is used. These sections took the
carmine stains nicely, but less so the nigrosin, Pal and Weigert
stains. The most serious action of the formalin on all of
these sections was a contraction, evidently of the neuroglia in
various regions of the cord, especially of the white matter result-
ing in the formation of open spaces or cavities. In the sections
hardened in the 10 per cent. solution these cavities were so large
as to destroy completely the slides for microscopical purposes. In
the 15 per cent. cord, the cavities were much smaller, but far more
numerous, and the dorsal white columns looked like a honey comb
or sieve. This action of the formalin was manifested in every
section examined and is therefore not of accidental, but of regu-
lar occurrence. The drawback of the formalin in preventing the
employment of the Pal and Weigert methods of staining has been
successfully overcome by Marcus (1), who after hardening the
cord for from two to four weeks in a one halt per-cent. solution

1, Neurologisches Centralblatt, Jan, 1, 1895,

318 PROCEEDINGS.

of Formalin, places small portions one-half cm. thick in Miller’s
fluid in a brood oven for seven days at a temperature of 37° C.
They are then dehydrated, imbedded and the cuts again placed in
Miiller’s fluid, for from two to seven days in a brood oven, quickly
washed in alcohol, then transferred into the Weigert stain.

The action of the formalin on the ganglion cells isa happy one,
swelling them and rendering their nuclei susceptible to very in-
tense staining.

The action of formalin on the brain has given very fine results.
Born succeeded in hardening the entire brain very quickly for
demonstrations, also small particles for microscopical purposes.
I have been equally successful and have some excellent speci-
mens, nicely hardened. I have not as yet tried any of the stain-
ing methods on brains thus hardened and cannot state what the
results would be, although I have some specimens under way.
From my experience with Formalin I can greatly recommend it,
for the hardening of the various organs and tissues for macro-
scopic as well as microscopic purposes, but would still cling to
the Miiller’s solution for hardening the spinal cord, even if the
time required for hardening be much longer than when Formalin
is used.

THE USE OF FORMALIN IN NEUROLOGY.

PIERRE A. Fis, D. Sc. Washington, D. C.

Formalin (HCHO) is the forty per-cent. solution of formic
aldehyde gas in water. The aldehyde is variously known as
formaldehyde, formol and formalose, and has, of late, come into
such a prominent degree of usefulness, that it might seem desir-
able to offer a brief, though inadequate survey of some of the
uses to which it has already been put, after a year’s experimenta-
tion upon neurologic material.

Formalin is prepared by subjecting methyl alcohol to oxida-
tion.

Methyl Alcohol + Oxygen = Formaldehyde + Water
CH,OH PEO HCHO + H,O.

Further oxidation will produce formic acid.

The method of its preparation in either large or small quan-
tities has been given by Stebbins (28). Its production in large
quantities is based upon the German patent No. 55,176, issued
to Auguste Trillat, Dec. 17, 1890.

It is miscible with water and alcohol in all proportions. It is
kept in darkened bottles, as the light may cause decomposition
or at least a separation of paraform, this may also sometimes be
seen as a white substance around the stoppers of bottles ;
exposed to a low temperature it is said, however, that this
separation has no influence on the action of formalin. Paraform
does not seem to appear so readily in weak as in the strong
solutions.

With various tissues, it is likely that different percentages will
be useful; the percentage suitable for one form of animal may be
inadequate for the proper preservation of another. Its utility as

*This article was prepared mostly in the Anatomical Laboratory at
Cornell University, Ithaca, N. Y,

320 PROCEEDINGS OF THE

a preservative for laboratory specimens has been pretty well
tested and found favorable. It has a neutral or slightly acid
reaction and an odor resembling that of Witch Hazel; but if
used in strong solutions the gas becomes very irritable to the
conjunctiva and to the mucosa of the respiratory passages-
Even when used in solutions diluted to two per cent.* some
discomfort is caused, unless the specimens be first rinsed or
soaked for a short time in water.

When the dilute solutions are used for hardening they should
be occasionally renewed and kept tightly covered to prevent de-
terioration. |

Formalin also has the advantage over alcohol of not being
inflammable and of not shrinking the tissue to the same degree,
nor does it destroy the natural color of the specimen so quickly;
but on the other hand it is not so suitable for museum prepara-
tions which may be exposed to cold temperatures, as the amount
of water present would invite freezing and consequent destruc-
tion of the jar and perhaps of the specimen.

On account of its penetrating action, large as well as small
organs or specimens may be hardened in it. Its cheapness is
another element in its favor; even at the rate of two dollars per
pound, at which it retails in this country, it is as cheap in dilute
solutions, as alcohol free of tax. In Germany it sells for four or
five marks ($1.00 to $1.25) per kilo (2.2 lbs.). It furthermore
possesses the advantage of dissolving certain salts more readily
than alcohol, and it may therefore have a wider range of applica-
tion as an adjunct in preservative methods.

A limited experience with the preparation introduced under
the name of formalose indicates that it has about the same per-
centage of formic aldehyde as formalin and may be used in the
same way.

In August, 1893, F. Blum (4) called attention to the use of
formaldehyde as an antiseptic in dilute solutions. In Septem-
ber of the same year, the same writer (5) speaks of its action as

*When the percentage is spoken of, it refers to the proportion of com,
mercial formalin present and not formic aldehyde,

AMERICAN MICROSCOPICAL SOCIETY. 321

a hardening medium. His attention was called to this feature
from the fact that the epidermis of his fingers became hardened
after working for a time with formaldehyde.

On immersing a field mouse and certain organs in a four per
cent. solution, he became convinced that they were hardened as
well and more quickly than when alcohol was used.

Hermann (16) finds no especial advantage nor disadvantage in
the use of formalin over other fixing agents; indeed he believes
that for section methods the after-treatment with alcohol is some-
what deleterious to the tissue.

F. Blum (6) discusses Hermann’s paper. J. Blum (7) used
formol in two per cent. solutions upon some fishes and a lizard
and found them to harden in a very short time and to preserve
their form and color unchanged; the latter condition being due to
the fact that the mucin of the mucus-secreting animals remains
transparent in formalin.

Alleger (1) states that attention was first called to the
germicidal action of formic aldehyde in 1886, by Low. Gelatin
is made insoluble by the formalin and this is found to be
of great advantage in bacteriology and histology; in the lat-
ter it is useful as a fixative in holding the sections to the
slide, by adding a few drops of the formalin for each gram of a
one-half to one per cent. gelatin solution. A gentle heat is
applied to the slide until the paraffin is softened and the superflu-
ous gelatin allowed to drain from the edge of the slide. Another
interesting recommendation of Dr. Alleger’s is that fresh tissues
may be placed directly in certain staining reagents to which have
been added five per cent. of formalin, and thus hardened and
stained in bulk at the same time.

Hoyer (18) has taken parts of the nervous system from corpses,
hardened them in formalin and then submitted them to the Golgi
method with good results.

Marcus (24) recommends hardening the spinal cord for two or
four weeks in a one-half per cent. solution of formalin, then small
pieces one-half centimeter thick are cut out and placed in Miller’s

fluid for a week in an oven at 37°C. The pieces are then dehy-
22

322 PROCEEDINGS OF THE

drated and imbedded in collodion, after the sections are cut they
are again placed in Miiller’s fluid and put back in the oven from
a day to a week. The sections are then quickly washed in
alcohol and put in the Weigert-Pal-hematoxylin solution for at
least two days. ; 3

No mention is made of the copper acetate bath, and the result-
ing stain is apparently due to the formation of a chromium lake
more or less modified by the use of the formalin.

Strong (30) advocates the following formula for the Golgi
method :

Potassium bichromate (84%-5%), 100 Vols. —
Formalin, 24-5 Vols.

After hardening several days the tissue is transferred to the
silver-nitrate solution (one per cent.). Or the tissue after one or
two days may be transferred from the above bichromate-formalin
mixture to the following :

Potassium bichromate (5%), 2 Vols.
Formalin, 1 Vol.

After twelve or twenty-four hours the tissue is put into the
silver solution. The advantages of this method are that it avoids
the use of osmic acid, and that the stage of hardening favorable
for impregnation lasts longer than when the osmium-bichromate
mixture is used. In other words the formalin-bichromate does
not over-harden. In this respect it is superior to the lithium-
bichromate method of the same author. For embryonic tissue
he finds the osmium-bichromate preferable.

Van Gieson (31) has used formalin in four per cent., six
per cent. and ten per cent. solutions for ordinary histologic
methods, followed by ninety-five per cent. alcohol and col-
lodion imbedding. Weigert’s hematoxylin method can be ap-
plied to such sections and gives very good results for the
plexus of fine fibers in the cortical and spinal gray matter.
The myelin of the fine fibers is well preserved and gives
the characteristic blue-black reaction with the Weigert hema-
toxylin stain, as in chrome-hardened preparatious. The back-
ground of the gray matter is especially clear and the fibers

——

—_

AMERICAN MICROSCOPICAL SOCIETY. 323

sharply delineated. The formalin-hardened sections should be
soaked in the neutral copper-acetate solution diluted one-half
with water for two hours, then thoroughly washed in water and
immersed in the Weigert lithium-carbonate-hazmatoxylin solution
from four to twelve hours. Weigert’s borax-prussiate-of-potas-
sium solution is useful for differentiation.

In the absence of any chrome salts in the above method, the
stain is the result of the formation of a copper lake. The method
has been confirmed in our own laboratory and the results were
all that could be desired.

Dr. Van Gieson also finds that formalin-hardened sections are
useful in Rehm’s modification of Nissl’s method, but that the
minute structure of the nucleus and cytoplasm is not quite so
sharply outlined as with fixation in absolute alcohol. The dura-
tion of the hardening period in formalin exerts an important and
varying influence upon the tissues. Further investigations upon
this matter are promised.

René Marie (25) uses a one per cent. solution of formalin and
allows it to harden the tissue for four or five days. The usual
staining methods are employed.

Lachi (21) finds that formalin in weak (one to two per cent.
solution) or in stronger solution (ten to fifteen per cent.) exerts an
injurious influence upon connective tissue by dissolving the
fundamental substance, especially in the elastic fibers and mucosa.
If used for such tissue some corrective should be employed.

He finds it of signal service in the nervous system, either in
the central or peripheral, or in the embryo or adult. Nerves
kept for a few days in from two to five per cent. solutions can be
treated with silver nitrate, and show the characteristic cross of
Ranvier. Pieces from the central nervous system treated from
five to nine days in a mixture of equal parts of twenty per cent.
formalin and six per cent. potassium bichromate, gave the black
reaction with silver nitrate equally as well as the osmium bichrom-
ate mixture with the advantage that the blackness of the tissue
was not so great as when osmic acid is used. Favorable results
were obtained from the myel and embryonic brain of cows, and

324 PROCEEDINGS OF THE

with the cerebrum, cerebellum and myel of the human adult.

The mixture indicated above is also recommended for the
Weigert method. Paraffin is not advocated for an imbedding
medium on account of friability.

Kenyon (20) records a free translation of J. Blum’s article in
the Bericht, iiber d. Senckenbergische naturf. Gesell. in Frankf.,
a. M., 1894, and adds valuable observations of his own.
Blum states that formaldehyde was discovered in 1863 by A. W.
Hoffman while passing wood spirit (methyl aleoholy and air over
a red-hot platinum spiral.

The vapor carried into water to the pbiak of saturation gives a
forty per cent. solution of formaldehyde. This Blum calls formal
because it was known under that name when it was first used in
aqueous solution for disinfecting, hardening and preserving.

A reference is made to Born: ‘‘ Demonstrationen einer Anzahl
in Formaldehyde (Formol) geharteter menschlisher Gehirne.
Mediz. Sektion der schlesisch. Gesell. f. vaterl. Kultur, 1894,”
stating that pieces or even the entire brain hardens quickly,
and the white and gray matter are sharply differentiated.

Blum also performed interesting experiments in preserving
hens’ eggs, certain invertebrates, vertebrates, fruits and plants.

Kenyon experimented upon a variety of forms and found a
four per cent. solution of formic aldehyde (ten per cent. of the
commercial formalin) best adapted to Salamanders.

Different percentages were tried; the lowest being one-fourth
per cent. and the highest twenty per cent., some of the dilute
solutions were found useful for some of the invertebrates. The
fact noted by F. Blum was verified, namely: that the vessels
containing blood lost color when hardened in formalin, but this
reappeared when treated by alcohol. This was explained by the
coagulation of the fibrin by the alcohol giving it a yellow color
and making it opaque and by bringing the corpuscles again to
view. In conclusion, Mr. Kenyon believes that a solution stronger
than two per cent. of the formalin is necessary to prevent the
swelling and decolorization of specimens, and that from four to
eight per cent. will give the best results,

AMERICAN MICROSCOPICAL SOCIETY. 325

To counteract the swelling caused by the weak solutions of
formalin, alcohol was added. For histologic purposes a mixture
of alcohol and formalin was found to act better than either one
used alone.

Durig (11) also employs formalin as a substitute for osmic
acid in the silver-impregnation method. His plan is to harden a
piece one-half centimeter square for three days in four to six per
cent. of formalin and three per cent. Potassium bichromate, then dry
off on filter paper and immerse in a three fourths per cent. silver
solution. After two days return to the first mixture—and lastly,
immerse it in silver containing a trace of formic acid.

Stebbins (28) gives a good description of the chemistry and
preparation of formalin. This agent has recently been intro-
duced into photography for hardening gelatine films, and found
to be of great service. Formalin is a powerful reducing agent,
In aqueous solution it reduces ammoniacal nitrate of silver to
metallic silver, forming a mirror on the sides of the vessel con-
taining the solution. It unites with bisulphate of soda or potas-
sium, to form a crystalline addition product. This reaction may
advantageously be used for separating formaldehyde, as well as
homologous aldehydes from mixtures of other bodies.

The combination of formalin with other hardening reagents
apparently has not, as yet, received much attention ; its use in
this connection will undoubtedly be of great value in macroscopic
as well as microscopic methods.

Experiments with formalin show that good results may be
obtained with nervous tissue when the following mixture is em-
ployed:

Water, : : 2000 cc.
Formalin, : C : 50 ce.
Sodium chloride, ’ 100 grams.
Zinc chloride, : : 15 grams.

The specific gravity should be about 1.05. In practice the
brain is left in this mixture for a week or ten days (a longer stay
is not detrimental) and when practicable the cavities and blood
vessels are injected with the same mixture in order to insure a

326 PROCEEDINGS OF THE

more uniform hardening. The specimen may then be trans-
ferred to 2% per cent. formalin (water, 2000 cc., formalin, 50 cc.)
and may remain in this solution indefinitely if the jar be kept
tightly covered ; o- if it is to become a museum specimen it may,
after a week in the second solution, be removed to fifty per cent.,
seventy per cent. and 90-95 per cent. alcohol for final storage.

An objection to the use of formalin solutions for museum
purposes would be the large proportion of water present, which
would freeze at low temperatures and cause injury to the speci-
men or jar containing it. Since formalin is readily miscible with
alcohol, as well as water, enough of the former might be added
to prevent the freezing, say equal parts of ninety-five per cent.
alcohol and two and a half per cent. formalin ; the exact propor-
tions have not as yet been determined.

After an immersion of two weeks in the formalin solutions, a
human brain lost only 6.8 per cent. of its weight; but after an
immersion in fifty per cent. alcohol for eight days and an immer-
sion for an equal length of time in seventy per cent. alcohol, a
total of sixteen days, it was found to have lost twenty-two per
cent. of its first weight. A monkey brain after an immersion of
eight days in the formalin mixture lost 5.4 per cent. of its weight;
continued immersion in the same fluid for eighteen days longer
caused a loss of less than two per cent. A fox brain was im-
mersed in a similar mixture for five days and lost 6.5 per cent. of
its weight ; it was left in the same mixture eighteen days longer
and lost 2.3 per cent. more of its weight. The brains were firm
and in excellent condition for dissection.

The second, or two and one-half per cent. formalin-solution
redissolves any of the sodium chloride that may remain in the
brain, which is an advantage if the specimen is to be treated with
alcohol, as the latter does not dissolve the salt. The brain
should not be put from the formalin solution immediately into
the strong alcohol as the tissue will shrink very materially.

Material treated in the way above described has yielded most
satisfactory results histologically. Portions of the central nerv-
ous system of an adult, after treatment with the above brain

AMERICAN MICROSCOPICAL SOCIETY. 327

mixture, have been later treated with other fixing reagents: e¢. ¢.
corrosive sublimate, picro-aceto sublimate and a chrom-acetic-
acid mixture with most excellent results. Equal parts of a two
and-one-half per cent. solution of formalin and the picro-aceto
sublimate proved very satisfactory. The same proportion of
formalin with the chrom-acetic mixture worked very well; but
the combined mixture turned green after a short exposure to the
light. |

On November 3, 1894, in working on the myel of a young
kitten, I substituted formalin for the osmic acid in the Golgi-Cajal
method, using the following formula :

Potassium bichromate, 37%, 4 Vols.
Formalin, 22, 1 Vol.

The-specimen was left in the mixture for nine days, then im-
bedded and cut; the results were very promising. In later ex-
periments I added the strong formalin directly to the bichromate
solution.

Formalin, 2 ce.
Potassium bichromate 34, 100 ce.

The specimens remained in the mixture for three days and an
equal length of time in the three-quarter per cent. silver solution.

The impregnated cells with their processes were particularly
' distinct, standing out like black diagrams ona clear back ground.
After a time, although the cells and processes remained distinct,
the action of the light caused the sections to lose their light
color and turn yellowish brown, resembling very much the sec-
tions prepared by the osmium-bichromate method.

The following mixture was also tried with even greater success
than the preceding :

Miillers fluid, 100 cc.
Formalin 10%, 2 cc.
Osmic acid 1%, 2cc.

The formalin-bichromate mixture was not tried upon embry-
onic tissue, nor the brains of low or generalised forms.

Care must be taken to keep the formalin bichromate mixture
in the dark to prevent its deterioration. Exposure to the light

328 PROCEEDINGS.

causes the solution to assume a dark muddy color, the result of
some chemical change. It is advisable, therefore, to mix the two
solutions as they are needed and keep the mixture concealed.

Gage (15) has tested formalin as a dissociating agent. He
recommends the following formula :

Normal salt solution, 1000 ce.
Formalin, 404, 2 ce.

His results were highly satisfactory. The sclution acts quickly
and yet retards deterioration for some time.

After three hours the ciliated cells from the trachea of a kitten
were easily separated upon a slide and almost as good prepara-
tions were obtainable after ten days. The endymal cells border-
ing the encephalic cavities were found to be very susceptible to the
mixture, also “some of the cerebral cortex from various regions
was tested, and it was found comparatively easy to obtain excel-
lent preparations in which many of the multipolar nerve cells
were wholly isolated.”

June, 1895.

vw

10.

IGE

12.

13.

14

15.

16.

AMERICAN MICROSCOPICAL SOCIETY. 329

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33°

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Anz., Vol. X., p. 494. (Science, Vol I., p. 166.)
Van Gieson, Ira.—(Weigert’s method, ete) Anat. Anz., Vol.
X., p. 494. (Science, Vol. I., p. 167.)

FORMALIN IN THE ZOOLOGICAL AND HISTOLOGICAL
LABORATORY.

D. 8. KeLticott, Columbus, O.

How often in the memory of most of us has the suffering
world been startled by the announcement, often by the highest
authority, of some wonderful medicine, specific or elixir. These,
one after another, have been put to the test and have taken their
places among the useful remedies, but on a plane below, often
far below, the first claim and the initial hope. Is it to be thus
with Formaldehyde as antiseptic, as remedial agent, as preserva-
tive, as a histological fixative? It can only find its true plane
among its kind by conscientious trials made and reported by in-
vestigators. During the last year it has been used in the labora-
tory of the Ohio State University quite extensively as a preserva-
tive and for histological purposes. I have brought together in
this paper the results and the reflections to which they obviously
lead. Papers have appeared in the last year or two giving results
from laboratories along similar lines. I may say at the outset that
my experience leads to conclusions essentially in accord with
results heretofore published.

In the statement of trials I shall by ‘“‘ per cent.” mean per cent.
by volume of the forty per cent. solution of formaldehyde in
water, procured of Scharing & Glatz of New York, and distilled
water. I have found this standard solution known as formalin
fairly constant, and so a reliable solution may be thus quickly made
of pretty constant strength.

I. As a preservative I have tried it on a variety of objects,
using varying strengths with results to be noted.

Experiment r. Small sunfish perfectly fresh was put into two
per cent. formalin Feb. 2; March 1 it was well hardened, without
shrinkage; colors fairly preserved compared with the best alcoholic

332 PROCEEDINGS OF THE

specimens; eyes better; fins extended. The fluid was filtered, the
bottle sealed and given a place on the shelf. July 1, no percep-
tible change observed.

Experiment 2. Two common eels from the market were im-
mersed in four per cent. March 11. July 1, examination showed
that they were in perfect condition as to color, absence of shrink-
age and firmness. The fluid had colored slightly and there was
a sediment. Filtering rendered the specimens fairly representa-
tive museum preparations, fit for either systematic or anatomical
purposes.

Similar trials were made with a variety of fish naked and
scaled, large and small, with uniformly good results. A stronger
solution, up to ten per cent., was found superior for larger speci-
mens; the colors are better preserved in solutions as strong or
stronger than three to four per cent.

Experiment 3. Amblystoma jeffersonianum killed with chloro-
form March I, put immediately into two per cent. formalin ; at
the end of four days it was found to be quite hard, color very
slightly changed, costal grooves plainer than in the living animal.
The fluid was filtered and was still clear July 1, while the prepara-
tion left nothing to be desired.

Experiment 4. Amblystoma tagrinum treated similarly but put
into four per cent. gave excellent results. So far as appears, for
museum, systematic or anatomical, purposes it was no better than
specimen in experiment 3.

Experiment 5. Amblystoma tigrinum was killed April 5, with
chloroform and the mucus washed off with water as in experi-
ments 3 and 4; it was then placed in equal parts of ninety-five per
cent. alcohol and two per cent. formalin. July 1 the specimen
was far superior in appearance and preservation than I have ever
been able to prepare by means of alcohol alone. I cannot say it
is superior to that of experiment 4.

Other Amphibia and several Reptilia have been preserved in
a water solution of formalin and in alcohol-water formalin with
excellent results so far as the time that has elapsed permits of a
demonstration. It seenis to be the best method for Ophidia.

AMERICAN MICROSCOPICAL SOCIETY. 333

Enperiment 6. A tongue of a lion was thoroughly washed in
water to remove blood, mucus and dirt and treated February 5 to
a five per cent. solution of formalin. July 1 it was found well
hardened, in clear fluid with but slight indications of shrinkage.

A camel’s tongue, larynxes of spider monkey, camel and
kangaroo were cleaned, washed and preserved in from two to four
per cent. during February and March; they are at the present
time fine preparations.

Experiment 7. Several trials were made to harden and pre-
serve brains, with different per cents. of formalin, with alcohol
and formalin, and with formalin and bichromate.

I will speak particularly of a few:

1. A brain of spider monkey removed twenty-four hours after
death was placed in four per cent. formalin. In a very few days it
was firm, remained perfectly white and did not noticeably shrink.
July 1, it was all that could be desired in a museum specimen.

2. The brain of a kangaroo was hardened in forty or fifty per
cent. alcohol and two per cent. formalin, equal parts ; the results
were no better ; the specimen was as good and prepared in less
time than by alcohol alone ; and of course much more cheaply pre-
pared.

3. Brains of dogs and cats were prepared for anatomical pur-
poses by adding to the usual bichromate solutions one or two
per cent. formalin, a smaller quantity of fluid was necessary, not
so many changes and a great gain in time; the material was
excellent, not brittle and with fine contrasts.

Before stating my general conclusions as regards this agent in
preserving museum and anatomical preparations, I would like
to add a note as to its usefulness in another direction, viz., in
preserving animals for dissection. If the subject is a small dog,
its blood vessels are washed out with salt solution and 250 cc.
more or less of five per cent. formalin injected. Thus prepared
and kept in a cool place, when not under examination, it may be
used by the dissector day after day and for several weeks without
unpleasantness. The brains of animals thus treated may
be removed for study much longer after death than in any

334 PROCEEDINGS OF THE

other way known to me, at the same cost of material and time.

The advantages of formalin in the lines discussed above
may be summarized briefly thus: It is cheaper than any other
method that gives good results; it gives results in much less time;
the colors are better preserved and there is less change of form
by shrinkage or by swelling; its penetrating power is excellent so
that insects, crustacea, etc., preserved in it are fit for work on the
internal organs; for mollusks and vermes it is also excellent.

Its disadvantages should likewise be stated. It is extremely —
volatile and the jars have to be sealed with care. I have not
had it in use long enough to decide how great an obstacle this
will prove in the museum. Again, the water solution will freeze
and not all museums are at all times above o°C. In such
cases, if not in all, hardening in formalin and transferring to
the lowest per cent. alcohol that will preserve would seem to be
the better and cheaper way. I have not yet had an opportunity
to demonstrate to what extent preparations thus preserved may be
treated to additional alcohol to replace that lost by evaporation,
without a precipitate.

II Experiments with formalin for histological purposes were
undertaken in February last. The visceral organs of Crypio-
branchus were placed while perfectly fresh in a suitable dilution of
borax carmine with five per cent. formalin; the third day the
pieces were washed, the stain fixed and dehydration in sixty per
cent. alcohol commenced; they were passed rapidly through alco-
hols of increasing strength, and at the end of seven to ten days
were infiltrated with paraffin and ready to cut. The liver, intes-
tines and kidney were sectioned, leaving nothing to be desired in
stain or character of the cells.

Parts were at the same time hardened in five per cent. without
bulk stain, also in dilute alcohol and formalin. We concluded
that the rapidity and less shrinkage were alone sufficient to recom-
mend the use of the new agent.

Mr. J. H. McGregor, who was conducting an investigation of
the central nervous system of Cryptobranchus in the same
laboratory, used the agent in combination with alcohol and other

AMERICAN MICROSCOPICAL SOCIETY. 335

agents to his entire satisfaction. He will doubtless give his results
when his paper is printed. Another student used it successfully
_ in preparing the eye.

I may say, in short, that the result of the few months of trial
warrant a high estimate of it for fixing animal tissues.

APPARATUS FOR ILLUSTRATING THE CIRCULATION OF THE
LYMPH.

G. S. Hopkins, D. Sc., Cornell University, Ithaca, N. Y.

In describing the lymph circulatory system before a class in
anatomy, the writer was enabled to present more clearly and
graphically than otherwise would have been possible, the
phenomena of the lymph circulation by means of the apparatus
described in this paper. But in order to appreciate more fully
the excellence of the apparatus as an aid in illustrating the cir-
culation of the lymph, certain points relative to the lymphatic
vessels, their function, origin and the course of the lymph through
them, should be kept clearly in mind. The function of the
lymphatic vessels is to convey the lymph from the tissues back
again to the blood from which it originally came; they also serve
as one of the main channels for conveying nutritive materials,
chiefly fat, from the enteron to the blood.

As to the origin of the vessels, there is considerable diversity
of opinion, but it seems safe to say that they originate as a net-
work of minute connective tissue spaces which so pervade all
living tissues and organs of the body that the individual elements
of these tissues and organs are surrounded by and thoroughly
bathed in the lymph. From these small interstitial spaces the
lymph is gradually collected into larger vessels of which the
largest empty into the great receptaculum chyli of the thoracic
duct, the latter terminating in the great veins at the base of the
neck. The flow of the lymph through the vessels is due to
several causes, the chief one of which is believed to be the differ-
ence in pressure of the lymph at the two extremities of the
lymphatic vessels. At the origin of the vessels the lymph in the
interstitial spaces of the connective tissue stands at a higher press-
ure than in any other part of their course, so that the vessels
lead from a region of high pressure—the lymph spaces of the

AMERICAN MICROSCOPICAL SOCIETY. 337

tissues—to a region of lower pressure, the veins. The high
pressure at the origin of the vessels 1s due, probably, to the fact
that the multitude of minute channels through which the lymph
must pass before reaching the larger vessels, impedes its flow, but
as the lymph is continually transuding through the walls of the
blood vessels, from the blood, it accumulates in the minute lymph
spaces or radicles, and soon stands at nearly as high a pressure
as the blood itself. This high pressure gradually forces the
lymph into the larger vessels, where less resistance to its flow is
encountered and therefore the pressure necessary to force it along
becomes less and less till finally at the terminal end of the thoracic
duct the pressure may be even negative. This difference in press-
ure alone would doubtless cause the lymph to flow in a con-
tinuous stream, but in addition to this other causes as the rhyth-
mical contraction of the walls of the lymph vessels ; contractions
of the muscles including respiratory movements and in some of
the lower vertebrates, lymph hearts as well, help maintain the
circulation.:

With this brief review of the lymphatic system we will pass to
the description of the apparatus used for illustrating some of the
points above mentioned. No claim to originality is made in con-
nection with the apparatus. It was devised by Dr. T.C. Charles
of St. Thomas Hospital, England, and was figured and _briefty
described by him in the Journal of Anatomy, Vol. XXII., p. 435.
The only modification made by the writer was to substitute a ves-
sel which opens at the top rather than the bottom, thus some-
what simplifying the construction of the apparatus from the
original. .

A glance at the figure will show the parts comprising it. The
jar, B, should hold from three to four liters, and should have a
cover that can be fastened on securely, making the jar air-tight.
In the cover were drilled three holes through which the glass
tubes a,b, c, were passed. A bit of rubber tubing was placed in
each hole in the cover, so that when the tubes were inserted the
joints were air-tight. To the lower ends of a and c were tied
Beces of prepared intestine, used by butchers as casing for saus-

338 PROCEEDINGS OF THE

age. The lower end of b opens directly into the general space
of the vessel. Attached toa, is a rubber tube with a syringe
bulb, H. From c extends another tube, v, which is joined at
any convenient point by the tube t. From b the tube x extends
to the vessel R. Some sponges, S, may be placed in the vessel
B, although this is not necessary.

To use the apparatus, the large vessel B is filled with water
and the rubber tubes connected as shown in the figure. By
means of the bulb, H, water is forced through the pieces of in-
testine, 1, and on through the tube v. As the water flows through
the pieces of intestine, some of it passes through their thin walls
into the space outside, and soon raises the pressure in B, so that
the liquid is forced through the openings of the tube b, and along
the tube x, into the vessel R. As the liquid accumulates in R
the pressure in this vessel is also raised sufficiently to force the
liquid out through the tube t. In this apparatus the vessel B
may be taken to represent the body—the simulation will be more
perfect if the vessel be partially filled with sponges to represent
the tissues of the body. The intestinal tubes, I, represent the
blood capillaries of the body. The heart is represented by H
the arteries by the tube extending from H, and the veins by the
tube v. The tube b represents the lymphatic radicles which, as
we have already seen, are believed to open directly into the

AMERICAN MICROSCOPICAL SOCIETY. 339

minute spaces of the body ; these lymph spaces are here repre-
sented by the spaces between and in the sponges. The tube x,
represents the lymph vessels extending from the lymph radicles
tothe receptaculum chyli. The vessel R represents the receptac-
ulum chyli. The tube t represents the thoracic duct which
extends from the receptaculum to the veins of the neck.

As illustrated by this apparatus the sole cause of the circula-
tion of the lymph is the higher pressure of the lymph at the
origin than at the termination of the vessels. In the living body
we have already seen that inequality of pressure is thought to be
the chief cause of the circulation, but in addition to it, muscular
contractions, respiratory movements, and in some of the lower
forms, lymph hearts as well, help to keep up the flow.

SOME NEW POINTS IN PHOTO-MICROGRAPHY AND PHOTO= —
MICROGRAPHIC CAMERAS,

W. H. Watms.Ley, Chicago, III.

Photography in connection with the microscope, Photo-Micro- |
graphy as universally termed, is now such an every-day affair that
one unacquainted with the facts can scarcely realize that only a
few years ago, its practice was confined to a very few enthusiasts
at home and abroad, and its results looked upon as interesting
and beautiful, but practically valueless. Yet such was the case
in the later ’70s, when Dr. J. J. Woodward was producing
his marvelous Photo-Micrographs at the Army Medical Museum
in Washington. His work was such a vast step in advance of
any that preceded it, as to attract the attention of the entire
scientific world, and in many respects it has never been excelled.
Being confined, however, almost exclusively, to the resolution
and delineation of difficult test objects, as diatoms and rulings on
glass, its sole practical value consisted in the improvements in
objectives, brought about by the efforts of many eminent opti-
cians, both American and foreign, to meet his exacting require-
ments. ‘‘ The Battles of the Lenses’ will doubtless be remem-
bered by most of you, and there can be little doubt that the
wonderfulimprovements in and perfection of modern objectives, are
due in a large measure to the impetus given by Dr. W oodward in
his efforts to obtain the best, for use in photo-micrography. Indeed,
Nobert saw for the first time the lines of his nineteenth band in a
photograph made by Dr. Woodward with one of these object-
olasses.

But even more marked in their effect upon photo-micrography,
than the improvements in objectives, have been the changes in
photographic methods, since Dr. Woodward's day. He worked
within his camera itself; his work-room constituting a gigantic
camera box, to which no ray of light was admitted during the

AMERICAN MICROSCOPICAL SOCIETY. 341

focusing of the object and exposure of the plate, save that which
passed through the microscope. The source of light varied
according to time and circumstances. Usually he employed that
of the sun through an immense heliostat, which is still in use at
the museum. But asa large portion of his work was done at
night, he also called in the aid of various artificial illuminants :
Magnesium ribbon, the lime light, and toward the end of his
work the electric are lamp, each with unvarying success. Not
being an expert photographer himself, this portion of his work
was done by a professional, and it may not be uninteresting to
know that collodion or wet plates alone were used. Gelatine
emulsions were as yet unknown, or practically unattainable.

It will thus be seen, that in addition-to his own wonderful skill
as a manipulator, Dr. Woodward had at his disposal, un-
limited Government resources, as aids to his researches and ex-
periments. Indeed it may be safely said, that no other worker in
the same field was ever so liberally provided with the means for
prosecuting it. The cost in every direction was deterrent to the
most of less fortunate mortals, and, as stated before, but for the
many radical changes since made in photographic methods,
photo-micrography would still be the recreation of the few, in-
stead of the practical realization of the many.

With the general introduction of gelatine dry plates, of such
exalted sensitiveness that the light of an ordinary lamp sufficed
for exposures with quite high powers; and portable cameras
adapted for use with any microscope having an inclinable body,
the making of a negative of almost any microscopical object was
brought within reach of every worker. The printing, however,
was not so satisfactory, especially where large numbers were
required in the illustration of papers or books, but, as in the past,
the steady advance in photographic methods speedily supplied
the existing need ; photo-gravure and other process-methods re-
produced the negative in positive form with wonderful exactness,
delicacy, and cheapness, so that at the present day, papers upon
any subject may be illustrated in a manner utterly unattainable a
short decade ago. By the same means the optical lantern has

342 PROCEEDINGS OF THE

been brought to the fore as one of the indispensable adjuncts of a
well appointed lecture room. Ready sensitized plates of thin
slass are now furnished at reasonable cost by several eminent
makers, by use of which one can make his own slides and from
his own negatives, either by contact printing, or by reduction in
the camera, if he is provided with one adapted to the latter pur-
pose. In short, the microscopist of the present day finds at his
disposal the ready means of illustrating his work at every stage ;
and one who publishes his notes without illustrations, finds him-
self at a disadvantage as compared with his more progressive
brother.

It is not the object of this paper to do more than glance at the
new points in photo-micrography which have fallen under the
notice of the writer during the past score of years, and to call
attention to a new form of camera combining some novel features,
which he has recently introduced under the name of the “‘ Auto-
graph.” It may be not uninteresting, however, if a very brief
allusion is made to his preceding work in this direction, as he
takes a perhaps pardonabie pride in the belief, that to his efforts
a considerable portion of the present acknowledged value and
popularity of photo-micrography are due.

Without the slightest previous knowledge of photography in
any form, I became greatly interested in its application to the
microscope by my friend and mentor, the late Dr. Woodward.
Many days passed in his work-room during my then frequent
visits to Washington, gave me a keen relish for and desire to en-
gage in this fascinating pursuit, without, however, the slightest
expectation of ever being able to do so. The costly and com-
plicated apparatus and appliances necessary placed it quite
beyond my reach. But in a few years, with the advent of port-
able cameras and gelatine dry plates, 1 became one of the numer-
ous army of amateur photographers, and very shortly afterward
by means of a make-shift attachment to my microscope produced
my first Photo-Micrograph, a little affair,on a plate scarcely three
inches square, and not at all well done, but esteemed as almost a
sacred treasure to the present day.

AMERICAN MICROSCOPICAL SOCIETY. 343

From this crude beginning was evolved the instrument known
by the lengthy title of the ‘“ Enlarging, Reducing and Copying
Photo-Micrographic Camera,” which I placed on the market
early in 1882. It met with instant and generous recognition,
and has maintained its popularity steadily ever since. So far as
I have been able to learn, it was the first American camera for
this purpose to be produced commercially.

As indicated by its title, this camera is adapted to a variety of
purposes. Any microscope withan inclinable body may be used
with it in making a Photo-Micrographic negative, with or with-
out an ocular. The latter is the usual method, since much
more light is transmitted by the objective alone, whilst the long
extension bellows permits a high magnification with any given
lens. Dr. Woodward always worked without an eyepiece.
With an ordinary photographic lens, the instrument may be used
for enlarging, reducing and copying; the very long bellows ren-
dering it particularly valuable for the latter purpose. It is, how-
ever, unnecessary to go into fuller details of the construction and
capacities of this camera, since it is already so widely known.

Certain defects or rather want of adaptibility to a// purposes in
this camera led to the designing and construction of my latest
box, the ‘ Autograph.” It was somewhat bulky, especially in
the larger sizes, which in the too often contracted work-room is
a hindrance to its habitual employment. It could be used only
in a horizontal position, and the microscope must have a joint
permitting inclination of the body, a feature not found in many
otherwise excellent instruments, especialiy those of German
manufacture. For use with these stands a vertical camera is of
course indispensable, as it is when the object is free in a fluid,
such as yeast spores, blood, pus, milk corpuscles, etc., etc.
But for the great majority of work, the horizontal position is the
better, especially where it is desirable or necessary to use the
direct rays of light from a lamp, without the intervention of the
mirror. To meet these varying demands, the “ Autograph ”
camera was designed, and it is believed successfully. It may be
described as follows, the dimensions given being those for a

PROCEEDINGS OF THE

344
camera carrying 4x5 plates, the only size so far constructed.

They would have to be proportionally greater for a larger sized

box.

4
—
=
—
~~,

The base or
wood twenty-six inches lon;
to insure steadiness on any table or other support, the front

platform is of polished mahogany or other hard
standing upon three very short feet,

ry
D>?

AMERICAN MICROSCOPICAL SOCIETY. 345

end being heavily weighted beneath. At the other end of the
platform a stout frame of japaned iron, twenty-four inches in
length, with joint close to its base is firmly bolted. This frame
carries the camera, which slides freely in two parallel grooves
milled in its upper surface and can be secured at any desired
point by a stout screw passing through a slot running the entire
length of the frame, in its centre. The joint permits the frame
carrying the camera to be placed and firmly held, in either verti-
cal or horizontal positions, or inclined at an angle of 45°. For
copying or making lantern slides from negatives by enlargement
or reduction, the latter position is almost indispensable and is one
of the most valuable ‘‘ New Points”’ embraced in the ‘“ Auto-
graph’ camera as will be seen presently.

The camera box is furnished with leather bellows of best
quality, extending twelve inches, which has been found to be the
most generally useful, though double that length can be employed
if necessary or desirable. It is fitted with a reversible back, car-
rying both focusing screen and plate holder, a most desirable
feature, as it greatly facilitates the proper arrangement of the
object in relation to its position on the plate, where the micro-
scope is unprovided with a rotating stage. The ground glass
focusing screen is mainly useful for arranging the illumination,
and the object in the field of view, its surface being too coarse to
permit fine focusing with high powers. It may, however, be
easily removed from its frame and replaced by a sheet of plate
class, when by means of a suitable lens the nicest adjustment can
be made. The plate holder is double, and fitted with inside kits
to carry 34% x 44%,2% x 2% or lantern plates, in addition
to those of its full size, 4 x 5 inches.

The front is fitted with a removable plain board, to which an
ordinary photographic lens may be attached, and an additional
board carrying an extension (which may be oblong or cone-
shaped as desired), with an opening in its front end to receive the
tube of the microscope. The flange of the photographic lens can
be attached to this extension front, if it be necessary to increase
the length of the camera in copying and enlarging. |

346 PROCEEDINGS OF THE

When the camera is used in the vertical or inclined positions,

both coarse and fine adjustment screws are within easy reach of
the hand and may be manipulated in connection with observance
of the focusing upon the screen. But when the horizontal posi-
tion is assumed, the distance is too great from screen to micro-
scope to permit this, and other means must be provided. A
short rod, turning freely in suitable bearings, is attached to the
base board on right hand side of the camera. To the end near-
est the observer is fitted a large milled head, and to the other a
pulley wheel, with V-shaped groove in its periphery ; a corre-
sponding groove being also turned in the Micrometer Screw of
the microscope. ‘This pulley-wheel slides freely upon the rod or
shaft, allowing it to be placed in line with the fine adjustment
screw, where it is firmly held by a small set screw. <A fine cord
passed around the two grooves, suffices to move the micrometer
screw when the milled head is revolved. This, of course, is an
old and well-known device, but being a good one has been
adopted in this case.

ee

AMERICAN MICROSCOPICAL SOCIETY. 347

The extension of the iron carrying frame beyond the end of
the base board, with the additional weight of the camera acting

’ asa lever, having a tendency to tip the front of the base upward,

a heavy iron bar forming one of the short tripod supports, is
fitted beneath the front of the board, entirely obviating any such
danger. The platform itself is of sufficient length to carry micro-
scope, lamps and bull’s-eye condensing lens on stand, the added
weight of which serves also to give increased steadiness to the
whole apparatus.

It is not within the scope of this already too lengthy paper to
say anything in regard to the making of a negative from a micro-
scopic object. This must be left to another occasion. But it
may not be amiss to glance for a moment at the source of light
for making the exposures. Diffused daylight reflected from the
mirror is probably the most generally useful illuminant, and the
various positions in which the “ Autograph’’ camera can be
placed give the day-worker many advantages in its use. But
most of us have, perforce, to do our work by night with artificial
light. Fortunately there are many of these, some one of which
is available to everyone. The lime light, the electric arc, the
Welsbach gas burner and the humble, omnipresent petroleum
lamp, are all good, varying mainly in the differing lengths of ex-
posure required with each. And finally we have the new
acetylene gas lamps, which place in the hands of every worker,
the itdeal light for Photo-Micrography.

A few words as to the value of the ‘“ Autograph” camera in
copying and in making lantern slides by enlargement or reduc-
tion, and J will tax your patience no longer. For both these pur-
poses, the camera, fitted with a photographic lens of not more
than nine inches focal length and inclined at the angle 45°, is to
be placed near a window and its base cleared of the microscope,
lamp, etc: A carrying frame with its upper surface parallel with
the camera front, takes their place upon the platform, to which .
the book or print to be copied is fastened. The lighting, focus-
ing and all such subsequent details are of course familiar to
every photographer. I cannot even hint at them here and would

348 PROCEEDINGS OF THE

suggest that if the copy is for lantern purposes, it would be well
to make it at once of the proper size to permit printing by con-

tact, thus effecting a considerable saving of time.

Negatives of microscopic objects are generally made consider-
ably larger than the dimensions of a lantern slide, though in some
cases, as a minute diatom for instance, they are much too small.
In either case the lantern slide must be made by reduction or
enlargement as necessary. For these purposes, the camera is
arranged precisely as for copying, except that its front end must
face the window and be close to the latter. A large sheet of
white paper is to be laid upon the platform as a reflector, and on
this the stand used in copying (and carrying a frame containing the
negative), must be placed. A focusing cloth or other covering

AMERICAN MICROSCOPICAL SOCIETY. 349

is then spread over the space between the frame and camera, so
that no light may enter the lens, save that which passes through
the negative. The camera is then moved to or fro upon its
ways, until the image projected upon the screen is of proper
dimensions, when it is to be fastened in that position, the focus
sharpened by moving the bellows, and the balance of the neces-
sary work, of exposure and development, done in the manner
familiar to all who have mastered the simple mysteries of photo-
graphic manipulations.

The accompanying cuts fully illustrate the various methods of
using the instrument.

THE QUESTION OF CORRECT NAMING AND USE OF MICRO
REAGENTS.

V. A, Laruam, M.D}, DUD:Se FRAMES.) Chicago, in

All technical workers in microscopy must have found much
annoyance in their experimental work. Teachers in the practical
laboratories of the medical and scientific schools all testify to
the great difficulty in securing reliable dyes and reagents. Few
technologists are such expert chemists as they ought to be, and
therefore must rely upon chemists and the manufacturers for their
information. This puts the technologist at a great disadvantage,
especially when he finds that the chemists do not always agree!
For example, in endeavoring to show the plexus of fibrils in the
nuclei of liver cells by maceration in Sulpho-Cyanide of Potas-
sium, I met with the greatest dissatisfaction. A large local drug-
house secured a sample, which I found to be the Sulpho-Cyanate
of Potassium, ‘‘ the same thing,’ I was assured. A well-known
chemist was questioned, and replied, ‘“‘ No, they are not the
same.’ A new sample, Merck’s, had an entirely different name,
yet I was told it was the article ordered. Two prominent im-
porting chemists disagreed as to the identity of the samples and
their correct names, and there the matter was dropped. If we
cannot trust expert chemists to know such reagents, how can we
amateurs know on whom to rely, with such a loose nomencla-
ture P

Let us look through a text-book on Microscopy, and note
the indiscriminate use of Benzol, Benzole, Benzine, Methyl
Blue, Methylene Blue; Vesuvin for Bismarck Brown; Iodine
Green for Methyl Green! These are serious errors, from the
stand-point of solubilities, if nothing else; some dyes being
soluble in water, others in alcohol. Indeed, different samples of
the same anilin dye will differ in their behavior to solvents, some
making a perfect solution, others leaving a more or less insoluble

AMERICAN MICROSCOPICAL SOCIETY. 351

residue. In some micro-books we find such statements as the
following: ‘Iodine Green—this is another name for Methyl
Green.” If we are lucky enough to. obtain a true, reliable Iodine
green, we find the price is much higher, and the color a lighter
Green. But the true dye has been largely adulterated with
Methy! Green, and is now very difficult to obtain. Indeed, the
terms Methyl! Green, Methyl Iodide, Methyl Chloride and Methyl]
Bromide are applied almost indiscriminately by dealers, who sub-
stitute one for the other. For double staining Iodine Green is
excellent when carefully used, and, according to Prof. H. Gibbes,
a most permanent stain. It certainly does not work on the same
tissues as the Methyl Green does. The differences will be found
in the Proceedings of this Society for August, 1892.

Again some authors include Gentian under the head of Dahlia
Violet ; but if we test these dyes by their reaction to certain cells,
we get different results, Methyl Violet being used for Amyloid
Degeneration, while only Dahlia will show the ‘“ Mastzellen.”
Dahlia has a red tint, while Violet has a strong blue tint. They
are both nuclear stains, but Gentain Violet requires the addition
of Glacial Acetic Acid to make it directly nuclear. And we must
also remember the stain mostly sold as Methyl Green is not pure,
but contaminated with Methyl Violet, giving many of the re-
actions of the latter, as for example, with amyloid.

Amongst the many formulas for Picro-Carmine we find a
variety of results which it is difficult to explain. It has always
been some source of surprise to me that, when ordering a certain
maker’s dyes, another’s should be sent out from reputable firms,
either openly, as a substitution, or wrongly under a false name.
These fraudulent imitations never give the results wanted. Per-
sonally, I cannot get the best counter-staining nor the true
double-reaction from any other Picro-Carmine than Ranvier’s. I
have Hoyer’s, Konig’s and some others, and, though taking
every care to follow the directions of each respective authority,
have not obtained satisfactory results. It is noteworthy that
the directions for using these dyes do not agree in the various
manuals and text-books extant. The student, after endeavoring

352 PROCEEDINGS OF THE

to follow first one and then another, with a like poor result each
time, is apt to grow skeptical with regard to the stain, and to
wonder whether anyone has succeeded with it! Who does not
read some article, bearing on a subject in which he has been
especially interested, and wishing to verify the author’s account
of the extra clear demonstration of cell varieties, etc., finds out
the name and sends for a sample of that particular stain or
reagent? Eagerly attending to every detail, in the hope of secur-
ing exact results, he too often finds that he has had his trouble.
for his pains, and has succeeded only in wasting. time, material
and patience! Failure may arise from any one of many causes.
Sometimes the author has not really meant anyone else to suc-
ceed, and has purposely omitted the designating letter or number
of the dye, or some trifling modification in the technique, or the
omission may arise through oversight or careless, ambiguous En-
clish! Or, again, the house from whom the reagent was ordered
may send a different dye of the same name, whose chemical con-
stitution may work havoc with the process in hand, for which it
is not adapted. For instance, the use of Ammonia Alum instead
of Potash Alum may seem to the theoretical histologist or pathol-
ogist of no great moment; but the technical laboratory worker
knows this is not so. In making a blood stain of Alum Eosin,
if Ammonia Alum be used in Alcohol, an almost solid residue is
obtained, showing incompatibility, and the result for Blood is
very poor if any at all. In Logwood, for Delafield’s formula, he
specifies Ammonia Alum—no other would do in that particular
case. In Klein’s Logwood, it is said, two points must be remem-
bered, viz.: Potash Alum, without Ammonia, and the English
Extract of Logwood, which mustbe used. When the logwood so-
lution, as made here from the chips, becomes reddish-brown, it is
usually found to contain some acid impurity which hinders stain-
ing. On the converse, in the Logwood solutions made from the
continental extracts, the reddish color is nearly always obtained
and gives the best results.

Eosin is said to bea blood-stain. Granted; but with the yel-
low, red, blue and orange varieties, some soluble in alcohol and

a

ad
AMERICAN MICROSCOPICAL SOCIETY. 353
some in water—‘ which shall we buy ?’”’ is the question. Profes-

sor Von Jaksch, I believe, is the only authority who comes forward
and specifies for his particular work, by quoting in his ‘‘ Chemical
Diagnosis’ and by giving in his lectures the name of the Eosin
number and firm, and insists upon that brand. Would that we
had more like him, thus saving time, money, and room in the
office or laboratory.

Picro-Carmine is a dye with which it is extremely difficult to
obtain the respective colors as well as the combination colors.
One has too much yellow remaining, and even using the carmine
will not control it; neither will the washing. Some say ‘Wash
in alcohol after staining, as the Picric Acid is so readily soluble
in water.’’ Others say ‘‘Use acidulated water, then alcohol and
then fresh alcohol.” Some say ‘Plain water,” and so forth. It
depends not so much on the dye as on the maker’s formula.
Ranvier’s has, until recently, held the chief place, and why it has
been superseded is a question. Principally, I believe, it is not so
readily obtained from the Gargon of the Laboratory as in years
gone by, and the quality is hard to obtain just right, and we are
now content with poor substitutes. Some one will say ‘ Well,
make your own dye, you have the formula in some book.” The
average worker with the microscope has neither time, money nor
laboratory, nor, worst of all, the requisite skill and experience to
make his dyes, some, as Picric Acid, being too dangerous to
work with, in the hands of every one. We perhaps do turn to
our book, for example ‘“‘Lee’s Vade Mecum,” and we get a rough
or not always reliable translation, or, as I said before, the minutiz
are not there. Lee carefully gives the authority, and we think
we can get the original article from some reference library. ‘‘Not
there” is usually the result. And this same formula, which is so
indefinite, is copied into future journals and text-books without
correction. ;

There are several kinds of Vesuvin in the market. Two kinds
only are of value, the rest being useless in staining tissues.
Vesuvin is freely soluble in water, while Bismarck Brown is very
slightly so, hence should not be substituted for it.

24

354 PROCEEDINGS OF THE

Thoroughly good staining requires :

1. Fresh tissues, properly fixed and hardened.

2. Good, reliable dyes; the identical ones recommended for
such use.

3. Knowledge of their acting properties, whether for direct or
indirect nuclear work or for plasma and ground staining.

4. Acquaintance with their power of substitution, as Malachite
Green combining with /chsin Hydrochloride ; this causes the
Tubercle Bacillus to lose its specific stain and color.

5. The various solubilities of the stains employed ;

(a) In washing and cleaning, whether acid, neutral or alco-
holic agents should be used.

(b) In mounting media, as Balsam in chloroform, etc.,
when we know that there are some dozen preparations of one dye
on the market, differing in color, solubility and histological action,
we must be careful to buy the one adapted to the case we wish
to investigate; and this is not an easy matter. Griibler and Min-
der supply at least fourteen varieties of Safranin, and in the two
houses the brands have different names and symbols. So that
only the most exact ordering will insure the proper specimen.
The student, directed to ‘“‘stain with Safranin’”’ may well feel at a
loss to know which Safranin to use. Has microscopical technique
no position, that it is placed so? Are the workers ignorant, or
what is the trouble? Few microscopists are the all-round scien-
tists that they should be, to do this work, and very few of them
are so careful as they should be to note the technical side, the
reaction to the tissues and the chemical combinations that result.
We do not know enough of Chemistry and Physiology before
we use methods of preparation. We do not always make our
comparisons under similar conditions as to the action of various
agents upon the same tissue. Who does not know how one
agent will shrink a tissue to one-quarter its size, and another
distend it to double? Still another will pick out one variety of
tissue from the rest, say the mucous, only, andso on. A student
is allowed a year or two of study in foundation principles, and
then may take a ‘‘Major”’ to investigate, after doing three to

AMERICAN MICROSCOPICAL SOCIETY. 355

six months’ Histology. He cannot give any truly reliable results,
and yet it is mainly such crude work that forms the basis on
which microscopy rests. To the true technologists we turn for
information, but alas! how few are they, and how scattered over
the world! So engaged in good work are they, and so trivial do
minor points seem to them, that their reports of methods and
results are brief and include only main points. We, at a dis-
tance from their laboratories, cannot tell what special dye or
reagent should be bought to secure a like result, because of the
unfortunate brevity, nay the zzaccuracy, of their report.

Some firms employ an expert chemist, for economy’s sake, to
test and analyze their dyes. He passes a given stain on to his
co-worker, the experimenting microscopist, who tests it, not only
in Histology, Pathology and Bacteriology, but botanically, so as
to define its best results; and as such a reagent it is then sold by
the firm, every sample being thus carefully tested to secure uni-
form, good results. Not many years ago I was told that enorm-
ous sums were spent in buying what seemed identical dyes, but
which, on testing, yielded different reactions, the firms themselves
not being able to secure the same dye in every case. Anilin
workers spend small fortunes in efforts to secure the true ““moon-
light color” for fashion purposes, as yet without success. But
so great is the hope of fame and fortune, a fabric worker told me,
“we are willing to try, and succeed some day.” So long as
fabric work, holds the premier place, so long will micro work
suffer, because of financial questions. Suppose a special blue is
wanted—a dye is made to match the sample in shade, irrespective
of the chemical combinations, provided the color is correct. It
is all one to the manufacturer whether it is passed over Sulphuric,
Nitric or Hydrochloric Acid; but to the microscopist such heed-
lessness would bring speedy disaster. In certain diseased or
normal tissues, the combination of the dye with the secretions or
fluids of the tissue would form a new chemical compound .
altogether. Indeed the chemistry of stains and their peculiar
combinations with individual tissues, if fully understood, would, I
believe, be of great value in throwing light on many intricate

356 PROCEEDINGS OF THE

problems in disease, by specifying the tissue in which the changes
occur. This is exemplified in the case of Amyloid Degeneration,
whose exact location in the vascular system is demonstrated by
means of its affinity for Methyl Violet and Methyl! Green. Al-
though its histological prototype is unknown as yet, since no
known normal tissue stains the same, its antecedent may yet
be discovered in some of the less complex substances of the
earlier degenerations, hyaline or fibroid, possibly. We should
then be able to trace its development from the normal tissue, as
well as its position by its specific reactions to certain dyes.

At present, perhaps the most fashionable dye in use is the
Triple Stain. A curious coincidence is the indefinite mode of
using the same. (Consult Lee, p. 161, 3d Edition.) There we
are told “it is troublesome to prepare, but may be readily obtained
prepared from Gribler. The receipt is as follows, ete.: The
method given is for making from the separate dyes. Happily,
Heidenhain tells us from which firm to buy, and specifies all
details, as to exact name of dye, and manner of washing, and so
forth. Lee says that it may be put into the hands of beginners.
This is hardly correct, in my experience, and in that of many
others more competent to judge. There are few stains so variable
on the market. Dr. Gribler has a mixture of the three colors
in the dry state. For this, you are told “to dissolve 0.4 grm of
the mixture in 100 cc. of water and add 7 cc. of a 0.5 per-cent.-
solution of Saurefuchsin.”” No further directions are given.
The majority of the text-books advise after staining sections to
wash in alcohol, xylol, and mount in Canada Balsam in xylol.
This stain is often very hard to work with, sometimes succeed-
ing only when the solution is old; sometimes only when freshly
made. Sometimes one color predominates over the others, and
we have to keep on adding and experimenting for some time. If
blood covers are examined by the washing in alcohol it will be
found that the corpuscles show only the Aurantia stain and what
seems to bea bluish deposit of pigment which will not easily
wash off. In blood-work, water only must be used, and that
very carefully. Some find Thayer’s formula by far the most

AMERICAN MICROSCOPICAL SOCIETY. 357

successful. The trouble with the Triple Stain, for blood, is: its
great fading, the indistinctness of the neutrophilic cells, especially
the green staining, and the great unreliability. of the stain.
Possibly if some successful worker would give the precise modes
of use, better results would be secured by others. The state-
ment so frequently made that-the Triple Stain is an easy, reliable
one to work with ought to be disputed. Out of some 5,000
specimes made by men in London, Vienna, Berlin, Paris and the
United States—mentioned in their work—showing Leukemia,
Plasmodium Malariae, tissue-sections of various kinds, Protozoa
of Cancer,—out of all these only five specimens showed to any
advantage, clearly and distinctly. A teacher in a medical school,
eminent in scientific work, assured me he never felt certain when
his work with the triple stain would be good, and that he always
took the precaution to stain some covers by other methods,
especially if valuable material. Not one of the many text-books
published within the last three years but has the same rough-and-
ready statement (copied from Lee) (?) withoutany remarks whatever.
And yet I have four bottles of the triple dye, bought from a most
reputable German firm, directly and indirectly, no two of which
have the same color in powder, solution or in results! Can we
wonder, then, from such instances, that so much discussion is
found, especially in such high-power work as that of the Proto-
zoan Theory for Cancers, in Bacteriology and blood-work ?
What is the use of publishing manuals for Clinical Diagnosis,
Microscopy and the like, if modes of preparation and using are
omitted? Every worker does not have the opportunity for study
he would wish; but he expects the details for practical work to
be clearly given in a book designed and published to aid him in
some special line of investigation. He is too often wholly dis-
appointed.

To further the microscopist’s work, would it were possible one
good journal could be published to cover Microscopy—or better °
four, one each for North, South, East and West. Then all matter
properly belonging to the subject could be published in the
journal of its own district, which would thus become a valuable

358 PROCEEDINGS.

medium of communication, and save its subscribers loss of time,
money and labor. Who would not infinitely prefer to subscribe
to one journal devoted exclusively to the subject he is turning
his attention to, instead of being compelled to pay for several
journals on general medicine or scientific subjects? The single
journal, when bound for reference, is a handy volume for the
library, quickly consulted. With the miscellaneous scientific
periodicals, one may waste hours in looking over the bulky files
to find—say a vaguely remembered article on the preparation of
Tubercle Bacillus in Milk. It may have been in “The American
Naturalist,” ‘‘ Journal of Hygiene,”
medical journals, or in ‘‘ Annals of Botany’’(! !) instead of in a

any one of a half dozen

Journal of Microscopy, or one specially belonging to that depart-
ment. Let us aim to secure concentration of the sciences, each
in a proper journal of its own, carefully edited, indexed and
published ; instead of having, as heretofore, scattered articles,
microscopical and of merit, appearing in cheap or questionable
papers all over the country. Few of us are rich enough to buy
many journals, or have the space to allow them to accumulate,
nor yet the time to clip valuable cuttings from poor journals.
Almost any one can afford ove good journal, giving him the new
methods, apparatus, bibliography, articles on his own study in
each of the scientific branches in which he is especially interested.
He would be glad to pay for the privilege of having his photog-
raphy in a journal on photography, anatomy in an anatomical
journal, and microscopy in a microscopical, more particularly in
that of the National Society.

A NEW WAY OF MARKING OBJECTIVES.

Wituiam C. Krauss, M.D., F.R.M.S., Buffalo, N. Y.

That every microscopist in demonstrating to his classes in
histology or pathology has been annoyed in determining the
focus of the various objectives when a nose-piece is used, no one
will dare contradict. The small letters or figures, designating
the focus, engraved on the body of the objective have often to be
sought for with great vexation, necessitating at times the removal
of the lens from the nose-piece, or in revolving the lens or nose-
piece so that the number will be discernible. Sometimes the
microscope must be upturned or the investigator is obliged to
place his head on the level with the table, thereby upsetting
reagent bottles or provoking other mirth and mischief before he
is abled to focus his tube correctly and with safety on some
valuable slide. This has been the writer’s experience, and now
that he has finally and so simply solved this perplexing question,
submits his discovery to the society, with considerable feeling of
pride and gratification.

On the diaphragm in the large part of the objective, or the
end that is screwed to the nose-piece, the designation of the
lens may be engraved, so that when the nose-piece is revolved the
designation of the various lenses will be at once visible. The
investigator with one eye at the ocular, need not change his
position in bringing all the lenses under the body tube, but can
with the other eye see the lens as it swings into place, and can
focus with coarse and fine adjustment accordingly. The writer
has been well pleased with the focal lengths of the Zeiss ob-
jectives, necessitating but one focusing for all the different lenses -
especially of the dry system. Working with these lenses, marked
as I have indicated, on a triple or Pein nose-piece, is not
only a pleasure, but a great convenience.

360 PROCEEDINGS.

The accompanying illustration which is purely diagrammatical,
represents a triple nose-piece with the objectives %, 34 and 1-5
attached, removed from the body tube. The nose-piece is so re-

volved that all the upper surfaces of the lenses are visible, disclos-
ing their designation.

———

IMPROVEMENTS IN OIL=SECTIONING WITH COLLODION.

Simon H. Gacg, Ithaca, N. Y.

Everyone who works much with any method must almost in-
evitably find out certain modifications which make the method
more efficient and more easily applied. If one has also to adapt
methods to the needs of laboratory students, modifications be-
come almost imperative, and frequently the necessary improve-
ments naturally grow up in aiding students to meet their special
requirements.

In the perfection of the collodion method of sectioning the
two greatest advances were made: (1) in learning to handle the
sections with paper and (2) in clarifying the tissue and the sur_
rounding collodion mass with an essential oil. Weigert (’85)
seems to have been the one to first publish the paper-method of
handling sections, and so far as I can judge, Bumpus (’92) first
published a practicable method of clarifying the imbedded tissue
and mass in oil, and of using oil to float the sections on the knife as
they were cut. In the “‘ oil method” of Bumpus the object must
be stained 27 foto and, after transferal to the slide and most of the
oil absorbed, mounted immediately in balsam. /y Zofo staining is
not by any means applicable to all tissues and for all work,
especially morphological work where large organs or entire
animals are sectioned. To obviate the defects in Bumpus’ method
Dr. P. A. Fish made two important improvements: (1) He
mixed with the essential oil (oil of white thyme) one-third its
bulk of castor oil, thus avoiding the rapid drying, and (2) he

‘fastened the sections to. the slide after the oil had been well re-

moved with absorbent paper by adding a small amount of a mixt-
ure of ether and alcohol. The ether and alcohol were allowed
to evaporate until the surface began to look glazed, then the slide
bearing the sections was placed in ninety-five per cent. alcohol
to remove the oil, then passed through seventy per cent. and

362 PROCEEDINGS OF THE

thirty-five per cent. alcohol to water, if a watery stain like hema-
toxlin was to be used. Finally after the stain was washed away
with water, the sections were dehydrated by again passing them
through thirty-five per cent., seventy per cent. and ninety-five per
cent. alcohol and cleared and mounted in balsam, thus rendering it
possible to use any stain desired after the sections were cut. This
method as perfected by Dr. Fish is truly admirable.

The improvements which have been evolved by my own ex-
perience are mostly in the direction of simplification and cheap-
ening. For the sake of those who may not be familiar with the
collodion method there will be given in very brief form the entire
method, pointing out at the end the special improvements which
it isthe purpose of this paper to put on record :

1. Fixing and Hardening—The tissue, organ, embryo or
animal of which sections are desired is fixed, and hardened by
any of thestandard or special methods.

2. Dehydration —The tissue is thoroughly dehydrated by using
one or more changes of plentiful ninety-five per cent. alcohol or
by the use of absolute alcohol. It is better not to dehydrate
more than twenty-four hours. By changing the alcohol three or
four times, two or three hours is sufficient for small pieces of
material, and five or six hours will suffice for the larger objects.

3. The tissue is thoroughly saturated with a mixture of equal
parts of sulphuric ether and ninety-five per cent. or stronger
alcohol. This requires two to five hours for small objects. Over
night doesno harm, The ether and alcohol complete the de-
hydration and prepare the tissue more perfectly than alcohol
alone for the infiltration with collodion.

4. Infiltration with Thin Collodion—The ether-alcohol is
poured off, and a mixture of thin collodion (ether and ninety-five
per cent. alcohol, equal parts, 100 cc.; soluble cotton one and one-
half grams). Two or three hours will suffice for objects two or
three millimeters in thickness. A stay of one or more days does
no harm. The larger the object the more time is needed.

5. Infiltration with Thick Collodion.—The thin collodion is
poured off and thick collodion added (ether and alcohol, equal

AMERICAN MICROSCOPICAL SOCIETY. 363

parts, 100 cc.; soluble cotton*, six grams). For very small objects
four or five hours will suffice to infiltrate, but for larger objects a
longer time is necessary. The tissue does not seem to be injured
at all in the thick collodion, and a stay in it during a day or even
of a week or more is more certain to insure a perfect infiltration.

6. Imbedding and Hardening the Mass.—The tissue may be
imbedded in a paper box such as is used for paraffin im-
bedding, or in any of the other boxes devised for paraffin. It is
better, if paper is used, to put a very small amount of oil on the
paper to prevent the collodion from sticking to it. Vaseline
spread over lightly and then all removed so far as possible with
a cloth or with lens paper gives the right surface. For small
objects it is more convenient to imbed immediately on a holder
that may be clamped into the microtome. Cylinders or blocks
of glass, vulcanite, wood and cork have all been recommended
and used. A cork of the proper size is most convenient and for
many purposes answers well. Some collodion is put on the end
of the cork and a pin put near one edge. The tissue is transfer-
red from the thick collodion to the cork and leaned against
the pin. Drops of the thick collodion are then poured on the
tissue and by moving the cork properly, the thick, viscid mass

*The substance used in preparing collodion goes by various names,
soluble cotton or collodion cotton is perhaps best. This is cellulose nitrate,
and consists of a mixture of cellulose tetranitrate C,, H,, (NO,)4 O,, and
cellulose pentanitrate, C,, H,; (NO,)5 O;. Besides the names soluble
and collodion cotton, it is called gun cotton and pyroxylin. Pyroxylin is
the more general term and includes several of the cellulose nitrates. Cel-
loidin is a patent preparation of pyroxylin, more expensive than soluble
cotton, but inno way superior to it.

Soluble cotton should be kept in the dark to avoid decomposition. After
it is in solution this decomposition is not so liable to occur. The decompo-
sition of the dry cotton gives rise to nitrous acid, and hence it is best to
keep it in a box loosely covered so that the nitrous acid may escape.

Cellulose nitrate is explosiye under concussion and when heated to 150°
centigrade. In the air, the loose soluble cotton burns without explosion.
It is said not to injure the hand if held upon it during ignition and that it
does not fire gun-powder if burned upon it. So far as known to the writer,
no accident has ever occurred from the use of soluble cotton for micro-
scopical purposes. I wish to express my thanks to Professor W. R. Orn-
dorff, organic chemist in Cornell University for the above information.

364 PROCEEDINGS OF THE

may be made to surround and envelop the tissue. Drops of
collodion are added at short intervals until the tissue is well sur-
rounded; and then as soon as a slight film hardens on the sur-
face, the cork bearing the tissue is inverted in a wide-mouth vial

Fig. 1. Wide-mouth vials for the purposes of Histology and Embryol-
ogy. These represent the two vials natural size that have been found
most useful. They are kept in blocks with holes of the proper size. They
are especially useful for objects treated by the collodion method as described
in this paper.

of considerably larger diameter than the cork (Fig. 1). The vial
should contain sufficient chloroform to float the cork. The vial
is then tightly corked. In imbedding somewhat larger objects
on the end of a cork or other holder it is frequently advantage-
ous to wind oiled paper around the holder or cork, tie it tightly
and have the projecting hollow cylinder sufficiently long to re-
ceive the object. The tissue is then put into the cylinder and
sufficient collodion added to completely immerse it. As soon as
a film has formed over the exposed end, the cork may be inverted
and immersed in chloroform as described above.

7. Hardening and Clarifying the Collodion —After a few hours

AMERICAN MICROSCOPICAL SOCIETY. 365

the collodion is hardened by the chloroform. If it acts long
enough the imbedding mass is rendered entirely transparent if no
water is present. Whenever the collodion is hard, whether it is
clear or not, the chloroform is poured off and the clarifier added
(the clarifier is made by mixing thoroughly xylene* three parts
with castor oil one part by volume.) In a few hours the imbed-
ded mass will become as transparent as glass and the tissue will
seem to have nothing around it. Sometimes the collodion remains
white and opaque for a considerable time. So far as the writer
has been able to judge, this is due to moisture. If one breathes
on the mass too much while imbedding, or if it is very damp in
the room, the opacity may result. Sometimes, in objects of con-
siderable size, this may remain for a week. This is the exception,
however, and if the mass seems sufficiently hard and tough the
cutting may proceed even if the clarification is incomplete.

8. Cutting the Sections—For collodion sectioning a long
drawing cut is necessary in order to obtain thin, perfect sections.
The object is therefore put in the jaws of the microtome at the
right level and the knife arranged so that half or more of the
blade of the knife is used in cutting the section. It is advan-
tageous also to have the object placed with its long diameter
parallel with the edge of the knife. The surrounding collodion
mass should be cut away so that there is not more than a thick-
ness of about two millimeters all around the tissue. This is to
render the diameter of the end to be cut as small as possible.
The smaller the object the thinner can the sections be made.
With an object two to three millimeters thick and not over five
millimeters wide and a good sharp knife, sections 5y to 6 can
be cut without difficulty. When knife and tissue are properly
arranged the tissue is well wet and the knife flooded with the
clarifier. (For wetting object and knife during the sectioning a
mixture of xylene four. parts and castor oil one part is rather
better than the ordinary clarifier as it is somewhat more fluid).

*The hydrocarbon xylene (C, H,,) is called xylol in German. In En-
glish, members of the hydrocarbon series have the termination ‘‘ene” while
members of the alcohol series terminate in ‘‘ol.”

366 PROCEEDINGS OF THE

Make the sections with a steady motion of the knife. Then
draw the section up toward the back of the knife with an artist’s
brush and make the next section. Arrange the sections in serial
order on the knife blade till enough are cut to fill the area that
the cover-glass will cover.

g. Transferring the Sections to the Shide—lIf the clarifier has
evaporated so as to leave the sections somewhat dry, on the knife,
add a small amount. Take a piece of thin absorbent, close-
meshed paper* about twice the size of a slide and place it directly
upon the sections. Press the paper down evenly all around and
then pull the paper off the edge of the knife. The sections will
adhere to the paper. Place the paper, sections down, on a slide,
taking care that the sections are in the desired position on the
slide. Use some ordinary lens-paper or any absorbent paper and
press it down gently upon the transfer paper. This will absorb
the oil, and then the transfer paper may be lifted from the slide.
The sections will remain on the slide.

10. fastening the Sections to the Shde—Drop just enough
ether-alcohol (equal parts of sulphuric ether and ninety-five per cent.
alcohol) on the sections to moistenthem. This will melt the collo-
dion and fasten the sections to the slide. Allow the slide to remain
in the air till the surface begins to look slightly dull or glazed.

11. Removing the Oi from the Sections—As soon as the
ether-alcohol has evaporated sufficiently to leave the surface dull,
place the slide in a jar of ordinary commercial benzin. It may
be left here a day or more without injury to the sections, but if
moved around in the jar the oil will be removed in three to five
minutes. From the benzin transfer to a jar of ninety-five per
cent. alcohol to wash away the benzin. One may use alcohol in
the beginning, but it dissolves the oil far less rapidly than the

“Various forms of paper have been used to handle the collodion sections.
It should be moderately strong, fine meshed and not liable to shed lint, and
fairly absorbent. One of the first and most successful papers recommended
is ‘‘closet or toilet paper.” Cigarette paper is also excellent. In my own
work the silky Japanese paper called ‘‘Usago” paper has been found almost
perfect for the purpose. Ordinary lens paper or thin blotting paper for ab-
sorbing the oil is used with it.

1

AMERICAN MICROSCOPICAL SOCIETY. 367

benzin. The slide may remain in the alcohol half a day or more
if one wishes, but a stay of five minutes or a thorough rinsing of
half a minute or so by moving the slide around in the alcohol
will suffice.

12. Staining the Sections.—(A), With an alcoholic stain. If
an alcoholic stain containing fifty per cent. or more alcohol (for
example hydrochloric acid carmine in seventy per cent. alcohol)
is used, the slide may be. removed from the ninety-five per cent.
alcohol, drained somewhat and then the stain poured upon the
sections or preferably the slide immersed in a jar of the stain.
The stain is finally washed away with sixty-seven per cent. or
stronger alcohol, the sections dehydrated in ninety-five per cent.
alcohol, cleared and mounted in balsam.

(B) With an aqueous stain like hematoxylin, etc.—If an aque-
ous stain is to be used the sections must first be rinsed with water.
In the past the plan for changing sections from ninety-five per
cent. alcohol to water for example, has been to run them down
gradually, using seventy-five, fifty and thirty-five per cent. alcohol
successively. Each percentage may vary, but the principle of a
gradual passing from strong alcohol to water was advocated.
On the other hand, I have found that the safest method is to
plunge the slide directly into water from the ninety-five per cent.
alcohol. The diffusion currents are almost or quite avoided in
this way. There is no time for the alcohol and water to mix, the
alcohol is washed away almost instantly by the flood of water.
So in dehydrating after the use of watery stains, the slide is
plunged quickly into a jar of ninety-five per cent. alcohol. The
diffusion currents are avoided in the same way, for the water is
removed by the flood of alcohol. This plan has been submitted
to the severe test of laboratory work and has proved itself per-
fectly satisfactory.

In staining with a watery stain then, the aa bearing the sec-
tions is transferred from the ninety-five per cent. alcohol and
plunged into a jar of water, and either allowed to remain a few
minutes or moved around in the water a moment. Then it is
placed horizontally and some of the stain placed on the sections,

368 PROCEEDINGS OF THE

or preferably it is immersed in a jar of the stain; in case of im-
mersion, however, the slide should stand vertically or nearly so,
then any particles of dust, etc., in the stain will settle to the bot-
tom of the vessel and not settle on the sections. When the sec-
tions are stained they are thoroughly washed with water either
by the use of a pipette or by immersing in a jar of water. They
may then be counterstained with some general dye like eosin or
picric acid or mounted with but the one stain.

13. Mounting in Balsam.—After the sections are stained they
must be dehydrated and cleared before mounting inbalsam. For
the dehydration the slide is plunged into a jar of ninety-five per
cent. alcohol. This removes the water so rapidly that the in-
jurious diffusion currents, which loosen or tear the sections,
are avoided. For clearing after the dehydration the slide is
drained of alcohol and put down flat and the clearer poured on,
or the whole slide is immersed in a jar of clearer (Carbol-turpen-
tine is a good clearer,—Crystals of carbolic acid melted, two
parts, turpentine three parts; or carbol-xylene clearer,—melted
crystals of carbolic acid two parts, xylene three parts). Clearing
usually is sufficient in a few minutes, a stay of an hour or even
over night does not injure most sections.

In mounting in balsam the clearer is drained away by standing
the slide nearly vertically on some blotting paper or by using the
waste bowl and standing it up in the little funnel. (Fig. 2.)
Then the balsam is put on the sections or spread on the cover-
glass and that placed over the sections.

The improvements here advocated are: (1) The use of
xylene and castor oil, the xylene taking the place of the
essential oils heretofore recommended for “oil-sectioning.”’ This
is much cheaper and in every way as good, perhaps better; it has
also the advantage of being nearly odorless. Imbedded tissues
have been preserved in the mixture nearly six months without
deterioration.

(2) The term clarifier is proposed to avoid using clearer,
which can then be restricted to its original use, viz.: to displace
the alcohol from sections and render them transparent before

AMERICAN MICROSCOPICAL SOCIETY. 369

adding balsam, the clearer being always quite freely miscible with
both alcohol and balsam.

(3) In the avoidance of diffusion currents in changing sec-
tions fastened to a slide or cover from liquids of greatly different
densities, as from alcohol to water, by plunging the slide directly
into the new liquid, the great flood of the new liquid serving to
remove the previous liquid so rapidly that there is no chance for
the destructive diffusion currents which tend to tear the sections
in pieces or loosen them from the slide.

Fig. 2. WASTE BOWL FOR HISTOLOGICAL WorRK.—(From the Reference
Hand-Book of the Medical Sciences, Supplement.) The glass rods are for
resting the slide horizontially and the funnel for draining them.

REFERENCES.

For articles and abstracts or references to articles on the col-
lodion method one can profitably consult the Journal of the Royal
Microscopical Society, the Zeitschrift fiir wissenschaftliche
Mikroskopie and the Proceedings of the American Microscopical
Society. Special references for the preceding article are:

’92. Bumpus, H.C. A new method of using collodion for

serial section cutting. American Naturalist, Vol. xxvi., 1892,
25

A [oh PROCEEDINGS OF THE

pp. 80-81. In this paper is given the method of sectioning by
the use of thyme oil for clarification and for floating the sections
on the knife. For Bumpus’ ‘‘oil-method,” see also Journal Roya]
Microscopical Society, ’92, p. 438 and Dr. Fish’s papers below.

’°93. Fish, P.A,—Recent Histological Formule. Reference
Handbook ofthe Medical Sciences, supplement, 1893 ,pp. 434-436.

’°93. Fish, P.A—A new clearer for collodionized objects.
Proceedings American Microscopical Society, vol. xv., 1893, pp.
86-89. It wasin this paper that Dr. Fish showed the advantage
of adding a fixed oil to the clarifier to be used for ‘‘oil-section-
ing,’ and also that sections made by this method could be
fastened to the slide and stained as desired.

‘91. Gage, S.H.—Albumenizing the slide for the more certain
fixation of serial collodion sections. Proceedings American
Microscopical Society, vol. xili., 1891, pp. 82-83. It is shown in
this paper that if the sections are to be subjected to long-continued
manipulation they may be more surely fixed to the slide by at
first albumenizing the slide with egg albumen, one to 200 of water
and allowing the albumen to dry on the slide.

93. Lee, A. B—The Microtomist’s Vade Mecum, 3d Ed.,
London and Philadelphia, 1893. In this work is given an excel-
lent account of the collodion and celloidin methods and the _his-
tory of the two methods, with modifications.

85. Weigert, C—Ueber Schnittserien von Celloidinprapara-
ten des Centralnervensystems zum Zwecke der Markscheidentar-
bung. Zeit. f. wiss. Mikr., Bd. 2, 1885, pp. 490-495.

In this article is described a method of using paper for the
handling of celloidin sections. On p. 491 he says, “closet
paper” had been used for a considerable time for this purpose in
the Pathological Institute of Heidelberg.

83. Viallanes—Rech. sur I’hist. et le dév. des Insects. 1883.
Viallanes recommends in this work the use of chloroform for the
hardening of the collodion.

THE IMPROVED SYRACUSE SOLID WATCH GLASS.

A..CLirFoRD Mercer, M.D., F.R.M.S., Syracuse, N. Y.

The original Syracuse solid watch-glass, described in the Pro-
ceedings of this society for 1884, on page 178, and shown here
in Fig. 1, had the concavity of a watch-glass, a shallower concave

bottom and a solid mass of glass between. It was soon widely
used in the microscopical laboratories of this country, and to a
small extent in Europe.

Use has suggested improvements. The capacity has been in-
creased. The new form makes a secure pile, Fig. 2, without the

uN

rs &*"eAa''d*

oe

5 .
(Cut two-thirds actual size ) (Cut two-thirds actual size.)

Fig, 2.

supporting frame required by the first form. All edges are
rounded and are, therefore, less liable to be chipped. When the

372 PROCEEDINGS.

rounded rims are accidentally wet they stick less than flat rims.
With pencil or ink the user can write or print on the smoothly-
ground bevelled surface, Fig. 3. The writing can be erased with
a wet cloth. What is written on the ground surface can be seen
when the user looks horizontally at or obliquely down upon the
pile, or vertically down upon a single dish. To the same surface
may be attached labels. When lifting a dish, a slight bevel given
the surface just beneath that ground furnishes a better hold than
the vertical surface of the first form, All the surfaces of the new
form are more smooth.

Like the first form in other respects; it rests solidly upon the
table or microscope stage ; is not liable to be overturned and its
contents spilled; is transparent and can be used over black,
white or colored paper, enabling the worker to use such back
grounds to his work as permit him to watch the progress of
staining, washing and the like to best advantage. In it, on the
microscope stage, can be examined from time to time, or dis-
sected and studied, transparent tissues while in water, alcohol,
oil of cloves or other bath, the worker being able to detect and
reject unsatisfactory specimens at any step in the process of prep-
aration. In it, when piled, specimens may remain for long stain-
ing or soaking without becoming dirty and with little or no loss
of fluid by evaporation. It is useful on the stage of a dissecting
microscope, and finds a place among the dishes of the bacteriol-
ogist. The new form is already widely known and used.

A METAL CENTERING BLOCK FOR MOUNTING.

M. Priaum, Pittsburgh, Pa.

The proper centering of objects upon slides has a practical
purpose outside of all considerations of mere beauty of appear-
ance. As most microscopes have, or should have, a stop upon
the stage against which the slide can rest, an object properly
centered thereon will therefore automatically be within the optical
axis of the lenses, and thus save time and patience in finding what
is wanted.

A card-board, having thereon a diagram of a slide is very
primitive in construction, and often exasperating in its use. The
card will warp, absorb and retain fluids, the slide must be held
in place, thus monopolizing one hand otherwise and better to be
employed, and still will often slip and spoil the centering. This
very inconvenience was useful ; it caused a desire for something
better. This was found in a metal block shown in the accom-
panying diagram. It is of brass, 2x4 inches in size, 3-16 inch
thick, with a depression for the slide 1-16 inch deep, and % inch
stop at one side.

Its advantages are numerous: being of metal, it is therefore
permanent and durable ; of considerable weight, therefore solid.
and firm; non-porous, therefore easily cleaned; and having a
groove for the slide, it is held in place, therefore convenient for
mounting and labeling.

A NEW METHOD OF MAKING AND FINISHING WAX-CELLS.

M. Priaum, Pittsburgh, Pa.

After several years’ testing, the following described method of
making wax-cells has answered every demand, whether for fluid
or dry mounting.

So that the wax may better adhere, a ring of asphalt (in benzole)
cement, wider than the intended ring, is first drawn upon the
slide. It is best to have such ringed slides in stock so that the
asphalt has thoroughly set and seasoned. A mixture of wax
and paraffin, in equal parts, is obtained by melting to a boil, and
with it, upon the turn table, a cell drawn of whatever depth re-
quired, and immediately well covered with the asphalt cement,
with special care to cover the inner and outer edges nearest the
glass, so that the wax is enclosed on all sides by the cement.
The paraffin hardening the wax, and the wax making the paraffin
less brittle, make together a cell which will resist any change of
temperature; the asphalt is used as an additional precaution in
that direction.

Such cells, of various depths, should be kept on hand for
thorough drying, the longer the better, to guard against any pos-
sible shrinkage; for which, however, there is in this cell very little
danger. For mounting, whether dry or fluid, the crest of the
cell should be covered with a very thin ring of the same mixture
of wax and paraffin, and the cover-glass firmly pressed down on
it. Mounts in such cells, with glycerin as a medium, have proved
of easy manipulation and in every respect satisfactory.

After the cover-glass is in position, the following method of
finishing the slide is recommended.

As the wax-cell has been enclosed with a benzole cement, the
cover-glass should be fastened with a cement having a different
solvent. Shellac (in alcohol) serves this purpose best. This

AMERICAN MICROSCOPICAL SOCIETY. 375

would finish the slide. If, however, it is desired to make the slide
still more permanent, as well as an object of beauty, the follow-
ing described process will well repdy the additional labor. After
the shellac has well dried, put on a ring of zink-white cement
entirely enclosing the shellac, and, within a few minutes, before
the zink has fully set, ring it with any color of King’s lacquer (I
have tried no others) in any manner taste might direct. The
lacquer unites with the zink, and gives it the appearence of porce-
lain. Around the cover-glass, and around the cell on the slide,
draw a ring of bronze paint. This will hide any defects in ring-
ing and give the slide a very handsome appearance, with, after
some practice, really little extra work.

Hints. In using shellac cement care must be taken that it be
free from bubbles. Their presence, if the cement has been stirred
shortly before use, has spoiled many slides. This, however, will
apply to the use of any cement.

It must be apparent that the use of many cements on one slide,
should make it almost indestructible. This is a decided advan-
tage, and will save the annoyance of periodical repairs to the
slide; but it requires care not to overload the cell with too many
layers of each cement. After the shellac on the cover-glass is
well set, the next cement, zink-white, should be well thinned. A
watch-glass with benzole, if it is solvent, will be a help in this
direction.

To obtain a variety of shades of the color of the lacquer used,
have a watch-glass, with its solvent, alcohol, handy. By its use
the color of the lacquer, whatever it may be, will yield an infinite
variety in the appearance of the slides, especially if aided by a
differenee in the size and location of the rings. I have used only
King’s blue lacquer, and have no two slides alike in appearance.
This color and the bronze, above referred to, compliment each
other and give beautiful results.

To finish a slide having no cell the same method, herein sug_.
gested, will be found very satisfactory. A single cement ring
may serve for a time; but as all cements have volatile solvents, it
is but natural that, with thorough drying, the remaining solid

376 PROCEEDINGS.

should become brittle. This is prevented by the use of several
layers of cement having different solvents. But it is claimed that
a mount prepared for the use of high powers should have no
ring to enclose the cover-glass, or else a very thin one. This
does not seem reasonable. Whatever the medium in such
mounts, usually balsam, it will certainly deteriorate if not pro-
tected by acement ring. The higher the power to be used the
more valuable the object mounted, and the more reason, there-
fore, to make the slide permanent. The heavy ringing, it is
claimed, obstructs the necessarily close approach of the lense.
This is true if asmall cover-glass is used. But there is no reason
why the smallest object, as one single diatom, well centered, can
not be*covered with a large cover-glass, and fastened with several
layers of cement. Thus every objection is removed, and a valu-
able mount amply protected.

CONSTITUTION.

ADOPTED AT ROCHESTER, N. Y. 1892.

ARTICLE OL,

This Association shall be called the AMERICAN MICROSCOPICAL
Society. Its object shall be, the encouragement of microscopical
research,

ARTICLE II.

Any person interested in microscopical science may become a
member of this Society upon written application and recommenda-:
tion by two members, nomination by the Executive Committee,
and election by a majority of the members of the Society present
at any regular session of the Society. Honorary members may
also be elected by the Society on nomination by the Executive
Committee.

ARTICLE III.

The officers of this Society shall consist of a President and two
Vice-Presidents, who shall hold their office for one year, and shall
be ineligible for re-election for two years after the expiration of
their terms of office, together with a Secretary and Treasurer, who
shall be elected for three years and be eligible for re-election.

ARTICLE IV.

The duties of the officers shall be the same as are usual in
similar organizations ; in addition to which it shall be the duty
of the President to deliver an address during the meeting at which
he presides ; of the Treasurer to act as custodian of the property
of the Society, and of the Secretary to edit and publish the Pro-
ceedings of the Society.

ARTICLE V.

There shall be an Executive Committee, consisting of the
officers of the Society, three members elected by the Society, and

AMERICAN MICROSCOPICAL SOCIETY.

the past Presidents of the Society, and of the American Society

of Microscopists.
PLR TICUEM de

It shall be the duty of the Executive Committee to fix the .

time and place of meeting and manage the general affairs of the
Society. ‘
ArTICLE VII.
The initiation fee shall be three dollars, and the dues shall be
two dollars annually, paid in advance.
ArticLe VIII.

The election of officers shall be by ballot.
ARTICLE IX.

Amendments to the Constitution may be made by a two-thirds’
vote of all members present at any annual meeting, after having
been proposed at the preceding annual meeting.

By-Laws.

I,

The Executive Committee shall, before the close of the annual
meeting for which they are elected, examine the papers presented
and decide upon their publication or otherwise dispose of them.

All papers accepted for publicalion must be completed by the
authors and placed in the hands of the Secretary by October Ist
succeeding the meeting.

ip

The Secretary shall edit and publish the papers accepted, with
the necessary illustrations, in four numbers. The first number
to be issued not later than October 1, after the meeting, and the
remaining three numbers at intervals of not more than three
months.

III.

The number of copies of Proceedings of any meeting shall be

decided at that meeting. Each author of papers accepted an

AMERICAN MICROSCOPICAL SOCIETY.

published shall be entitled to twenty-five separates of his paper
and as many more at the cost of publishing, as he may request
at the time that his manuscript is sent to the Secretary.

IV.

Papers accepted for publication by the Society shall not be
published elsewhere until after they have appeared in the Proceed-
ings of the Society, except by consent of the Executive Com-
mittee.

V.

No applicant shall be considered a member until he has paid
his dues. Any member failing to pay his dues for two consecu-
tive years and after two written notifications from the Treasurer
shall be dropped from the roll, with the privilege of re-instatement
at any time on payment of all arrears. The Proceedings shall
not be sent to any member whose dues are unpaid.

VE

The election of officers shall be held on the morning of the
last day of the annual meeting. Their term of office shall com-
mence at the close of the meeting at which they are elected, and
shall continue till their successors are elected and qualified.

VIL.

Candidates for office shall be nominated by a committee of five
members of the Society. This committee shall be elected by a
plurality vote, by ballot, after free nomination, on the second day
of the annual meeting.

Vit.

All motions or resolutions relating to the business of the So-
ciety shall be referred for consideration to the Executive Com-
mittee before discussion and final action by the Society.

EX.

- Members of the Society shall have the privilege of enrolling
members of their families (except men over 21 years of age) for
any meeting on payment of one-half the annual subscription (1.00).

Approved by the Society, August 11, 1892.

~]

LIST OF MEMBERS.

The figures denote the year of the member’s election, except ‘78, which marks an original
member. The Proceedings are not sent to members in arrears, and two years’ arrearage
forfeits membership. (See Article VII. of the Constitution.)

[embers Elected at Ithaca, N. Y., 1895.

For addresses see regular list.

BuscuH, FREDERICK CARL. HumpHREY, O. D.

BusH, BertHa E., M.D. KERR, ABRAM TUCKER.
CONSER, HARRY NEWTON. KRAFT, WILLIAM.

Dorr, 8S. HOBART. LANDACRE, FRANCIS L.
EGELING, B. F. GUSTAVUS. McGREGOoR, JAMES H.
EIGENMANN, C. H. MERCIER, A.

ELuiotT, L. BAYARD. SECOR, JABIN.

GREEN, DUDLEY T. Wison, Mrs. Mary R.
ABERDEEN, ROBERT, M.D., F.R.M.S., 782 ; J ‘ Syracuse, N. Y.
ACKER, GEO. N., M.D., ’91, : 913 11th St., N. W., Washington, D. C.
AINSLEE, CHAS. N_, 92, : : j Rochester, Minn.
ALLEGER, WALTER W., M.D., 94, . 906 S, St. N. W., Washington, D. C.
ATWOOD, E. S., 79, 3 : : - 242 South St., New York.
AtTwoop, H. F., F.R.M.S. 78, ; 5 3 > 3 Rochester, N. Y.
AYRES, MorRGAN W., M.D., ’87, —.. : ‘ Upper Mont Clair, N. J.
BAHRENBURG, W.N., M. D., ’89,_ . 919 Washington St., St. Louis, Mo.
BALL, MICHAEL, VALENTINE, 793, . 1182 Spruce St., Philadelphia, Pa.
Banks, NATHAN, 791, ‘ ; ; : Sea Cliff, Queens Co., N. Y.

BARNSFATHER, JAMES, M.D., ’9i,
Cor. Fairfield Av. and Walnut St., Dayton, Ky.

BaRR, Prof. CHas. E., ’90, 3 : : Albion College, Albion, Mich.
BARTGES, ARTHUR F., 90, . 3 j A : Akron, Ohio.
BasseETT, CHAS. H., 82, . ; : , 40 Bedford St., Boston, Mass,
BauscH, EDWARD, ’78, : : 179 N. St. Paul St., Rest stan Ney:
BauscuH, Gro. R., 80, , ; - . 20 Arcade St., Rochester, N. Y.

BauscH, Henry, 86, . ; : , : i aie rochester, N.Y.

AMERICAN MICROSCOPICAL SOCIETY.

BauscH, WILLIAM, 88, . x ; : . Rochester, N. Y.
BELL, CLARK, 792, : : 3 ; ; BT Ereaiwar, New York City.
BENNETT, HENRY C., 793, 4 , . 256 W. 42d St., New York City.
BIGELOW, EH. F., 7925 9 : . : . Portland, Conn.
Biscog, Prof. THomas D., ’91, : ; 404 Rront St., Marietta, Ohio.
BuEILeE, A. M., M.D., ’81, . 2a Es . 218 King Av., Columbus, Ohio.
BootH, AUGUSTINE RUE, M.D., ’87, . 520 Market St., Shreveport, La.
Booru, Mary A., F.R.M:S.,°82; — . 982 Byers St., Springfield, Mass.
Boyce, JAMES C., Es@., ’86, : : Carnegie Bi'dg., Pittsburgh, Pa.
BoYER, C. S., A.M., 792, : é . 38228 Clifford St., Philadelphia Pa.
BROMLEY, ROBERT INNIS, M.D., 93, . : ; ; : Sonora, Cal.
BROWN, Miss L. S., 792, : ; ; : Angelica, N. Y.
Brown, N. Hownanp, 791, : : 33 Ss. 10th St., Philadelphia, Pa.
BROWN, ROBERT, ’85, : . Observatory Place, See Haven, Conn.
BRUBAKER, J. C., M.D., ’91, E 1501 Jackson Boulevard, Chicago, Ill.
BRUNDAGE, A. H., M.D., 794, us . 1158 Gates Av., Brooklyn, N. Y.
BULL, JAMES EDGAR, EsqQ., ’92, 253 Broadway, New York City, N. Y.
BULL, JAMES; ’88;. {unis . . . Hanging Rock, Ohio.
BURRILL, T. J., Ph D., F.R. M. S., ‘ . Champaign, IIL.
Burt, Prof. E>Dwarp A pols : "vagal College, Middlebury, Vt.
BUSCH, FREDERICK CARL, 95. : 179 Richmond Av., Buffalo, N. Y.
Busi, BertHa E., M.D., 95’ P : 808 Morse Av., Rogers Park, Il.
BUTTOLPH, Harry, T., C.E., 80, . ‘ 13 City Hall, Buffalo, N. Y.
CAMPBELL, D. P., M.D., ’88, ; é : . Green Springs, Ohio.
CARTER, JOHN E., 86, : : ; Garinantowe Philadelphia, Pa.
CASSATT, M., M.D., ’86, : : ; . 818 Elm St., Cincinnati, Ohio.
CHEESEMAN, FE. L., 84, . ; é Knowlesville, N. Y.
CHESTER, ALBERT H., A.M., ’88, eh alleen New Brunswick, N. J.
Cuapp, GEo. H., ’86, : 3 ; , . 116 Water St., Pittsburgh, Pa.
Cuapp, W. A., M.D., ’80, ‘ : . 11 E, Main St., ie Albany, Ind.
CLAYPOLE, AGNES M., ’94, : ; : Akron, Ohio,

CLAYPOLE, EpitH J ANE, Phebe ove Pi 98,

Wellesley College, Wellesley, Mass.

CLAYPOLE, EDWARD, W., B. Sc., F.G.S.,, ’86, ; : . Akron, Ohio.
Cops, Cuas. N., A:M., ’86, : i : 117 Lake Av., Albany, N. Y.
CONSER, HARRY NEWTON, 95 y 29 N. 4th St., Sunbury, Pa.
Coon, H. C., A.M., M.D., ’82, Alfred University, Alfred Centre, N. Y.
Coopst, A. F., M.D., ’86, ‘ ; ; : P Oil City, Pa.
CoucH, FRANCIS G., ’86, . : : 846 Braise ay, New York City.
Cox, CuHas, F., F.R.M.S., 85, —.. Grand Central Depot, New York City.
Cox, Jacos D., LL. D., F.R.MS., '82, : : : Cincinnati, Ohio.
CRAIG, CHARLES FRANCIS, M.D., ‘94, . 21 Balmsforth Av., Danbury, Conn.
CRAIG, THOMAS, ’93, ; . 259 Water St., Brooklyn, N. Y.
CRANDALL, RAND PERCY, ae D., 1, Navy Pay Office, San Francisco, Cal.
Curtis, Lester, M.D., : : 35 University Place, Chicago, TIL

ae

AMERICAN MICROSCOPICAL SOCIETY.

DANIELS, C. M., M.D., 83, : ; : 315 Jersey St., Buffalo, N. Y.

Davis, W. Z., ’86, , F : : ‘ : s ‘ Marion, Ohio.
Deck, Lyman, L., M.D., ’90, ; ‘ ; . Salamanca, N. Y.
DENISON, C. H., 791, : ; P 146 Willow St., Brooklyn, N. Y.
Dops, A. WILSON, M.D., ’90, : : : Fredonia, N. Y.
Dorr, S. Hopart, Ph.G., 95 . : 887 eeoeperk Av., Buftalo, N. Y.
DOUBLEDAY, Henry, H., Esq., 790, lade ltSime IN We S Wasting. DRG:
DRESCHER, W. E., ’87, 3 ‘ a 1033, Rochester, N. Y.

DunuaM, E. K., M.D., _—_Bellevue Eecnital Med. College, New York City.

Eastman, Lewis M., M.D., F.R.M.S., 82, 772 Lexington St., Baltimore, Md.

EGELING, B. F. GUSTAVUS, ‘95 Apartado, No. 4, Monterey, Mexico.
EIGENMANN, C. H., 95, : : ? , Bloomington, Ind.
ELLIOTT, Prof. ARTHUR H.. 91, . : 4 Invi ing Place, New York City.
Euiott, L. BAyarp, 95, Bausch & Lomb Optical Co., Rochester, N. Y.
ELLIS, SYLVANUS A., 92 : : E 13 Clifton St., Rochester, N. Y.
ELSNER, JOHN, M.D., ’83, ; : . P.O. box 454, Denver, Col.
ELWELL, A. T., 89, . ; : . 16 Pearl St., Council Bluffs, Iowa.
ENTRIKIN, F. W., M.D., ’87,_ . : : Lock Box W., Findlay, Ohio.

EWELL, MARSHALL D., LL.D., M.D., F.R.M.S., ’85,
59 Clark St., Chicago, Ill.

FEIEL, ADOLPH, M.D., ’81. : P : 520 Main St., Columbus, Ohio.
FELL, Gro. E., M.D.. F.R.M.S., 778, . . %2 Niagara St., Buffalo, N. Y.
FELLOWS, CHAS. S., F.R.M.S., ’838, P. O. box 966, Minneapolis, Minn.
Freips, A. G., M.D., ’82, F : : . . Des Moines, Iowa.
FIELDS, Miss Eva H., 94 , ; Des Moines, Iowa.
FISH, PIERRE A.,D .Sc., 790, Bureau Patina acute Washington, D. C.
FISHER, Max. ’98, : ; ; Zeiss Optical Works, Jena, Germany.
FLINT, JAMES M., M.D., 791, . . 1117 Vermont Ay., Washington, D. C.
Fox, Oscar C., 92, _ . : : ; Washington, D. C.
ORD SD Mhe DD: 781, momele College, Elmira, N. Y.
FosTER, AGNES WINSLOW, 98, : . Brewster, Mass.
FRANCIS, MARK, D.V.M., ’87, } iGalloge Station, Brazos Co., Tex.
FRENCH, GALE, M.D., ’86, : : 5219 Center Av., Pittsburgh, Pa.
FRENCH, S. H.’ M.D., ’82, Z f 40 Church St., ei tae WL, AL

FULLER, CHAS. G., M.D., F.R.M.S, ’81,
39 Central Music Hall, Chicago, III.

GAERTNER, FRED., M.D., 87, 3 f 3613, Penn. Av., Pittsburgh, Pa.
GAGE, SIMON H., B.S., Prof., ’82, . Cornell University, Ithaca, N. Y.
GAGE, Mrs. SUSANNA S. PHELPS, ’87, ; ; P Ithaca, N. Y.
GILBERT, JOHN L., M.D., 94, _—.. ; 7 17 ane Ay., Topeka, Kansas.
GLEASON, S. O., M.D.,’80,_ _.. : : , : : Elmira, N. Y.
GOETZ, Rev. GEORGE, 791, : : ‘ . Rochester, Beaver Co., Pa.

GREEN, DUDLEY T., 95. : ; : 11 Mill St., Binghamton, N. Y.

AMERICAN MICROSCOPICAL SOCIETY.

GREGORY, JAMES C., M.D., 98, . . Nyack-on-Hudson, N. Y.
GRIFFITH, BENJ. W., 92, . 3 1241 W. State St., Los Angeles, Cal.
GRIFFITH, J. D., M.D., ’87,

Rialto Bldg. cor. 9th and Grand, Kansas City, Mo.

GUTTENBERG, Prof. GUSTAVE, ’91, ; High School, Pittsburgh, Pa.
HaaG, D. E., M.D., F.R.M.S., 86, . 1121 Washington St , Toledo, Ohio.
HANAMAN, C. E., F.R.M.S , 779, : : : Box 527, Troy, N. Y.
Hanks, HENRY G., 86, . . 718 Montgomery St., San Francisco, Cal.
HARDING, LAWRENCE A., B. Sce., Ph.D., 90, . 167th St., St. Paul, Minn,
HATFIELD, JOHN J. B., 82, s 189 Arsenal Av., Indianapolis, Ind.
HAYS, JOS cass hans i Joule 98 William St., New York City.
HEINEMAN, H. Newton, M. D., "91, 6 : 60 W. 56 St., New York.

HERVEY, Rey. A. B., Ph.D., 78, ‘ Canton, St. Lawrence Co., N. Y.
HILu, Rev. ELLSwortu, J., 78, 7100 Eggleston Ave., Sta. O, Chicago, IL.

HILL, HERBERT M., Ph.D., 5 . University of Buffalo, Buffalo, N. Y,
HoEnny, A. J. 87, : , . . 8681 N. Grand Ave., St. Louis, Mo.
HOLBROOK, M. L., M.D., ’82, c ; 46 E. 21st St., New York.
Ho.umss, E. S., D.D.S., ’84, : 5 2 . Grand Rapids, Mich.
Hoop, THOMAS B., M.D., ’91, , 1009 O St., Washington, D. C.
HOPKINS, GRANT S., D.Sc., 90, . : : = & Ithaca, N. Y.
Hoskins, WM., 79, : . . ‘Bt S. Clark St., Chicago, Ill.
HOwarkD, Curtis C., M.D., "83, Beanie Medical College, Colamiie Ohio.
Howe, Lucien. M.D., F.R.M.S., 78, . 87 W. Huron St., Buffalo, N. Y.
HuBER, Rev. E. D., ’82, . . 1800 KE. Fayette St., Baltimore, Md.
HuBER, G. CARL, M.D., F.R.M.S., ’90, 24 E. Ann St., Ann Arbor, Mich.
HUMPHREY, O. D., M.D:, 95, . : : : 227 W. 8th St., Erie, Pa.
Hunt, JOSEPH H., M.D., 794, F 1085 Bedford Ave , Brooklyn, N. Y.
BYATION, DS 78: Se : ; : : : 69 Burling Lane, N. Y.
JACKSON, CHEVALIER Q., M.D, ’87 ‘ : 63 6th Ave., Pittsburgh, Pa.

JAMES, BUSHROD W., M D., 794,
N.E. Cor. Green and 18th St., Philadelphia, Pa.

JAMES, FRANK L., Ph.D., M.D., ’82, : 615 Locust St., St. Louis, Mo.
JAMES, GEO. W. 92, i ; : : 108 Lake St., Chicago, Ill.
JOHNSON, FRANK S., M.D., 83, : 416th St., Chicago, Ill.
JOHNSON, H. L. E., M.D., 91, i 1400 L St. N. W., Washington, D. C.

KeLLicoTT, D. 8., Ph.D., F.R.M.S., ’7%, State University, Columbus, Ohio.

KELLOGG, J. H., M.D., ’78 - . ; : ; Battle Creek, Mich.
KENNEDY, THOMAS, ’90, ; : P ; ; ; New Brighton, Pa.
KENNEY, HERBERT EASTMAN, 91, . : ; : Littleton, N. H.
KERR, ABRAM TUCKER, Jr., ; : : "1921 Main St., Buffalo, N. Y.
KINGSBURY, BENJAMIN F., A.B., M.S., 94, . . ~ . Ithaca, N. Y.
KIRKPATRICK, T. J., '83, ; : ; Springfield, Ohio.

Knap, Wm. H., M.D., ’98, P 106 Wabash Ave., Chicago, Ill,

0?
AMERICAN MICROSCOPICAL SOCIETY.

Kom, A. ., M:D:, 791; , 2 . ‘ . 58S. 4thSt., Easton, Pa.
KRAFT, Wacnrax: 795 Wing . ; . 411 W. 59th St., New York.
Krauss, Wm. C., B.S., M.D., F.R. M. S., 790, 382 Virginia St., Buffalo, N. Y.
KUHNE, F. W., 79, é : ‘ 5 79 Court St.; Fort Wayne, Ind.
Lamp, J. MELVIN, M.D., ’91, ; 906 G St., N.W., Washington, D. C.
LANDACRE, FRANCIS L., A.B., 795, 5 Columbus, Ohio.
LANDSBERG, A., 779, : E 145 Woodwadd Ave., Detroit, Mich.
LASCHE, ALFRED, 792, . “ . Lake and Franklin Sts., Chicago, Il.
Last, Louis, Ph.G., ’90 : : : 3 319 Reed St., Moberly, Mo.
LatTHAM, Miss V. A., M.D., D.D.S., ’88, . 808 Morse Av., Chicago, II.
LAWTON, EDWARD P., ’88, 4 : : 3 Linden Ave., Troy, N. Y-
LEBER, FREDERICK C., M.D., ’90, 550 E. Jefferson St., Louisville, Ky.
Lewis, Mrs. KATHARINE B., ’89, “Elmstone,” 656 7th St., Buffalo, N. Y.
Lewis, Ira W., ’87, : : : i j : : . Dixon, Ill.

Liwis, Wm. J., M.D., ’83,
. 812 Bennett Building, cor. Fulton and Nassau Sts., N. Y.

LIBBEY, WM., JR., D.Sc , ’88, 2 ; ? ‘ Prinicetar: IN: oe
Ling, J. Epwarp, D.D.S., F.R.M.S., ’82, 39 State St., Rochester, N. Y.
Lirron, A., M.D., ’78, : is : 2220 Eugenia St., St. Louis, Mo.
Locks, JOHN D., ’93, : : : 8600 Chestnut St., Philadelphia, Pa.
Loms, ADOLPH, 792, _ . A ; . 48 Clinton Place, Rochester, N. Y.
Lome, Car. F., ’79, : : . IiSINe St, Paul St: Rochester, N. ¥.
Lome, HENRY, ’84, : . : . 48 Clinton Place, Rochester, N. Y.
Kove, Prof. E.G: 9t, —. ; : : 69 E. 54th St., New York City.
Lyon, Howarp N., M.D., ’84,_ . : 34 Washington St., Chicago, Ill.
MALLONEE, JOSEPH D., ee ; : ; 84 Fargo Ave., Buffalo, N. Y.
MARSHALL, WM., JR., 792, . . Coudersport, Pa.

MATHER, ENOCH, M.B., MD., DDS. Ph.D., Dens LL.D., ’93,
124 Hamilton ser Patterson N. J.

McCaLLa, ALBERT, Ph.D., ’80,_ . ; 414 Monadnock St., Chicago, IIL
McGReEG@oR, JAMES H., ’95, ; : : F : Bellaire, Ohio.
McKay, JOSEPH, 84, . ; ; f : : * 9455 8th St., Troy, N. Y.
McKim, Rev. HAsLett, ’85, : : 30 West 20th St., Neve York City.
MACKAY, A. E., M.D., ’91) . Oregonian Building, Portland! Oregon.
MANnNrTON, W. P., M.D., F.R.M.S., ’85, 02 Adams Ave. W., Detroit, Mich.
MEEKER, J. W., M.D., 91, 4 ; : ; Nyack-on-Hudson, N. Y.
MELLOR, CHas, C., ’85, : : ; : 77 5th Ave., Pittsburgh, Pa.

MERCctgER, A., M.D., 95, Taulensee Bad at Spier, Canton Berne, Switzerland.
MERCER, A. CLIFFORD, M.D., F.R.M.S., ’82,
324 Montgomery St., Syracuse, N. Y.
MERCER, FREDERICK W., M.D., F.R.M.S., ’83,
2600 Calumet Ave., Chicago, IIL
MILLER, JOHN A., F.R.M.S. ’89,_. Niagara University, Buffalo, N. Y.
MILNOR, CHAS. G., 86, 4 : 5 318 Hiland Ave., Pittsburgh, Pa.
genie E. KENNARD, 792, . : : 326 N. Water St., Chicago, II.

AMERICAN MICROSCOPICAL SOCIETY

Moopy, Mrs. Mary B., M.D., *83,
P. O. box 206, Annex, New Haven, Conn.

Moopy, Rosert O., ’91, : Fair Haven Heights, New Haven, Conn,
Moors, V. A., M D., ’87, i Dept. of Agriculture, Washington, D. C.
Nonny, RICHARD, J., M.D., 83. ; ‘ 119} York St., Savannah, Ga.
OERTEL, T. E., M.D., 792, f - . P.O. box 482, New York City.
OuuER, W. H., 791, 2 : , ; . 18 Locust St., Portland, Me.
OLIN, R. C., M.D., 90, ; : ; ‘ 110 Henry St., Detroit, Mich.
PAQUIN, PAUL, M:D., “915 ; : . 3536 Olive St., St. Louis, Mo.
Park, ROSWELL, A.M., M.D., ’84, , 510 Delaware Ave., Buffalo, N. Y.
PATCHEN, YORK, M.D., ’84, : : : Westfield, N: We
PATRICK, FRANK, Ph.D., ’91, ; 601 arene Ave., Topeka, Kansas.
PEASE, FRED N., 87, . ; Box 210, Altoona, Pa.
PENNOCK, ED., *79, : 5029 ree St., Germantown, Philadelphia, Pa,
PERRY, STUART H., ’90, : stele /\. X. House, Ann Arbor, Mich.
PrFLAUuM, Maanus, Hsq., 791, : 415 Grant St., Pittsburgh, Pa.
PLars, Orro B:, M.D.791, N. W. cor. 8th and State Ave., Cineeaetal Ohio.
PortTE, JAMES H., EsqQ., ’87, ; ; f 409 Grant St., Pittsburgh, Pa.
PRENTICE, Wm. J., 87. : : ; 1009 Liberty Ave , Pittsburgh, Pa.
PRESTON, W.N.. 792, ; : : : 940 W. Adams St., Chicago, Ill.
PYBURN, GEORGE, M.D., ’86, ; : 11th and H Sts., Sacramento, Cal.
Rau, EUGENE A., 786, : f : : Bethlehem, Pa.
REMINGTON, Jos. P., M. te ROT é 1832 Pine St., Philadelphia, Pa.
REYBURN, ROBERT, M. D., 790, 2129 F St., N. W., Washinenees D. C.
REYNES. PLACIDE, ’87, 5 5 _ 204 Baronne St., New Orleans, La.
Rice, GEO. W., 90, : 5 : : 186 E. High St., Detroit, Mich.
Ropsins, HENRY A., M.D., 91, 1750, M St., N. W., Washington, D. C.
RoGers, Wm. A., A.M., Hon. F.R.M.S., ’82, C . Waterville, Maine.
ROWLEY, WILLARD W., 90, . ; : ; : Ithaca, N. Y.
Rust, GEOo., ‘ f ; , : P. O. box 25638, Denver, Col.
SCHAUFELBERGER, F. J., M.D., ’90 : é 713 2d St., Hastings, Neb.
SCHRENK, HERMANN, 93, ; Manual Training School, St. Louis, Mo.
SEAMAN, WM. H., M.D., ’86, 1424 11th St., N. W., Washington, D. C.
SECOR, JABIN, 795, : : : F ; Elmira, N. Y.
Srymour, Prof. M. L., ’85, ‘ : State Nurmed School, Chico, Cal.
SHEARER, J. B., 88, __.. ; : ‘ 809 Adams St., Bay City, Mich.
SHERMAN, WALTER N.,M.D.,’90 . : : ; . . Merced, Cal,
ScHULTZ, CHAS. S., ’82, ‘ ¢ . ; . Hoboken, N. J.
SHURLEY, E. L., M.D., ’81 : . 544 Jeftarect Ave., Detroit, Mich.
Sr1emon, RUDOLPH, ’91, t 22 B. Jefferson St., Fort Wayne, Ind.

SLOAN, JOHN, M.D., ’80 : . E. 6th and Main St., New Albany, Ind

3%]
4

AMERICAN MICROSCOPICAL SOCIETY.

Stocum, CuHas. E., Ph.D., M.D., ’79, : : 5 ; Defiance, Ohio.
SMITH, JOHN B., M.D., 91, ; : ; ; . New Brunswick, N. J.
SPENCER, HERBERT R., ’85, : ; ; 367 7th St., Buffalo, N. Y.
STEDMAN, J. M., ’87, x . " ; : . . - Columbia, Mo.
STERNBERG, GEO. M., M.D., F.R.M.S., ’87, , ‘ Washington, D. C.
STILLSON, J. O., A.M., M.D, ’80, 245 N. Penna. St,, Indianapolis, Ind.

Stimson, JAMES, M.D., 791, Lock box 66, Watsonville, Santa Cruz Co., Cal.
STOWELL, THOMAS B., A.M., Ph.D., ’82, Potsdam, St. Lawrence Co., N. Y.

Summers, Prof. H.E., 86, ... University of Illinois, Champaign, Il.
SyLvesteR, WILLIAM H., M.D., 90, —.. 6 Clarendon St., Natick, Mass.
TAYLOR, Rev. F. W., D.D., ’82, : Speingfield, Il.
TAYLOR, THOMAS, M.D., ’79, Dent of er fenttare, Washington, D. C.
TERNAN, JAMES C., 93, Bausch & Lomb Optical Co., Rochester, N. Y.
THEILE, OTTO A., M.D., ’84, , , , 287 3d St., Milwaukee, Wis.
THOMAS, Prof. MASON B., ’90, : Wabash College, Crawfordsville, (nd.
TIFFANY, FLAVEL B., M.D., ’86, ‘ 1235 Grand Ave., Kansas City, Mo.
TINGLEY, J., M.D., ’89, : : : . P. O. box 425, Pittsburgh, Pa.
BEITYs Wi. EL., (Sls. | ; 4 207 N. 2d St., St. Louis, Mo.
TOLMAN, Henry L., F.R.M.S., 88, 923 Opera House Src, Chicago, IIL.
TURNER, HENRY H., ’84, : ; ; . 45 Stone St., Rochester, N. Y.
howe. AWA, Ph.D., HRMS anos 5 University of Virginia, Va.
TWITCHELL, GEO. B., ’86 5 j 556 Freeman Ave., Cincinnati, Ohio.
‘VVANDERPOEL, FRANK, M.E., ’87, é 191 Roseville Ave., Newark, N. J.
VEEDER, ANDREW T.., M.D., ’88, Hamilton Building, Pittsburgh, Pa.
VEEDER, M. A., M.D., ’85, : ; : ; duyonsN. Y.
Vorcs, C. M., F.R.M.S., 778, ; . 5 Rouse Block, Cleveland, Ohio.
VREDENBURGH, E. H., ’84, : . 12258. Fitzhugh St., Rochester, N. Y.
WAGENHALS, Rev. SAMUEL, ’82, . 5 ; ° Fort Wayne, Ind.
WALL, JOHN L., F.R.M.S., 78, f , 338 6th Ave., New York City.
WALMSLEY, W. H., F.R.M.S., ’78, 5 134 Wabash Ave., Chicago, Ill.

WARD, HeEnRY B., A.M., Ph.D., ’87.
Professor of Zodlogy, Nebraska University, Lincoln, Neb.

MAR. hte H., M.D.) BER. M.S, 778; A 5 : 53 4th St., Troy, N. Y.
WEBER, HENRY A., Ph.D., 86, : 1342 Forsyth Ave., Columbus, Ohio.
WEIGHTMAN, CHAS. H., ’86, ; : 5859 Michigan Av., Chicago, IIL.
WELCH, Geo. O., M.D., ’91, F : Box 416, Fergus Falls, Minn.
WENDE, ERNEST, M.D., 91, ‘ . 71 Delaware Ave., Buffalo, N. Y.
WERuUM, J. H., 93, : : ; Toledo, Ohio.
West, CuHas. E., LL. D., F.R. MS., E 76 eerrepont St., Brooklyn, N. Y.-.

WESTOVER. H. W., M. D, (SGian ; ; . St. Joseph, Mo.
WHELPLEY, H. M., M.D., Ph.G., F.R.M. S., 90,

: 9342 Albion Place, St. Louis, Mo.
WHITE, JONATHAN, Esq., 91, _ . : . 254 Main St., Brockton, Mass.

{o*

AMERICAN MICROSCOPICAL SOCIETY. <

i a

Waite, MosesC., M.D.,’85, . +. Box 1674, New Haven,
WHITLEY, James D., M.D., ’85, F F { ate sbu
WIARD, MarrTIN S., ’86, ae 21 Walnut St., New Britain,
WIEGAND, Karu McKay, 794, : : Ithaca, 4
WILBER, GEO. D., M.D., 98, . . ~.. " 3108 Gilpin St., Denver, ¢
WILLICH, CHAS., JR., 790, : : 696 President St., New Yorks
WILLSON, LEONIDAS A., ’85, : . — 13 Public Sees Cleveland,
WILsoNn, Mrs. Mary R., M.D.,’95_ ; : Ithaca,

Woopwarp, ANTHONY, 85, . . 206 W, 128th St., New York City.
WooLMAaN, Geo. S., ’79, : : : 116 Fulton St., New York City.

Youne, Aveustus A., ’92, , =a Newark, Wayne Co., N.
YznaGa, José M., Esq., 90, . : May Building, Washington, D.

ZENTMAYER, FRANK, "91, . . . 2098, 11th St., Philadelphia, I

Honorary [lembers.

Crisp, FRANK, LL.B., B.A., F.R.M.S., 5
5 Landsdowne Road, Nottingham Hill, London, England.

DALLINGER, Rev, W. H., F.R.S., F.R.M.S., :

Toei Lee, S. E., London, Englar

Hupson, Rev. C. T., A.M., LL.D., F.R.MS., _ ay

6 posal York Crescent, Clifton, Bristol, England, —

2ST OLIN CE Xt Boast i
OF THE

KIGHTEENTH ANNUAL MEETING

OF THE

The American Microscopical Society,

Pe Eee Sree IME SORK,

August 21, 22 and 23, 1895.

OFFICERS OF THE AMERICAN MICROSCOPICAL SOCIETY FOR 1895.

PRESIDENT,
PROFESSOR SIMON HENRY GAGE, Ithaca, N. Y.
VICE-PRESIDENTS,
Dr. V. A. MOORE, Washington, D. C. H.C. HANKS San Francisco, Cal.
SECRETARY,
Dr. WILLIAM H. SEAMAN, Washington, D. C.
TREASURER,
MAGNUS PFLAUM, Pittsburg, Pa.
EXECUTIVE COMMITTEE,
Dr. ROBERT O. MOODY, New Haven, Conn.
CHARLES S. SCHULTZ, Hoboken, N. J.
PROFESSOR HENRY B. WARD, Lincoln, Neb.

EIGHTEENTH ANNUAL MEETING
OF

THE AMERICAN MICROSCOPICAL SOCIETY.

PLACE OF MEETING.

The Eighteenth Annual Meeting of the American Microscop-
ical Society will convene in McGraw Hall, at Cornell University,
Ithaca, N.Y., Wednesday morning, August 2Ist, 1895, and continue
in session three days.

If one considers the distribution of the members, Ithaca is as
central a point for meeting as can be found. Moreover those who
desire to attend the meeting of the American Association for the
advancement of Science the following week will find little incon-
venience in attending the Ithaca meeting.

The acccmmodations afforded by the University buildings, and
their equipment for carrying on all lines of microscopical work, add
very materially to the attractiveness of Ithaca as a place of meeting.
Add to this the richness of both terrestrial and aquatic fauna and
floya, and it is almost an ideal place, both to the student of natural
history and to those who love beautiful scenery.

It has often enough been demonstrated that local interest con-
tributes a great deal toward the success of a meeting. Much inter-
est in the coming meeting has already been shown by the people of
Ithaca. <A large local committee has been formed and everything
will be done within its power, that will contribute to the comfort and
enjoyment of the members and their friends.

In most of the scientific departments of the University there are
already members of the Society, and in all departments there will be
a most hearty welcome, and every reasonable aid will be furnished
for the success of the meeting. Finally and not least, the President
of the University extends to the Society a most cordial welcome.

The University buildings, which will be at the disposal of the
Society, are especially adapted for the formal presentation of papers,
blackboard illustration, hanging of diagrams, etc., as well as for any
demonstration that authors may desire to make. ‘The Armory is
very conveniently located for the University and for the city, anda
soiree there can hardly fail to be a success.

The University possesses one of Roger's dividing engines and
the department of Physics has kindly promised to show the members
exactly how micrometers are made. ‘There is also a large compara-
tor for carefully testing micrometers after they are made. ‘This one
was actually used in determining the exactness of the rulings of our
standard centimeter.

AMERICAN MICROSCOPICAL SOCIETY.

HOW TO REACH ITHACA BY RAILROAD.
The local time-table is as follows :

LEHIGH VALLEY.—Northward—*6:41 A. M.,8:20A M., *4:30P. M.,5:45P. M.
Southward —8:25 A. M., 11:45 A. M., 8:00 P. M., *10:15 P. M.; also train
from the north arrives at 12:40 P. M., daily.
Southward—11:30 A. M., Sunday only.

AUBURN BRANCH, L. V.—Leave—8:30 A. M., 1:00 P. M., 4:05 P. M.
Arrive—8:10 A. M., 12:35 P. M., 7.55 P. M.

DELAWARE, LACKAWANNA & WESTERN.—Leave—8:55 A. M., 12:00 M.,
7:00 P. M.
Arrive—6:55 A. M., 1:28 P. M., 4:55 P. M.
ELMIRA, CORTLAND & NORTHERN.—Eastward—8:56 A. M,, 3.22 P. M., 6:17
P. M., Sunday, 9:50 A. M.
Westward—g:42 A. M., 3:58 P. M., 9:03 P. M., Sunday, 4:53 P. M.
*Sunday.

Ithaca is upon the main line of the Lehigh Valley Railroad,
about four hours ride from Buffalo, and ten hours from New York
and Philadelphia. The Elmira, Cortland and Northern Railroad passes
through the city connecting, at Elmira with the New York, Lake
Erie, and Western Railroad, the Delaware, Lackawanna and West-
ern Railroad, and other roads ; also connecting in Canastota with
the New York Central Railroad and the West Shore Railroad. The
Delaware, Lackawanna and Western Railroad has a line into Ithaca
connecting with the main line at Owego. A branch line of the
Lehigh Valley Railroad connects with the Auburn branch of the
New York Central Railroad at Auburn and Owego.

Steamboats connect daily with both east and west bound trains
on the New York Central Railroad (Auburn branch).

A representative of the local committee will meet each incom-
ing train Tuesday and Wednesday, in order to give information to
arriving members. He will wear a conspicuous badge bearing the
initials of the Society, A. M. S. }

RAILROAD RATES.

Arrangements have been made with the ‘Traffic Association
whereby a reduction to one and one-third fare for round trip for per-
sons attending the meeting will be made.

INSTRUCTIONS TO PERSONS ATTENDING THE MEETING.

1. The reduction is made upon all railroads comprised in the the Trunk-
Line territory and also the territory of the Central Traffic Association. The
membership is almost entirely within the limits of the territory covered by
these organizations. :

2. The reduction is fare and a third on Committee’s certificate, condi-
tional on there being an attendance at the meeting of not less than 100 per-
sons who traveled thereto on some legitimate form of railroad transportation.

The reduction applies to persons starting from Trunk Line or Central
Traffic Association territory who have paid 75 cents or upwards for their

ANNOUNCEMENT OF THE

going journey. Each person availing of it will pay full first-class fare going
to the meeting and get acertificate filled in on one side by the agent of whom
the ticket is purchased. Agents at all important stations and coupon ticket
offices are supplied with certificates.

4. Certificates are not kept at all stations. If, however, the ticket agent
at a local station is not supplied with certificates and through tickets to place
of meeting, he can inform the delegates of the nearest important station
where they can be obtained. In such a case the delegate should purchase a
local ticket to such station and there take up his certificate and through ticket
to place of mreting.

5. Going tickets, in connection with which certificates are issued for
return, may be sold only within three days (Sunday excepted) prior to and
during the continuance of the meeting ; except that, when meetings are held
at distant points to which the authorized limit is greater than three days,
tickets may be sold before the meeting in accordance with the limits shown in
regular tariffs.

6. Deposit the certificate with the secretary or other proper officer of
the organization at the meeting, for necessary endorsement and visé of
special agent.

7. Certificates are not transferable, and return tickets secured upon cer-
tificates are not transferable.

8. On presentation of the certificate, duly filled in on both sides, within
three days (Sunday excepted) after the adjournment of the meeting, the ticket
agent at the place of meeting will return the holder to starting point, by the
route over which the going journey was made, at one-third the highest lim-
ited fare by such route. The return ticket will iu all cases be closely limited
to continuous passage to destination.

9. No refund of fare will be made on account of any person failing to
obtain a certificate.

WHERE THE SESSION WILL BE HELD.

The regular daily sessions will be held in McGraw Hall in the
rooms occupied by the Department of Physiology and Vertebrate
Zoology. The evening sessions will be held in the Botanical Lecture-
room in Sage College. The soiree will be in the University gym-
nasium. Members of the local committee will be in attendance at —
the places of meeting to give all desired information. Badges will
be given each member as he arrives and registers for the meeting.

HOTELS.

Ithaca Hotel—Corner of State and Aurora streets. Rates, $2.00
and $2.50 per day.

Clinton House—Cayuga street, one block from the street cars.
Rates, single, $2.00 per day ; double, $1.50 per day.

Tompkins House—Seneca street, one block from the street cars.
Rates, $1.25 per day.

Hollister House—Seneca street, a block and a half from street
cars. Rates, single, $1.50 per day ; double, $1.25 per day.

Boarding Houses—There are many private boarding houses up-
on and near the campus, and near the street cars, where board and
rooms can be secured at very reasonable rates. Suites of rooms can
be secured for from $.50 to $1.00 per day, and table board at from
$.75 to $1.00. These are both pleasant and convenient. Single

AMERICAN MICROSCOPICAL SOCIETY.

meals can be gotten for 25 cents each. Any one desiring accommo-
dations at boarding houses will find it to their advantage to inquire
of members of the local committee either before coming or imme-
diately upon arriving.

Boarding places will be secured for those who fill out and return
the enclosed blank. Such persons will be notified of the quarters
secured for them.

STREET CAR SERVICE.
&

Ithaca has ample and very efficient street car service. There
are two main lines, one through the main street of the city (State
street), connecting the railroad depots with the University campus,
and within one minute walk of both places of meeting. Cars run
upon these lines every ten minutes. ‘The other is from the business
portion of the city to Renwick Park, a delightful resort upon the
shore of Cayuga Lake.

EXCURSIONS.

An excursion upon Cayuga Lake will be given the members of
the Society by the people of Ithaca on the afternoon and evening of
the second day of the meeting. The street car company will give a
ride to Renwick Park and return, from which point the party will
take boats for the excursion upon the lake.

SOIREE.

Microscopes will be furnished from the University laboratories
for exhibitors to use at the soiree, thus doing away with the expense
and trouble of transporting microscopes. ‘Those who desire to make
special demonstrations will need to bring their own instruments.

EXPRESS.

Members desiring to send material intended for exhibition or
use at the meeting, should address it to Ithaca, N. Y., care of Mc-
Graw Hall. All such packages will be delivered by the express
company and cared for until called for by the owner.

MEMBERSHIP.

Blanks for membership are enclosed and others may be had
on application to the Secretary, Professor W. H. Seaman, Washing-
LOD Der Cc:

ANNOUNCEMENT OF THE

GENERAL PROGRAMME.

Tuesday, 8:00 Pp. M.—Meeting of the Executive Committee,
Botanical Lecture room.

FIRST DAY—WEDNESDAY.

Morning, 9:30—McGraw Hall, Address of welcome by Hon. D.
F. Van Vleet. Response by the President of the Society. Read-
ing of papers.

Afternoon—Inspection of the Library and other University
buildings. Illustration of methods of marking micrometers upon a
ruling engine at Franklin Hall (Physical Building. )

Evening, 8:oo—Botanical Lecture room, President Gage’s
address. Subject : The Processes of Life Revealed by the Microscope.

SECOND DAY——EHUR SD AGW.

Morning, 8:30—McGraw Hall, Meeting of the Executive Com-
mittee. 9:30—Reading of papers.

Afternoon and Evening—Excursion on Cayuga Lake.
THIRD DAY—FRIDAY.

Morning, 8:30—McGraw Hall, Meeting of Executive Commit-
tee. 9:30—Demonstrations and reading of papers on methods.

Afternoon—Business meeting of the Society.

/vening—Soiree in the University Gymmasium.
PRELIMINARY LIST OF PAPERS PROMISED FOR THE MEETING.

Up to the present time the following titles of papers have been
received. They are arranged alphabetically under the authors.

It is hoped that the blanks for additional titles sent out with this
announcement will be used and titles sent as long before the meet-
ing as possible so that the program may be arranged in a way to
make the most advantageous use of the time available for the ses-
sions.

1. Comparison of the Fleischel, the Gower and the Specific
Gravity Method of Determining the percentage of Hemoglobin in
Blood for Clinical Purposes. F. C. Buschand A.T. Kerr, Jr., Buffalo,
Pe,

2. Cocaine in the study of pond-life. Professor H. N. Conser,
Sunbury, Pa.

3. Paraffinand Collodion Embedding. Professor H. N. Conser.

4. The Fruits of the Order Umbelliferee. Dr. E. J. Durand,
Ithaca, N. Y.

AMERICAN MICROSCOPICAL SOCIETY

5. The history of the Sex-Cells from the time of segregation
to sexual differentiation in Cymatogaster. Proiessor C. H. Eigen-
mann, Bloomington, Indiana.

6. ‘The use of Formalin in Neurology. Dr. P. A. Fish, Ithaca,
ae

7. The action of strong currents of electricity upon Nervous :
Massue. Dr. P. A. Fish, Ithaca, N: Y.

8. The morphology of the Brain of the Soft-Shelled Turtle and
the English Sparrow compared. Susanna P. Gage, Ithaca N. Y.

g. Improvements in the Collodion Method. Professor S. H.
Gage, Ithaca, N. Y.

1o. A fourth study of the Blood showing the relation of the Col-
orless corpuscle to the strength of the constitution. Dr. M. L. Hol-

brook, New York City.
11. The Lymphatics and the Lymph Circulation with demon-
stration of specimens and apparatus. Dr. Grant S. Hopkins,
ithaca, N. Y.

12. Some peculiarities in the structure of the Mouth Parts and
Ovipositer of Czcada septendecim. Professor J. D. Hyatt, New Ro-
chelle, N. Y.

13. The Lateral Line System of Sense Organs in Amphibia.
Dr. B. F. Kingsbury, Defiance, O.

14. The Spermatheca and Methods of Fertilization in some
American Newts and Salamanders. Dr. B. F. Kingsbury, Defiance,O.

15. The Segmentation of the Vertebrate Head. Dr. J. S.
Kingsley, Tuft College, Mass.

16. Formalin asa Hardening Agent for Nerve Tissue. Dr.
Wm. C. Krauss, Buffalo, N. Y.

17. Anew Way of Marking Objectives. Dr. Wm. C. Krauss,
Buffalo, N. Y.

18. Demonstration of Histological Preparations by the Projec-
tion Microscope. Drs. Krauss and Mallonee, Buffalo, N. Y.

19. Secondary Thickenings of the Rootstalk of Spathyema.
Mary A. Nichols, Ithaca, N. Y.

20. A Practical Method of Referring Units of Length to the
Wave Length of Sodium Light. Professor Wm. A. Rogers, Wa-
terville, Me.

21. The Chlorophyll Bodies of Chara coronata. Professor
W. W. Rowlee, Ithaca, N. Y.

22. The Fruits of. the Order Composite. Professor W. W.
Rowlee and K. M. Wiegand, Ithaca, N. Y.

23. Corky Outgrowth of Roots and their Connection with Res- .
piration. H. Schrenk, Cambridge, Mass.

24. New Points in Photo-micrographs and Cameras, W. H.
Walmsley, Chicago, Il.

25. Some Experiments in Methods of Plankton Measurements.
Professor Henry B. Ward, Lincoln, Neb.

ANNOUNCEMENT OF THE

26. The Primitive Source of Food Supply in the Great Lakes.
Professor Henry B. Ward, Lincoln, Neb.
27. Two Cases of Intercellular Spaces in Vegetable Embryos.

K. M. Wiegand, Ithaca, N. Y.
EXHIBIT OF MICROSCOPES, MICROTOMES AND OTHER APPARATUS.

The firms named below in alphabetical order will be represented
at the meeting with the products of their manufacture as follows :

The Bausch & Lomb Optical Company of Rochester and New
York, N. Y., with a full line of their Microscopes and Microtomes
of New construction. Also their Photo-micrographic Camera and
various Accessories.

E. Leitz, of Wetzlar, Germany, represented by Wm. Kraft, of
New York, will exhibit six grades of Microscope Stands ; a Mechan-
ical Stage ; Dissecting Microscope with Camera Lucida ; Edinger’s
Projection Apparatus with Photographic Camera ; Photo-Micro-
graphic Apparatus ; Microtomes of the Schanze and Thoma patterns.

Walmsley, Fuller & Co., of Chicago, will exhibit Walmsley’s
new ‘‘Autograph’’ Photo-micrographic Camera, the Improved Handy
Camera, and Ross New Eclipse Microscopes.

Joseph Zentmayer of Philadelphia, Pa., will exhibit his Colum-
bian and Continental Microscope stands and the Ryder Microtome.

These exhibits of the newest forms of microscopes, microtomes,
photo-micrographic and other apparatus and accessories, together
with the various apparatus in the laboratories of the University, it is
believed, will help the members in selecting the apparatus likely to
be most useful and best adapted to their special needs.

LOCAL COMMITTEE

PROFESSOR W. W. ROWLEE, Chairviman.

Professor L. H. Bailey,
Eugene Baker, M.D.,

H. Bergholtz,

H. B. Besemer, M.D.,

C. P. Biggs, M.D..,

Chas. H. Blood, Esq.,

C. D. Bostwick,
Professor F. D. Boynton,
A. B. Brooks,

Maj. D. W. Burdick,
Professor G. C. Caldwell,
E. M. Chamot,

Wm. A. Church,

Professor J. H. Comstock,

Dr. E. J. Durand,
Professor W. F. Durand,
Professor H. W. Foster,
Professor E. A. Fuertes,
Chas. W. Gay,

Professor A. C. Gill,
Professor E. Hitchcock,
Dr. G.S. Hopkins,

D. F. Hoy,

Chas. A. Ives
Professor H. S. Jacoby,
B. F. Kingsbury,
Professor James Law
A. D. MacGillivray,
Professor G. S. Moler,
Professor J. L. Morris,
V. D. Morse,

J. T. Newman, Esq.,
Jacob Peters,

F, W. Phillips,

GG. Platt

Professor A. N. Prentiss,
Jacob Rothschild,

Belle Sherman,

M. V. Slingerland,

D. B Stewart,

C. D. Stowell,

Professor R. S. Tarr,
Professor R. H. Thurston,
Professor E. B. Titchener,
Mayor L. G. Todd,

Hon. D. F. Van Vleet,
K. M. Wiegand,

Dr. A. C. White,

Chas. H. White,
Professor B. G. Wilder,
E. lL. Williams, Esq.,
Geo. R. Williams, Esq.
R. B. Williams.

Professor S. G. Williams,
Professor H. H. Wing,

J. Winslow, M.D.

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A3 | Transactions
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PLEASE DO NOT REMOVE
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