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i Bom eke 2

A COURSE

OF

ELEMENTARY

PRACTICAL PHYSIOLOGY
AND HISTOLOGY. —

oe Mav A COURSE

1:

OF

ELEMENTARY

PRACTICAL PHYSIOLOGY
AND HISTOLOGY,

BY

M. FOSTER, M.D., F-RBS.,

PROFESSOR OF PHYSIOLOGY IN THE UNIVERSITY OF CAMBRIDGE.

AND

J. N. LANGLEY, M.A... F.RS,,

FELLOW OF TRINITY COLLEGE, CAMBRIDGE,

SIXTH EDITION.

Dondon:
MACMILLAN AND CO.
AND NEW YORK.

1888

[The Right of Translation is reserved.)

First Edition, Crown 8vo., Fuly 1876.
| April 1877, December 1877, October 1880,
November 1880, 1881, 1882, 1883.
| Revised Cs Napa Edition February 1884.

Ps cseecad 1887, 1888.

én

’ .
e er oe

Shae

(on,

Soe wey

CONTENTS.

LESSON I. Dissection of a Rabbit and of a Dog .
LESSON II. Structure of Blood

LESSON II. eee of Blood. Characters of
Proteids ‘ : : :

LESSON IV. Hyaline Cartilage . , ° ° °
LESSON V. Connective Tissue . 4 ¢ ; ?

LESSON VI. Modification of Connective Tissue and
Hyaline Cartilage

LESSON VII. Bone, Ossification, Teeth

LESSON VIII. Structure of Contractile Tissues .
LESSON IX. Properties of Contractile Tissue . °
LESSON X. Structure of Nervous Tissues

LESSON XI. General da sa of Nervous Tissue.
Automatic Actions : .

LESSON XII. Structure and Properties of Blood-Vessels
LESSON XIII. Structure and Action of the Heart
LESSON XIV. Blood Pressure .

PAGE
1—34

— 8543

44—51
52—57
58—67

68—71
72—79
80—89
90—104
105—115

116—123
124—135
136—146
147—1é4

CONTENTS.

LESSON XV. Salivary Glands and Pancreas. Saliva .
LESSON XVI. Stomach. Gastric Juice. Milk .
LESSON XVII. Intestine.
LESSON XVIII. The Lymphatic System . ° °
LESSON XIX. Structure of Liver. Glycogen .

LESSON XX. The Structure of the Lung. The Me-
chanics of Respiration . 2 ‘ A ‘

Bile. Pancreatic Juice

LESSON XXI. The Colour of Blood. Respiration
LESSON XXII. Structure of the Kidney .

LESSON XXIII. Urine ; ‘ ; ‘ ; “
LESSON XXIV. Skin and Touch . ; .
LESSON XXV. Taste and Smell .

LESSON XXVI. The Eye

LESSON XXVIII. Vision

LESSON XXVIII. The Ear.

LESSON XXIX. The Spinal Cord

LESSON XXX. The Brain

LESSON XXXI. Dissection of the Larynx . F .
LESSON XXXII. Tissues of Reproduction . ; :
APPENDIX . % eae .

AppiTIons To APPENDIX 7 2 >
INDEX . : . : é a

Appitions To InpEx .

PAGE
155—166

167—177
178—190
191—203
204—210

211—218
219—227
228—236
237—245
246—256
257—262
263—274
275—290
291—300
301—307
308—330
331—335
336—344

345—386

387—403

404—412
413

LESSON I.

DISSECTION OF A RABBIT AND OF A DOG.

In the following, the descriptions in large type apply more particularly
to the rabbit, but the general directions for dissection serve also
for the dog: some points in which the two animals differ, and
some which are better seen in the dog, are printed in small type.

A. 1. Make a median incision through the skin, down

h L.

the whole length of the front of the body from

the neck to the pubis, and reflect the skin as far

as possible on both sides.

In dissecting a female rabbit note, just under-
neath the skin, the thin arborescent mammary
glands, one to each mamma.

Observe the thin, pale, abdominal muscles.
It is better to dissect out the individual mus-
cles in the dog as below, but the dissection
may be done on the rabbit.

In the dog observe
a. The tendinous aponeuroses of the abdominal
muscles forming in the middle line the linea

alba,
1

ELEMENTARY PHYSIOLOGY. [i

b. The obliquus externus abdominis, a thin muscle,
with descending fibres; it arises from the ribs
by separate bundles, from the back by a broad
tendon and runs to the linea alba and to the
pubis.

c. The recti abdominis, one on either side of the
middle line, covered by the tendon of the exter-
nal oblique.

If b be carefully reflected, there will be seen under-
neath :

d. The internus obliquus abdominis, with ascending
fibres, it arises from the pubis and lumbar
fascia and runs to the linea alba and lower ribs;
and underneath this

e. The transversalis abdominis, it arises from the
lower ribs, the dorso-lumbar fascia and the
pubis, and runs to the linea alba.

Lift up the abdominal wall and cut it through
in the middle line from the sternum to the
pubis, being careful to avoid puncturing the
intestine. From the middle of this cut make
transverse cuts nearly as far as the spinal
column. Hook or pin back the four flaps.
Simply turning the parts over without cutting
or tearing anything, trace out as far as possible
the alimentary canal, noting the narrow ceso-
phagus entering into the stomach about the
middle of its concave upper portion, the pyloric
end of the stomach placed on the right side and
continuous with the small intestine which is
not distinctly divisible into duodenum, jejunum
and ileum, the large dark thin-walled cecum

Bal

DISSECTION OF A RABBIT AND OF A DOG. 3

having a shallow spiral constriction around. it,
the rather thick-walled, light coloured appendix
proceeding from the end of the cxcum, the
large intestine of much smaller diameter than
the cecum, much puckered in the first part of
its course, less puckered in its median portion,
and becoming soon quite smooth and passing
without change into the rectum. The latter
part of the large intestine and the rectum usually
contain balls of faeces,

Trace out thé mesentery which supports the
intestine; observe its continuity with the peri-
toneum or membrane lining the abdominal
cavity, note the manner in which the blood-
vessels run in it.

Observe in the dog the loose fold of mesentery
loaded with fat, hanging from the lower border
of the stomach and forming the great omentum.

Observe the spleen, an elongated dark red body
lying near the broad end of the stomach to
which it is attached by a mesenteric fold (gastro-
splenic omentum).

Turn the stomach oyer to the left’, gently stretch
out the duodenum and observe in the mesentery
belonging to it, the diffuse, pale-red pancreas ;
trace the entrance of the pancreatic duct as a
pale thin band into the duodenum: this occurs
rather more than a foot below the pylorus, where
the duodenum turns back on itself to form a loop.

1 Right and left are used throughout for the right and left of the
animal,

1—2

4 ELEMENTARY PHYSIOLOGY. [1.

In the dog the pancreatic duct is close to the
entrance of the bile duct (see § 14),

Observe the mesenteric lymphatic glands,
small greyish white lumps, more abundant
in the duodenal mesentery than elsewhere.

7. Turning the stomach and intestines over to the
right side observe the dorsal aorta‘ and inferior
vena cava lying close together in the median
line, trace the aorta upwards to the point where
it descends through the diaphragm, tearing
through the mesentery as little as is consistent
with tracing the aorta,

8. Note the right suprarenal body, small, ovoid
and yellowish white, lying close to the aorta,
carefully tear away the connective tissue above
and medially of this and note the solar plexus
consisting of three or more greyish semi-trans-
parent ganglia connected by bundles of pale
nerve fibres. Into the laterally placed ganglion
runs the main branch of the splanchnic nerve,
trace this up alongside the aorta as far as the
diaphragm,

9, Note the celiac artery given off by the aorta
a little below the diaphragm, and the superior
mesenteric artery given off somewhat lower
down, possibly underneath the suprarenal body,
and a little farther down, the renal artery, run-

* When an artery and a vein run together, as here, they may be
distinguished by the artery haying thicker walls and containing less
blood than the vein; the artery too has generally a bluish-white tint
whilst the vein has generally a dark red tint with a tinge of blue.

1.]

10,

iY

DISSECTION OF A RABBIT AND OF A DOG. 5

ning to the hilus of the kidney: note the renal
vein running parallel to the renal artery into
the vena cava. Follow the superior mesenteric
artery a short distance and observe the branches
given off to the pancreas, these are more easily
seen when the intestines are turned to the left.

Tearing through the mesentery around the
lower part of the cesophagus, observe the right
and left pneumogastric nerves (cp. C §§ 17,
24,) dividing into several fibres which spread
out over the stomach. One or more branches
may be traced to the solar plexus. Observe
the number of pale nerves which are given off
by the ganglia of the solar plexus; bundles °
of them may be followed along the celiac,
mesenteric and renal arteries.

Then turning the stomach and intestines over to
the left side, carefully tear away the mesentery
over the aorta and note the right splanchnic
nerve close beside it, trace the nerve on its course
(being careful not to puncture the vena cava)
past or underneath the right suprarenal body
into a ganglion a little removed from the rest
of the solar plexus.

Lift up the stomach, and viewing from the right
the mesentery below it, note the portal vein,
a large vein dividing close to the posterior
surface of the liver and running into it. This
vein is formed by the union of the lieno-gastric
and mesenteric veins, the former is much the
smaller and joins the latter close to the liver;

13.

_ ELEMENTARY PHYSIOLOGY. [I.

follow for a short distance the course of the
mesenteric vein, noting the small numerous
branches received by it from the pancreas.
Viewing the mesentery from the left side note
the juncture of the splenic and gastric veins to
form the lieno-gastric.

Trace out the branches of the celiac artery; it
first gives off the splenic artery which besides
giving off a row of smaller arteries to the spleen
sends several branches to the greater curvature
of the stomach and some small branches to the
pancreas, it then gives off at short intervals
branches to the lower part of the cesophagus,
the stomach and the upper part of the duode-
num and a branch, the hepatic artery, which
runs to the liver.

In the dog, pull the spleen downwards and to the
left away from the stomach, a branch of the
lieno-gastric artery will be seen sending branches
to the spleen and to the greater curvature of
the stomach ; the corresponding veins are best
seen on turning the spleen over towards the
stomach. Double ligature and cut through
these vessels, pull. the spleen downwards as
before, a smaller branch of the lieno-gastric
artery and vein will be seen; centrally of the
lieno-gastric vessels will be seen two or more
gastric and pancreatic arteries and veins, Pull
the pancreas to the left over the spleen and
note the junction of the lieno-gastric and me-
senteric veins, Then pull the duodenum over
the part of the pancreas previously showing and

1]

DISSECTION OF A RABBIT AND OF A DOG, 7

note the fairly large vein from the pancreas
and the upper part of the duodenum joining
the previously mentioned vein to form the portal
vein; note also the branch from the celiac
axis dividing into the hepatic artery and an
artery supplying the greater part of the
pancreas and the upper part of the duodenum ;
it then gives off branches to the lower part of
the esophagus and the stomach and finally
divides into two branches, one the hepatic
artery going to the liver, the other going to
the lower part of the stomach and the upper
part of the duodenum (with branches to the
pancreas).

14. Turning the liver up towards the diaphragm, the

gall-bladder will be seen in a hollow on the
under surface of the posterior right lobe: trace
the cystic duct or duct from the gall-bladder to
the point where it joms the hepatic duct,
proceeding from the liver itself; trace the united
duct or common bile duct into the duodenum,
close to the pylorus.

. Ligature the cesophagus and the rectum ae cut

through both, the former above the ligature the
latter below it. Turning the intestine to the
right, cut through the mesentery close to its
abdominal attachment and remove from the
abdomen the alimentary canal and its appen-
dages except the liver. Observe now the posi-
tion and form of the liver, especially in relation
to the diaphragm.

16. Pull the liver down from the diaphragm,

17.

18.

19.

ELEMENTARY PHYSIOLOGY. [I.

through the transparent tendon of the diaphragm
the lungs will be seen in close contact with it.
Puncture the tendon on the right side and note
the collapse of the right lung as soon as air
enters the pleural cavity.

With the liver still pulled down, note the short
hepatic veins proceeding from the liver to join
the vena cava inferior just below the diaphragm.
Cut through the hepatic veins as close to the
liver as possible and remove the liver.

Cut open one of the hepatic veins and trace it
in this way back into the substance of a liver
lobe. Observe on its inner surface the opening
of numerous smaller veins; cut through the lobe
near its base, and try to distinguish the portal
veins from the hepatic by the small bile duct
and small thick-walled artery running alongside
the former.

Cut away the mesentery from the alimentary
canal, and trace out the latter along its whole
length, observing more fully the features men-
tioned in § 3, and noting in addition one or more
white patches (Peyer’s patches) on the free sur-
face of the ileum, due to clumps of lymph-follicles;
also note the connection of the caecum with the
small and large intestine, the thin walls of the
eecum and the thicker spotted walls of its
appendix.
Note in the dog, the wider esophagus entering into
the stomach nearer the cardiac end than is the
case with the rabbit; note also the shorter

23.
24,

20.

21.

22.

DISSECTION OF A RABBIT AND OF A DOG, 9

length of the intestine, the small cecum, and
the less difference between the large and small
intestines.

The small intestine may be washed out by tying a
funnel into the duodenum, and letting water
from a tap stream down the funnel. The large
intestine may be similarly treated.

Cut through the stomach along the lesser curva-
ture, throw away its contents and wash the
mucous membrane. Note that the mucous
membrane of the greater curvature is pale red,
that of the pylorus is greyish-white and semi-
transparent. The contrast is more marked
when the whitish superficial layer of mucous
cells is removed.

The mucous membrane may be used to prepare a

glycerine extract of pepsin (cp. Lesson xv1.),

Wash out the duodenum, its inner surface has
a velvety look which is characteristic of the
mucous membrane of the small intestine; it is
caused by the villi, examine these with a lens.
Observe the openings of the biliary and pancrea-
tic ducts, and carefully pass bristles through
them into the ducts.

Cut open a piece of the large intestine, wash it,
and with a lens examine its inner surface ; it has
no villi,

Note again the position of the suprarenal bodies.
Note the position of the kidneys, the left being

much nearer the pelvis than the right; observe
on either side the ureter, a pale semi-transparent

10

26.

27.

28.

ELEMENTARY PHYSIOLOGY. [T.

duct passing downwards from each kidney over
the muscles of the back towards the middle line;
trace them to their entrance into the urinary
bladder.

. Trace out the renal artery and vein noted in § 9,

follow them into the substance of the kidney.
Divide one kidney longitudinally, note the single
pyramid opening into the pelvis.

In dissecting a female rabbit, observe the uterus,
with its two cornua, from each cornu proceeds a
Fallopian tube which taking a winding course
upwards for some little distance ends in a clump
of processes or fimbriz. Near the end of each
Fallopian tube a little below the kidney will be
seen a small, ovoid spotted body, the ovary.

In dissecting a male rabbit, observe in each side
of the lower part of the abdominal cavity a white
convoluted tube the vas deferens. Cut through
the symphysis pubis with bone forceps, stretch
the halves apart and cut away as much bone on
each side as may be necessary. Trace the vasa
deferentia downwards cutting open the scrotal
sacs; each vas deferens is continuous with a
coiled mass of tubes, the epididymis, attached
to one side of the testis. Note that the smooth
membrane, tunica vaginalis, lining the scrotal
sacs is continuous with the peritoneum.

Lay open the bladder, observe its neck ending in
the urethra, note the openings of the ureters into
the dorsal part of the bladder and in the male
the openings of the vasa deferentia near its neck.

I] DISSECTION OF A RABBIT AND OF A DoG. 11

B. 1. Make a median incision over the skull from the
nose to behind the level of the ears. Reflect the
skin on each side. Cut away the attachment of
the muscles of the neck to the occiput until the
occipito-atlantoidean membrane between the
occiput and the atlas is laid bare. Carefully
divide this with scissors and observe the medulla
oblongata.

2. With a trephine saw through the roof of the
skull in its broadest part, a little behind the
orbits, working very carefully when the bone is
nearly sawn through. With a lever raise the
circular piece of bone and remove it. Then
with the bone forceps cut away piecemeal the
rest of the roof of the skull.

3. Note the thickish membrane, the dura mater
covering but not attached to the brain, it dips
down between the cerebral hemispheres as the
fale cerebri and between the cerebrum and
cerebellum as the tentoriuwm ; cut away the dura
mater and observe the very thin vascular mem-
brane, the pia mater, nl Si to the surface of
the brain.

4, Make a rough sketch of the exposed cerebrum,
cerebellum and medulla oblongata for com-
parison with the same parts in the dog. Note
particularly that in the rabbit the cerebral
hemispheres are smooth and that the olfactory
lobes are directly in front of the cerebral
hemispheres, being separated from them by a
constriction only.

ELEMENTARY PHYSIOLOGY. [t.

5. In the dog

a, The dura mater is much thicker and the pia
mater more obvious.

b. The cerebral hemispheres have deep fissures.

ce. The pia mater dips down into the fissures,
above the pia mater and bridging over the
fissures may be observed the thin transparent
arachnoid membrane, also distinctly visible as a
covering to the pia mater at the base of the
brain. In the space between the arachnoid and
pia mater is contained the clear watery sub-
arachnoid (or cerebro-spinal) fluid. A smaller
quantity of fluid also exists between the
arachnoid and dura mater.

d. Compare the exposed surface with the sketch
made of the surface of the brain of the rabbit,
noting the relative sizes of the cerebrum and
cerebellum in each,

With a scalpel divide the front of the cerebral
hemispheres from the olfactory lobes. Lift up
with the handle of a scalpel the extreme front of
the cerebrum, and turning it backwards bring
into view the optic nerves. Cut these through
with a sharp pair of scissors close to the skull.
Still turning the brain back cut through succes-
sively all the other cranial nerves. A little
behind the optic nerve is the small but evident
third nerve (motor oculi), close behind this the
considerably smaller fourth nerve (trochlear),
farther back in the hollow behind the attach-
ment of the tentorium lies the thick fifth nerve,
to the median side of which the small sixth

1] DISSECTION OF A RABBIT AND OF A DOG. 13

(abducens) is fairly conspicuous, A little behind
and to the outside of the fifth, in the hard
petrous bone are seen together the seventh
(facial) and eighth (auditory). Some distance
back and nearer the middle line come the ninth
(glossopharyngeal), tenth (pneumogastric), and
the small eleventh (spinal accessory). Lastly,
still farther back is the twelfth (hypoglossal).
Cut through the spinal cord below the medulla
oblongata, and remove the brain entirely. The
outlying lateral portions of the cerebellum will
probably be left in the skull. Do not injure the
skull in attempting to get these out’.

7. Cut and scrape away the tissue above the
cervical vertebre ; with bone forceps remove the
arches of the vertebre and cut them away at
the sides piece by piece so that the spinal cord
is well exposed. Pull the cord a little to one side
and note the nerves running into it, one between
each pair of vertebre. Carefully cut through
the dura mater and pull it up with forceps, a row
of fine nerve fibres will be seen issuing from the
spinal cord; they converge and form one bundle the
posterior root of the spinal nerve. Cut through
these filaments, and pull the dura mater a little
farther from the spinal cord; ventrally of the
above set of fine nerve fibres will be seen another
similar set which unite and form the anterior

1 The brain may be placed in spirit to harden and be dissected
later: most of the points of structure of the dog’s brain given in Lesson
xxx. can also be made out on the rabbit’s brain.

14

ro

' ELEMENTARY PHYSIOLOGY. [1.

root of the spinal nerve. Observe carefully the
roots on the outside of the dura mater, they join
almost immediately forming the nerve trunk, on
the posterior root at or a little before its junction
with the anterior root note the swelling caused
by the spinal ganglion.

Examine again the diaphragm (cp. A § 16).

Observe the large central tendon, with the vena

cava and cesophagus passing through and tightly
attached to it. The muscular part of the
diaphragm consists of a costal and vertebral
portion. The former is attached by short
tendons to the ribs and sternum. The latter
is attached to the upper lumbar vertebre; it is
a somewhat thick mass of muscle divided into
right and left portions by the descending aorta,
the right is much the larger; the two form the
pillars of the diaphragm.

Pull down the diaphragm by its pillars, on its
unpunctured side the lung will follow it.
Observe the pectoral muscle proceeding from
nearly the whole length of the sternum to the
humerus, cut it through together with the
vessels and nerves going to the arm and note
its attachments. ;

Several muscles will now be exposed, note the
serratus anticus major proceeding from the
lower part of the internal border of the scapula
to the 3rd to 9th ribs inclusive. Cut it through
and reflect the parts.

1]

DISSECTION OF A RABBIT AND OF A DOG. 15

Note the scalenus medius running from the
neck to the upper ribs (2nd to 5th); cut this
through where it is inserted into the ribs and
turn it forward, the scalenus anticus will be
seen attached to the Ist rib at its junction with
the costal cartilage.

The serratus anticus minor running from the
upper part of the internal border of the scapula
to the lower cervical vertebra and Ist and 2nd
rib.

The serratus posticus, a thin inconspicuous
muscle proceeding by rather a long broad tendon
from the cervical vertebre and dorsal fascia.
It is inserted into the 4—12th ribs about the
middle part of their course.

These muscles having been cut through the
small scalenus posticus will be seen running
from the neck to the Ist rib laterally of the
scalenus anticus. The three scaleni originate
from one or more of the transverse processes of
the 4th to 7th cervical vertebre.

Note the thick muscle the longissimus dorsi
covering the ribs dorsally ; cut away this and the
adjoining muscles and note the inconspicuous
levatores costarum proceeding from the trans-
verse processes of the dorsal vertebre to the ribs
below.

Clear away all muscles and tendons attached to
any two of the ribs (say 4th and 5th) except the
intercostal muscles joining them. Note the
external intercostal muscle, the fibres run

16

10,

12.

ELEMENTARY PHYSIOLOGY. [1.

downwards and ventrally, and are absent between
the costal cartilages, here the internal inter-
costal muscle is seen; carefully remove the
external intercostal, and so follow the internal
intercostal towards the vertebre ; the fibres run
downwards and ventrally and near the vertebre
are scanty or absent.

Observe more closely the costal cartilages and
their connection with the ribs and sternum,

11. The above mentioned muscles, especially the
thinner ones, should also be observed in the
dog, where they are larger. There are some
differences in arrangement,

The pectoral has an upper portion which runs
not to the scapula but to the humerus,

The serratus anticus runs from the whole length
of the internal border of the scapula to the
lower cervical vertebre and first seven ribs,
The origins and insertions of the scalent are
somewhat different.

The serratus posticus is divided as in man into
an upper and a lower portion (s. p. superior
and s. p. inferior.)

Cut through the costal cartilages on either side
close to the sternum, cut through the muscles be-
tween the 2nd and 3rd and the 8th and 9th ribs,
with bone forceps cut through the 3—8th ribs
dorsally and remove them. The pleural cavities
will be seen to be separated from one another by
the median parietal portions of the pleure,
between these is a space, the mediastinum.
From the surface of the lungs a shred of a fine

]

13.

14,

16.

DISSECTION OF A RABBIT AND OF A DOG. 17

membrane, the visceral portion of the pleura,

- may be torn; note that at the base of the lungs

this is continuous with the ‘parietal portion of
the pleura attached to the walls of the chest
and bounding the mediastinum. Note the
position of the heart.

In the mediastinum attached to the pleura note
on either side the phrenic nerve distributed to
the muscular fibres of the diaphragm.

With fine forceps tear off the membrane over
the phrenic nerve in the middle part of its
course; another membrane will be seen under-
neath, outside of which the phrenic runs, this is
the parietal layer of the pericardium; cut it
through, the heart will be seen to lie in a bag
formed by it. Remove the middle and posterior
portions of the sternum. Trace the connection
of the parietal layer of the pericardium with the
covering of the heart and of the roots of the
great vessels.

15. Turning in the dog the heart and lungs over to
the right, pull up the large aortic trunk, and
note the almost transparent thoracic duct,
lying alongside the esophagus; trace it up to
its termination into the venous system (at the
junction of the left jugular and left sub-clavian
vein, cp. § 20). With a little care the thoracic
duct may also be traced in the rabbit.

Prolong the median skin incision to the chin and
reflect the skin as far as possible, Observe on
2

18

17.

18.

ELEMENTARY PHYSIOLOGY. [T.

each side the external jugular vein arising an-
teriorly from two branches: avoid puncturing it.

Cut through in the middle line the thin super-
ficial muscle (platysma); draw it to one side,
clearing away the connective tissue. Lying on
either side of the muscles immediately surround-
ing the trachea will be seen the sterno-mastoid
muscle (cp. § 28) diverging from the lower part
of the neck. Cut through the connective tissue
on the inner side of one sterno-mastoid and draw
the muscle outwards; there will be seen the
common carotid artery, and, running along
the outer side of this, the pneumogastric nerve.
Free in one place the carotid, and lift it up with
a hook. In the underlying connective tissue
will be seen two nerves more or less closely
united by tissue ; the larger is the sympathetic,
the smaller the superior cardiac (depressor).

Clear away the connective tissue from the artery.
Draw the larynx from the carotid by means of a
hook to which is tied a string having a weight
at the end. Passing over the carotid at the
level of the larynx will be seen the descendens
noni, a branch of the 12th nerve. Cut this
through and remove it entirely. Passing under-
neath the carotid nearly at the same level is the
superior laryngeal branch of the pneumogastric.
Trace this with especial care; soon after it leaves
the pneumogastric it will be seen to give off a small
nerve, the depressor. Follow this down the neck,
separating it from the sympathetic. Sometimes

19.

20.

21.

DISSECTION OF A RABBIT AND OF A DOG. 19

the depressor receives a branch direct from the
pneumogastric; occasionally this is its sole
origin.

Remove the first rib and the remains of the
sternum, avoiding any injury to the tissues
below. Observe the thymus, a fatty looking
body covering the roots of the great vessels.
It may be torn away.

Trace out on each side the junction of the
external jugular and subclavian veins to
form respectively the right and left vene
cave superiores: near the junction ends the
internal jugular vein, this brings blood from
the brain and may be traced from the foramen
jugulare (cp. E. § 21) down the neck laterally of
the common carotid and vagus.

Observe the right vena cava superior passing
straight down to join the right auricle; the left
vena cava superior passing obliquely downwards
underneath the left auricle to join the right
auricle; and the inferior vena cava passing
upwards from the diaphragm to join the right
auricle.

Trace up one phrenic nerve. It makes its way
out of the thorax by the side of the superior
vena cava, and then passes beneath it. Place a
double ligature round the vein and divide be-
tween the ligatures. Follow up the phrenic to
its origin from the 4th and 5th (and also from
the 6th and 7th) cervical nerves,

2—2

20

23.

24.

ELEMENTARY PHYSIOLOGY. [r

Trace out the arch of the aorta by clearing away
the tissue from its upper surface. Take care
not to injure the pneumogastric nerves (see
next section). Observe on the right side the
innominate artery, which gives off first the
left common carotid, and then divides into
the right subclavian and right common
carotid; on the left side the left subclavian.
Note the vertebral artery on either side pro-
ceeding from the subclavian. On a level with
the anterior part of the larynx, note the division
of the common carotid into external carotid
and internal carotid. The former curls round
the angle of the jaw, the latter enters the skull
a little in front and to the median side of the
tympanic bulla.

Trace both pneumogastric nerves downwards,
observing the recurrent laryngeal branches
passing on the right side round the subclavian
artery, and ou the left round the aorta. Place
a double ligature round the innominate artery
and divide between the ligatures. Trace the
recurrent laryngeal nerves along the back of the
trachea to the larynx. Pursue the main pneu-
mogastric trunks on the cesophagus to the point
where they were seen in A, § 10,

. Trace the sympathetic nerve downwards to the

inferior cervical ganglion lying a little above
the subclavian artery, and close to the vertebral
artery; follow it thence to the first thoracic
ganglion.

I] ‘DISSECTION OF A RABBIT AND*OF A DOG. 21

Observe the branches going from these ganglia
towards the heart. Observe also the depressor
nerve passing to the heart.

From the first thoracic ganglion trace down the
thoracic sympathetic nerve trunk lying on the
heads of the ribs with the ganglia (twelve in all)
and the rami communicantes connecting each
ganglion with its corresponding spinal nerve.

26. Trace out the splanchnic nerve on one side; it
will be found to separate from the sympathetic
at the 8th, 9th, or 10th thoracic ganglion. At
first sight it appears to be the continuation of
the sympathetic instead of a branch of it; since
the sympathetic at its lower part becomes more
transparent, and running in a groove between
two muscles, is rather easily overlooked. The
splanchnic receives branches from each of the
thoracic sympathetic ganglia below its origin.

27. Tie a tube in the trachea and distend the lungs,
note the appearance of the distended lungs.
Cut out the heart’ with the lungs attached, and
trace the pulmonary arteries and veins.

28. Having reflected on either side the skin of the
neck of the dog, and cleared away the fascia of
connective tissue, observe the muscles under-
neath.

a. The sterno-hyoid close to the median line.
It runs from the sternum to the hyoid bone.

1 The heart may be dissected in the manner given for the sheep’s
heart in Lesson x11,

22

ELEMENTARY PHYSIOLOGY. [1

b. The sterno-thyroid lying laterally of (a) and

for the greater part of its course close to it,
it runs from the sternum to the thyroid
cartilage of the larynx.

c, The thyro-hyoid, a small muscle running
from the thyroid cartilage to the hyoid bone,
in the upper part of its course it lies lat-
erally of (a).

d. The sterno-cleido-mastoid lying laterally of
(6) and covering it near the hyoid bone,
thence it proceeds outwards, and disappears
under a white oval mass, the submaxillary
glands.

These muscles may be dissected in the rab-
bit also, the representative of the sterno-
cleido-mastoid has however no clavicular
attachment and hence is called the sterno-
mastoid, it does not come in contact with
the submaxillary gland.

29. Carefully separate the sterno-mastoid from the
sterno-thyroid; the sympathetic-pneumogastric
trunk and the carotid artery will come into
view.

Observe the following points in which the dog
differs from the rabbit :

a. There is but one superior vena cava form-
ed by the junction of the two innominate
veins.

(The arrangement of the main arteries is usually
that described above for the rabbit, but con-
siderable variations occur.)

-

ar |=,

1.]

30.

DISSECTION OF A RABBIT AND OF A DOG. 23.

B. There is in the neck no separate nerve corre-
sponding to the depressor in the rabbit.

y. The sympathetic and the vagus run in the neck
in a thick sheath common to both. At the lower
end of the neck, the sympathetic joins the infe-
rior cervical ganglion. From the ganglion run
several pale nerves to the heart and lungs, and
receives two white ones—the annulus of Vieus-
sens—from the first thoracic ganglion. The
latter receives rami from the lower cervical and
first two dorsal nerves, of these the 2nd dorsal
only (the 10th spinal nerve) gives an obvious
white as well as a grey ramus to it.

Clear away any muscles that may remain around
the lower part of the larynx; on either side
of it is attached a thin, dark red lobe of the
thyroid gland, the lobes run a short way down
the trachea, and there join over the ventral
surface of the trachea by a very thin connecting
piece.

Cut through the skin in the front of the thigh
and turn it back on either side; in the upper
median part blood-vessels will be dimly seen
through the thin sartorius muscle; cut through
this muscle and note the femoral (crural)
artery and vein, and the crural nerve run-
ning side by side; trace the artery upwards,
it unites with other arteries to form the common
iliac, which with the common iliac of the other
side forms the abdominal aorta; trace similarly
the femoral vein to the.common iliac vein and

24

ro

ELEMENTARY PHYSIOLOGY. [I.

the inferior vena cava. Follow the crural nerve
up to the spinal cord, it arises chiefly from the
5th lumbar nerve (receiving branches also from

the 6th and 7th).

Remove the skin from the back of the thigh, cut
through the tendonous line seen over the femur
and pull the outside mass of muscle outwards,
the large sciatic nerve will be seen, trace this
to the top of the thigh, then turn the rabbit
over and follow the nerve to its origin from the
spinal cord; it arises chiefly from the 7th
lumbar and Ist sacral nerve (receiving branches
from the 6th lumbar and 2nd and 3rd sacral
nerves.)

The Student should have a rabbit’s and a dog’s
skull before him, and make out the several

openings by which the nerves spoken of below
issue from the skull.

Carry up to the chin the median skin cut and
reflect the skin, place the head on one side; just
in front and ventrally of the base of the ear will
be seen the thin dorsal part of the parotid
gland, often much hidden by fat tissue; the
gland stretches ventrally a little past the angle
of the jaw.

From the anterior border of the parotid gland,
issues the greater part of the facial nerve
(7th) dividing into several branches which run

I.]

DISSECTION OF A RABBIT AND OF A DOG. 25

forwards across the masseter muscle to their
endings in certain muscles of the face.

The duct of the parotid (duct of Stenson)
runs forward with the facial nerve, from the
gland; it is small, thin-walled and inconspicuous,
it may sometimes be made evident by pressing
on the gland and so forcing some fluid into it.
The branches of the facial should be carefully
isolated close to the gland, the connective tissue
being cut through with a fine pair of scissors as
close as possible to the nerves lest the duct be
inadvertently severed; on pulling the nerves to
one side the duct will be seen, follow it forwards
to the anterior edge of the masseter where it
dips down to the mouth; make a small cut in it
with scissors and pass a bristle down it.

In the dog the duct is much more obvious, the
facial nerve does not accompany it.

Cutting through the parotid gland, trace the
facial nerve to its exit from the skull by the
stylo-mastoid foramen; observing the branches
going to the muscles of the ear.

Behind the parotid gland will be seen a nerve
running from the under surface of the sterno-
mastoid muscle (cp. C. § 21) dividing into two
branches, and passing up the ear. This is the
great auricular, which arises from the 3rd
cervical nerve, and is the main sensory nerve
for the ear. Trace as far as possible its course
in the ear.

26

~]

ELEMENTARY PHYSIOLOGY. [1

In the dog, reflect the skin of the head, note
again the position of the submaxillary gland
(cp. C. § 28 d.) as seen from the surface ; it lies
between two large branches of the jugular vein ;
attached to the inner part of the posterior
extremity of the lower jaw will be seen the
digastric muscle; clear away the connective
tissue surrounding it, cut it through, taking
care not to injure the parts beneath, and
reflect the cut ends; the submaxillary duct
(duct of Wharton) will be seen running from
the gland, trace it forwards, it runs underneath
(dorsally of) a muscle with transverse fibres, the
mylohyoid, cut through this, turn the lateral
part as far back as possible, taking care that
the fascia on its lower surface is not attached
to it and follow the duct forwards.

Attached to the anterior end of the submaxillary

gland and stretching for some little way along
its duct will be seen the smaller sublingual
gland, from this runs the sublingual duct,
alongside and laterally of the duct of the
submaxillary gland.

A short distance from the lower border of the
mylohyoid inuscle the lingual nerve will be seen
crossing the ducts and running on to the tongue:
pull the tissues on which the lingual rests well
away from the jaw, about three quarters of an

inch centrally of the point where the lingual

crosses the ducts, it will be seen to give off a
small nerve the chorda tympani. This curves
towards the ducts and then runs alongside them
towards the sublingual and submaxillary glands.

1]

10.

Rie

DISSECTION OF A RABBIT AND OF A DOG, 27

9. Trace the ducts peripherally, they unite and
open underneath the tongue ; trace the lingual
nerve peripherally, it supplies chiefly the tip of
the tongue.

In the rabbit there will be seen lying between
the angles of the lower jaw the tolerably com-
pact but soft submaxillary glands touching
one another in the median line. Each gland is
laterally in contact with the ventral lobe of the
parotid, its tint is redder than that of the
parotid; pull the submaxillary gland laterally
and backwards, its smalh duct will be seen
running from it over the muscle attached to the
inner surface of lower jaw, and then underneath
(dorsally of) the digastric muscle, which here
has a conspicuous tendon; cut through the
digastric and trace the duct forward underneath
the mylohyoid muscle; a short distance from
the lower border of the mylohyoid this duct is
covered by the lobules of the small sublingual
gland, turn this back, the lingual nerve will be
seen crossing the duct, with care in dissection
fine nerve fibres, chorda tympani fibres, may
be seen running from the lingual nerve to the
sublingual gland and to the duct of the sub-
maxillary gland, the latter fibres are too small to
follow towards the gland itself.

Now follow up the pneumogastric nerve from
the place where it was left in C.§ 18. A little
above the superior laryngeal branch will be seen
the pharyngeal nerve, and higher up still a

28

12.

13.

16.

ELEMENTARY PHYSIOLOGY. [L.

fusiform enlargement, the ganglion of the
trunk.

Note the hypoglossal, a large nerve running
across the pneumogastric a little centrally of its
ganglion. Trace it forwards to the muscles of
the tongue.

Follow up the sympathetic nerve, it has, at about
the level of the ganglion of the pneumogastric, a
considerable enlargement, the superior cervi-
cal ganglion; observe the fibres which run
from this along the carotid artery and its
branches.

14. In the dog the sympathetic and pneumogastric
nerves which run in a common sheath in the
neck (cf. C., § 28 (y)) separate from one another
a little distance from their respective ganglia.

. Partly saw through the symphysis menti, then

use a lever and force the rami asunder, and in
the following dissection cut through or remove
any muscles necessary.

Trace the lingual backwards. It will be found
to join the inferior dental (a large nerve
entering into the lower jaw), to constitute, with
other branches, the inferior maxillary nerve.
Trace this back to the front edge of the tympa-
nic bulla.

17. Note in the dog the small nerve, chorda tym-
pani, which joins the lingual soon after the
latter branches off from the inferior dental;
trace the chorda tympani centrally, it will be
found to make its exit from the tympanic bulla

1.]

19.

20.

21.

DISSECTION OF A RABBIT AND OF A DOG. 29

close to the Glaserian fissure. Break through
the bulla, and observe the chorda running
across the tympanic cavity over the handle of the
malleus (cp. Lesson xxvii). This course of the
chorda tympani may be followed in the rabbit,
but the dissection is not easy.

18. Note also in the tympanic cavity the very small
nerve running over the promontory or projec-
tion of the cochlea. This is Jacobson’s nerve,
a branch of the 9th.

Trace up the pneumogastric beyond its ganglion,
to its exit from the skull by the foramen jugulare.
Note, passing from the skull with the pneumo-
gastric, the small spinal accessory nerve
behind and the glosso-pharyngeal in front;
the communicating branches between these
nerves may be neglected.

Trace the glosso-pharyngeal forwards to the
tongue and pharynx. It runs nearly in the
same direction as, but at a higher level than, the _
hypoglossal, and may be traced to the hinder
part and to the sides of the tongue.

Cut through the above three nerves, a little
distance from the skull, break away with small
bone forceps the tympanic bulla, and trace more
thoroughly the exit from the skull of these
nerves and of the hypoglossal. The latter issues
through the condyloid foramen, which is separ-
ated by a distinct width of bone from the
foramen jugulare, through which the other three
issue,

30

22.

23.

ELEMENTARY PHYSIOLOGY. [I.

Saw through the base of the skull and the face,
from the occiput to the nose, a little on one side
of the median line.

The nasal septum will be seen dividing the
nasal cavities except posteriorly. Note the
anterior and posterior turbinate bones both
consisting chiefly of thin folded laminz, pass a
bristle through the anterior nasal opening into
the nasal cavity, using bone forceps and scissors
trace the passage from the nasal cavity through
the posterior nasal opening into the pharynx
and trachea; note that the posterior turbinate
bones are not in the direct course between the
anterior nares and the trachea. Cut through
the septum nasi dorsally close to the nasal
bones, and remove the nasal bones, note that
the posterior turbinate bones and the posterior
dorsal part. of the septum are covered with a
yellowish mucous membrane which is thicker
than that lining the rest of the nasal cavities ;
this is the olfactory part of the mucous mem-
brane (Schneiderian membrane). Trace the
olfactory nerve forwards from the brain; it
divides into a number of fibres which run to the
Schneiderian membrane.

Looking down into the pharynx, observe the
epiglottis and the way in which it when pushed
backwards folds over the opening to the larynx.

Put the larynx of the dog into weak spirit for
dissection later, (Lesson xxx11.)

24. Look at the side of the pharynx for the opening

26.

DISSECTION OF A RABBIT AND OF A DOG. 31

of the Eustachian tube, pass a probe up it into
the tympanic cavity, Pass another probe down
the meatus externus and, rupturing the mem-
brana tympani, make sure that the first probe
has entered into the tympanic cavity.

. Remove one eye from its orbit, cutting through

the tissues close to the eye. In the anterior part -
of the orbit note the white Harderian gland;
in the anterior lower part the pale red infra-
orbital gland, the duct of which opens into the
mouth near the upper molars; and the lachry-
mal gland pale-red like the infra-orbital in
the posterior part of the orbit. Observe the
point of entrance of the optic nerve into the
orbit,
In the dog the muscles of the globe of the eye may
be dissected out, after removing with bone
forceps the roof of the orbit.

Immediately below, and in front of the eye,
the superior maxillary nerve will be found
issuing from a foramen in the superior maxillary
bone, to supply the skin of the face, &c., with
sensory fibres. Cutting away the bone with a
small pair of bone forceps, trace this nerve back

_ along the floor of the orbit.

27. In the upper part of the orbit of the dog note
the ophthalmic nerve. It passes from the front
of the orbit to the forehead,

The superior and the inferior maxillary nerves
and the ophthalmic, when traced back, will be
found to unite into one large nerve, the fifth or

32

29.

ELEMENTARY PHYSIOLOGY. [1.

trigeminal. Observe on the nerve at the junc-
tion of the three branches, the swelling of the
Gasserian ganglion.

Observe also that the nerve in leaving the brain
has two roots, a small and a large, that the
small root passes beside the ganglion on the
large root, without entering into it, and that the
fibres of the small root are, beyond the ganglion,
almost entirely confined to the third or inferior
maxillary branch.

Cut out the tongue taking care to remove the
whole of it; on either side of the posterior upper
surface, will be seen a small oval patch, the
papilla foliata or lateral taste organ; note the
parallel ridges running at right angles to the
long axis of the papilla.

As an introduction to the methods of preserving
and hardening tissues, the following should be
done by each student. The tissues should be
removed from the rabbit as soon as possible
after it has been killed, and sections should be
cut when the Lessons dealing with the several
tissues are being worked through.

Cut out from the greater curvature or fundus of
the stomach a piece about 1 c.m. square, wash
it for a moment in NaCl. ‘6 p.c. to remove any
acid or any food substance on the surface of the
mucous membrane; with hedgehog quills or
small pins fasten it out on a-piece of cork with

DISSECTION OF A RABBIT AND OF A DOG. 33

the muscular surface downwards, stretching it
slightly, and place it in alcohol about 75 p.c. for
about an hour, then remove to 95 p.c. alcohol for
a fortnight; keep in 75 p.c. alcohol.

Cut out of the small intestine a piece about two
inches long; tie into each end a short glass tube
with lumen about 5 m.m. in diameter, over the
free end of each glass tube slip a piece of india-
rubber tubing; by means of a syringe wash out
the piece of intestine with NaCl’6 p.c. for
about twenty seconds, then inject chromic acid
‘2p.c. When the salt solution has been displaced
by chromic acid tie or clamp the peripheral piece
of tubing, inject a little more chromic acid to
distend slightly the intestine, and tie or clamp
the central piece of tubing. Place the distended
intestine in ten times its bulk of chromic acid
‘2 p.c. In two to three days cut off both ends
of the intestine, cut it open longitudinally, and
place it in fresh chromic acid ‘2 p.c.; in about
ten days place it in water for some hours, and
then in alcohol 30 p.c.; on the next day transfer
it to alcohol 50 p.c. renewing the alcohol as long
as it becomes coloured and finally place the tissue
in strong spirit.

Take a piece of costal cartilage about 5 m.m.
long and place it in about 10 c.c. of saturated
aqueous solution of picric acid; after about ten
days wash it well with water and place for a
day in 50 p.c. alcohol, then transfer to strong
spirit.

3

34

ELEMENTARY PHYSIOLOGY. [I.

Take of the sciatic or other large nerve a piece
about 10 m.m. long and place it in about 10 c.c.
of ammonium bichromate 2 p.c. In a week
renew the ammonium bichromate; in this fluid
it may be kept until sections are required, or in
a month or more it may be washed with water
and placed in spirit as in § 2.

LESSON IL.

STRUCTURE OF BLOOD.

A. Buoop or Frog or Newt.

Having destroyed the brain and spinal cord of a
frog‘, cut through the skin in the median ventral
line, cut transversely through the lower part
of the sternum just above the epigastric vein,
and expose the heart. Cut off the tip of the
ventricle ; with a glass rod transfer a small drop
of blood to a glass slide and place on it a cover-
slip.

Examine it under the microscope with a low
magnifying power® and observe the numerous
corpuscles floating in the plasma.

Examine it with a high magnifying power’ and
observe the red corpuscles; if a large drop of

1 Cp. Appendix,

* For convenience the term ‘low power’ will be used throughout
for a combination of lenses which magnifies less than 100 diameters,
and the term ‘high power’ for a combination of lenses which mag-
nifies more than 300 diameters. In Zeiss’ microscope, objective A
with ocular 2 magnifies 55 diameters, with ocular 3 it magnifies 75
diameters; objective D with ocular 3 magnifies 320 diameters, with
ocular 4 it magnifies 440 diameters. If the tube be drawn out the

3—2Z

36

ELEMENTARY PHYSIOLOGY. [11.

blood has been taken the corpuscles will pro-
bably form a continuous layer, in which case a
drop of °6 p.c. sodium chloride solution should be
made to run under the cover-slip (cp. § 4).

a.

d.

The red corpuscles are flattened ellipsoids ;
note their spindle shape as they roll over.

They appear homogeneous; if however the
specimen be not carefully prepared a certain
number of the corpuscles will be altered and
show a central oval nucleus.

A single corpuscle is pale yellow, the colour-
ing substance being equally diffused through-
out it; when several corpuscles lie over one
another they together appear red.

The great majority are of the same size and
tint.

2. Examine the colourless corpuscles in parts of
the specimen where the red are not very nume-
rous.

a.

b.

Cc.

d.

They are much fewer than the red.

They are smaller than the red, but vary
considerably in size.

Most have an irregular form, some are
spherical.

They are colourless and granular; the gra-
nules vary greatly in distinctness and size.

magnification is of course greater. The 3 inch and ¢ inch objectives
of English make correspond respectively to the A and D objectives of
Zeiss. With Hartnack’s microscope the nearly corresponding lenses
are oc. 2 or 3, obj. 3 (low power) and oc. 3 or 4, obj. 7 (high power).

IL]

STRUCTURE OF BLOOD. 37

e. The nucleus can seldom be made out,

except when the corpuscle is very extended.
Do not confound a heap of granules or a
protuberance with the nucleus.

Choosing a corpuscle either elongated or
having several processes, watch carefully its
amceboid movements; make half a dozen
drawings of its outline at intervals of about
twenty seconds.

g. When a drop of blood is first mounted the
colourless corpuscles are usually spherical,
they soon begin however to put out pro-
cesses; if it is desired to watch the move-
ments for any length of time a fresh drop
should be mounted and protected from eva-
‘poration in the following manner. With
a morsel of blotting-paper dry if necessary
the slide at the edges of the cover-slip.
Keep the cover-slip in place by gently
holding a needle against one edge, and,
with a small brush, brush carefully the
melted paraffin A.’ (which melts at 39°C.)
over the edges all round. The paraffin need
not extend more than } or } inch over the
cover-slip.

. With the aid of a camera lucida’ make an

outline drawing of two or three red corpus-
cles; substitute for the specimen a stage

‘micrometer’, and being careful that the mi-

croscope and the drawing-pad are in the

1 Cp. Appendix.

38

ELEMENTARY PHYSIOLOGY, [II.

same positions as before, make a drawing of
the micrometer lines over the previously
made drawing of the corpuscles; then, the
real distance between the micrometer lines
being known, the diameters of the corpuscles
can be at once read off; thus if the micro-
meter lines are ;4, mm. apart and in the
drawing a corpuscle exactly occupies one
division its diameter in that direction is
evidently ;4; mm.

The drawing of the micrometer lines may be
kept as a scale, and any object drawn under
the same magnifying power and with the
pad and microscope in the same relative
positions may be directly measured by it.

Substitute for the ordinary eye-piece of the
microscope one which has a ledge for sup-
porting an ocular micrometer’, the values of
which have been determined, the size of the
corpuscle can then be at once read off.

Mount another small drop of blood, place a
small drop of ‘1 p.c. acetic acid on the glass
slide so that it just touches the edge of the
cover-slip; place a piece of blotting-paper on
the opposite side just touching the fluid at the

- edge of the cover-slip, the acetic acid will then
run under the cover-slip and mix with the blood.
Note the changes which take place.

a. In the colourless corpuscle, the cell sub-

stance becomes more transparent. but shews

1 Cp. Appendix.

ee

ee —— -_
aS

eS ee

f

STRUCTURE OF BLOOD. 39

several dark granules; a granular nucleus,
often irregular or lobed, comes into view,
usually more than one nucleus will be seen.

In the red corpuscles the nucleus be-
comes obvious; it is when first seen nearly
homogeneous, and oval in outline, later
it becomes granular and usually irregularly
rod-shaped.

The red corpuscles swell up owing to absorp-
tion of water, most after a time become
spherical (if strong acid be used the cor-
puscles usually preserve their shape).

They become colourless, the colouring matter
being dissolved; occasionally the colouring
matter is massed round the nucleus before
complete solution takes place (effect of water)
and occasionally the nucleus becomes stained
yellow by the colouring matter (effect of acetic
acid).

Finally the outline of the corpuscles is seen
as a faint line at some distance from the
nucleus. Observe the not infrequent excen-
tric position of the nucleus,

Some corpuscles are much more readily
acted on than others.

Irrigate with a strong aqueous solution of
Spiller’s purple or magenta,

a.

The outline of the red ecorpuscle becomes
distinct, its nucleus stains deeply, around
the nucleus a little faintly stained granular

40

ELEMENTARY PHYSIOLOGY. [i qs

substance is seen which often stretches out
to the periphery of the corpuscle in the form
of a star. The nuclei of the colourless cor-
puscles also stain deeply.

Place several very small drops of blood two or
three mm. apart on a slide and leave for a few
minutes, then cover with a cover-slip, and put
under a high power. Take a little blood from a

freshly killed frog and establish a current under-

neath the cover-slip from one side of it to the
other (cp. § 4). The first small drops will have
partially clotted and will serve as an imperfect
barrier to the corpuscles in the current; in such
places note that the shape of the red corpuscles

is easily changed and recovered, and that the ©

colourless corpuscles stick to one another and to
the glass more than do the red. After the
current has passed a short time largish clumps
of colourless corpuscles will be seen.

Having destroyed the brain and spinal cord of a
frog, expose the heart and cut it across, suck up
a little blood in a clean pipette and add it to
about five times its volume of 2 p.c. boracic acid,
stirring gently. Mount a drop of the mixture at
once and observe the red corpuscles with a high

power.

The nuclei scarcely visible at first become in a
short time rather deeply stained with hemo-
globin ; small spheres of hemoglobin appear also
in the body of the corpuscle; occasionally the
hemoglobin may appear to stretch in rays from

STRUCTURE OF BLOOD. 41

the nucleus through the body (if the rays are
not seen irrigate with 2 to 5 p.c. salt solution,
but in this case be careful not to mistake foldings
of the corpuscle for rays). Later the corpuscle
becomes spherical and its body colourless. Whilst
the earlier changes are taking place some of the
corpuscles may be seen to extrude their nuclei.

Dilute a little fresh blood with twice its volume of

“6 p.c. salt solution ; mount a drop of the mixture

and place it aside for an hour or so to clot;
urigate it with 30 p.c. alcohol and then with

-§piller’s purple dissolved in water or in dilute

alcohol. Note the deeply stained network of
fibrin fibrils and the numerous long threads of
fibrin running from the broken-down colourless
corpuscles.

B. Btoop or MAN.

With a needle prick the end of a finger, and

squeeze out a small drop of blood and mount it

(cp. A.§ 1). Observe the red corpuscles.

a. They roll about readily, when the cover-slip
is lightly touched.

b. Soon after being taken from the body they
stick to one another, and, owing to their

- shape, usually in rouleaux.

c. They are biconcave discs. Note that on fo-
cussing down on the circular face a darkish
centre and a light rim is first seen and then

ELEMENTARY PHYSIOLOGY. [IT.

a light centre with a darkish rim: when
viewed in profile and the centre focussed
they appear somewhat dumb-bell shaped.

d. They appear homogeneous, their colour is
like that of the red blood corpuscles of the
frog (ep. A. § 1, c).

e. Towards the outside of the drop, where
evaporation is going on, many of the red
corpuscles are crenate,

f. They are much smaller than the red corpus-
cles of the frog. Measure them (cp. A. § 3).

Observe the colourless corpuscles. They are
larger than the red, they resemble the white
corpuscles of the frog (A. § 2, ¢. d. e.); to observe
their amceboid movements a drop should be
protected from evaporation (A. § 2, g) and, pre-
ferably, warmed to the temperature of the body.

Irrigate with °5 p.c. acetic acid (ep. A. § 4).

a. The red corpuscles swell up and become
spherical, their hemoglobin is dissolved,
leaving the hardly visible stroma. (Effect
of water.)

b. No nucleus is brought into view.

c. The white corpuscles behave like those of
the frog (A. § 4, a).

Count the red corpuscles with Gower’s hemato-
cytometer in the following manner.

Fill the larger pipette with sodium sulphate
solution of Sp. Gr. 1025 up to the mark on the

OO ae

STRUCTURE OF BLOOD. 43

stem, it then contains 995 c.m.; empty it into
the measuring glass. [ill the small pipette
with freshly drawn blood up to the line marked
5 c.m.; empty it into the measuring glass, and
with the fluid in the measuring glass wash out
the blood sticking to the inside of the tube;
thoroughly mix the blood and salt solution with
the glass spatula, place a small drop of the
mixture in the centre of the glass cell and
over it lay a cover-slip, arrange the springs on
the cover-slip to keep it in position, and under
a high power count the number of red corpuscles
in ten of the squares which are marked at the
bottom of the glass cell.

Since the depth of the cell is £ mm. and the
side of each square is ;4, mm., there is beneath
each square 5}, c.m. of the mixture, 7.¢. 53555
e.m. of blood, hence the number of corpuscles
in 10 squares multiplied by 10,000 gives the
number of corpuscles in 1 ¢. m. blood.

DEMONSTRATIONS.
The method of using the simple and Stricker's
warm stage.
The ‘platelets’ of frog’s or newt’s blood (ef.
p- 387).
Specimens to show the chief stages of indirect
nuclear division (cf. p. 395).

Nore. If the brain of a frog be destroyed, a drop of curari injected
under the skin, and the frog be left for a day in about } inch of water,
the lymph sacs will become filled with lymph containing numerous
white corpuscles, many in a state of active amoeboid movement.

LESSON III.

COAGULATION OF BLOOD. CHARACTERS

OF PROTEIDS.

Observe the coagulation of freshly shed blood’;
it is at first fluid but soon passes into a jelly
which gradually becomes firm; if then placed
aside for some time, drops of clear serum will,
by the shrinking of the fibrin, be pressed out on
the surface of the clot; later the clot shrinks
more or less completely from the vessel squeez-
ing out more and more serum.

With a feather stir slowly about 10 c.c. of freshly
shed blood’; a considerable portion of the blood
will form a clot on the feather; squeeze out the
clot under a stream of water from a tap; the
clot shrinks considerably and a small quantity
only of fibrin is. obtained,

Repeat § 2, but.this time stir quickly, filaments
of fibrin will be obtained ; note that the fibrin is
extensible and. elastic; leave the defibrinated
blood for a day, no further clot is produced.

1 This will be obtained by the Demonstrator.

ee eee Ce eee ee ee ee ee eee

Er

— ee ae

111. ]

or

~]

8.

COAGULATION OF BLOOD. 45

Place a small drop of fresh blood on a piece of ©
glazed neutral litmus paper, in about ten seconds
wipe off the drop, a blue spot will be left showing
that the blood is alkaline. Test also the reaction

of serum.

Apply the Xanthoproteic and Millon’s test for
proteids (cp. § 16) to fibrin chopped up and
suspended in water.

Take two test-tubes and in each place a few

flocks of fibrin.

a. Add water and place in y wats bath at about
39° C. for a day; the fibrin does not dissolve
(it thus differs from albumin and peptone).

b. Treat similarly but with dilute (1 p. c¢.) solu-
tion of sodic chloride; the fibrin does not
dissolve (it thus differs from globulin),

Place two or three flocks of fibrin in a test-tube
containing a few c.c. of ‘2 p.c. HCl, the fibrin
soon swells up and becomes transparent; neu-
tralize the acid with Na,CO,, the fibrin shrinks
to its original size. Ifthe fibrin is warmed with
the acid, solution slowly takes place, acid-albu-
min being formed (cp. Lesson IX.).

Examine the plasma of horse’s blood kept, by
means of cold, from coagulating’.

1 The blood is allowed to run from the animal into a tall narrow
vessel contained in a much larger one packed with ice, a little salt
may be mixed with the ice, but of course not enough to reduce the
temperature so much that the blood is frozen; sometimes also a vessel

46 ELEMENTARY PHYSIOLOGY. [11d

a. Transfer with a pipette 2 or 3 cc. of the
plasma into a small test-tube. Observe the
coagulation and compare it with that of § 1.
Avoid shaking. Probably the fibrin will
adhere so strongly to the sides of the tube
that little contraction will take place. On
being freed from the glass it will contract.
If the clot has already shrunk away from
the sides of the vessel, it may since it is
colourless be overlooked unless the fluid be
carefully examined.

b. Dilute 1 cc. of the plasma with 50 cc. of
distilled water or normal saline solution.
Carefully avoid shaking and leave it till
the next day. Observe the fine delicate
fibrils of fibrin which are formed.

9. Examine the plasma of blood prevented from
coagulating by the presence of neutral salts’.

a. Remove 1 or 2 cc. carefully with a pipette,
avoiding blood-corpuscles as much as possi-
ble, and. dilute five to tenfold with water.

filled with ice is placed in the one which receives the blood. Horse’s
blood is preferable to bullock’s or dog’s, since it clots less readily and
the red corpuscles sink more quickly.

1 In preventing coagulation by neutral salts, blood is collected in
a vessel containing a saturated solution of magnesic sulphate; as the
blood runs in, it must be mixed well with the salt solution, preferably
by stopping the flow of blood now and then and turning the vessel
upside down. There should be about 1 vol. of the salt solution to
4 vols. of blood. The vessel may advantageously be surrounded by
ice or by ice and salt. On either method (§ 9 or § 10) clotting some-
times takes place, but the remaining fluid may still give a clot on
appropriate treatment.

—— |

III. ]

COAGULATION OF BLOOD. 47

The mixture will clot very speedily if placed
in the warm chamber; less speedily if left at
the ordinary temperature.

. Remove about 10c.c. into a small conical glass.

Add powdered sodic chloride to excess, stirring
but not more than is necessary to assist the
salt to dissolve. As the point of saturation
is reached, a flaky precipitate makes its
appearance. If the precipitate be plentiful,
remove it with a spatula, put it on a small
filter wetted with a saturated solution of
sodium chloride and wash with small quan-
tities of the same: if the precipitate be
small, decant it and the fluid from the
undissolved salt; filter, and wash the pre-
cipitate on the filter paper with small quan-
tities of a saturated sodic chloride solution.
Dissolve the substance so obtained, the plas-
mine of Denis, in a small quantity of dis-
tilled water, and filter. Probably a portion
of it will not dissolve, having already coagu-
lated. The clear, colourless fluid filtrate will,
if set on one side, clot. Avoid shaking after
filtration. If a small quantity only of the
fibrin factors be present, the fine threads of
fibrin, as they are formed, are loosened by
the shaking, and contract; thus the more
easily recognized gelatinous stage is lost.
This operation is the more successful, the more
rapidly it is carried. on.

10. To 2c.c. of hydrocele or other serous fluid which

48

11.

12,

13.

14.

ELEMENTARY PHYSIOLOGY. (111.

has been ascertained not to coagulate, nor to have
coagulated spontaneously, add 2c.c. of fresh blood-
serum, gently mix, and put on one side.

After a while, possibly not until after twenty-
four hours, the mixture will have coagulated.
The coagulation will be more rapid in the warm
chamber. :

Take 10c.c. of fresh blood-serum and saturate it
with magnesic sulphate by adding the salt in
powder. Paraglobulin will be precipitated,
since like other globulins it is insoluble in a satu-
rated solution of a neutral salt; filter (before
filtering the precipitate may be allowed to settle
and most of the fluid removed by decantation),
wash on the filter with a saturated solution of the
salt, then add 5 c.c. water to the precipitate, the
salt solution clinging to the precipitate will be
diluted and the paraglobulin dissolved. It does
not coagulate spontaneously.

‘Add a small quantity of paraglobulin solution to

hydrocele fluid. Coagulation will result.

Treat 10c.c. of hydrocele or pericardial fluid with
sodium chloride to saturation, and proceed as in
§ 11; a precipitate of fibrinogen will be ob-
tained, its solution does not coagulate sponta-
neously.

Add to 1 c.c. of a strong solution of fibrinogen an
equal volume of blood-serum, and set aside;
coagulation will take place.

5. Take 2c.c. of plasma (§ 9), add to it 16 cc. of

111]

16,

ways;
a.

COAGULATION OF BLOOD. 49

water, and determine that the mixture coagu-
lates very slowly.

Take another 2 c.c. of plasma, and add to it 16 c.c.
of an aqueous solution of prepared fibrin fer-
ment’; coagulation will quickly take place.

Dilute serum ten-fold with water and with it
observe the following general reactions of pro-
teids. (If sufficient serum is not obtainable,
take the white of an egg, cut through the
membranes in several places with scissors, add
50 vols, of water, beat up well, filter through
flannel and then through filtering paper.)

a. Xanthoproteic reaction. Take a little of the

1 The student may prepare fibrin ferment in one of the following

Let blood run into 10 times its volume of water, tilt it upside
down once or twice so that the fluids are well mixed, and let it
stand for a day. Filter through muslin and squeeze the excess
of fluid out of the clot, chop it up and wash with water until
all or nearly all of the colouring substance is removed, place it
in 10 times its bulk of 8 p.c. NaCl solution, and warm for one
to two days; filter; the filtrate contains fibrin ferment.

Add alcohol in abundance to serum until no further precipita-
tion takes place, filter, and dry the residue over a water-bath
at 35°C.; place the residue in a bottle containing an excess of
absolute alcohol, and leave for a month; at the end of this
time decant as much alcohol as possible, evaporate the rest
at a low temperature (under 40°C.); extract the residue with
200 times its volume of water, and filter, The alcohol will
have coagulated the greater part of the paraglobulin and albu-
min, etc., and so rendered them insoluble in water, hence the
aqueous filtrate will contain little else than fibrin ferment, the
more so the longer the alcohol has been acting; the compara-
tive absence of proteids should be tested by the reactions given
in § 16,

a +

ELEMENTARY PHYSIOLOGY. [ITl.

dilute serum, add a few drops of nitric acid,
and boil. The white precipitate of proteid
material at first formed becomes yellow and
partially dissolves, forming a yellow solution.
If the quantity of proteids present is small,
the yellow solution only will be obtained.
Place the test-tube in a stream of water from
a tap to cool and when cold add ammonia;
the yellow is turned to orange.

b. To another small quantity of the serum add
a few drops of Millon’s re-egent’. <A pre-
cipitate will be formed which turns pinkish
on boiling; if the amount of the proteids
present be small, no distinct precipitate will
be formed but the fluid will turn pink on
boiling.

c. Add a drop of cupric sulphate solution to an
excess of sodium hydrate. To the blue solu-
tion so formed add a little dilute serum, the
fluid will become violet (cp. reaction of pep-
tone, Lesson xvi). The violet colour be-
comes deeper on warming.

d. Add strong spirit, a precipitate is formed
(peptone in solution is precipitated with
difficulty).

e. Add excess of acetic acid and a few drops of
a strong solution of potassium ferrocyanide, a
precipitate is formed (peptone is not thus
precipitated).

17. With the diluted serum or white of egg observe
1 Cp. Appendix.

tL]

18.

i)

COAGULATION OF BLOOD. 51

in the following manner the coagulation of
albumin and globulin by heat. The white of
egg contains a small quantity only of globulin,
Place a test-tube containing 10 c.c. of the fluid
in a water-bath at about 50°C. and heat the
bath not too slowly up to about 80°C. Note on
a thermometer placed in the bath or in the
test-tube the temperature at which coagulation
begins (fluid slightly milky) and that at which
coagulation is complete (precipitate with clear
fluid). If the fluid is very slowly heated the
coagulate may be small owing to the formation of
alkali albuminate (cp. Lesson 1x. § 16); to avoid
this the serum should be carefully neutralized
with acetic acid before dilution.

Add 2¢.c: of serum drop by drop to about 100 c.c.
of water in a glass beaker. A faint cloudy
precipitate of globulin, which is insoluble in
water, will be seen (cp. Lesson Ix. § 14),

DEMONSTRATIONS,

Coagulation of blood over mercury without
access of air,

Fluidity of blood in living blood-vessels,

Coagulation of blood within the vessels during
life on foreign bodies or in injured blood-vessels.

a

LESSON IV.

HYALINE CARTILAGE.

1. Snip off a piece of the free edge of any of the
thin cartilages projecting from the sternum or
shoulder-girdle of a freshly killed young newt’.
Gently scrape away with a scalpel any tissue
attached to it. Mount it in normal saline solu-
tion? and with a high power note that
a, The matrix is studded at tolerably regular

intervals with cartilage cells or corpus-
cles.

b.. Each corpuscle consists of a spherical or ovoid
mass of cell-substance, in which lies a
relatively large nucleus. Both of these are
fairly clear and transparent, though usually
showing a variable number of fine granules.

c. Most of the cells entirely fill up the cavities
in which they lie.

1 Instead of this, sections of the head of the humerus or femur of
any young animal may be made.

2 i.e, *6 p.c. aqueous solution of sodium chloride. In this and the
following Lessons whenever a tissue is directed to be mounted in
normal saline solution it is to be understood that if either fresh
aqueous humour, or fresh blood-serum is obtainable, it is to be
preferred.

a a

Iv.]

HYALINE CARTILAGE. 53

d. Along the cut edge some cavities from which
the cells have fallen out may generally be
seen.

e. Except at the free edge, the cells are two or
more layers deep.

f. The matrix is hyaline or faintly granular,
and is comparatively (cp. infra §§ 4, 7) in
small quantity.

If the newts are not quite young the cartilage

will vary from the above in the quantity of ma-

trix, the shape and arrangement of the cells, and
in the cells possessing small fat globules and
often two nuclei.

Trrigate the piece with acetic acid 1 p.c.
The following changes will be seen as the irriga-
tion proceeds :

a. The nucleus becomes much more granular
and distinct.

b. Granules appear also in the cell-substance,
and more or less hide the nucleus.

c. The cell-substance becomes transparent owing
to the disappearance of the granules.

d. The cell-substance shrinks from the matrix,
and presents a granular-looking finely ser-
rated border. Note the space thus formed
between the cell and the matrix.

Place a small piece in gold chloride solution *5
p.c. for about half an hour, (until it is of a light
yellow colour,) then wash well with water, and
place it in a vessel containing water just acidu-

54

ELEMENTARY PHYSIOLOGY. [Iv.

- lated with acetic acid; leave it exposed to the

light. In one to two days, when it has become
a red-purple colour, mount it in glycerine.

Observe the cell-substance well coloured, and
hardly at all shrunken, the nuclei deeply coloured
and having a sharp outline, the matrix coloured

very slightly.

Take a piece of rib cartilage from a recently
killed adult animal and with a razor cut thin
transverse sections’ and transfer them with a
camel-hair brush to a watch-glass containing
normal salt solution. Mount one section in
normal salt solution and another in osmic acid
1 p.c.; in the former observe under a high
power, that

a. The cells frequently shew signs of partial
degeneration, containing fat globules which
may be nearly as large as the cell; these are
very highly refractive and so have a very
distinct outline.

b. In places the matrix may be fibrillated, and
in places semi-opaque owing to calcareous
deposit; irrigate with 1 p.c. hydrochloric
acid, the calcareous deposit is dissolved, but
the fibrillation is not affected (it differs thus
from the fibrillation of white fibrous connec-
tive tissue, cp. Lesson v. § 2 6).

c. Indications of primary and secondary capsules
may be seen (cp. § 6).

1 The sections may be cut with a freezing microtome, ef. Appendix.

CEE

IV.] HYALINE CARTILAGE, 55

5. Examine the section which has been mounted in
osmic acid; the fat globules are stained a deep
brown-black, the cell-substance, the nuclei and
the matrix are but slightly stained.

6. Prepare transverse sections of costal cartilage

which has been preserved in picric acid (cp.
Lesson 1 F. § 3), the sections may be made as in
§ 4 or the tissue may be imbedded (cf. Lesson VI.
§ 1). Stain the sections by placing them in a
2 p.c, solution of picrocarmine for 5 to 10 minutes
or in a very dilute solution (e.g. 02 p.c.) for a
day; if placed in the strong solution they should
be looked at now and then to see that they are
not overstained. Wash the sections with water
and mount them in glycerine.
Observe that the cells are arranged in groups
(each group having arisen by division from a
single cartilage cell), note the outline of the thin
layer of newer cartilage (capsule) around each
cell, sometimes the whole of the cells in a group
may be seen to be also surrounded by a thin
layer just marked off from the rest of the matrix,
Towards the outside of the cartilage the cells
become flattened in a direction parallel with the
surface, This specimen should be preserved* for
examination later (Lesson vy. B. § 7).

1 With a small brush or glass rod spread a fairly fluid solution of
Canada balsam over the edges of the cover-slip and the adjoining part
of the glass slide; the balsam will dry in a day or so and the specimen
can be moved about without fear of the cover-slip being displaced,
If the glycerine in which the section is mounted does not stretch to
the edges of the cover-slip the balsam will run underneath it and

56 ELEMENTARY PHYSIOLOGY. [IVv.

7. Take a piece of cartilage from the head of a
cuttle fish which has been preserved in picric acid
and subsequently in alcohol. Cut sections, im-
bedding if necessary (Lesson vi. § 1). Select a
thin section and immerse it in hematoxylin for
a few minutes watching that it does not stain
too deeply, place it in a watch-glass containing
spirit to remove the colouring matter simply ad-
hering to the tissue (if it is teo deeply stained,
place it in a little 1 p.c. acetic acid and when it
is suificiently decolourised, replace it in the
spirit), then mount it in glycerine. Observe

a. The groups of cartilage cells.

b. The marked processes proceeding from the
cells of one group, running towards and
anastomosing with the processes from the
cells of other groups.

8. Take a small piece of the ear of a freshly killed
mouse or young rat, remove the skin and scrape |
away the tissue surrounding the ear-cartilage.
Mount the cartilage in normal saline solution.
Observe that the cavities of the cells, generally
polygonal in form, are separated by very narrow
partitions of matrix. From many of the cavities
the corpuscles have disappeared altogether,
leaving simply a honeycombed matrix. This

probably spoil the section; on the other hand if the glycerine spreads
beyond the cover-slip the balsam will not stick to the glass, in this
case it is generally less trouble to remount the specimen than to wipe
away the superfluous glycerine. The Student should make one or
two trials to find out the proper quantity of glycerine to take with the
cover-slips he uses,

ern) ee eee

rv]

HYALINE CARTILAGE. 57

form of cartilage is frequently called “ parenchy-
matous.”

If it is desired to preserve this specimen or one
of newt’s cartilage (§ 1), place it in 75 p.c. alcohol
for about half-an-hour, stain with haematoxylin
(cp. § 7) and mount in glycerine.

LESSON V.

CONNECTIVE TISSUE.

A. CONNECTIVE-TISSUE FIBRES.
1. Blastic Fibres.

a, Tease out.in water a morsel of ligamentum
nuche. It is almost entirely made up of
rather large branching, and anastomosing
fibres having distinct outlines, and curling at
their ends.

b. Irrigate with acetic acid (1 to 5 p.c.); the
fibres are unaffected.

2. White Fibres.

a. Place on a slide a small piece of a thin
tendon, e.g. from a digit of a frog or from
the tail of a mouse (cp. B. § 3) and, keeping
it in position at one end with a needle,
separate as completely as possible the fibres
from one another at the other end by passing
a needle through it in the direction of its
length. If the unteased end of the tendon
is so thick that it would tilt up the cover slip,
separate with needles a small outer portion

CONNECTIVE TISSUE. 59

and tease this out as above; the tissue should
be kept moist with normal salt solution but
no more than is necessary should be added.
Place a small drop of salt solution on a cover
slip and let this fall gently on the tissue,
Observe the wavy bundles of fibrille; the
outlines of the fibrillz are not very distinct
but they may be seen to run parallel to one
another; owing to their small size and to
the cementing substance between them
special preparation is required to isolate the
individual fibrillz completely.

Irrigate with acetic acid (1 to 5 p.c.), The
fibrillze disappear, and the whole tissue swells
up into a transparent gelatinous mass, in

which hardly anything can be distinguished

except a few elastic fibres similar in general
characters to those of A. 1, but much finer,

Pull up with fine forceps the thin connective
tissue lying between the muscles of the thigh
of a rabbit or frog, cut off with scissors a

- small piece and tearing it as little as possible

spread it out on a slide with the aid of needles,
breathing on it if necessary to prevent it from
drying. Mount as in § aor press a dry cover-
slip gently on the film and allow saline
solution to run under the cover-slip; thus
the film is kept extended.

It is chiefly composed of wavy bundles of fibril-
lz similar to those seen in (b); the bundles
are of various sizes and run across one another

60

ELEMENTARY PHYSIOLOGY. ia

in all directions. Some small elastic fibres
running singly will also be seen, their out-
lines are more distinct than those of the bun-
dles of white fibres; they sometimes branch
and sometimes anastomose where they cross
one another. Where the film is well stretched
the elastic fibres run for the most part quite
straight, elsewhere they are curled at their
ends and have a more or less sinuous course.

If there is any doubt as to which fibres are

elastic, irrigate with acetic acid, cp. (0).

d. To make evident the corpuscles, take a
rather large piece of the film and let the
edges partially dry so that it may stick to
the glass slide ; add a drop or two of a strong
solution of Spiller’s purple in water or in 30
p.c. alcohol, in a few minutes wash away the
excess of the purple with water from a pi-
pette, and mount in water.

B. CONNECTIVE-TISSUE CORPUSCLES.

iP

Take a piece of the thin expansion of a tadpole’s
tail preserved in chromic acid ‘2 pc. Gently
break up a piece in glycerine, Where the hex-
agonal cells of the epidermis have broken away,
the homogeneous matrix may be seen, imbedded
in which are many blood-vessels and a number
of stellate cells.

The larger dark stellate cells are pigment-cells,
the smaller stellate cells are connective-tissue
corpuscles, each of which consists of a nucleus
and a branched protoplasmic body, Their pro-

v.] CONNECTIVE TISSUE. 61

cesses divide much, but irregularly, and commu-
nicate with the processes of other cells.

Take a frog, of which the brain and spinal cord
have been destroyed’, squeeze the side of the
head gently to make the eye bulge out, then
slice boldly at the edge of the cornea; if it is not
completely separated, take up the edge with
forceps, and cut through the cornea with scissors
at its junction with the sclerotic; in doing this
remember that the success of the preparation
greatly depends upon the cornea being placed
in gold chloride without having been dragged
or pulled about; any blood which may be on
the cornea should be removed by placing it in
a watch-glass containing normal saline solution
and very gently brushing it with a camel-hair
brush. Put it in gold chloride ‘5 p.c, solution
for twenty to forty minutes, wash well with
water, transfer to water just acidulated with
acetic acid, and expose to light”, When the
cornea has become of a red- or blue-violet colour,
put it on a slide with glycerine, and brush or
scrape both surfaces to remove the epithelium,
then mount in glycerine and examine it under
a high power. Where the hexagonal surface
epithelium has been removed, there will be seen
stained connective-tissue corpuscles, with numer-
ous fine branching processes which anastomose
with the similar processes of neighbouring cells,

1 See Appendix.

2 Probably it will not be stained until the following day. By the
tartaric acid method (cp. Appendix) the staining is quicker.

ro

62

ELEMENTARY PHYSIOLOGY. [v.

This preparation should be preserved to shew the
nerves of the cornea (cp. Less. x. C. § 2).

Break off the tip of the tail of a mouse or young
rat just killed, and pull out from the stump a
bundle of fine tendons. Gently stretch this over
a glass slide from edge to edge, and hold it in
this position till the edges are dry, putting a
drop of normal saline solution in the middle
only; the fibres are thus kept extended. Cover
with a cover-slip. On examination the bundles
of wavy fibrillze will be seen, but no corpuscles.
Slowly irrigate with acetic acid 1 p.c., watching
the bundles the while under a high power. Be-
tween the bundles of fibrillz will be seen rows
of cells. Note in each the round or oval nucleus,
the rectangular outline of the cell, and the
granular cell-substance. Indications of flange-
like lateral processes may be seen as lines run-
ning along the cells in the direction of the tendon.
Afterthe acetic acid has been added for some
little time the cell-substance becomes very indis-
tinct, and between the swollen bundles scarcely
anything is seen but rows of elongated irregu-
larly shaped nuclei. (cp. Action of acetic acid
on cartilage cells, Lesson Iv. § 2.)

Cut transverse sections of a tendon which has
been placed in 30 p.c. alcohol for a few hours
and preserved in 75 p.c. alcohol, imbedding (cp.
Less. VI. § 1) if satisfactory sections cannot
otherwise be obtained: mount in dilute glycerine.

Observe

v.]

or

CONNECTIVE TISSUE. 63

a. The fasciculi separated from one another by
a small amount of connective tissue.

b. In each fasciculus the thin branched tendon
cells between the bundles of fibrillz of the
tendon. The processes of neighbouring cells
not infrequently join.

Dissect back a piece of skin of a recently killed
young and fasting rat or other animal, and into
the subcutaneous tissue attached to this inject
a ‘5 p.c. solution of gold chloride with a Pravatz
syringe, until the fluid begins to exude. Let
this stay for a few minutes, and before the
swelling produced subsides snip off with a sharp
pair of scissors one or two pieces as thin as
possible, and place them in a watch-glass con-
taining three parts of water and one part of
formic acid 1°06 sp.gr. When the pieces are well
coloured, which will probably be in two or three
hours, shake them gently in a glass containing
distilled water, then mount in the manner given
in A. § 2 c, but in acid glycerine’. The pre-
paration will improve in a day or two. Observe
a. The leucocytes scattered about between the
bundle of fibres; the fibrous bundles will be
more or less swollen up by the formic acid,
and so not offer sharp outlines.

b. Connective-tissue corpuscles rather larger
than the leucocytes, and having processes

1 Glycerine containing 1 p.c. formic acid.

64

ELEMENTARY PHYSIOLOGY. [v.

which, in some cases, are seen to be connected
with the processes of other similar cells. (cp.
B. §§ 1, 2.)

c. Larger, more rectangular cells, generally
without evident processes, with oval nuclei,
lying over the bundles of fibrous tissue often
in rows or patches; on a side view these
appear as thin long cells.

This preparation should be preserved for the
observation of fat tissue (D. § 5).

Snip off a small piece of the jelly-like subcuta-
neous tissue in a foetal or new-born mammal’,
and mount as in A. § 2, ¢.

Note the pale granular cells of various shapes,
some not unlike those seen in B.§ 1. To make
the cells and their nuclei more distinct stain
with Spiller’s purple in the manner given in A,
§ 2 (d), or irrigate with dilute acetic acid watch-
ing closely the early changes produced.

This may also serve to shew the development of
fat-cells (D. § 6).

Note in the section of costal cartilage prepared
in Lesson Iv. § 6 the connective tissue layer out-
side and closely attached to it forming the peri-
chondrium ; in places sections of tendons running
into the cartilage will probably be seen.

1 An animal just killed serves best, but one preserved in picric acid

or Miiller’s fluid will answer the purpose ; if it is desired to observe
the forming fat-cells the animal must not have been transferred to
alcohol stronger than 50 °/.

v.]

CONNECTIVE TISSUE, 65

C. PIGMENT-CELLS,

Pin out on a frog-board one of the webs between

the toes of the frog used in B, § 2, and observe
first under a low and then under a high power.
There will be seen large corpuscles loaded with
dark pigment, and possessing numerous branched
processes. In some places the pigmented cells
will appear as round dots, the processes in this
case having been retracted.. Every intermediate
stage between these two states may be observed
by watching from time to time.

E. Fat-Cetts.

1.

bo

Cut out a small piece of the omentum, from a
part containing comparatively little fat. Spread
it out on a slide and mount it in normal saline
solution. Observe

a. Under a low power, the groups of highly
refractive fat-cells.

b. Under a high power, the variable size of the
fat cells, the apparent absence of a nucleus,
the connective tissue passing between and
over the cells,

Place a similar piece of omentum in osmic acid
1 p.c. for about half-an-hour, wash with water
and mount in dilute glycerine. Observe that
the fat-cells are stained a deep brown-black (cp.
Less. Iv. § 5). If the tissue after washing be
placed in alcohol (preferably 75 p.c.) the tint will
become much darker.

5

66

ELEMENTARY PHYSIOLOGY. [v.

Take a small piece of omentum, which has been
kept for some time in alcohol, and place it in
hematoxylin until it has become well stained.
Wash it with spirit, place it on a glass slide, and
tease it-out if it is too thick. Remove as much
of the spirit as possible with blotting-paper.
Cover the tissue with a mixture of creosote
(1 part) and turpentine (4 parts), and let it
remain until it is completely transparent, chang-
ing the fluid and gently warming if necessary.
Remove the excess of turpentine, cover the tissue
with a drop of Canada balsam, and put on a
cover-slip.

Observe the groups of cells from which the fat
has been thus removed. Note the shrunken
outlines of the cells, the marked membrane, and
the presence in each cell of a deeply stained
nucleus. A small amount of protoplasm, also
stained, may be seen as in § 4.

In the gold chloride preparation of sub-cutaneous
tissue made above (B. § 5), observe
The network of capillaries in a small collection

" of fat-cells.

The large, flat, connective-tissue corpuscles (B.

-§ 5, c) in greater quantities near the groups.

When the fat-cell group is spindle-shaped, these
may often be seen proceeding in rows from the
ends.

In some fat-cells a nucleus and a small amount
of protoplasm surrounding the fat may be seen.
Do not mistake a deposit of crystals in the fat
for a nucleus.

CONNECTIVE TISSUE. 67

Take a piece of tissue as in B. § 6 but just
outside the border of a small fatty clump visible
to the eye.

Observe the transitional forms between the
granular cells containing a few small fat
globules and cells in which hardly anything
but fat is to be seen.

LESSON VI.

MODIFICATION OF CONNECTIVE TISSUE

AND HYALINE CARTILAGE,

A. 'TRANSITION TO FIBRO-CARTILAGE.

:

Take a piece of intervertebral cartilage’, which
has been treated with chromic or picric acid and
subsequently with alcohol, and imbed it in the
following manner. At one end of a small oblong
block of paraffin B* scoop out a small hole; with
blotting-paper remove the excess of alcohol from
the piece of cartilage and place it in the hole so

- that the plane of the uppermost surface is at

right angles with the plane of the vertebral face.
Pour over it a little of the paraffin mixture heated
so as to be just melted ; remove any bubbles that
may be present and keep the tissue in its proper
position by means of a heated needle; when the
whole is thoroughly solid pare down the paraffin

1 Cut off from one face of a vertebra of a rabbit a thin slice about
1 mm. thick, leaving the intervertebral cartilage in connection with it;
divide the more or less circular piece so obtained into four parts, and
treat with chromic or picric acid to dissolve the salts (ep. Appendix).
2 Cp. Appendix.

:

VL] MODIFICATION OF CONNECTIVE TISSUE, ETC. 69

to the surface of the cartilage, at the same time
bevelling the edges. In cutting the sections,
cover the tissue and the blade of the razor with
spirit (for convenience from a wash-bottle) and
remove the sections by means of a camel-hair
brush from the razor to a watch-glass. With
the platinum “lifter” transfer them to hzema-
toxylin and treat as in § 7, Lesson Iv.

It is to be remembered that if the chromic acid’
has not been completely removed from the tissue
the staining will be very imperfect; in this case
the sections should be placed in a little Na,CO,
1 p.c. for a few minutes and then washed with
water before staining. Observe in the sections

a. The softened bone passing into

b. The thin layer of hyaline cartilage; outside
this

c. The arrangement of the cartilage cells in
rows, and the concomitant appearance of
fibres in the matrix, this gradually but
rapidly merging into

d. Fibro-cartilage consisting of bundles of white
fibrous tissue and of ratherlargecellsarranged
in rows between the fibrous bands; these cells
are like the cells of hyaline cartilage and
each has a thin hyaline capsule around it.

e. Towards the outer part of the cartilage, the
capsules and cells become smaller and more

1 Miiller’s fluid, potassium bichromate and ammonium bichromate,
must similarly be completely removed from tissues which are to be
stained,

70

ELEMENTARY PHYSIOLOGY. [VI.

elongated, the capsules disappear, and so the
fibro-cartilage merges into
f. Connective tissue.

The fibro-cartilage does not form one continuous
band connecting the vertebra, but passes in
parallel bundles, between and at right angles
to which are tendinous bundles which thus
will be seen in cross sections; the cartilage

. cells may in some bundles also be absent.

Prepare through the junction of the round liga-
ment and head of the femur in a young animal
a section parallel to the direction of the fibres.
If the tissue has been treated with picric acid,
‘stain the section in picrocarmine ; if in chromic
acid, with hzmatoxylin. Observe as the ten-
don approaches the cartilage the transition of
the flat rectangular branched cells of the ten-
don to oval fuller cells with round nuclei. Note
the gradual disappearance of distinct fibres and
their replacement by a hyaline matrix, the cells
at the same time becoming scattered. Thus
the tendon passes into fibro-cartilage, and fibro-
cartilage into hyaline-cartilage.

B.. TRANSITION To ELASTIC CARTILAGE.

Dissect out the arytenoid cartilage of a sheep,

preserved in alcohol, and cut out a small piece
of the tissue immediately above the hyaline
cartilage, and together with a little of the
hyaline cartilage itself. Imbed, and cut sec-

v1]

MODIFICATION OF CONNECTIVE TISSUE, ETC. 71

tions, including the hyaline cartilage and the
tissue above it. Stain with picrocarmine and
mount in glycerine. Observe

a. That at the upper surface of the hyaline
cartilage, the matrix becomes granular, and
the granules are arranged in rows, which then
often have less the appearance of rows of
granules than of granular fibres; these may
be traced into ordinary fine elastic fibres,
which increasing in quantity, form -

b. The elastic cartilage in which the cells
stained red with carmine are similar to
those of hyaline cartilage; they have thin
clear outlines around them, the capsules, and
these are surrounded by a meshwork of elastic
fibres stained yellow with picric acid lying in
a varying quantity of matrix. This meshwork
may have not so much the appearance of
distinct elastic fibres, as of a system of thick
bars, recalling in general feature parenchy-
matous cartilage.

c. Above (b) ordinary loose connective tissue,
note the transition of the elastic cartilage
into this by the elongation of the cells and
the breaking up of the elastic fibres into
loose bundles and the appearance in those
of white fibrous tissue staining red.

LESSON VIL.

BONE, OSSIFICATION, TEETH. |

A. STRUCTURE OF BONE.

1,

Examine under a low power prepared transverse
sections’ through the shaft of a long bone.
Most of the smaller spaces (i.e, Haversian canals,
lacune, canaliculi) will be filled with air or
debris and will therefore appear dark. Observe

a. The central cavity surrounded by a small

amount of spongy bone which is arranged in

_ a network with rather large irregular spaces

(Haversian spaces); externally the spongy

bone passes into the compact bone: note the

transition from the Haversian spaces to the
Haversian canals,

b. In the compact bone the Haversian systems,
each consisting of a Haversian canal sur-
rounded by concentric lamell# which are
chiefly marked off from one another by the
lacune which lie between them.

1 It is simplest to buy these sections,

VIL]

Lo

C.

BONE, OSSIFICATION, TEETH. 73

The intersystemic lamelle between the
Haversian systems; in the outer part of the
bone these run for the most part parallel
with the surface.

In the spongy bone the lamellz run in the
main concentric to the spaces.

Examine the specimen under a high power.
Observe

a.

The irregularly fusiform lacune giving off
numerous wavy branches, the canaliculi,
which run across the lamellze to join the
similar branches of neighbouring lacune.

In the compact bone, canaliculi open into the
Haversian canal; in the spongy bone into the
Haversian spaces and central cavity.

Examine with a low power prepared longitudinal .
sections of the shaft of a long bone. Observe

a.

The Haversian canals running in the main
parallel with the surface of the bone, they

_ have connecting branches and in places open

on the surface and into the central cavity.

The lamellz running for the most part
parallel with the Haversian canals. Here
and there a Haversian system may be cut
obliquely, in such cases the lamellz will be
concentric to the canal.

Examine the specimen under a high power.
The individual lacunz are much as in the trans-
verse section, but rather longer; observe the
arrangement of the canaliculi (ep. § 2).

74

ELEMENTARY PHYSIOLOGY. [vu

Prepare transverse sections through the shaft of
a long bone which has been decalcified’. Stain
with picrocarmine (cp. Lesson Iv. § 6). Wash
with water and mount in glycerine. Observe
that

a. Each lacuna is occupied by a cell or bone
corpuscle.

b. The canaliculi are scarcely visible.

c. The Haversian canals and spaces are occupied
by cells, connective-tissue fibres and blood-
vessels (red blood corpuscles if present will
be stained yellow).

d. If the tissue of the central cavity has not
fallen out of the section, it will be seen to
consist of fatty tissue.

e. The periosteum is closely attached to the out-
side of the bone; it consists externally chiefly
of white fibres, internally largely of small
elastic fibres some of which may be seen to
run into the bony matrix.

Take a bone (e.g. the parietal) which has been
decalcified by nitric or hydrochloric acid, well
washed with water and preserved in spirit;
remove the periosteum, tear off from the surface
thin strips of the bone matrix and mount them
with the inner side uppermost in water. Under
a high power note the perforating fibres pro-
jecting from the surface and the apertures
through which similar fibres have passed.

1 Cp. Appendix for methods of decalcification.

|) ee

————— a ——

se ee ee ——

VII.] BONE, OSSIFICATION, TEETH. 75

Examine carefully the thinnest part of the strip
(containing fewest lamellz) for the fine decus-
sating fibres of the matrix; add acetic acid, both
the perforating and decussating fibres swell up
and become indistinct or lost to view.

B. OSSIFICATION.

1. Cut a longitudinal section through the head of
the femur of a newly-born rabbit, cat, ete. The
femur should have been divided longitudinally
into four parts, and treated with picric or
chromic acid to harden it and to dissolve the
salts. Stain the sections with eosin or picro-
carmine, wash with water, place first in dilute
and then in strong spirit, clear with creosote and
turpentine’ (or turpentine alone) and mount in
Canada balsam (cp. Lesson Iv. footnote). Observe

a. The normal hyaline cartilage.

b. The cartilage cells arranged in rows. Note
that many of the cells in section are tri-
angular, the broad side of one lying above
the angle of the other; this indicates an

1 The creosote and turpentine is used since it dissolves any
paraffin that may be attached to the tissue (from the imbedding),
makes the tissue transparent, and mixes well with the Canada balsam;
earbolic acid may be used instead of the creosote, it is cheaper but
does not dissolve the paraffin mixture quite so readily. Clove oil
causes less shrinking in many tissues than creosote and turpentine,
but it is much dearer than either of them and does not dissolve the
paraffin mixture in the cold, The clearing of the sections takes place
more readily and certainly if they are placed for a few minutes in
absolute alcohol before they are placed in the clearing agent.

76 ELEMENTARY PHYSIOLOGY. [VIt.

origin of two cells from one by oblique
division.

e. A layer of cells much larger than the previous
ones, with clear cell-substance (in the speci-
men probably shrunken) and a distinct
nucleus; they are also arranged in rows and
have only a small amount of matrix between
them.

d. The large irregular cavities below this layer
surrounded by bone matrix, and containing
many osteoblasts, cells very similar in ap-
pearance to white blood corpuscles, but
larger, most of them are in contact with
the bone matrix. In the centre of some of
the cavities may also be seen a blood-vessel
with surrounding connective tissue.

e. Osteoclasts may be seen here and there in
contact with the bone matrix ; they are large
multinuclear cells, and where they touch the
bone are often striated at right angles to the
surface.

2. Imbed a piece of the shaft of any long bone’ of a

foetal mammal that has been preserved in picric’

or chromic acid, any remaining salts in the bone
may be extracted by placing it for a day in spirit
containing ‘5 p.c. nitric acid, Cut transverse

1 Instead of these, sections may be made of the lower jaw of a
foetal mammal; the developing sub-maxillary bone shews admirably
the osteoblasts and osteoclasts but the section is confusing to
beginners owing to the number of other tissues present.

——E

— eee

ee

VI1.] BONE, OSSIFICATION, TEETH. 77

sections and stain them with picrocarmine, and
mount in glycerine or Canada balsam. Observe

a. The periosteum consisting externally of
coarse connective tissue, internally of fine
connective tissue with numerous cells.

b. The trabecule of the bone matrix, exter-
nally these give off numerous small projec-
tions into the inner layer of the periosteum.

c. The osteoblasts forming a layer on the
outer surface of the bone matrix; some will
also be seen on the trabeculz throughout the
section; note the fine connective tissue and
the blood vessels in the trabecular spaces.
Many of the osteoblasts especially in the ex-
ternal layer are elongated and their smaller
ends appear to run into the matrix.

d. Some osteoclasts as in § 1, e. may be seen.

C. STRUCTURE OF TEETH.

1. Examine under a low power longitudinal sec-
tions of teeth prepared in the same manner as
bone in B. Observe the dentine surrounding
the pulp cavity, the cement or crusta petrosa
covering the dentine of the fangs, and the
enamel covering the dentine of the crown.
Note the general arrangement of the dentinal
tubules.

i)

Examine under a high power the same section,
and study in detail

78

ELEMENTARY PHYSIOLOGY. [VIL

The Dentine.

a.

In the matrix, apparently homogeneous, are
numerous dentinal tubules which run in a
wavy course from the pulp-cavity outwards.
These, dividing as they go, and giving off
many anastomosing lateral branches, finally
end either in loops or in small irregular
cavities, the so-called interglobular spaces,
on the surface of the dentine.

In some places the dentinal tubules are cut
transversely. Here the central dark spot in-
dreates the space formerly occupied by the
dentinal fibre, the ring round this is the
dentinal sheath,

The Cement or Crusta Petrosa.

a.

B.

This differs little from bone, but Haversian
canals are generally absent. .
The canaliculi of the lacunz, next to the in-
ter-globular spaces, open into these, thus
bringing the dentinal fibres into connection
with the bone-corpuscles.

Where the cement is massive, wavy ‘contour’
lines may be seen, indicating the successive
deposits.

The Enamel.

The striated enamel fibres or prisms are
arranged in sets, perpendicular to the sur-
face of the dentine. In mounted specimens
the indications of the separate fibres are
generally not well marked, but are visible at

Vil] BONE, OSSIFICATION, TEETH. 79

places, The line of junction of the enamel
and dentine is generally very conspicuous, in
consequence of a change of level in the
specimen, the dentine having been ground
thinner than the harder enamel. The enamel
is frequently split or cracked.

3. Examine prepared transverse sections of Teeth,
taken at different heights, and compare with the
above,

LESSON VIII.

STRUCTURE OF CONTRACTILE TISSUES.

1. Cut off the head of a frog which has just been
killed, remove carefully the lower jaw and upon
the mucous membrane of the roof of the mouth
pour a little osmic acid 1 p.c. (before doing this
a small piece of the membrane may be cut out
to serve for § 2). Cover up the preparation and
leave it for about half an hour’ then scrape a
small area of the mucous membrane and tease
out the flakes so obtained in water or in dilute
glycerine, putting a small piece of paper under-
neath the cover-slip to avoid pressure. Examine
under a high power. Groups of ciliated cells
intermingled with mucous (goblet) cells will be
seen; neglect the latter, note in the former

a. The shape of the cell varies, it often branches
irregularly at its attached end.

1 Or the mucous membrane after treatment with osmic acid as
above may be gently washed with water to get rid of the fumes of the
acid, removed with forceps and scissors, pinned out with hedgehog
quills and placed in osmic acid 1 p.c. for 1 to 24 hours as convenient.

The oesophagus and the mucous membrane of the floor of the mouth -

which also have cilia may be similarly treated.

|
|
.
.
|

Vu]

STRUCTURE OF CONTRACTILE TISSUES. 81

b,

The free surface is densely studded with
cilia. The cilia seen in profile appear to
form a row, but when a cell is seen in face,
they are foreshortened into mere dots and
are seen to be spread over the whole surface,

The nucleus is not very obvious but may,
with its nucleolus, be made out; the cell-
substance is somewhat granular but has a
non-granular, hyaline border just below the

cilia,

Cut out a small piece of the same membrane in
a recently killed frog, place it on a slide, from a
portion of it scrape off the epithelium and tease
out the scrapings in normal salt solution; with
this mount the unteased portion of membrane
putting a fragment of paper underneath the
cover-slip, Note

a.

In the unteased portion the shimmering ap-
pearance caused by the movements of the
cilia,

The movements of the cilia in the isolated
cells or clumps of cells. Observe carefully the
cilia which are moving slowly; it may be
seen that the down-stroke (contraction) takes
place more quickly than the return (relaxa-
tion); there is no perceptible pause between
the two movements,

The results of ciliary action. Granules and
blood-corpuscles are driven along; detached

6

82

ELEMENTARY PHYSIOLOGY. [ VIII.

cells may also be seen carried about by the
action of their own cilia.

d. The contracted, almost globular form of the
cells which have been set free.

Tear off the smallest possible strip of a frog’s
muscle, which has been kept extended and
preserved in alcohol, and tease it out as finely as
possible in dilute glycerine. Note

a. The varying size of the fibres,

b. The striation of the fibres: alternate dim and

bright cross-bands passing through the whule
thickness of the fibre.

c. The breaking up of the fibre into fibrille,
the latter also striated. Try to obtain as
fine fibrillee as possible. The fibres sometimes
split transversely into discs; this is usually
the case when the muscle has been preserved
in picric acid; the surfaces of the discs ap-
pear dotted.

Cut through the skin of the front of the thigh
of the frog used in § 2, note the sartorius muscle
running somewhat obliquely across the thigh
from top to bottom, seize with forceps the con-
nective tissue lying along its inner border in the
upper part of the thigh and tear this back to
the knee, remove similarly the connective tissue
at its outer border, thus the tissue covering the
muscle will also be removed. Take up with fine

forceps a few fibres at one end of the muscle,and —

—

vu]

STRUCTURE OF CONTRACTILE TISSUES. 83

gently pull them out to the opposite end. Lay
them on a glass slide, and separate them a little
in the centre, place a bristle across them and
press gently: remove the bristle, put on a cover-
slip and let a drop of normal saline solution run
underneath it (cp. method, Lesson V. A. § 2 a,c).
Where the bristle was pressed on a fibre the
muscular substance will be seen in many cases
to be broken across leaving its delicate trans-
parent sheath, the sarcolemma, stretching across
the gap; an indication of the sarcolemma as a
fine line bulging out from the muscle substance
may also be seen where a fibre is bent.

Trrigate with acetic acid ‘5 p.c. and observe

a. The fibres at first become cloudy and semi-
opaque.

b. The precipitate causing this cloudiness is
after a while dissolved, and the fibres be-
come more transparent than normally.

c. The nuclei of the fibres come into view scat-
tered throughout the muscle substance ; they
are elongated in the direction of the fibre:
lines of granules proceeding from their ends
may usually be seen.

Cut through the skin in the mid ventral line in
a small frog, lift up the flap of skin on one side
over the middle of the sternum, a thin band of
muscle will be seen running from the skin down-
wards towards the lower part of the sternum.
6—2

84

“I

ELEMENTARY PHYSIOLOGY. [VIIL

- Cut through the skin above and below the muscle,

and tear away the connective tissue around it,
then, keeping it stretched, pour a little 1 p.c.
osmic acid over it. In a minute or so the muscle
will be fixed; with fine forceps tear away carefully
from the muscle any connective tissue that can
be seen on its surface and cut it out being care-
ful to cut the upper end as close as possible to
the skin ; place it in osmic acid for a few minutes,
wash with water, mount it in either dilute
glycerine or after treatment with alcohol ete.
in Canada balsam, keeping that surface upper-

most which was next the body. (This specimen

should be preserved for examination later, Lesson
x. C.§ 1.)

Observe at the upper end of the muscle the
endings of the muscular fibres, these will prob-
ably be covered by a good deal of connective
tissue, but the rounded or conical ends of the
muscle-substance of the fibres and the continua-
tion of the sarcolemma into the connective tissue
(tendon) will be seen.

Remove with as little injury as possible one of
the thin muscles from the leg of Hydrophilus’,
tease it out a little without adding fluid and
mountasin$4. The fibres will for a brief period
be seen in a normal condition. In many, the
alternate dim and bright striz will be very dis-

1 If not obtainable, take the large common water-beetle (Dytiscus
marginalis), or failing that a cockroach or a fly,

VIII. ]

STRUCTURE OF CONTRACTILE TISSUES. 85

tinct; in others the transverse markings will be
more or less obscured by an appearance of longi-
tudinal fibrillation. Occasionally fibres are met
with, having only a confused granular aspect.
If the preparation has been successfully made,
waves of contraction may, at times, be seen to
travel along the fibres.

Observe in the fibres with distinct transverse
striation, that,

a. The dim band has a faint longitudinal stria-
tion, as if made up of small rods.

b. The bright band shews a dotted line running
across its centre, thus dividing it into two
parts. If the preparation be good, and a
high: magnifying power be used, this may be
seen to consist of two rows of dots.

c. The usually round nuclei, imbedded in a
granular mass of protoplasm; the whole
forms a band or rod running a variable
distance in the substance of the fibre (cp.

§ 5c).

Imbed a small piece of mammalian muscle,
hardened in chromic acid ‘5 p.c., and cut trans-
verse sections, stain with hematoxylin, and mount
in glycerine. Observe

a, The connective tissue (perimysium) around
the bundles of fibres and around the whole
muscle, from this runs a small amount of
connective tissue (endomysium) between the
muscle fibres,

86.

ELEMENTARY PHYSIOLOGY. [ VIII.

6, The cut ends of the fibres are finely dotted
corresponding with the fibrille; sometimes
the cut ends are divided into larger polygonal
areas (areas of Cohnheim), this appearance is
produced by the hardening agent.

c. The nuclei lying just beneath the sarcolemma,
few or none being imbedded in the muscular
substance of the fibres (cp. § 5 ¢).

d. In some part of the section, the fibre-frag-

- ments may be turned over and lie’ length-
ways; in these the striation and tendency
to break into fibrille may be noticed as in
frog’s muscle,

Remove the peritoneum from a part of the
intestine of a recently killed mammal, and tear
off with fine forceps a strip as thin as possible of
the longitudinal muscular coat. Place this in
potassium bichromate ‘1 p.c. or in 30 p.c. alcohol
for two days, wash with water, stain with picro-
carmine, wash and tease out in dilute glycerine.
Note

a. The isolated fibre-cells, These are long, and,
when not broken off in the preparation, fusi-
form masses of hyaline or faintly granular
protoplasm, They appearvery much thinner
when seen in one direction than in another;
that is to say, they are much flattened. In
the middle of each fibre an elongated nucleus
will be seen stained deeply. In the neigh-
bourhood of the nucleus, especially at its

VIII. ]

10.

11,

STRUCTURE OF CONTRACTILE TISSUES, 87)

ends, granules are more abundant in the
protoplasm.

b. The arrangement of the fibre-cells in bands
and sheets: the fibres may frequently be seen
projecting like a palisade from the torn end
of a band,

Take a small strip of unstriated muscle as in § 9
and tease it out in salt solution.

The sheets and bands of the tissue are fairly
transparent, the individual fibre cells are scarcely
visible. Add acetic acid, ‘1 to ‘5 p.c., the cells
become for a short time somewhat more obvious,
and the nuclei very distinct, often showing one
or two nucleoli,

Make longitudinal vertical sections of the mus-
cular coats of the cardiac end of a cat’s or dog’s
stomach. The tissue should have been pinned
out, placed in Miiller’s fluid for two to four weeks
and then treated with alcohol, Stain with
carmine or hematoxylin and mount’, Observe

The cells are arranged in bundles, probably
this will be distinct in the inner (circular)
coat only, the outer (longitudinal) coat being
cut parallel to the direction of its bundles,
Note the nuclei of the cells in the inner coat;
these will be seen in some only of the cells
and will appear as largish deeply stained cen-

1 When it is indifferent whether a tissue be mounted in glycerine
or in Canada Balsam, neither is mentioned; if balsam is used, of
course the tissue must be treated with creosote, etc. (cp. p. 75).

88:

ELEMENTARY PHYSIOLOGY. : [ VIII.

tral spots. (This section should be preserved
for examination later, Less. xvi. A. § 3.)

The reduction of nitrate of silver by the cement
substance between the muscle cells will be seen
later, Less. x1. C. § 4.

12. Tease out thoroughly a small portion of cardiac
“muscle preserved in potassium bichromate’.
Note that.

a. The striation is less distinct than in skeletal

b.

muscle.

‘No sarcolemma is present.

¢. At short intervals along the fibres are not

very distinct transverse bands of hyaline
cement substance, this joins together the
muscle cells of which the fibres are composed.
At about the centre of each muscle cell is
a nucleus. (The nuclei will’ become more
evident if the tissue be stained with alum-
heematoxylim or picrocarmine.)

The isolated muscle cells may frequently be
seen to have a short obliquely running pro- ~
cess ; in the fibres the processes of the cells
are joined to other similar processes by a
little cement substance, forming thus the
anastomoses of the fibres,

- 1 Good preparations can also be obtained by the following method
(Ranvier). Thin sections in the direction of the fibres are cut from a
fresh heart, placed in chromic acid -02 p.c, for one day, then washed
well with water, placed in picrocarmine for one or more days, washed
and mounted in acid glycerine,

viil.]

STRUCTURE OF CONTRACTILE TISSUES. 89

DEMONSTRATIONS.
The structure of a living muscular fibre of an
Insect as seen under a high power.

The appearances of muscular fibre under polarised
light.

LESSON IX.
_ PROPERTIES OF CONTRACTILE TISSUE.

1. Constant Current.

Destroy the brain and spinal cord of a frog and
make a longitudinal incision through the skin of
the back of the thigh. With fine-pointed scissors
cut through the connective tissue between the
large semi-membranosus muscle and the small
biceps: the former lies on the inside of the thigh,
the latter is seen along the outer and lower
border of the semi-membranosus. The sciatic
nerve and the femoral artery will come into view;
at about the upper third of the thigh, the artery
will be seen to give off two small transverse
branches running, in the position of the frog, over
the nerve; from this point down to the knee,
isolate the nerve by tearing away with a “seeker”
the connective tissue around it; if the tissue is
resistant it is better to lift up the nerve with a
seeker and to cut through the connective tissue
with fine scissors, Be careful not to, pinch the
nerve with the forceps, nor to put more strain
upon it than is absolutely necessary, and further
not to puncture the artery.

Ix.]

PROPERTIES OF CONTRACTILE TISSUE. 91.

Take a pair of electrodes which have their pla-
tinum points exposed on one side only, and
connect them with two Daniell’s cells arranged
in series (Le. with the copper of one joined by
means of a wire with the zinc of the other) in-
terposing a key’ to short circuit the current,
Instead of Daniell’s cells any other cells of cor-
responding strength may be used.

The key being shut’, place the electrodes under

the nerve so that the nerve, and the nerve only,

touches the platinum points, then open the key,
so as to allow the current to pass into the nerve,
and after a few seconds shut it again. A move-
ment in the leg caused by contractions in the
muscles to which the nerve is distributed, will
be seen when the key is either opened or shut,
or on both occasions. Save in exceptional cases,
there is no movement during the passage of the
current through the nerve, but only when the cur-
rent ts thrown into or shut off from it (the current
being supposed to be tolerably constant), Do
not repeat this experiment more than once or
twice on the same nerve, lest it be too much ex-
hausted for the succeeding observations,

1 Cp. Appendix for a description of the instruments and for the
methods of using them.

2 For convenience of description the terms ‘shut’ and ‘open’
which apply to a Du Bois Reymond key are used, but if the student
remembers that to shut the key is to let the current pass through it,
and to open the key is to break the current passing through it, he will
have no difficulty in using a Morse or other key in the place of that
of Du Bois Reymond,

92.

ELEMENTARY PHYSIOLOGY. [Ix.

Single Induction Shocks.

Now connect a Daniell’s cell with the top screws
of the primary coil interposing a key to break
the current, connect the electrodes with the
secondary coil of a Du Bois Reymond’s in-
duction machine’, place the marker of the se-
condary coil at 10 on the scale, and, the frog’s
leg being at rest, shut and open the key several
times.

At each shutting and opening of the key, ie. at
each momentary induced current, a single sharp
movement of the leg will take place. The ‘single
induction shock’ causes a single contraction or
spasm of the muscles of the leg. If the nerve
slips off the electrodes during the movements,
be careful not to pinch it in replacing it.

If good contractions are not obtained with the
secondary coil at 10, push it up gradually over
the primary coil, one division at a time.

Shut and open the key as rapidly as you can,

for afew seconds. The contraction of the muscle
taking longer time to develop than the induced
current, each contraction will take place before
the preceding has disappeared, and the leg will
remain rigidly contracted, or nearly so, as long
as the breaking and making of the primary cur-
rent is kept up. In other words, tetanus is pro-
duced. This is however more easily brought
about as follows.

1 Cp. Appendix.

1x.]

A.

PROPERTIES OF CONTRACTILE TISSUE. 93

Interrupted Current.

Introduce into the secondary circuit a key to
short circuit the current, and remove the wires
of the Daniell’s cell from the screws at the top
of the primary coil to those at the base’. The
machine should work in such a way that im-
mediately the key in the primary circuit is put
down the hammer is at once thrown into oscilla-
tions (as indicated by its noise). The key in the
secondary circuit being open, shut the primary
key. Immediately that the sound of the hammer
is heard, the leg is thrust out straight by the.
tetanic contractions of the muscle, and remains
so as long as the current continues to be thus
made and broken. Open the primary key; at
once the limb becomes flaccid and quiet. The
‘interrupted current’ should not be applied for
more than a few seconds,

Chemical Stimulation.

Expose the heart (cp. Less. 11. A. § 1), and bleed
the frog by cutting across the aorta. Cut through
the tissue above the sciatic nerve up to a quarter
of an inch beyond the end of the urostyle, and
there cut it across, with scissors and seeker isolate
the nerve up to the cut. Let the extreme end
of the nerve dip into a saturated solution of
sodium chloride; watch the leg.

In a variable time twitchings of the toes will be
seen, and after a while these will increase in

1 Cp. Appendix.

94

ELEMENTARY PHYSIOLOGY. [Ix.

vigour and will extend over the limb until it
becomes almost as rigid as in § 4.

Mechanical Stimulation.
Cut off the part of the nerve which has been

_ dipping in the salt solution; the chemical sti-

mulus being thus removed, the leg will become
flaccid again.

Pinch the remaining nerve sharply with a pair
of forceps several times. At each pinch the
muscles of the leg will contract. The pinching
kills or injures the part of the nerve pinched;
hence it is necessary to begin at the cut end,
and work down towards the muscle.

The gastrocnemius muscle of this leg may be
used for § 13, b.

Curve of Single Muscular Contraction.

Place the frog on a glass or porcelain plate; on
the left side cut through the skin at the back of
the thigh and carry the skin cut half way up

the back. Lift up the urostyle and cut through ©

the muscles attached to its lower half, then
holding it well up cut through the muscles at-
tached to its upper half: it is as well to avoid
cutting the 10th nerve which runs along the
under surface of the muscles. Note the 7th, 8th,
9th and 10th nerves which unite to form the
sciatic; with strong scissors cut away the uro-
style and cut through the vertebral column
above the 7th nerve: lift up the lower piece of
the vertebral column and without injury to the

REED IEE NI £ EE

IX]

PROPERTIES OF CONTRACTILE TISSUE. 95

nerves of the left side cut away the rest of the
tissue attached to it. Then lifting up the piece
of bone without stretching the nerves, cut
through with scissors the tissue attached to
them, follow the nerves on to the sciatic and cut
through the tissue above and attached to the
sciatic including the branches given off by the
nerve ; isolate thus the sciatic down to the knee.
Letting the nerve rest on the muscles of the
thigh, cut through the skin of the upper part of
the thigh; seize the flap of skin with stout
forceps, and tear it away from the leg as far as
the foot. Placing the nerve on the muscle
below the knee cut away the muscles attached
to the lower half of the femur, and with a strong
pair of scissors cut through the middle of the
femur. Cut through the lower (Achilles) tendon
of the gastrocnemius muscle close to its attach-
ment to the foot, and seizing the tendon with for-
ceps tear the gastrocnemius away from the other
muscles up to its attachment with the femur;
cut through the tibia and fibula just below the
femur. The preparation so obtained is called a
nerve-muscle preparation’.

Now pass a hook through the Achilles tendon,
clamp the femur, and taking up the preparation

1 When two nerve-muscle preparations are to be obtained from one
frog, the lower part of the vertebral column should be bisected
longitudinally. It is perhaps somewhat better not to bleed the frog
before commencing the dissection, but in this case considerable skill
and patience are required to avoid cutting small vessels and so covering
the nerve with blood.

96

ELEMENTARY PHYSIOLOGY. —~ on

by the clamp and by the piece of vertebral
column, arrange the preparation in the moist
chamber as in Fig, 2 Appendix. Load the lever
with 15 or 20 grammes, and bring the lever
to mark on the revolving cylinder.

Connect the electrodes with the induction ma-
chine arranged for single induction shocks as in
§ 2.

Set the cylinder rotating at its swiftest speed and
with the marker of the secondary coil at 10 take
a tracing of the contraction caused by a break in-
duction shock (the make shock is prevented
from reaching the nerve by shutting the key in
the secondary coil, arranged for short circuiting,
before that in the primary is shut; on opening
in the reverse order the breaking shock passes
into the nerve). Observe the rapid but steady
rise to a maximum, passing almost immediately
into a similar but rather less rapid fall; the later
stages of the fall are distinctly less rapid than
the earlier ones.

Take another tracing over the previous one with
the secondary coil at 5; the height of the con-
traction is somewhat greater.

Place the secondary coil at 25, the key in the
secondary circuit being open, shut that in the
primary circuit and after a short interval open
it; if no contraction follows shift the secondary
coil nearer and nearer the primary, stimulating
at each shift,

Since the induction shock caused by breaking

IX.]

10.

PROPERTIES OF CONTRACTILE TISSUE. 97

the current is stronger than that caused by
making it, a contraction will be obtained earlier
on opening than on shutting the primary key.

Tetanus.

Now arrange the induction machine for an inter-
rupted current (§ 4), the secondary coil being at
15 or 20 to begin with.

With the cylinder moving slowly, take a tracing
of a tetanic contraction of a muscle, stimulating
for not more than three or four seconds.

Observe the rise to a maximum at first rapid
but afterwards more and more slow, the mainte-
nance of the maximum during the application of
the current, and the fall, at first rapid but after-
wards more slow, after the removal of the current.
Watch during the tetanus the muscle itself as
well as the curve.

Connect one wire from the Daniell’s cell with
one of the top screws of the primary coil, and
the other to the binding screw of the oscillating
rod (cp. Appendix), connect a wire with the
other top screw of the primary coil and bending
the wire place its opposite end in the mercury
cup. A key for short-circuiting should be kept
in the secondary coil. Allow the full length of
the rod to oscillate, open the key in the secon-
dary circuit and take a tracing of the muscular
contractions; then take successive tracings with
the rod shorter and shorter.

Observe the gradual fusing of a series of single

contraction curves into the curve of tetanus.

7

98

11.

ELEMENTARY PHYSIOLOGY. [Ix.

Load the muscle with 50 grammes, allowing the
lever to write on the stationary cylinder. Note
the elasticity of the muscle. The weight when
applied stretches the muscle to a certain extent,
but when raised so as not to drag on the muscle,
the latter returns, at once to ;almost, and even-
tually to quite, its former length. Stimulate
with an interrupted current (coil say at 20).
Note the amount of contraction. Repeat this
with 100, and then with 200 grammes, using in
each case the same strength of current for the
same time, moving the cylinder with the hand
so as to expose a fresh black surface for each
observation.

Observe that with medium weights the work
done (the product of the weight into the height)
is greater than with very light or very heavy
weights.

Observe also that the muscle gradually becomes
exhausted, ie. the contractions caused by the
same stimulus gradually become less and less.

2. Probably the preparation will be nearly exhaust-

ed; take as in § 7 a tracing of a single muscular
contraction, and note the difference in the form
of the curve; the rise is less rapid, the fall
more gradual, the muscle may take some time
to regain its original length.

13. Reaction of Muscle.

a. Remove the muscle from the moist chamber,
place it on a clean glass slide, and apply
the electrodes, connected with the induction

Ix.] PROPERTIES OF CONTRACTILE TISSUE, 99

machine arranged for an interrupted current,
directly to the muscle.

Observe that the current applied directly to
the muscle produces the same effect as when
applied indirectly through the nerve. To
completely eliminate the effect of stimu-
lating the nerves urari should be given toa
frog the brain of which has been destroyed.
Tetanize the muscle with currents of increas-
ing strength until it is completely exhausted,
pushing the secondary coil right up over the
primary.

Cut the muscle across in half, with a quite
clean scalpel, and press on one section a piece
of faintly blue, on the other a piece of neutral
litmus paper. Both will be turned red where
they come in contact with the muscle,

b. Dissect out the gastrocnemius from the leg
used in §§ 1—6, and place it for five minutes
in normal saline solution heated to 50° C.
Observe that it has become contracted and
opaque, le. rigor mortis has set in.

Cut the muscle across, and test with litmus

- paperasina. The muscle will be distinctly
acid, even more so than the tetanized muscle
in a.

c. Take a fresh, living gastrocnemius from a frog
just killed, cut it across as in a and J, and
test the sections with neutral litmus paper.
The muscle will be found neutral or faintly
alkaline,

7—2

100

ELEMENTARY PHYSIOLOGY. [1x.

14, Myosin.

Strip off the muscles from a rabbit or other
animal and chop them up as finely as possible,
place them in a large jar filled with water and
stir; in about a quarter of an hour place a piece
of muslin over the top of the jar and pour off the
water ; fill up the jar again with water and let it
stand for an hour, then pour off the water and
refill the jar as before. When this has been
repeated once or twice the greater part of the
substances soluble in water will have been re-
moved ; it is best however to let the muscle stay
in water for a day adding a little thymol or
salicylic acid to prevent putrefaction. The
filtrate from a small quantity of muscle which
has been in an equal bulk of water for an hour
should give no proteid reaction if the muscle
has previously been thoroughly washed.

Collect the muscle on linen, squeeze out the
water, grind it up with clean sand and add 5
times its bulk of 10 p.c. ammonium or sodium
chloride, and stirring occasionally place it aside
for an hour or so (naturally if it is left longer |
more myosin will be obtained). Filter through
muslin,* through linen, and then through coarse
filter paper. A somewhat viscid fluid is ob-
tained; pour this into a tall vessel containing
about a litre of water and observe the pre-
cipitate of myosin which takes place (or the

1 The residue may be again treated with 10 p.c. ammonium
chloride.

IXx.]

PROPERTIES OF CONTRACTILE TISSUE, 101

fluid as it filters may be allowed to drop into
water).

After a short time decant or draw off with a
pipette as much fluid as possible, shake up the
rest and pour about 5c¢.c. with the myosin
suspended in it into each of three test-tubes.

a. Add drop by drop a strong (e.g. 20 p.c.) salt
solution, the precipitate soon dissolves ; then
place it in a water bath at about 50°C. with a
thermometer and heat; at about 57°C. the
fluid becomes milky (or a precipitate is
formed) owing to the coagulation of the
myosin.

b. Add powdered salt, the precipitate soon
dissolves, but when the fluid is saturated
with salt is reprecipitated (the precipitate
may be in the form of fibrous-looking clumps).

c. Dissolve the suspended precipitate in salt
solution, and test for proteids, e.g. with Xan-
thoproteic reaction (cp. Less. 111. § 16 (@)).

Myosin belongs to the class of globulins; it will
have been seen that it is insoluble in water and
in saturated neutral salt solutions, soluble in not
too dilute solution of neutral salts, its solution
coagulating on boiling (cp. also Less. ur. §§ 11,
13, 17, 18).

. Syntonin or Acid Albumin.

Treat a portion of muscle washed as in § 14
with ten to twenty times its bulk of HCl *1 p.c.
and place in the warm chamber at about 40°C,

102

16.

ELEMENTARY PHYSIOLOGY. [1x.

frequently shaking. After 3 or 4 hours most of
the myosin of the muscle will have been con-
verted into syntonin or acid-albumin, and dis-
solved, Syntonin may be prepared in a similar
way from white of egg or from serum.

Filter and carefully neutralize the filtrate with a
weak solution of sodium carbonate (1 c.c. of HCl

_ ‘Lp.c. requires about 1°5 c.c. of Na, CO, ‘1 p.c. to

neutralize it), a copious bulky precipitate of
syntonin takes place; if too much alkaline salt
is added the syntonin will be converted into

alkali albuminate and redissolved (cp. § 16).
Filter and wash the precipitate once on the filter

with water, then break through the filter paper
and wash the precipitate into a beaker with

_ water. The syntonin is thus obtained suspended

in water. Place 2 c.c. in each of 2 test-tubes.

. a. Add a little HCl ‘1 p.c. the precipitate at

once dissolves and is not precipitated on
boiling; cool under a tap, and test for pro-
teids e.g. with acetic acid and potassium
ferrocyanide (cp. Less. m1. § 16).

b. Boil, cool under a tap, and add a little HCl
‘1 p.c.; the precipitate suspended in the water
has been coagulated by boiling and is no
longer soluble in dilute acids.

Alkali Albuminate.

Treat a little washed muscle as in § 15 but with
NaHO ‘1 p.c. instead of with HCL On neutral-
izing a precipitate of alkali albuminate will be

Ix.]

PROPERTIES OF CONTRACTILE TISSUE. 103

- obtained like that of ‘syntonin ; if too much acid

is added the precipitate will be converted into
acid-albumin and re-dissolved. Take in separate
test-tubes a little of the precipitate suspended in
water.

a. Add a little NaHO ‘1 p.c. the precipitate at
once dissolves and is not precipitated by
boiling ; with the proteid tests a reaction is
obtained.

b. Boil, cool under a tap and add a little NaHO

‘1 p.c., the precipitate suspended in water has
been coagulated by boiling and is no longer
soluble in dilute alkalies.

c. Adda little NaHO ‘1 p.c. and a little sodium
phosphate, with a drop of litmus solution; add
drop by drop dilute HCl: on neutralizing no
precipitate takes place, add a little more
acid a precipitate is obtained, if still more is
added the precipitate is re-dissolved.

It will have been seen that proteids in solution
(except peptone) on warming with dilute acids
or alkalies give rise to acid albumin or alkali
albuminate respectively, these substances do not
coagulate on boiling and so differ from albumin
and globulin, and are insoluble in water and so
differ from peptone (cp, Less, xvi. C. § 5).

DEMONSTRATIONS.

The electric currents of resting muscle removed
from the body (demarcation currents)

104

ELEMENTARY PHYSIOLOGY. [1x.

The negative variation of the currents of the
muscle during a contraction (action currents)

The rheoscopic frog

Action of heat and cold on the single muscular
contraction

Action of veratrin on the single muscular con-
traction

Curve of elasticity of muscle

Measurement of latent period of contraction ~
of muscle by means of pendulum myograph,

LESSON X.
STRUCTURE OF NERVOUS TISSUES.

A. Spryat NERVEs.

1. Cut off about one-third of an inch of a small
perfectly fresh nerve (e. g. a branch of the sciatic
of a frog), and place it on a glass slide without
any fluid. Fixing one end by pressing on it
with the blunt end of a scalpel, pass a needle
through the other end in the direction of the
nerve fibres, and so spread it out into the shape
of a fan; add a drop of normal saline solution,
and cover with the cover-slip. Observe
a, The medullated nerve-fibres of variable

size.

b. In each fibre the double contour, due to the
medullary sheath or white substance of
Schwann.

c. The primitive sheath; this is seen with
difficulty except at points where the me-
dullary sheath has been displaced in mount-
ing,

d. The connective tissue (endoneurium) running
amongst and round the nerve-fibres.

106

ELEMENTARY PHYSIOLOGY. [x.

e. Drops and fragments of the medullary sheath,
extending from the cut ends of the fibres and
shewing a double contour.

Some non-medullated nerve-fibres will also be
seen (cp. B).

Cut off a similar piece of nerve and treat it in
the same way, but instead of adding a normal
solution, add a drop of chloroform, and irrigate
with chloroform as evaporation goes on.

In the middle of the nerve-fibre the pale granu-
lar axis cylinder will be seen running through
the swollen and partially dissolved medullary
sheath.

Take another piece of fresh nerve, and place it in
a small quantity of osmic acid 1 p.c. for ten
minutes, covering it up to prevent evaporation.
Remove it to a slide, and separate a small bundle
of fibres, putting the rest back in osmic acid;
isolate the fibres as much as possible, add a drop
of water, and put on a cover-slip. If it is desired
to preserve the specimen the nerve should be
washed with water before teasing and mounted
in dilute glycerine. |
Select a nerve fibre which is isolated for a con-
siderable part of its length, and observe

a. The medullary sheath, stained black with
osmic acid,

b. The nodes of Ranvier. These will be
seen as short but distinct breaks in the me-
dullary sheath; with Zeiss obj. D. oc. 2 they

STRUCTURE OF NERVOUS TISSUES. 107

occur usually at intervals of about twice the
diameter of the field, varying considerably,
however, in different nerves. The axis cy-
linder may be seen continuing across the
nodes.

c. The nuclei of the primitive sheath; one to
each internodal nerve portion situated about
halfway between two nodes, and appearing
as a small transparent elongated body pro-
jecting usually into the medulla.

The nuclei of the primitive sheath may be
stained by placing a piece of the nerve, after
treatment with osmic acid, in Frey’s carmine
for two or more days. Insuch a preparation,
the cells of the fine connective-tissue around
the nerve fibres will also be well seen.

Stain with carmine a short piece of a rather
thick nerve which has been preserved in ammo-
nium bichromate 2 p.c. (cp. Less.1. F. § 4 and
footnote p. 69). Imbed, and cut thin transverse
sections, treat these with creosote and turpentine
(cp. p. 75 footnote) and mount in Canada
balsam. Observe

a. The cut ends of the nerve-fibres varying in
diameter, and in each

b. The section of the stained axis cylinder sur-
rounded by

c. A transparent ring indicating the former
position of the medullary sheath, which has
been dissolved or made transparent in the

108

B. SymMpatHetTic NERVES,
1,

d. The primitive sheath as a limiting circle,

é.

Cut out from the fresh spleen of a large animal
(e.g. ox) a small piece of one of the large sympa-
thetic nerves running alongside the blood-vessels.
Remove the connective-tissue sheath, and tease
out the nerve carefully in normal saline solution.
Note

a.
b.

Take a small piece of the sympathetic nerve
from the neck of a freshly-killed rabbit, place it
in osmic acid 1 p.c. for five or ten minutes, wash
and stain with strong Frey’s carmine or with
picrocarmine, Tease out in dilute glycerine.

ELEMENTARY PHYSIOLOGY. [x4

process of mounting. If the nerve has been
kept long in ammonium bichromate the
medulla will shew an indistinct network
stained yellow.

The arrangement of the fibres in bundles.
The perineurium around each bundle con-
tinuous with the endoneurium between the
fibres.

The scanty medullated nerve fibres,

The non-medullated nerve fibres consti-
tuting the bulk of the nerves: they are
fairly transparent, and often fibrillated, some
give off fibrillae. Add acetic acid to bring
out more distinctly the small elongated nuclei
which lie on them at short intervals.

~

—

x.] STRUCTURE OF NERVOUS TISSUES. 109

Numerous small medullated fibres will be
seen mingled with the non-medullated fibres.
Compare the size of the medullated fibres
here and in the sciatic nerve (A, § 1).

C. PERIPHERAL CouRSE OF NERVES.
1. In the specimen prepared Less. VulI. $6: observe

a. Under a low power, the nerve running across
the lower part of the muscle and sending off
fibres or bundles of fibres at intervals and so
spreading out over it.

b. Under a high power, that where the lateral
bundles (especially the smaller ones) are
given off, one or more of the nerve fibres
divide into two fibres, the division taking
place at a node. Trace a small bundle of
nerve fibres, the nodes are very close together.
Each nerve fibre apparently ends abruptly
over a muscle fibre; in this specimen it can
only be traced as far as the blackened me-
dulla extends.

2. Make a gold chloride preparation of the cornea’

1 The mesentery of the frog may be taken instead of the cornea,
but it is more difficult to obtain a good preparation. The mesentery
of the lower coil of the small intestine is pinned out with the
intestine, and placed in gold chloride °5 p.c. for about half-an-hour,
it is then well washed, the intestine cut away, and the mesentery
exposed to light in acidulated water. The larger nerve bundles,
containing a few medullated fibres, accompany the larger arteries
and form coarse and fine plexuses of non-medullated fibres as in the
cornea, but there are many more nuclei, even very small fibres
having nuclei on their course ; varicose fibrils like those of the cornea
will probably not be seen.

110

ELEMENTARY PHYSIOLOGY. [ke

of a frog as in Less, v. B. § 2 and mount in
glycerine a sector of it, including its junction
with the sclerotic. Note

a. The small separate bundles of nerve fibres
entering the cornea at its periphery; the
medullated fibres on account of their me-
dulla are more deeply stained than the non-
medullated.

b. Trace as far as possible the course of one of
the nerve bundles; the medulla soon dis-
appears, the fibres, still showing nuclei at
intervals, join with the fibres from other
bundles to form a coarse plexus; from this
proceeds a plexus of smaller bands which
have few nuclei; finally from this plexus run
very fine varicose non-nucleated nerve-fibrils
in straight lines across the cornea (these fine
fibrils may also be seen forming part of the —
finer bands of the plexus). :

D. SprnaAL GANGLIA.

2:

Take a mammal’s spinal ganglion (e.g. a dorsal
spinal ganglion of a cat or dog) which has been
preserved in ammonium bichromate 2 p. c. about
three weeks, and subsequently in spirit. Prepare
sections passing through the trunk of the nerve, —
the ganglion, and the anterior and posterior
roots of the nerve. Stain with hematoxylin or
carmine, clear (cp. p. 75) and mount in Canada
balsam.

x.]

STRUCTURE OF NERVOUS TISSUES. lll

Observe with a low power that

a.

The fibres of the anterior root mix with
those of the posterior root below the gan-
glion.

Most of the ganglion cells lie outside the
fibres of the posterior root; some lie in rows
between the fibres.

Observe with a high power

a.

In the ganglion cells the rather granular
cell-substance, containing a large spherical
nucleus which often has one or more distinct
nucleoli.

The many nuclei of the capsule of each
ganglion cell.

The stained axis cylinders of the nerve fibres.
(The medulla may show an irregular network
of substance hardened by the ammonium
bichromate.)

The nerve fibres coiled amongst the cells,
and fibres running more or less transversely
from the lateral mass of cells to the central
bundle of nerve fibres,

Destroy the brain and spinal cord of a frog and
cut off its head. Remove the lower jaw and
divide the skull in two down the middle line.
Tear away the mucous membrane covering the
roof of the mouth; the inner and lower part of
the eye will be laid bare. Just behind the eye is
a depression ; from this two or three small fila-
ments, the branches of the fifth nerve, will be

112

ELEMENTARY PHYSIOLOGY. [x.

seen issuing. Cut through the bone from the
middle line to the place where the nerves issue,
turn the piece of bone forward, and gently scrape
out the brain.

A filament (the fifth nerve) will then be seen
going towards the branches spoken of above.
Between the two is a slight enlargement, the
Gasserian ganglion, surrounded by tough con-
nective tissue and by connective tissue attached
to the bone. Cut away this tissue and excise
the ganglion.

Tease it out a little in ‘25 p.c. osmic acid’, leave
it in the acid for about half-an-hour, then wash,
and place it in strong Frey’s carmine for a day;
wash, and tease out in glycerine, during the
teasing examine from time to time under a low
power and throw away the fragments which

‘consist of connective tissue only.

Instead of the Gasserian ganglion a spinal gan-
glion from a frog or mouse may be taken. In
the frog the spinal ganglia are surrounded by a
chalky mass; when this is removed the ganglion
can be seen as a greyish semi-transparent bulg-
ing on the nerve, the 7th, 8th, 9th are easiest
to obtain.
Examine carefully the more isolated cells; here
and there one will be seen with its cell substance
prolonged into a process; from most of the cells
the process will probably have been torn away
in teasing.

1 Or it may be placed in potassium bichromate °2 p.c. for two or
three days and then teased out.

x.] STRUCTURE OF NERVOUS TISSUES. 113

[E. SYMPATHETIC GANGLIA.
Place a sympathetic ganglion e.g. the superior
cervical sympathetic ganglion of a rabbit in
1 p.c. osmic acid for a day. Wash it with water
and leave it in water for some hours. Transfer to
50 p.c. spirit for an hour, then to strong spirit for
a week, Cut longitudinal sections and observe

a. The large number of nerve-cells of various
sizes.

b. Thesmall number of medullated nerves com-
pared with the spinal ganglion (D § 2a).

c. The small size of nearly all the medullated
nerves (cp. D § 2d), when these are in
bundles they are usually at the periphery
of the ganglion.

d. The bundles of non-medullated nerve-fibres
at the ends of the ganglion; the distinctness
with which these are seen will of course vary
in different sections.

Dissection of Sympathetic Ganglia in the Frog. Lay
open the abdomen of a recently killed frog, pick up
the intestine and cut through the mesentery just
above the kidneys; pulling up one kidney, cut
through the peritoneum along its edge, and turn
over the kidney to the opposite side of the body.
Gently stretch the spinal nerves away from the
spinal column, a row of small nerve fibres will be
seen running transversely from the spinal nerves
(one from each nerve) to the chain of pigmented,
semi-transparent sympathetic ganglia which will be
seen lying over the spinal column.

L. 8

114 _ ELEMENTARY PHYSIOLOGY. [x.

F. CELts oF SPINAL CorD.

Cut transverse sections of the spinal cord of a
large mammal (e.g. calf or ox) which has been
hardened in ammonium bichromate 2 p. c. and ©
subsequently treated with alcohol. Stain with

carmine or dilute picrocarmine (it is better still
to stain the small piece of cord before cutting
sections by placing it in strong Frey’s carmine for
several days); clear and mount in balsam. Note

a. In the white substance the cut ends of the
nerve-fibres differing very greatly in size
(cp. with A § 4).

b. The small amount of connective tissue (neu-
roglia) between the nerve fibres; this is finely
dotted (transverse sections of semi-elastic
fibrils), and is more abundant near the grey
substance. |

c. In the grey substance of the anterior cornu,
the large multipolar nerve-cells each with a —
large nucleus and distinct nucleolus. Select a
conspicuous cell and note that the processes
run off in all directions; many of them will be
seen to branch and these branches to branch
again and so on (protoplasmic processes) ;
from some of the cells a rather large process —
which does not branch may be seen to run
outwards through the white substance (axis
cylinder process).

d. In the grey substance of the posterior cornu,
the much smaller usually fusiform nerve-cells —
with branching processes at either end.

x.]

re)

3.

STRUCTURE OF NERVOUS TISSUES. 115

DEMONSTRATIONS.

Preparations to shew the method of ending of
motor nerves in muscle.

Preparation.to shew the nerve-cells of the plexus

of Auerbach in the intestine.

Mounted transverse section of a nerve which has
been treated with osmic acid.

Nore to D §3. Isolation of nerve cells. The ganglion may be
placed in osmic acid 1 p.c. for half an hour, washed in water, placed
in a mixture of equal parts of glycerine and strong acetic acid for
three or four days, and then teased out,

LESSON XI.

GENERAL PROPERTIES OF NERVOUS TISSUE.
AUTOMATIC ACTIONS.

A. ReEFiLex ACTION.

1, Place on its belly a frog, which has been pre-
viously deprived of its brain* by the demonstrator.
Observe that its hind-limbs are drawn up under
the body; but that it differs from the normal
frog in the following respects.

a. Its head is depressed, instead of being erect.

b. Its fore-limbs are spread out, or flexed, instead
of being held nearly vertical; thus the angle
which the body makes with the table is
diminished.

1 This is done by cutting across the spinal cord just below the
skull and thoroughly destroying the brain with a seeker; in order to
ensure the complete removal of the medulla oblongata, it is advisable
to destroy the parts a short distance down the'spinal canal. The
frog should be left in a moist place for a day. Instead of this the
frog may be chloroformed the skull cut open and the brain re-
moved; or the upper jaw with the anterior part of the skull may be
cut off with strong scissors and the medulla removed with forceps,
with this method the reflex action is often weak owing to the
considerable loss of blood.

XL]

GENERAL PROPERTIES OF NERVOUS TISSUE. 117

c. There are no respiratory movements, either
of the nostrils, or of the throat.

Gently pull out one of the hind-limbs, until it
becomes quite straight, and then let it go. It
will be immediately drawn up into its old posi-
tion under the body. If this experiment be
made soon after the operation of removing the
brain, or if much blood has been lost, the leg
may be drawn up slowly instead of sharply.

Gently tickle one flank with a feather or a blunt
needle; a contraction of the flank muscles of
that side will be observed.

Pinch the same spot rather sharply with a pair
of forceps; the leg of the same side will be first
extended, and then drawn up and swept over the

’ flank, the movement tending to thrust away the

points of the forceps.

Pinch with the forceps the skin round the anus;
both legs will be drawn up and thrust out again;
the movement tending as before to sweep away
the points of the forceps.

Leave the animal alone for five minutes and
watch it carefully: if no disturbing circumstances
are brought to bear on it, it will remain perfectly
motionless,

Place the animal on its back; it will make no
effort to regain its normal position, 7.e. all sense
of equilibrium has been lost.

118

10.

11.

_ Take two glasses, fill one with dilute sulphuric

ELEMENTARY PHYSIOLOGY. [ XI.

Pass a hook through the lower jaw, and fasten it
to the cross-bar of a stand so that the body can
be raised up and down. The hind-limbs, after a
few movements of flexion and extension, will
remain pendant and motionless.

Gently pinch the tip of one of the toes of either
leg; that leg will immediately be drawn up.

acid (‘1 p.c.) and the other with water: lower
the frog until the tip of one of the toes just
touches the dilute sulphuric acid. In a short
time the foot will be withdrawn. At once let
the foot dip into the second glass containing
water, in order to wash away the acid. Measure
with a rapidly beating metronome, the time
between the moment when the toe comes into
contact with the acid and the moment when it
is withdrawn. Make, at intervals of a few
minutes, three such observations, and take the
mean of the three.

Cut a small piece of blotting-paper one or two
mms. square, moisten it with strong acetic acid,
and place it on the flank of the animal. The
leg of the same side will be speedily drawn up
and swept over the flank as if to remove the
piece of paper.

Place similar pieces of paper dipped in acetic
acid on different parts of the body; different
movements will be witnessed in consequence; —

XI.] GENERAL. PROPERTIES OF NERVOUS TISSUE, 119

12,

13.

14,

all however tending to remove the irritating
substance,

Wash off all the acid from the frog, and when it
has become perfectly quiet, place it in a basin of
water; it will sink to the bottom (unless the
lungs be accidentally much distended with air),
and no movements of any kind will be witnessed.

Observe that all the movements produced in the fore-
going observations, although complicated, co-ordinated,
and purposeful in character, are partial, and only by
accident bring about locomotion. However stimulated,
the animal never springs or leaps forward.

In order that the same frog may serve for observations
on the lymph-hearts, B, I. § 1 should be performed

here.

Make a small cut through the skin of the back’,
and with a fine glass tube inject one drop of a
1 p.c. solution of strychnia. In a few minutes
the slightest stimulus applied to any part of the
animal will produce violent tetanic spasms of the
whole body. A preliminary stage of increased
reflex action may also be observed.

With a straight seeker or a piece of stout wire
destroy the whole of the spinal cord. Observe
that the spasms immediately cease,

1 Immediately underneath the skin of the back of the frog is the
dorsal lymphatic sac, and any fluid placed in this rapidly makes its
way into the blood. -

ioe. ELEMENTARY PHYSIOLOGY. [ XI.

15. Repeat any of the above observations (§ 2—13).
No reflex actions will now be produced.

B. AvtTomMAtTic ACTION. .
I. The Lymph-Hearts.

1. Placing the animal on its belly watch the move-
ments of the posterior lymph-hearts. They may
be seen beating on either side of the extremity
of the urostyle, in a depression between that bone
and the hip-joint. The contractions are generally
visible through the skin, but become more evident
if the skin be removed, care being taken not to
injure the lymph-hearts themselves.

2. Observe that after destruction of the posterior
part of the spinal cord the lymph-hearts cease
to beat. |

IJ. The Heart.

1. Lay the frog on its back, make a median incision
through the skin, and from the middle of this
make transverse incisions. Raising up the end
of the sternum with a pair of forceps, cut it
through a little above the end so as to avoid
cutting the epigastric vein. Then again lifting
up the sternum cut it through longitudinally
with a strong pair of scissors; pin back each
part of the sternum, cutting through such
muscles as may be necessary; the heart will

XL] GENERAL PROPERTIES OF NERVOUS TISSUE. 121 .

be seen in the thin membranous pericardium
beating with considerable regularity and force.
Pinching up the pericardium with a fine pair of
forceps, cut it away from the surface of the heart
then tilt up the apex of the ventricle; a small
band of connective tissue will be observed passing
from the posterior surface of the ventricle to the
adjoining wall of the pericardium. Seizing this
band with the forceps, divide it between the
forceps and the pericardial wall. Lift up the
apex of the ventricle, by means of this band,
and with a sharp pair of scissors cut through the
aorte, the superior venz cave, the inferior vena
cava, and the surrounding tissue. Take care not
to injure the sinus venosus. Place the heart in
a watch-glass, moistening it when necessary
with normal saline solution. The beats will
either not be interrupted at all or for a very
short time. |

In cold weather the heart may stop on being
removed from the body, but if the heart be
warmed by putting the watch-glass’ containing
it in the palm of the hand, the beats will be
resumed, |

2, Lifting up the apex of the ventricle by means of
the same band as before, cut through the ventricle
with a sharp pair of scissors at its upper third.
The lower two-thirds of the ventricle will remain
motionless without any spontaneous beat; the
upper third and the auricles will continue to
beat with regularity.

III.

bo

‘ELEMENTARY PHYSIOLOGY. [XI

By means of a longitudinal incision divide the
auricles with the attached portions of ventricle
into two lateral halves. Each half will continue
to beat.

Cilia.

Placing the frog on its back, cut through the
lower jaw, in the middle line, and carry the
incision down the oesophagus as far as the
stomach. Pin back the parts and moisten the
mucous membrane, if it is at all dry, with
normal saline solution. Place on it, a little
below the orbits, a small thin piece of cork.
The cork will be seen to be driven by ciliary
action down towards the stomach,

DEMONSTRATIONS.

The electric currents of nerves removed from the
body (demarcation currents).

The negative variation attending a nervous
impulse (action current).

The effect of a constant current on demarcation
currents (electrotonus).

The effect of a constant current on the irritability
of nerves (electrotonus as affecting irritability).

The law of contraction,

. -i
= é t i

a= te ll iy
5 % 7 = Ain aan

=

5 OF NERVOUS TISSUE. 123
Es, EEN

The function of the anterior and Shere roots
of the spinal nerves.

The peristaltic movements of the intestines sacl =
of the ureters :

;
: ‘

LESSON XII.

STRUCTURE AND PROPERTIES OF BLOOD-

VESSELS.

A. THE LARGER ARTERIES.

i.

Cut transverse and longitudinal sections of a_
small piece of the aorta or carotid artery of a

dog (or other animal), which has been preserved

in potassium bichromate 1 p.c.; stain with picro-

carmine, Note

a. Thethin inner coat thrown into longitudinal
folds by the contraction of

b. The much thicker middle coat, consisting
of alternating layers of elastic and circularly
arranged muscular tissue. Both of these are
stained of a yellower tint than the white
fibrous tissue. The disposition and relative
amount of the elastic and muscular tissue
will be found to vary much in different
arteries. |

c. The external coat (tunica adventitia), con-
sisting mainly of white fibrous connective

x11.]

STRUCTURE OF BLOOD-VESSELS, 125

tissue, with some elastic tissue not infre-
quently arranged in layers, especially at the
inner part of the coat: Note the nuclei of
the muscle cells (cp. Lesson vit. § 11).

As a rule in the more peripheral arteries, the
middle coat is formed of bundles of muscular
fibres separated from one another by a small
amount only of connective tissue with elastic
fibres.

Take the jugular vein from a freshly-killed rab-
bit, cut it open longitudinally, and pin it out
with hedgehog quills with the inner surface
uppermost on a thin piece of cork ; stream water
gently over it for a moment, then pour over it a
‘2 p.c. solution of nitrate of silver and leave it for
five minutes, wash well with distilled water, and
expose to light, also in distilled water, until it
becomes of a brownish colour: spread it out on
a slide with the inner surface uppermost, treat
it successively with 50 p.c, 75 p.c, 95 p.c.
alcohol, clear and mount it in Canada balsam.

Observe the rather jagged dark lines of the
cement substance between the cells; the cells
form a continuous layer, are more or less elon-
gated in a longitudinal direction, and are flat;
indications of the nuclei of the cells may be
seen, and in some places indications also of the
fibres of the muscular coat, as transverse or
longitudinal markings caused by a deposition of
silver in the cement substance between the muscle
cells, If the vein has been left too long in the

ELEMENTARY PHYSIOLOGY. [xian

silver nitrate solution, or exposed too long to
light, silver will be deposited in the substance
of the cells also.

_ A vein is here taken as being a somewhat easier

preparation for a student than an artery; the main
features in either case are the same.

Cut open longitudinally a large artery, e.g. carotid
of sheep, from a freshly killed animal, and very
gently scrape the internal surface. Tease out
the material so obtained in normal saline solu-
tion. |
Observe in surface and profile the cells of the
lining epithelium, with their elongated nuclei.
Probably there will also be seen fragments of
the elastic fenestrated membrane. Add acetic
acid 1 p.c. to distinguish more clearly the elastic
from the white fibrous tissue. (The perforations
in the fenestrated membrane are rendered more
distinct by staining.)

Tear off a strip from the inner coat of a medium-
sized artery which has been in potassium bichro-
mate ‘2 p.c. for two to six days; tease it out in
the fluid ; it will be found to consist almost en-
tirely of elastic lamine.

Observe the gradations from an almost homo-
geneous elastic perforated membrane to a mesh-
work of elastic fibres.

. xix]

STRUCTURE OF BLOOD-VESSELS. 127

B. THE LARGER VEINS.

.

bo

Cut transverse sections of a fairly large vein, as in
A. §1. Relatively the muscular element will
be found to be less (though varying considerably
in different veins); the elastic element also to
be less, and the white fibrous connective tissue
more plentiful.

Attempt to make fluid pass along a vein of some
large animal in the wrong direction, in order to
shew the action of the valves. Cut open the
vein and examine the structure and position of
the valves.

C. CAPILLARIES AND SMALL BLOOD-VESSELS.

¥,

Place on a slide with a drop of glycerine the tail
of a tadpole which has been hardened in Muller's
fluid or in chromic acid ‘2 p.c. and subsequently
treated with alcohol. Brush off the surface epi-
thelium or if this is unsuccessful break it up
into not too small fragments with needles (in
doing so try to scrape off the epithelium). The
tissue may be previously stained by placing it
for a day in dilute picrocarmine. Note

a. The network of capillaries; in some, blood-

corpuscles will probably be seen.

b. The nuclei in the walls of the capillaries, es-
pecially at the points of bifurcation.

c. The branched connective-tissue corpuscles, in
places partially surrounding the capillaries,

128 ELEMENTARY PHYSIOLOGY. [ XII.

and constituting the simplest form of a
tunica adventitia.

d. The connection of some of the processes of
the connective-tissue corpuscles with the ca-
pillary walls.

2. Remove a small portion of pia mater from a per-.
fectly fresh brain of a chloroformed cat or dog
allowed slowly to bleed to death, and spread it
out carefully in normal saline solution, washing
it gently with the same, and mount. Note

a. The capillaries, with their nuclei bulging into
the channels.

b. The small arteries, their scanty external con-
nective-tissue coat or adventitia, and the
indications of their muscular coat composed
of a single layer of fibres wrapped round the
intima.

c. The small veins generally filled with blood-
corpuscles, with their muscular coat absent
or developed only to a slight degree.

3. Gently irrigate with acetic acid 1 p.c. Note
a. In the capillaries, the more distinct nuclei.
b. In the small arteries,
(a) The fusiform cells and nuclei of the

adventitia, arranged beng sana with
elastic fibres.

(8) The nuclei of the muscular coat ar-
ranged transversely. By watching the

XI. ]

STRUCTURE OF BLOOD-VESSELS. 129

earlier stages of the action of acetic
acid, the outlines of the muscle fibres,

- as well as of their nuclei, will be dis-
tinctly seen.

(y) The thin inner elastic layer, internal to
the muscular fibres, seen as wrinkled.
longitudinal markings.

(6) The oval nuclei of the lining epithelium
arranged longitudinally.

c. In the small veins, appearances similar to ),
consistent with the less development of mus-
cular and elastic tissue.

A piece of fresh pia mater may be stained with
aqueous hematoxylin to bring out more clearly some
points of the above; the following will, however, be
found a better method. Place a fresh brain, with
the dura mater at least in part removed, in potas-
sium bichromate 1 p. c. for two or three days (or
longer), then wash it with water, and place it in
75 p.c. spirit for half an hour to twenty-four as
convenient, cut off then a small piece of pia mater,
stain with hematoxylin, or picrocarmine, and mount.
Care should be taken to drag the membrane as little
as possible, and to avoid creases.

Destroy the brain and spinal cord of a frog, lay

bare the heart, cut through the whole length of

the muscles of the abdomen a little.on one side

of the epigastric vein. Cut off the ventricle

and cut across the venous sinus (cp. Less. XI.

B. I), sop up with a sponge the blood which
9

130

_ ELEMENTARY PHYSIOLOGY. [XI

flows out, when the bleeding has ceased take
a small glass cannula having a piece of india-
rubber tubing over one end and filled with
normal salt solution and insert it into the aortic
bulb, tie it near or at the branching of the
aorte with a silk thread. Then take a syringe
full of normal salt solution, and the tubing being
also full of salt solution gently force the nozzle
of the syringe into the tubing, being careful
that no air-bubbles get in at the same time.
Then slowly inject the salt solution: this should
be repeated until the fluid issuing from the
venous sinus is perfectly clear; sop up the fluid
and inject with water, then inject once or twice
a ‘2 p.c. solution of nitrate of silver, sopping up
the fluid which issues from the sinus; inject
water to wash out the nitrate of silver, place the
frog in a basin of water and cut out the intes-
tine ; cut through the mesentery at its junction
with the intestine and lay open the intestine,
brush away its epithelial coat and expose it to
light in water or in 30 p.c. alcohol. The out-
lines of numerous vessels will soon be visible;
then cut out a piece, spread it out on a slide
with the inner surface uppermost, wash with
alcohol of increasing strength, clear and mount
in Canada balsam’.

1 If the tissue be left too long before it is mounted the cells as
well as the cement substance will become black. From the frog
injected as above a piece of the mesentery, or of the lungs, or of the
bladder, may be mounted.

XII.] STRUCTURE OF BLOOD-VESSELS, 131

Note with a low power the small arteries branch-
ing into capillaries and the uniting of the capil-
laries to form veins which again unite to form
larger veins.

Note under a high power the outlines of the flat
elongated epithelium cells of the small arteries,
of the capillaries and of the small veins; the
outlines in the capillaries are more irregular than
in the arteries or veins; in the veins the cells are
rather broader than in the arteries.

In the small arteries the cementing substance
between the cells of the muscular coat (here
usually a single layer) will also shew as black
transverse lines, the small veins are without
these transverse markings.

D. CIRCULATION OF BLoop. INFLAMMATION.

Take a frog with a pale web which a day or two
previously has been deprived or its brain by the
Demonstrator (cp. p. 116, footnote) and kept in
a moist place. With scissors make a small cut
through the skin of the back, with a fine pipette
introduce underneath the skin one drop of a 1 p.c.
solution of urari, leave the frog under a glass
jar until it does not move on being pinched;
this should be the case in about three-quarters
of an hour, if it occurs earlier too much urari has
been given and the blood-vessels will be abnor-
mally dilated. Lay the frog on its belly on the
frog-board, and tie, not too tightly, a piece of soft
cotton round the end of the (e.g.) 3rd and 4th

9—2

132

ELEMENTARY PHYSIOLOGY. [ XII.

digits. Stick two pins into the board a little
distance from the hole; by twining the cotton
round them the web may be stretched out level
above the hole (or the toes may simply be pinned
out). Surround the web, and cover the leg and
body of the frog, with moist blotting-paper.
Put a small drop of water upon the web, and
cover it with a triangular piece of cover-slip,
taking care that the glass does not cut into the
digits and that no fluid flows over it.

Examine, first with a low and then with a high
power. Note

‘a. The course of the blood from the arteries to
the veins. A slight pulsation may be ob-
served in the larger arteries and sometimes
in the smaller ones.

b. The greater velocity of the blood in the
arteries (owing to their smaller size) than
in the veins; probably in neither can the ~
individual corpuscles be made out.

c. The axial and peripheral zones in the arteries
and veins; the peripheral zone is small, and
under a low power appears free from corpus-
cles; under a high power one or two white
corpuscles may, if the current is not very
fast, be seen in the peripheral zone of the
arteries; in that of the veins a few white
corpuscles and occasionally a red one will be
seen moving along comparatively slowly.

d. The passage of corpuscles usually in single
file through the capillaries ;

xIL.] STRUCTURE OF BLOOD-VESSELS. 133

e. The elasticity of the red corpuscles, observing
the way in which they bend and become
deformed, and then regain their normal shape.

6. Remove the cover-slip, and soak up the fluid on
the web; touch the middle of the web with
the tip of a pointed glass rod that has been
dipped in creosote’, so as to leave a minute
drop on the web, and replace the cover-slip.
The early stages of inflammation may be
seen. Observe in succession

a. The dilatation of the arteries, the much more
distinct appearance of the capillaries, and the
enlargement of the veins, accompanied by a
quickening of the current.

b. A slowing of the current, the vessels remain-
ing dilated.

c. The white corpuscles increase in number in
the peripheral zone of both arteries and veins,
in the latter the white corpuscles begin to
cling to the sides, at first for a time only,
then being carried off, later permanently
until the veins have a more or less complete
lining of white corpuscles, In the capillaries
the white corpuscles and also, though less
frequently, the red corpuscles, stick to the
capillary walls, partially or completely block-
ing the way. Later the way becomes com-

1 Instead of this, olive oil containing 1 to 2 p.c. of croton-oil may
be used. In either case the irritant should be washed off if stagnation
sets in rapidly,

134

ELEMENTARY PHYSIOLOGY. [ XII.

pletely blocked and stagnation sets in. Note
in the stagnated vessels the gradual oblitera-
tion of the outlines of the corpuscles.

d. The emigration of the white corpuscles from
the capillaries and veins. This begins as
the circulation becomes slow; a white cor-
puscle which is adhering to the wall of a
capillary or vein should be noted at intervals
of 10 to 15 minutes.

e. The diapedesis of the red corpuscles from the
capillaries; this is best seen in the capillaries
in which the current has almost stopped.

Notice that these effects are local, that they are
of greatest intensity in the spot touched, that
they extend for some distance round, but that
the circulation in the rest of the web is normal.
If the injury to the vessels from the creosote has
been slight, the circulation may be seen to be
re-established in the stagnated spots, the corpus-
cles gradually recover their outline and are carried
off by the current; this is not seen if stasis has
set in i.e. if the blood has clotted,

Observe now the circulation in the tongue; the
frog being on its belly, draw forward the tongue
over the hole in the stage and pin out the two

_cornua; the tongue at first pale soon becomes

flushed and its vessels full of blood. With a low
power the peripheral layer in the arteries and
veins will probably be seen better than in the
web. The phenomena of inflammation (§ 6) can

xII.]

STRUCTURE OF BLOOD-VESSELS, 135

be well seen in this or in the mesentery, in these

further the cedema will be more obvious. To
observe the circulation in the mesentery, place
the frog on its back, cut through the skin and
abdominal muscles on one side, draw out and
pin over the stage a loop of the mesentery.

DEMONSTRATION.

Circulation in the mylo-hyoid muscle of the frog.

LESSON XIII.

STRUCTURE AND ACTION OF THE HEART.

A. HEART OF SHEEP’,

1, Observe the attachment of the parietal pericar-
dium to the roots of the great vessels.
Remember that the parts of the heart which are
right and left in the body are called right and
left after removal. The front of the heart may
be recognized by a groove filled with fat, the
interventricular sulcus, which runs from about
the middle of the base of the ventricles to rather
below the middle of the right margin of the
heart. The front is also more convex than the’
back. Holding the heart with the front towards
you, note that the right ventricle, which will be
on your left hand, is much more yielding than
the left ventricle, which will be on your right
hand, Note also the pulmonary artery arising
nearly in the middle line of the heart at the
upper part of the ventricles, and immediately
behind this the aorta.

1 The heart should be obtained from the butcher with the
pericardium; to secure this it is advisable to purchase the ‘bag,’
i.e. the heart with the lungs still attached to it,

J

XIIl.] STRUCTURE AND ACTION OF THE HEART. 137

2.

Tie a short glass tube into the superior vena
cava and connect with it a piece of india-rubber
tubing. Ligature the inferior vena cava and the
left vena azygos which opens close beside it.
Tie a glass tube about two feet in length into
the pulmonary artery. Fill the india-rubber
tubing with water, and squeezing it press the
water onwards. The water will amount in the
tube connected with the artery, and will only
descend a little way on unclasping the india-
rubber tubing. Pour water into the long glass
tube by means of a funnel, and observe the
column of water which the semilunar valves will
sustain. Note the distension of the arterial
walls and the bulging at the attachment of the
valves. When the pressure of the column of
fluid is removed the artery by its elasticity
returns to its previous dimensions,

Repeat the above observation with the pul-
monary veins and aorta.

Compare the united sectional areas of the
superior and inferior venze cavze when distended,
with the area of the aorta below the origin of
the innominate artery.

Having removed the tubes, lay open the superior
and inferior vene cave, and bring the incisions
to meet in the front of the auricle. Note

a, The size and form of the auricular cavity.

b, The auricular appendage with its muscular
fretwork,

138

ELEMENTARY PHYSIOLOGY. [x11 I

c. The septum auricularum.

d. The fossa ovalis, or expression of the fcetal
foramen ovale, which is early closed by the
growth of the septum auricularum.

e. The Eustachian valve, a slightly projecting
membranous fold, immediately beneath the
entrance of the inferior vena cava, and again
beneath this.

f. The opening of the comparatively large left
azygos vein.

g. The auriculo-ventricular orifice.

Cut open longitudinally the azygos vein, and
observe the coronary vein opening into it a
very short distance from the heart.

Cut away most of the auricle, and holding the
ventricle in the left hand, pour water suddenly
into the auriculo-ventricular orifice. The auri-
culo-ventricular valve will float up and close the
orifice. Note the star-shaped junction of the
valve-flaps. ‘

Introduce a pair of scissors between two of the
valves, and cut through the wall towards the
apex. Having arrived at the bottom of the
ventricular cavity, turn the scissors sharp round
and carry an incision at an acute angle with the
previous one, alongside the septum, towards, but
not into, the pulmonary artery. Lifting up the
flap, note

XIII. ]

10.

11.

STRUCTURE AND ACTION OF THE HEART. 139

a. The thickness of the ventricular wall, the
projections of its inner surface or columnz
carnee. The> band of muscle (moderator
band) running from wall to wall of the ven-
tricle across its cavity.

b. The ventricular cavity does not extend to
the apex.

c. The tricuspid valve, its form, and attach-
ment to the auriculo-ventricular ring, the
chorde tendinee, and their attachment to
the summits of the papillary muscles.

Holding the heart vertically, pour water into the
pulmonary artery; observe from below the form
of the semilunar valves, and their mode of
closing.

To observe the valves from above, insert into the
pulmonary artery a short wide tube, fill it with
water, and cover it with a piece of glass, exclud-
ing air-bubbles,

Prolong the incision of § 8 so as to lay open the
pulmonary artery. Note

a. The form and attachment of the semilunar
valves.

b. The small nodule of tissue in the middle of
the free edge of each valve, the corpus
Arantii.

c. The slight depressions in the arterial walls
opposite each valve, the sinuses of Val-
salva.

140

13.

14,

ELEMENTARY PHYSIOLOGY. [ XIII.

Lay open the left auricle in a manner similar to
that employed for the right. Note that the
left auriculo-ventricular valve, the bicuspid or
mitral has but two flaps. Observe its manner
of closing (cp. § 7).

Lay open the left ventricle in a manner similar
to that employed on the right side, carrying
the incision at first along the extreme left of
the heart. Note the thick walls, the mitral
valve, &c.

Lay open the aorta, and examine its semilunar
valves, -corpora Arantii, and the sinuses of
Valsalva, which are here very distinct. Note
that the coronary arteries open respectively
into the two anterior sinuses. —

B. Heart oF FRoc.

1.

Expose the heart of a just-killed frog in the
manner directed in Lesson x1. B. II. With the
pericardium intact, observe the pulsations of the
heart, noting the alternate beats of the auricles
and the ventricle; and the synchronous beats of
the two auricles,

Lay open the pericardium and observe

a. The synchronous contractions of the two
auricles, followed almost immediately by

b. The contraction of the ventricle, note that

ge, ee eee

ik ih

XIIL] STRUCTURE AND ACTION OF THE HEART. 141

the ventricle during its contraction or systole
becomes pale and conical, and that its apex
is thrown forwards and upwards.

The slight contraction of the bulbus arteri-
osus immediately succeeding the ventricular
systole.

The pause, or diastole, which follows before
the auricle again beats.

The increased redness and distension of the
ventricle after the auricular, and immediately
preceding its own systole.

3. Divide the band attaching the ventricle to the
posterior pericardial wall, and turn the apex of
the ventricle over. Observe

d.

The junction of the two superior ven cave
with the inferior vena cava to form the sinus
venosus.

The white line, roughly y -shaped, marking
the junction of the sinus venosus with the
right auricle,

The cardiac branches of the pneumogastric,
running along each superior vena cava and
then plunging into the interior of the
heart.

The wave of contraction; it starts in the
vena cava, spreads to the sinus venosus,

142

ELEMENTARY PHYSIOLOGY. (xm, 9

almost immediately after the auricles con-
tract, then the ventricle, and finally the bulbus
arteriosus.

’ Make now a transverse cut through the skin of

the frog just below the jaw, and carry the cut as
far as the vertebral column; cut through all the
muscles proceeding from the head of the hume-
rus and from the part of the sternum left attached
to it, to the hyoid bone or to the angle of the
jaw. |

Coming up from underneath the angle of the
jaw and stretching towards the lower extremity
of the hyoid bone will be seen a thin narrow
band of muscle and two small white fibres, one, —
the glossopharyngeal nerve, running along its

upper border, the other, the pneumogastric nerve, —

running along its: lower border. Very carefully
separate the pneumogastric from the surrounding
tissue, and place a loose ligature around it, it
will be seen to divide into two branches, the
smaller branch, the laryngeal, may be cut through.
It will be safer not to attempt to dissect out the
pneumogastric close to the heart.

The nerve may be ligatured close to the skull
(some care is required in doing this) and cut
above the ligature.

_ Pass an interrupted current through the pneumo-

gastric nerve. It is well to place a small piece
of thin india-rubber membrane underneath the
nerve to prevent contraction of the neighbouring
muscles by an escape of current.

--

XIIl.] STRUCTURE AND ACTION OF THE HEART. 143

a. Observe that during and for a short time
after the passage of the current, the heart
remains with all parts in diastole. (If no
effect is produced push the secondary nearer
the primary coil.) |

b. The period of rest (inhibition) may be followed
by a period (reaction) in which the beats are
quicker and more forcible; and then the
previous normal condition is regained.

Now stimulate the pneumogastric nerve, indi-
cating on the curve by means of a time-marker
(cp. Appendix) the moment at which stimulation
begins, and ends. Note that the heart does not
stop immediately after the current is sent into
the nerve.

Place the frog on a stage and arrange a lever
(cp. Appendix) so that it presses lightly on the
ventricle, and bring the end of the lever to mark
on the revolving drum.. Take a tracing of the
ventricular pulsations with the drum at a mode-
rate speed.

Note the rise and fall of the lever, indicating
the change of form during contraction. The
rise increases at first rapidly, then more slowly,
to a maximum, and the fall is similarly at first
slight, then more rapid, but finally slow again.

Turn the heart over and stimulate the line of junc-
tion of the sinus and right auricle; the heart
will stop beating just as it did when the pneumo-
gastric was stimulated.

144

ELEMENTARY PHYSIOLOGY. [ XIII.

Carefully cut away the connective tissue around
the great vessels, tie the superior ven cavz
close to the heart, pass a silk ligature under-
neath the bulbus arteriosus and two underneath
the inferior vena cava. Make a loose knot in
them so that they can be tightened at any
moment. Cut across the aorta and sop up the
blood. With a fine pair of forceps raise up the
wall of the inferior vena cava close to the liver.
With fine pointed scissors, make a cut in the
vein and with the lower of the two ligatures tie
in it a small cannula, wash out the heart with
salt solution in the manner given in Lesson XI.
cp.§ 4. Inject a°5 p.c. solution of gold chloride
until the solution begins to issue from the aorta.
Then ligature the bulbus arteriosus. Again in-
ject, and whilst the heart is distended tie the
remaining ligature round the inferior cava just
beyond the end'of the cannula. Cut out the
heart, immerse it in °5 p.c. gold chloride solution
and leave it for ten to fifteen minutes. Then
remove it to water, cut open the auricles and
shake so that the gold chloride is thoroughly
washed out of it. Transfer to water acidulated
with acetic acid and expose to light for a day;
when it is well stained, observe the septum be-
tween the auricles, cut away the auricles from
the septum and cut away the ventricle, in re-
moving the last portions of the auricles it is well
to examine the septum from time to time under
a low power. Mount the septum in glycerine.
Observe

iin os eel

ee

XII] STRUCTURE AND ACTION OF THE HEART. 145

a.

é.

The two bundles of nerve fibres running
down the septum and having clusters of
nerve cells on them (this is best seen with a
low power); a few of the nerve fibres are
medullated.

From the nerve bundles, nerve fibres extend,
forming a plexus over the septum; in this
plexus nerve cells occur either singly or in
small groups.

Not infrequently a cell may be seen to give
off a process, sometimes a spiral process
may also be made out.

The septum is chiefly composed of a thin
plexus of fibres made up of faintly-striated
muscle cells, the cells and their nuclei are
elongated and more nearly resemble in form
the cells of unstriated muscle than the cells
of mammalian heart muscle (cp. Lesson VII.
§ 12). The outlines of the individual cells
are not distinct.

No capillaries are present.

Numerous nerve fibres and cells may also be
seen in the sinus and at the junction of the
septum with the ventricle (Bidder’s ganglia); a
few occur in the auricles and the basal portion
of the ventricle. A piece of the auricles should
be mounted in glycerine to observe the inter-
lacing bundles of fibres, the further development
of which gives the ventricle the spongy appear-
ance shown in a section; as in the septum (and
rest of the frog’s heart) no capillaries will be seen.

10

ee See den

DEMONSTRATIONS.

Experiment of Stannius.

The antagonistic effects of muscarin leetlbeaeae
and atropin on the heart.

The action of the mammalian heart.
Pneumogastric inhibition in the mammal.
Sounds of the heart.

Endocardial pressure of frog.

Effect of constant current and of successive in-
duction shocks on the ventricle apex.

Rhythmical contraction of ventricle apex under
pressure.

Latent period of ventricle apex.

i.

LESSON XIV.

BLOOD PRESSURE.

A. Minor ARTERIAL SCHEME’.

Z.

bo

Clamp the india-rubber tube at its proximal end
close to the pump, and leave the glass tube open
so that all the water flows through the latter.
Work the pump with a uniform force at about
30 to 40 strokesaminute. To ensure regularity,
the strokes had better be timed with a metro-
nome. The water will flow from the open mouth
of the glass tube in jerks, corresponding to the
strokes of the pump. At each stroke as much
will issue from the distal end as enters at the
proximal end.

Introduce into the open mouth of the glass tube
a fine nozzle, so as to offer considerable resistance
to the outflow of fluid. Work the pump with
the same force and frequency as before. The
outflow will still be intermittent, though less
fiuid will issue from, and consequently less enter
into, the tnbe at each stroke.

1 See Appendix,
10—2

148

ELEMENTARY PHYSIOLOGY. [ XIV.

Clamp the proximal end of the glass tube and
unclamp the elastic tube. Let the distal end of
the latter be quite open. Work the pump as
before. There being little resistance to the
outflow, the elasticity of the tube is not called
into play, and consequently the flow will be, as
in the case of the glass tube, intermittent.

Working the pump as before, insert the fine
nozzle into the open mouth of the tube. Con-
siderable resistance will now be offered to the
outflow of fluid, the elasticity of the walls of the
tube will be called into play, and the water will
issue from the end of the tube in a continuous
instead of an intermittent stream. If the tube
be sufficiently long and sufficiently elastic in
proportion to the force and frequency of the
strokes, the flow will be absolutely continuous.

B. Masor ARTERIAL SCHEME’.

The pump represents the heart; the small tubes
represent the resistance of the small arteries and
capillaries.. The tubes on the proximal side of
this resistance represent the arteries, those on
the distal side the veins.

The Mercurial Manometer.

The manometer A is connected with the arterial,
V with the venous tubes.

1 See Appendix.

BLOOD PRESSURE. 149

. Open the clamps marked ¢, c’ and ce”, so that

as little resistance as possible intervenes be-
tween the arterial and venous tubes. Bring
the manometers to mark on the revolving
cylinder, placing V about an inch under A,
in the same vertical line. Work the pump
steadily, regulating the time with the metro-
nome.

In A, the mercury rises at each stroke, and
in the interval between each two strokes
falls again to its previous level. (The mo-
mentum of the mercury frequently carries it
below this level, and the descent may be
followed by one or more oscillations.)

In V, a similar rise and fall is observed, of
nearly if not quite the same extent. —

Close the clamps c, c’ and ce”, so that the
capillary resistance becomes very considerable.
In A, the mercury rises rapidly at the first
stroke, and at the end of the stroke begins to
fall again, but more slowly than was the case
ina. It has not fallen far before the second
stroke raises it to a higher level than before.
On falling still again, it is once more raised
to a yet higher level, but the increase is not
so great as before. Each succeeding stroke
has a similar effect. Thus at the end of a
few strokes, the mean arterial pressure is
reached, marked only by comparatively small
oscillations corresponding to the strokes of
the pump. ~

ELEMENTARY PHYSIOLOGY. . xv.

On the strokes ceasing, the mercury gradually
falls until the previous level is reached.

In V the mercury rises to a much less extent
than was the case in a, a slight mean pressure
much less than in A is established, marked
either with no oscillations at all or such as
are much less conspicuous than those of A.
Owing to the presence of the resistance, a mean
pressure (arterial blood pressure) is established on
the proximal (arterial) side of the resistance.
This pressure is marked by oscillations syn-
chronous with the strokes of the pump. On the
distal (venous) side the mean pressure is much
less and the oscillations are either slight or
altogether absent.

Flow from Arteries and Veins.

Remove the clamps from the fine nozzles a and ».
Let the clamps c, c’ and ce’ remain closed. Set
the pump going. The flow from @ on the
proximal (arterial) side is in jets; that from v
(venous) side is uninterrupted or nearly so.

Sphygmograph.
Bring the levers S, (arterial side) and S, (venous

side) to write on the revolving drum, one under
the other.

a. Open the clamps ¢, c’ and ¢”, and set the pump
going. The two levers describe two nearly —
straight lines, a slight rise only being evident —
(and that to about the same extent in both)
at each stroke.

—

-‘XiIv.)

BLOOD PRESSURE. 151

When there is little or no resistance in the capil-
laries, comparatively little distension of the arterial
walls is produced at each stroke of the pump.

b. Close the clamps ¢, c’ and c”.
The lever S, now describes a well-marked
curve with each stroke of the pump.
Observe the sudden rise to a maximum, the
commencing fall, the break in the fall, fol-
lowed by a slight rise (dicrotic wave) and
the final descent.
The lever S, describes now a straight line.

The rise in pressure at each stroke indicated by
the mercurial manometer is accompanied by a
distension of the proximal (arterial) part of the
tubing, indicated by the rise of the lever. This
is the pulse.

On the distal (venous) side of the resistance no
pulse is visible.

Progression of the Pulse-wave.

Place two levers, one S,, as near as possible to
the pump, the other S’,, as near as possible to
the resistance. Bring the two levers to mark on
the cylinder the one exactly beneath the other.
(The pressure exerted by the two ievers must be
as nearly equal as possible.)

Observe that each rise of S, begins a little before,
and is over a little before that of S’,. In other
words, the pulse of S’, is a little later than that
of S,.

(By means of a tuning-fork this interval may be

152 ELEMENTARY PHYSIOLOGY. [XIv.

measured, and the length of tubing between the
two levers being known, the rate of progression
of the pulse-wave ascertained.)

5. While the pump is working, the clamps being
closed and the manometers A and V tracing
their curves, gradually diminish the resistance
by opening slowly first c’ and then c”.

The arterial pressure curve will gradually fall,
still marked by the pulse oscillations; the venous
curve will gradually rise.

Diminution of capillary resistance lowers arterial, but
INCTEASES VENOUS PTessure.

6. Close the clamps c’ and ce”, and take tracings
with the manometers, then gradually reduce the
strength of the strokes of the pump.

Both arterial and venous pressure will diminish.

7. Theclampc being closed, the main arterial trunk
of the scheme divides into two chief branches, X
and Y, each with its own resistance and venous
tube.

Leave the clamps c’, c” closed, and put clamps
on the tubing immediately beyond a and y.

a. Work the pump with great regularity, and
measure the quantity of fluid which escapes
during a given time (say ten seconds) from
the venous tube of X, and from that of Y, by
the side tubes x and y. ve

b. The clamp c” of X remaining closed, open

XIV. ]

BLOOD PRESSURE. 153

that c’ of Y, and the pump working exactly
as before, measure again the outflow during
ten seconds.

The outflow of Y will be increased. That of
X on the other hand will be diminished,
though the resistance in X is the same as
before.

The flow of blood through an artery is dependent
not only on the resistance offered by ws own
small arteries and capillaries but also on that of
other arterves.

DEMONSTRATIONS.

The effects in the rabbit on the temperature of
the ear, and on the calibre of its blood-vessels,
following

a.

Stimulation of the central end of the great
auricular nerve.

Section of the sympathetic nerve in the neck.

Stimulation of the peripheral end of the
sympathetic.

Normal kymographic tracings of the blood-pres-
sure of a mammal obtained by the use of a
mercurial manometer.

The effects on the arterial blood-pressure, as
indicated by the tracing, produced by

a. Inhibition of the heart through stimulation

of the peripheral end of the vagus.

154

Di . pieallle poe |
: Te, 2 nfs ,

Pres ee eR ne eee ,
ad ie —- —s we ae ee = yee :
" eee ite ~} ac Aaa > } j ;

ELEMENTARY PHYSIOLOGY. [XIV.

b. Dilatation of the small blood-vessels through

stimulation of the central end of the depressor
nerve.

Methods of measuring the velocity of the blood-
current in large vessels.

Comparison of venous and arterial pressure.
Method of using the sphygmograph.
Method of using the cardiograph. -

LESSON XV.

SALIVARY GLANDS AND PANCREAS.
SALIVA.

A. Mucous SALIVARY GLAND.

1. Prepare sections of a dog’s submaxillary gland
which has been placed in 75 p.c. alcohol for an
hour and then in absolute alcohol. Stain the
sections with carmine (best dilute) or hema-
toxylin, and mount them in glycerine.

Under a low power observe

a. The division of the section into irregular
angular areas, by connective-tissue septa,
which, if the section includes the circum-
ference, will be seen to proceed from the
sheath of the gland. These are the primary
lobules; they may be seen to be divided
into smaller lobules, but probably not dis-
tinctly.

b. The alveoli, appearing as small roundish
bodies closely aggregated together to form
the lobules; each will be seen to consist of

156 ELEMENTARY PHYSIOLOGY. [Xv.

a group of cells more or less surrounded
by connective tissue, continuous with that of
the septa.

c. At intervals oblique and transverse sections
of the small ducts. They are usually stained
darker than the alveoli, are not surrounded
by a sharp ring of connective tissue, and
have a well-defined lumen.

2. Under a high power observe that

a. The alveoli vary considerably in size, and
frequently have no obvious lumen; when
visible the lumen is usually an irregular
central space between the cells.

b. The mucous cells are comparatively large ;
most have a disc-shaped nucleus situated in
the outer part of the cell near the basement
membrane; in some, the nucleus is spherical
and lies farther from the basement mem-
brane. Around the nucleus is a small amount
of stained cell-substance, from this a stained
network may more or less clearly be seen
stretching throughout the cell, the rest of
the cell-substance stains little or not at all.

c. The demilune cells, occurring often in
groups, lie immediately beneath the mem-
brana propria, and are stained throughout:
they are generally half-moon shaped, and
often have two nuclei; they frequently send
processes in between the mucous cells, and
then appear simply to fill up the spaces

ie

XV.] SALIVARY GLANDS AND PANCREAS. SALIVA. 157

)

between the mucous cells and the membrana
propria,

gw

The epithelium of the small ducts consists of
a single row of slender columnar cells, the
inner borders of which apparently coalesce
and form a distinct ring bounding the lumen:
there is no such distinct boundary to the
outer (circumferential) part of the cell, which
especially in hematoxylin specimens has a
well-marked: striation. Each cell contains
an oval nucleus, situated a little on the inner
side of the centre of the cell.

Take a small piece of a dog’s submaxillary gland
which has been for three to six days in a 5 p.c.
solution of neutral ammonium chromate, and
tease it out in the same fluid. Observe the
isolated mucous and demilune cells, noting in
the mucous cells that the deep-seated end, in
which the nucleus lies, is prolonged into a process,
and that this, together with a varying amount
of the cell-substance around the nucleus, is more
granular and opaque than the rest of the cell.
The cell-network may also be obvious.

Examine a mounted specimen of a dog's sub-
maxillary gland which has been taken after
prolonged secretion’. Observe under a high
power that

1 In a dog under morphia and chloroform the chorda tympani (or
chorda tympani and sympathetic) is stimulated with a fairly strong
interrupted current for alternate minutes during six hours about.

158

ELEMENTARY PHYSIOLOGY. [xv.

a. The mucous and demilune cells are much
more alike.

b. The mucous cells are smaller, and a much
larger part of their cell-substance is stained,
the unstained part being as before next
the lumen. Their nuclei are spherical in-
stead of disc-shaped, lie more towards the
centre of the cells, and have conspicuous
nucleoli.

Some alveoli and patches of alveoli shew the
changes mentioned above much more distinctly
than others, in some the only changes observable
are that the demilunes are more conspicuous,
and the nuclei of the mucous cells spherical.

B. Serous SALIVARY GLANDS.

1,

Prepare specimens of a mammalian parotid gland
or of a submaxillary gland of a rabbit. The
tissue may have been preserved in alcohol (ep.
A. § 1) or in ‘2 p.c. chromic acid. Compare
with the section of the mucous gland (A).
Note that in the alveoli

a. The cells are more or less polyhedral, having
less rounded outlines than the cells of the
mucous gland. One kind of cell only is
present.

b. The cells stain fairly equally throughout
and appear to be densely granular.

c. The nuclei are spherical (unless shrunken

XY.] SALIVARY GLANDS AND PANCREAS, SALIVA. 159

-

by the reagents used) and are placed nearly
in the centre of the cells (they are usually a
little nearer the outer side).

bo

Tease out in normal salt solution a small piece
of the parotid of a rabbit (preferably one which
has been killed eight to ten hours after a full
meal).

Observe the numerous granules in the cells.

C. PANCREAS.

1. Cut sections of ‘resting’* pancreas (dog, rabbit or
frog) which has been preserved in osmic acid;
mount the sections in dilute glycerine. Observe

a. Under a low power; the gland consists of
lobes, lobules and alveoli like the serous and —
mucous glands.

b. Under a high power; the cells contain a
great number of granules which stretch
nearly or quite to the outer border; the
nuclei are more or less completely hidden by
the granules.

2. Cut sections of active’ pancreas and compare
with the above.

1 By a ‘resting’ gland is meant one which has not been secreting, or
has been secreting slightly only, for several hours ; by an ‘active’ gland
is meant one which for an hour or more has been secreting rapidly.
In both cases the animal is understood to be in a good state of nutri-
tion; the pancreas of an animal which has long fasted is not a ‘resting’
pancreas. With the dog or rabbit the gland should be taken five to

160 ELEMENTARY PHYSIOLOGY. [Xv.

a. The granules are fewer, being absent from
the outer part of the cell, thus forming an
inner granular and an outer non-granular
zone.

b. The spherical nucleus is more obvious (partly
in consequence of the disappearance of the
granules) and is situated chiefly or entirely
in the non-granular zone, the nucleolus is
usually distinct.

c. Ifthe gland has been taken after very active
secretion the cells may be nearly free from.
granules, the remaining granules will be
smaller and the lumina will be more obvious.

3. Prepare sections of an active pancreas which has

been hardened in spirit, stain with carmine and
mount in acid glycerine.
The division of the cells into granular and non-
granular zones will be seen as in the osmic acid
preparations ; the inner zone however appears as
a confusedly granular mass instead of as an
aggregation of separate spherical granules; the
outer zone appears homogeneous and stains more
deeply than the inner zone.

six hours after a full meal for the active state; about twelve hours
after a full meal for the resting state. With the frog the pancreas
should be taken in eight to ten hours after a full meal for the active
state, in one to two days for the resting state. Worms serve best for
feeding frogs. In unhealthy frogs and in frogs which have long
fasted, the pancreas-cells have usually a distinct outer non-granular
zone. In all cases the pancreas should be placed in osmic acid 1 p.c.
for two to twenty-four hours as convenient, then washed, and trans-
ferred to 75 p.c. alcohol.

XV.] SALIVARY GLANDS AND PANCREAS. SALIVA. 161

D. SALIVA.

i

Look at a little fresh saliva under the microscope
with a high power. Disregarding the flat epi-
thelial cells from the mucous membrane of the
mouth, note the salivary corpuscles; they are
larger than the ordinary white blood-corpuscles,
but in other respects closely resemble them. In
many, a very active Brownian movement of the
granules within the corpuscle may be observed.

Test with neutral litmus paper the reaction of a
drop of saliva, it will be found to be alkaline.

Induce the secretion of saliva by chewing a small
piece of india-rubber tubing, by filling the mouth
with ether vapour, or by rubbing the tongue
with a crystal of tartaric acid.

If time allows let the saliva stand until the
turbidity has settled down into a sediment. To
a few c.c. of the fluid add strong acetic acid;
mucin will separate out as a stringy mass,
which does not dissolve in excess of acid.
Shake gently, or stir it with a glass rod the
mucin will form a clump, remove it, and if the
fluid is cloudy, filter.

To the clear fluid add a drop or two of a strong
solution of potassium ferrocyanide, The slight
precipitate which results indicates the total
quantity of proteids present (cp. Lesson III.
§ 16). If the reaction is not obvious, test
another small portion with Millon’s reagent.

11

162 . _ ELEMENTARY PHYSIOLOGY. [xXv.

5. To a few cc. of starch mucilage’ 1 p.c. ina
test-tube add a drop or two of a moderately.
strong solution of iodine ; an indigo-blue colour
will be produced, if the colour is very dark fill up
the test-tube with water.

6. To5dc.c. of a5 p.c. aqueous solution of dextrin’
~ add a strong solution of iodine drop by drop.
A deep brown-red colour will be produced.
Warm; the brown-red colour will rapidly dis-
appear, a light brownish-yellow tint due to
the iodine remaining; on cooling the dextrin
colour returns. Now add water; as the dextrin
solution becomes more dilute, the red tint be-
comes less obvious, the fluid appears yellow-
brown. That this colour is not due to the iodine
can be seen by warming the fluid.

7. To5cc. ofa ‘1 p.c. solution of dextrose (grape-
sugar) add an excess of a strong solution of sodium
hydrate and a couple of drops of a 1 p.c. solution
of cupric sulphate; the precipitate of hydrated
cupric oxide at first formed will dissolve, giving |
a blue solution. Boil; the cupric oxide will be
reduced and a yellow or red precipitate of

1 To prepare the starch mucilage, take 1 gram of starch, and rub
it into a thin paste with cold water. Pour it into a beaker containing
one hundred c.c. of boiling water, boil for a few minutes and place it
aside to cool. It should have no lumps in it and should be thin
enough to be measured out readily with a pipette.

2 This may be bought at a chemist’s or it may be prepared by
boiling a little starch with sulphuric acid about 3 p.c., until a drop
of the fiuid gives a red-brown colour with a drop of iodine.

XV.] SALIVARY GLANDS AND PANCREAS. SALIVA. 163

cuprous oxide will be produced (Trommer’s
test). When a very small quantity of sugar
is present no distinct precipitate is obtained, but
the fluid is decolourized or turns faintly yellow.
Repeat this, adding half a dozen drops of a
strong solution of cupric sulphate; the reaction
will be much less obvious, partly owing to the
blue colour of the dissolved hydrated cupric
oxide and partly to the brown-black precipitate
of anhydrous cupric oxide,

8. In this and the following experiments the saliva’
used should be diluted 5 to 10 times.
Mix equal quantities (say 5 c.c.) of starch and
saliva in a test-tube and place in a water bath at
about 37°C. At short intervals (1 to 3 minutes)
take a drop of the mixture and add it to a drop
of iodine on a porcelain plate. The colour pro-
duced at first blue will later become a blue-
violet, a red-violet, a red-brown and a light-
brown yellow, according to the relative amounts
of starch and dextrin present, finally there will
be no colouration, no more starch or dextrin
(erythrodextrin) being left. Then divide the
fluid into two parts.

1 An aqueous extract of a ptyalin-containing gland may be used
instead of saliva. To prepare the extract take (e.g.) the parotid
glands of a rabbit and having removed the connective tissue around
them chop them up well and place the pieces in about 200 c.c. of
water; leave in the warm for an hour or two and filter. The aqueous
extract thus prepared contains much proteid material, and this
obscures the reducing action of sugar on cupric hydrate in Trommer’s
test when a small quantity only of sugar is present.

11—2

164

10.

ELEMENTARY PHYSIOLOGY, [Xv.

a. Add iodine; no colouration is produced (there
may be a little tint from dextrin since in
mixing the drops a faint colour may escape
notice which in a larger quantity of fluid is
obvious).

b. Add an excess of sodium hydrate and a drop
of 1 p.c. cupric sulphate and boil; the fluid
turns yellow and a yellow precipitate will be
formed showing the presence of sugar.

Boil a little saliva, add it to starch in a test-tube
and warm. In half-an-hour divide into two
parts and test as in § 8a,6. The blue colour
from starch will be as distinct as at first, no trace
of sugar will be found; hence boiling destroys
the ferment (ptyalin) which converts starch into
sugar.

If the saliva used in § 8 converts starch into
sugar very rapidly dilute it still further for the
following experiment. Into each of three test-
tubes pour equal quantities of saliva and starch.
Place A in a water bath at about 37°C., leave B
at the temperature of the room (noting it) and
place C in a vessel with ice (it is best to cool
the starch and saliva before mixing them). At
short intervals take with a glass rod drops from
each and add them to drops of iodine on a
porcelain plate and so compare the rate of dis-
appearance of starch (cp. § 8) in the three
mixtures. It will disappear much more quickly
in A than in B; in C there will be very little
change.

7:

XV.] SALIVARY GLANDS AND PANCREAS, SALIVA. 165

11.

12,

When no starch is left in A, remove C from the
ice and place it in the warm chamber and test
at intervals as before, the starch soon disappears.
Hence a temperature of 0°C. arrests the action of
saliva but does not destroy it.

Neutralize a small quantity of saliva; to 5 c.c.
of this add 5 «ec. of HCl ‘2° p.c., the mixture
thus contains ‘1 p.c. HCl. Place at 37°C. for

-ten minutes, add 3°5 cc. Na,CO, ‘4 p.c. and

complete the neutralization with a more dilute
solution. Add a few cc. of starch and place at
37°C. In half-an-hour test for starch and sugar;
starch will be found but no sugar, hence the
acid has destroyed the ptyalin.

Place in one dialyser* (A) 15 c.c, of starch and
in another (B) 10 cc. of starch with a little
saliva.

Test from time to time the external water in
each. That from (A) will give no trace of
starch or sugar. That from (B) will contain
sugar, but no starch. Sugar dialyses, but starch
does not.

1 Pure strong commercial hydrochloric acid contains about 33 p.c.

HCl.

? In these and in similar experiments (Less. xv1.) 37° C. is taken

since that is very nearly the normal body temperature of man, but a
rather higher or a rather lower temperature will serve equally well.

® A very convenient dialyser may be made from a short length of

parchment paper tubing (Papier-Diirme) sold by Carl Brandegger,
Ellwangen, Wiirttemberg (cp. also Gamgee, Physiological Chemistry,

Vol. 1. p. 6).

166. ~ELEMENTARY PHYSIOLOGY. [Xv.

13. Add a little raw starch* to saliva and place in
the warm chamber, shaking frequently. The
raw starch is converted into sugar very slowly, it
may be an hour or more before any sugar can be
detected,

DEMONSTRATION,

- Effect on the submaxillary gland of the dog of

stimulating the chorda tympani and the sympa-
thetic nerves.

1 Arrow-root will do very well.

+

LESSON XVI.

STOMACH. GASTRIC JUICE, MILK.

A. STRUCTURE OF THE STOMACH.

1. Make transverse vertical sections from the fundus
or greater curvature of a rabbit’s stomach which
has been preserved in spirit (cp. p. 32, F, § 1) or
in chromic acid ‘2 p.c. Stain the sections with
carmine or with aniline blue (the sections are
best stained if they are left in a dilute solution
of the staining agent one to two days). Observe
under a low power

a. Externally, the thin connective-tissue layer

b.

of the peritoneum.

The muscular coat, consisting of an outer
longitudinal and an inner circular layer of
unstriped muscle. If the sections are ac-
curately transverse, the former will appear
as a cross section of a number of bundles
with connective tissue running in between
them from the peritoneum, the latter as a
continuous layer. On the inner side of this
may also be seen a much thinner oblique
muscular layer.

168 ELEMENTARY PHYSIOLOGY. [XvVI.

c. The submucous coat of connective tissue.
If the mucous membrane is in folds the sub-
mucous but not the muscular coat will be
seen to run up in the folds.

d. The muscularis mucose, or thin stratum
of unstriated muscle fibres a little below the
glands, this is divided more or less distinctly
into an outer longitudinal and an inner
circular layer.

e. The mucous coat. Note in this
The gastric glands with their openings and
the ridges between the openings. The
bifurcation of some of the glands may pro-
bably be made out.

2. Observe under a high power

a. The columnar mucous cells, lining the
‘mouths of the glands and covering the free
surface of the gastric mucous membrane
between the glands; they are long slender
cells becoming shorter in passing down the
mouths of the glands; the upper third of
the cell is usually much more transparent
than the remaining portion, and the nucleus
lies at about the lower third. These cells
may have become detached if the tissue has

not been removed from the animal soon after
death.

b, The large deeply stained ovoid or border
cells with ovoid nuclei,and the short columnar
or polyhedral central cells with spherical

XVI]

STOMACH. GASTRIC JUICE. MILK. 169

nuclei. At the base of the glands the central
cells are usually most numerous, the ovoid
cells being placed between them and the
basement membrane; towards the neck of
the glands the ovoid cells usually increase
in number, in the neck the majority of the
cells are ovoid; they are however considerably
smaller than in the body of the glands.
The ovoid cells frequently cause a bulging
outwards of the basement membrane, this is
especially the case if the animal has been
killed soon after it has fed.

c. The fine connective tissue immediately inter-
nal to the muscularis mucosee, surrounding
the bases of the glands, and sending up pro-
cesses between them, Towards the surface
the fibres have a much closer arrangement,
and appear as a number of slender, compara-
tively dark, bands, which stain deeply. Mark
the scarcity of leucocytes. ,

Compare the longitudinal iectical section pre-
viously made (Less. vil. § 11) of the muscular
coat of a dog’s stomach with the transverse
vertical section of the muscular coat of the
rabbit's stomach (§ 1), in the former the muscular
coats are much thicker.

Take a small piece of the fundus region of a
rabbit’s gastric mucous membrane and prepare a
section parallel to the surface through the bodies
of the glands. Observe

170

Or

ELEMENTARY PHYSIOLOGY, [ XVI.

a, The central cells forming a. tube with very
small lumen.

b, The comparatively rare ovoid cells outside
the central cells,

From the pyloric end of the stomach prepare
vertical sections and stain them with hzema-
toxylin, compare these with the sections made
of the cardiac end. Note

a. The greater thickness of the longitudinal
and circular muscular layers.

b. The wider and longer mouths to the glands,
their more frequent branching and the
absence of ovoid cells (if the section passes
through the upper part of the pyloric region
a few ovoid -cells may be seen). ' The cells
below the mouths of the glands (pyloric
gland cells) resemble in general appearances
the central cells of the cardiac end of the
stomach.

B. STRUCTURE OF THE CSOPHAGUS,

1. Make transverse vertical sections from the lower

third of a rabbit's cesophagus, which has been
hardened in potassium bichromate 1 p.c., and
compare them with the corresponding sections
of the stomach. Note the ‘following points of
contrast :

a. The external muscular layers contain striped
as well as unstriped muscular fibres; sections

XVI]

2 &

STOMACH, GASTRIC JUICE. MILK. 171

from the upper part of the cesophagus show ~
no unstriped fibres,

The submucous tissue contains small serous
and mucous glands (cf. Lesson Xv.). Each of
these consists of a duct, dividing and ending
in dilatations, the alveoli.

Traces of the muscularis mucose,
The papillz of the mucous membrane.

The epithelium forming a layer several célls
deep, the deeper being columnar or sphe-
roidal, the superficial cells flattened (cf. Epi-
dermis, Lesson XXIV.),

C. GASTRIC JUICE.

1.

b.

Artificial Gastric Juice.

a.

Tear off the mucous membrane from the
stomach of a mammal, cutting away the

pyloric region (the stomach of a pig obtained

from the butcher’s will serve). Mince it
finely. Put it in a flask with two hundred
times its bulk of hydrochloric acid ‘2 p.c.,
and place the flask in a water bath at about
40°C. After some hours a considerable part
will be dissolved. Decant, and filter the
decanted fluid, A solution of pepsin in
hydrochloric acid will be obtained; it will,
however, contain a considerable quantity of
peptone,

Mince another gastric mucous membrane;

172 ELEMENTARY PHYSIOLOGY. [XvI.

remove with blotting-paper the excess of
fluid, add five times its bulk of glycerine
and place aside stirring occasionally. It is
best to leave the mixture for some days
before use, it may be kept almost indefinitely.
When required for use, filter through muslin,
add to the fluid ten to twenty times its
volume of HCl ‘2 p.c. and filter.

Action of Gastric Juice.

2. a. Take four test-tubes. In A place 5c.c. of
' - hydrochloric acid ‘2pc. In B bee. of
artificial gastric juice. In C 5c. of the

same juice, carefully neutralized with dilute
Na,CO, In D 5cc. of the same juice,
thoroughly boiled. Add the same quantity

of fibrin’ to each, and place in a water bath

at about 37°C. Examine from time to time.

A, the fibrin will swell up and become trans-
parent, but will not be dissolved; on neu-
tralization it will appear unaltered,

BD, the fibrin will be digested.
C, the fibrin will be unaltered.
D, the fibrin will be like that in A.

1 Raw fibrin digests more easily than that which has been boiled
or kept in alcohol, it often however contains traces of pepsin so that
a slow digestion may take place when acid only is added to it. When
it is required to measure accurately the amount of fibrin added, raw
fibrin finely chopped up should be placed in dilute HCl until it is
well swollen, the excess of acid poured off and the fibrin measured in
small tubes containing (e.g.) 2 ¢.c

XVL.]

STOMACH. GASTRIC JUICE. MILK. 173

These experiments show that acid alone (A)
and pepsin alone (C) will not digest fibrin,
and that pepsin loses its power on being
heated to boiling point (D). Now add acid
again to C, and place it in the warm chamber.
Digestion will take place. The neutraliza-
tion has only suspended, not destroyed, the
action of the pepsin.

0b, Take two test-tubes, with 5c.c. of gastric

juice and a morsel of fibrin in each.

Place A in the warm. Surround B with
ice, or put it in a cold spot.

The fibrin in A will be digested rapidly ;
that in B very little or not at all.

Take 5 cc. of artificial gastric juice which has
been found to digest fibrin rapidly, neutralize it,
filter and add an equal bulk of Na,CO, 2 p.c.
thus obtaining pepsin in the presence of a small
quantity of an alkaline salt. Place at about
40° C. for half-an-hour to an hour. Then add
HCl until the mixture is distinctly acid (or
neutralize and add an equal volume of HCl -4
p.c.). Add a flock or two of fibrin and warm.
Little or no digestion will take place. The
pepsin has been destroyed by the alkaline salt.

Place 50 cc. of artificial gastric juice together
with some fibrin or other proteid in a beaker
and leave in the warm until a small part only
of the proteid remains undissolved. Filter and
neutralize carefully, a precipitate of acid-albumin

174 _ ELEMENTARY PHYSIOLOGY. a

(parapeptone) will be obtained (ep. Less. rx. § 15).
Filter off the acid-albumin, the filtrate contains
peptones.

5. Determine the following characters of peptones
with the solution obtained in § 4.

a. Apply the tests for proteids (Lesson 111. § 16),

- Millon’s and the xanthoproteic reaction are
obtained, but no precipitate is produced with
acetic acid and potassium ferrocyanide.

b. Boil; it does not coagulate.

cc. Add excess of sodium hydrate and a drop or
two of dilute cupric sulphate, a rose colour is
produced which becomes violet when more
cupric sulphate is added. Compare the
colour reaction with that given by diluted
serum or white of egg (Lesson I. § 16 c).

d. Pour into one dialyser (A) a solution of
peptone and into another (B) diluted serum
or white of egg. Leave for an hour or
longer, then apply the xanthoproteic test to
the fluid outside the dialyser, a reaction will
be obtained from (A) only, ie. the peptone
has dialysed, the albumin has not.

D. MILK.

1, Examine a drop of fresh cow’s milk under the
microscope with a high power. It consistsofa
clear fluid containing a large number of highly _
refractive fat globules of varying size. Adda

XVI.]

e2

STOMACH. GASTRIC JUICE. MILK. 175

drop of osmic acid; in a short time the globules
become stained brown-black.

Test the reaction of fresh cow’s milk with litmus
paper. It will be found to be alkaline: oc-
casionally it is acid owing to the presence of
free lactic acid.

Dilute a little milk five to ten times with
water; neutralize it with dilute acetic acid, no
precipitate will fall. Continue to add the acetic
acid drop by drop, a copious precipitate of casein
will occur carrying down with it nearly all the
fat. When there is a distinct flocky precipitate
no more acid should be added as casein is soluble
though not very readily in excess; it is not
precipitated on merely neutralizing since alkaline
phosphates are present in milk (cp. Lesson Ix.
§ 16, c). To precipitate the whole of the casein
the milk must be much diluted.

Filter off the precipitate. The filtrate should be
clear; if it is not, either too little or too much
acetic acid has been added; in this case add
either a little more acetic acid or a little dilute
sodium carbonate and filter again.

Boil the filtrate; a precipitate of albumin (with
a little globulin) takes place. Filter, and to the
filtrate

Apply Trommer’s test (Lesson xv. D, § 7), a yellow
precipitate will be obtained showing the presence
of milk-sugar.

176

or

ELEMENTARY PHYSIOLOGY. [ XVI.

Place a small quantity in a warm place for one
or two days; then test the reaction, it will be
found to be acid; this is due to fermentation,
in the process of which the milk-sugar is con-
verted into lactic acid.

Action of Gastric Juice on Milk.
Neutralize with dilute Na,CO, a little artificial

- gastric juice prepared by method (a) § 1, filter

and add 5 «.c. of the filtrate to 5 ¢.c. of fresh milk,
place in the warm.

Observe at short intervals the condition of the
milk, it will soon form a firm clot so that the
test-tube can with safety be held upside down,
Jater the clot shrinks and presses out a nearly
clear fluid; the clot continues to shrink for
some time.

The rennet-ferment in the extract has coagu-
lated the casein, and this has carried with it
the greater number of the fat globules.

If the amount of rennet-ferment contained in
the extract is large the clotting may be almost
instantaneous ; in this case the experiment should
be repeated taking a smaller quantity of the
extract and without warming. The extract is
neutralized since (cp. § 4) excess of acid of itself
precipitates casein.

To the milk clotted by rennet-ferment add
5 ec. HCl ‘4 p.c. and warm for an hour or so, the
casein will be converted into peptone by the
pepsin of the extract in the presence of acid.

eeu)!

eae —_

(
or
?

Baia. “MILK. Ps:

ke Ae a tas ice more in the fluid
but it is much less white than originally.)

j AS TE

DEMONSTRATION.

# Separation of casein and fad trom silk by filtra-
! tion under pressure through a porous cell,

+ al

~Y

*
7
*

12

LESSON XVII.
INTESTINE. BILE. PANCREATIC JUICE.

A. STRUCTURE OF INTESTINE.

The outer coats of the intestine have the same general
characters as those of the stomach (Lesson xvi. A, § 1,
a—d), except that there is no oblique muscular layer.

1. Prepare vertical sections of a cat’s or dog’s small
intestine hardened in chromic acid ‘2 p.c. (ep.
p. 33 F § 2). Stain with hematoxylin (the
tissue may be stained before sections are made
by placing it in dilute hematoxylin for a day).
Observe in the mucous coat

a. The projections of the mucous membrane, or
villi, either extended and long, or contracted
and short with the surface thrown into folds.
Note

a. Theepithelium, consisting of rather long
columnar cells, each with a clear free
border more or less distinctly striated
with vertical lines, rather granular cell-
substance, and oval nucleus placed at
about the lower third of the cell; the

XVII] INTESTINE. BILE. PANCREATIC JUICE. 179

b.

/-

clear free borders of the cells frequently
appear to have coalesced into’a narrow
highly refractive band, which may be
traced over the whole villus.

The mucous or goblet cells, irregularly
scattered among the former, sometimes
abundant, sometimes scanty or absent ;
they have an upper ovoid portion which
has sharp outlines, is transparent and
may be empty, and a lower basal granu-
lar portion containing the nucleus,

The adenoid tissue, forming the sub-
stance of the villus: this consists of a
fine meshwork of fibres with nuclei or
flattened cells at some of the nodal
points. The meshes are seen to be
crowded with leucocytes.

There may also be seen with varying dis-
tinctness

5.

Capillary blood-vessels with the nuclei
of their component cells; they may be
filled with blood-corpuscles.

The ‘lacteal radicle’ as a space in the
centre of the villus.

Unstriped muscular fibres as narrow
bands running up the villus.

The rather deep depressions of the mucous
membrane, the intestinal glands or glands
of Lieberktihn. Note that

12—2

180

C.

ELEMENTARY PHYSIOLOGY. [XVI

a. The epithelium lining them consists of
short columnar cells. Observe their
gradation into the cells covering the
villi; usually they have a clear free
border like that of the columnar cells
of the villi. |

8. There is usually a distinct basement
membrane immediately beneath the epi-
thelium. This is formed of connective-
tissue corpuscles very much flattened
and fused together into a membranous
sac; the outlines of the cells are not
seen in the section, but the nuclei are
fairly conspicuous.

y. The lumina of the glands are in thin
sections distinct. :

The adenoid tissue around the bases of the
glands of Lieberkiihn and between them and
the muscularis mucose. This, unlike the
corresponding tissue in the stomach (Lesson
xvi. A, § 2, c), has a large number of leuco-
cytes in its meshes.

The lymph follicles; cither isolated, or ag-
gregated into Peyer’s patches ; the follicles
are round or oval masses of adenoid tissue
crowded with leucocytes, lying immediately
beneath the surface epithelium and usually
stretching down into the submucous tissue.
They are in the midst of the glands of Lie-
berkiihn, and the villi are absent over them.

, = =—— 7 ©. >

————— 7

XVII] INTESTINE. BILE. PANCREATIC JUICE. 181

They will be more fully studied under Lym-
phatics (Lesson xvull. §$ 1, 2, 3).

2. Snip off a few villi from a fresh intestine; tease

out some in normal salt solution; place others in
osmic acid 1 p.c. for half an hour, then tease
out in water or in dilute glycerine.
Observe more closely the characters of the cells
(§ la (a) (@)). It will be seen that some of the
isolated columnar cells branch at their attached
ends; and that the goblet cells have usually a
tapering process. Where a surface view of a
portion of mucous membrane is obtained the
goblet cells will appear as clear round spaces.

3. Prepare sections of the ileum or jejunum through
the glands of Lieberkiihn parallel to the surface,
and compare them with the vertical sections

(§ 1, b, c).

4, Prepare vertical sections of the duodenum at its
commencement close to the pylorus. In addition
to the villi and intestinal glands, note |
The glands of Brunner’. Each has a duct
with basement membrane, short columnar epi-
thelium and usually distinct lumen; the duct
runs down into the sub-mucous tissue and there
divides and sub-divides, the end tubes enlarging
slightly, like small alveoli; the number of sub-

1 Brunner’s glands stretch some little distance from the pylorus in
ruminants and in the pig; in carnivora and rodents they are close
to the pylorus and usually small; in the mole they form a marked
bulging ring just below the pylorus.

_ ELEMENTARY PHYSIOLOGY. [XVII.

- divisions varies greatly in different animals, there

is also considerable variation in the form of the
tubes and in the appearance of the cells in diffe-

‘rent animals, generally speaking the cells are

much the same in the ducts and throughout the
tubes; the nuclei are placed near the basement
membrane.

Prepare vertical transverse sections of the large
intestine. Observe

a. The longitudinal muscular coat, thin except
where the section has passed through one of
the three conspicuous bands.

b. The circular coat, thick and well developed.

c. The mucous membrane, frequently thrown
into longitudinal ridges, the sub-mucous tissue
running up into the ridges.

d. The absence of villi.

¢. The intestinal glands (glands of Lieberkiihn);
they are larger than in the small intestine,
and their features, owing to the absence of
villi, much more easily seen. The epithelium
covering the free intestinal surface or the
ridges between the glands consists of long
columnar cells with usually a striated clear
free border, in the glands the cells are
shorter, have no clear border, and the nuclei
are nearer the basement membrane; in some
animals (e.g. dog) there are many distinct
goblet cells.

XVII.] INTESTINE. BILE. PANCREATIC JUICE. 183

6.

“I

Examine sections of a small intestine in which
the blood-vessels have been injected, and note
the capillary network round the glands of Lie-
berkiihn, and the small artery running up each
villus and dividing into a capillary network just
below the cells.

Feed a frog with a small piece of bacon; on the
next day’ kill the frog, remove the stomach and
intestine, pin the tube out on cork, cut it open
along its whole length, and gently wash it with
salt solution.

Note that the mucous membrane of the stomach
has a yellowish semi-transparent look, whilst the
mucous membrane of the intestine is of an
opaque white, this is more marked in the upper
than in the lower part of the intestine; the
rectum is greyish and semi-transparent. Teaze
out a small piece of the opaque white mucous
membrane in normal salt solution; the epithe-
lium cells are crowded with fat globules, scarcely
anything but these being visible.

Fat is absorbed by the cells of the small intestine, and
is absorbed little or not at all by the cells of the
stomach.

Pin out pieces of the intestine; place some in
75 p.c. alcohol for an hour, and then in strong

1 The difference in the tint of the stomach and intestine is still
more obvious if the frog be fed again after two days and killed on the
subsequent day. The frog is fed by placing the piece of fat in the
upper part of the wsophagus, the fat is then usually swallowed at
once. For liardening the intestine ep. § 2. p. 33,

184

ELEMENTARY PHYSIOLOGY. [XVII.

spirit; place others in osmic acid 1 p.c. for half
an hour, wash and place in 75 p.c. alcohol.

In sections of these pieces note that there are no
villi and no proper glands of Lieberkiihn. The
mucous membrane is however thrown up into
considerable folds. In the osmic acid specimens,
the cells will probably be so full of deeply
stained fat globules that little structure can be
seen in them except the hyaline free border; in
the sub-mucous connective tissue few or no fat
globules are seen.

In the alcohol specimens the cell substance will
be seen as a distinct sponge-work or network,
the fat globules having been dissolved.

B. BIe.

1.

Test the reaction of bile* with litmus paper. If
fresh it is slightly alkaline or neutral.

Toasmall quantity add strong acetic acid drop by
drop. A curdy precipitate of mucin coloured with
the bile-pigment will be thrown down. Since the
mucin of bile is not formed in the liver but in the
mucous glands and cells of the gall-bladder and
duct, the longer the bile has been in the gall-
bladder the greater the precipitate which will
be obtained.

For the following tests (§§ 4, 5) it is best, although
not necessary, to precipitate the mucin with acetic

1 Ox-gall or sheep's gall may be obtained from a butcher’s,

|
:

_ iets “

XVII.] INTESTINE. BILE. PANCREATIC JUICE. 185

acid, to filter and use the filtrate; before filter-
ing, the bile may be diluted four or five times
with water.

The mucin may also be removed by adding an excess
of alcohol, the filtrate from this should be evaporated
to dryness, and the residue dissolved in water.

3. Gmelin’s test for bile-pigment. To a small
quantity in a test-tube add drop by drop, nitric
acid, yellow with nitrous acid, shaking after each
drop; the yellowish green colour becomes first a
dark green, then blue, then violet, then red, and
finally a dirty yellow. The blue and violet
colours are less obvious than the rest. Repeat
the test in the following form; place a drop of
bile on a porcelain slab, and place a drop of
yellow nitric acid so that it runs into the drop
of bile; where the fluids mingle, zones of colour,
green, blue, violet, red and yellow, from the bile
to the acid, are seen.

4. Pettenkofer’s test for bile-acids’. To a little bile
in a test-tube, add one drop of a 10 p.c. solution
of cane-sugar (or a small particle of sugar) and
shake. Add strong sulphuric acid to nearly the
same amount as the bile taken, inclining the

1 Bile-salts may be prepared in the following manner. Rub ox-
gall with animal charcoal into a thin paste. Evaporate on a water
bath to complete dryness, and extract with absolute alcohol. The
alcoholic filtrate should be colourless, Add to it anhydrous ether as
long as any precipitate is produced, and let it stand. The precipitate
either crystallizes out or falls to the bottom as a thick viscid syrup;
it is a mixture of sodium glycocholate and taurocholate.

186

ELEMENTARY PHYSIOLOGY. [XvII.

test-tube so that the acid settles at the bottom.
Gently shake the test-tube from side to side,
when the fluids have nearly mixed a deep purple
colour is produced. If too much sugar is added,
the fluid will turn brown or black; if too little
sulphuric acid is added the proper temperature
(about 70°C.) for the production of the colour
will not be obtained.

Add a few drops of oleic acid to 10 ¢.c. of bile in
a test-tube, shake well, and at once mount a drop
and observe in it under the microscope the
numerous fatty globules. Place the test-tube
with the bile in a warm bath for an hour or so,
then shake and mount a drop of the fluid;
comparatively few fatty globules will be seen
in it under the microscope. The oleic acid has
combined with the base of the bile-salts to form
a, soap (cp. C. § 4). 7

Place in separate test-tubes 10 c.c. of bile and a
couple of drops of oleic acid (a); 10 c.c. of bile
(b); 10 cc. of water; to each add 2°5 c.c. of
melted fresh butter’, shake well, and place in
the warm bath. The emulsion will last much
longer in (a) than in (0); it will last much longer
in (6) than in (c). The emulsifying power of bile

as slight; but in the presence of fatty acids tt

forms soaps (cp. § 6) which have a much greater
emulsifying power.

1 Tf olive oil is used instead of melted butter, it will depend upon

the amount of fatty acids contained in the olive oil whether any
difference is observed in (a) and (b).

i

XVIL] INTESTINE. BILE. PANCREATIC JuIcr. 187

7. Mount a few crystals of cholesterin‘ in water
and examine them under a microscope, they
consist of rhombic plates.

8. Irrigate the crystals with strong sulphuric acid ;
they turn red,

9. Toa small quantity of chloroform in a test-tube
add a little cholesterin and shake, the cholesterin
will dissolve; add strong sulphuric acid and
gently shake, the upper (chloroform) layer will
turn bright red.

10. Digest a little fibrin in 10 c.c. of artificial gastric
juice; when the fibrin is dissolved add drop by
drop, bile which has been decolourized by filtering
through animal charcoal; a precipitate will be
formed consisting of parapeptone, peptone and
bile acids. (If excess of bile is added especially if
it contains taurocholic acid, the peptone and bile
acids will be more or less completely dissolved.)

11. Add to the preceding 5 c.c. of *4 p.c. HCl and a
few flocks of swollen fibrin; the fibrin will shrink
and will be digested slowly or not at all. Bile
acids prevent gastric juice from digesting proteids.

1 Cholesterin may be prepared from gall stones (those which have
a soapy feel) in the following manner. Powder the gall stones and
add a small quantity of strong spirit (or absolute alcohol) and boil;
filter hot, using a hot-water funnel; on cooling, cholesterin crystals
will separate out. Collect the crystals, place them in a small
quantity of spirit containing a little sodium hydrate and boil. On
cooling, purified cholesterin crystallizes out; wash the crystals with
water,

188

ELEMENTARY PHYSIOLOGY. [ XVII.

C. PANCREATIC JUICE.

i

bo

Artificial Pancreatic Juice.

Mince finely a pancreas from a just killed animal,
pound it well with clean sand and add about
100 vols. Na,CO, ‘2 p.c. and a little thymol.
Place it in the warm for some hours to a day,
strain through muslin, filter through lmen and
then through filter paper.

Leave a pancreas moistened with water for a
day, then mince it well and add 10 vols. of
glycerine. When required for use add to a
small portion of this glycerine extract 10 to
20 vols. of Na,CO, 1°5 p.c., shake, strain through
muslin and filter,

Mince a pancreas and pound it with sand, for each
gramme of gland-substance add 1 c.c. of acetic
acid 1 p.c. and mix thoroughly in the mortar for
ten minutes ; add ten times its bulk of glycerine.
In a day or two a little strong solution of sodic
carbonate should be added to make the fluid
slightly alkaline. When required for use add
sodium carbonate as in § 2.

Properties.

To about 5 c.c. of distilled water in a test-tube
add a drop of oleic acid and shake, the fatty
globules soon rise to the surface; add 5 cc.
Na,CO, 1 p.c.; a white precipitate of soap
forms; shake, the precipitate partially dissolves
and more completely or wholly on boiling.

——e,. =o

XVII.] INTESTINE, BILE. PANCREATIC JUICE. 189

or

~y

Examine a drop of the fluid under the micro-
scope, no fat globules will be seen,

Place in a warm bath two test-tubes, each con-
taining 5 ¢.c. Na,CO, 1 p.c.; melt a little fresh
butter in a porcelain dish over a flame and with
a warm pipette add an equal quantity (about
2°5 c.c.) of the melted butter to the fluid in each
test-tube. To one of these (a) add a couple of
drops of oleic acid. Shake the test-tubes and
replace them in the warm bath, examining
them from time to time; the fine emulsion
formed on shaking the fluids will last much
longer in (a) than in (6). The emulsion in this
case is much greater than with bile (cp. B, § 6).

With either of the extracts of § 1 or § 2, make
observations on the amylolytic ferment of the
pancreas similar to those made in Lesson Xv.
§$ 8—13 on the amylolytic ferment of saliva.

Test the proteolytic action of either of the
extracts § 2 or § 3 in a similar manner to that
in which the action of artificial gastric juice was
tested (Lesson xvi. C, § 2) substituting 1 p.c.
Na,CO, for 2 p.c. HCl.

In test-tubes A and D the fibrin will be una!ter-
ed, in C it will be very slowly dissolved, in B it
will be rapidly dissolved, hence sodium carbonate
alone does not digest fibrin (A), trypsin alone
digests it very slowly (C), trypsin in the presence
of sodium carbonate dissolves it rapidly (B), the
digestive power of trypsin being destroyed by
boiling (D).

190

19

ELEMENTARY PHYSIOLOGY. [ XVII.

Experiments corresponding to those of §$ 4, 5,
Less. XVI. may also be made; in this case alkali-
albuminate instead of acid-albumin is formed.

To 10 c.c. of extract § 1 add 5 ¢.c. of an emulsion
of oil of almonds and a little litmus solution.
Place in the warm. In a short time the litmus
solution will be turned red. The fat-ferment of
the pancreas has split up the neutral fat into
fatty acid and glycerine.

DEMONSTRATIONS.

The appearance of the chyle in the lacteals of
the mesentery of a rabbit a few hours after a
meal,

The flow from the thoracic duct.

LESSON XVIII.

THE LYMPHATIC SYSTEM.

A. lLywMpHATIc GLANDS,

1.

PREPARE vertical sections of a rabbit’s Peyer’s
patch which has been hardened in ammonium
bichromate 5 p.c., and stain them with carmine.
When stained, mount a section at once, tu observe
the immense number of leucocytes; shake up
the others in a test-tube with water. Look at
them under a low power to see if the leucocytes
are for the most part shaken out; if so, mount
them in glycerine. Select a comparatively iso-
lated follicle, and observe

a, The adenoid tissue of the follicle (cp. Less.
xvut. § 1, @ (y)), continuous more or less
distinctly with the neighbouring adenoid
tissue, and resembling it in all respects ex-
cept that the fibres are, as a rule, finer, and
the meshes smaller.

b. Around parts of the follicle, narrow spaces
between it and the surrounding tissue ; these
represent the lymph-sinus on the outside of
the follicle,

192 ELEMENTARY PHYSIOLOGY. [XVII.

c. The leucocytes scattered on the outside of,
but especially abundant within, the follicle,

bo

Examine prepared sections of a Peyer's patch
with the blood-vessels injected. Note the capil-
lary network in the follicle showing usually a
radial arrangement.

i)

Examine prepared sections of a Peyer’s patch

- with the lymphatic system injected. Note that
the injected material envelops to a greater or
less extent the separate follicles; it occupies the
lymph-sinus spoken of above around each follicle
and does not penetrate into the interior of the
follicle.

4. Take a small lymphatic gland (e.g. one of those
lying near the sub-maxillary gland in the cat or
dog) which has been preserved* in ammonium
bichromate 2 to 5 p.c. and cut sections passing
through the whole gland and including the hilus.
Shake the sections in a test-tube with water to
get rid of most of the leucocytes, stain with
carmine or picrocarmine and mount in glycerine.

1 Good specimens are more certainly obtained by the following
method. A cat or dog is killed (best by bleeding after chloroform has
been given) and warm salt solution is injected into a carotid for a
quarter to half an hour, the lymphatic glands of the neck are then
cut out and placed in ammonium bichromate 5 p.c. for a few days,
sections are cut with a freezing microtome and shaken. In sections
so prepared the lymph channels are almost completely free from
leucocytes and by careful and more prolonged shaking they may
be removed very largely from the follicles and medullary cords.

re

XVIII. ]

THE LYMPHATIC SYSTEM. 193

a. Observe under a low power

a.

B.

The connective-tissue of the capsule
surrounding the gland and sending in

The trabeculz which divide the outer
portion of the gland, the cortex, into
compartments the alveoli, and which
then in the inner portion, split up into
bands forming a network with rather
narrow, elongated meshes, the medulla
of the gland,

In the alveoli of the cortex the roundish
masses of tissue crowded with leucocytes,
the follicles of the cortex; in the inter-
trabecular spaces of the medulla the
elongated masses of similar tissue the
medullary cords. Note that the folli-
cles are continuous with the medullary

cords.

Around the follicles and around the me-
dullary cords and separating them from
the trabecule the lymph-channels
comparatively free from leucocytes.

b. Observe under a high power

a.

The connective-tissue of the capsule and
trabecule (in some animals e.g. ox, this
contains unstriped muscular fibres) con-
tinuous with

The reticulum of the lymph-channels.
13

194

ELEMENTARY PHYSIOLOGY. [XVUI.

y. Limiting the follicles and medullary
cords may usually be seen a fine line
with nuclei at intervals, indicating the
flat cells bounding the lymph channel.

6. The adenoid tissue of the follicles and
cords, with finer fibres and smaller meshes
than that of the lymph channels; unless
the section has been well shaken this
will be largely hidden by leucocytes.

Examine sections of glands with the blood-vessels
injected.

The arteries enter at the hilus surrounded by
connective tissue and branch in the trabeculae
of the gland. From the smaller of these branches
fine arteries run to the follicles and medullary
cords and form in them a capillary network.
The veins have a distribution similar to that of
the arteries.

B. SPLEEN.

1.

Take the spleen of a cat which has been hard-
ened in ammonium bichromate 5 p.c. and cut
out a piece at right angles to the long axis of
the spleen. Prepare sections of this with the
freezing microtome (cp. Appendix) or if that is
not available take a portion of the piece, imbed,
and cut by hand. Stain some sections with
picrocarmine and mount in glycerine. Observe
under a low power

XVIII. ]

a.

THE LYMPHATIC SYSTEM. 195

Externally the broad fibrous sheath, the cap-
sule sending in

Large and conspicuous trabecule; these
run throughout the spleen branching as they
go into roundish bundles which are connected
with other similar bundles and so form an
irregular trabecular network throughout the
spleen: the bars of the network cut in all
directions will be seen scattered about the
section.

In many of the trabecule largish central
spaces the veins, which may contain blood
corpuscles, will be seen, devoid of any proper
muscular and connective tissue coats; in the
centre of the section large trabecule cut
transversely or obliquely will probably be
seen containing both arteries and veins; if
the section passes through the point of en-
trance of the vessels these will be seen run-
ning towards the centre surrounded by tissue
continuous with the capsule.

The splenic pulp occupying the spaces of
the trabecular network; it resembles some-
what the follicular. substance of the lymphatic
glands but has a yellowish-mottled appear-
ance, in it will be seen roundish masses of
tissue, the Malpighian corpuscles more
deeply stained than the splenic pulp, the
small arteries are surrounded by a variable
amount of similar tissue.

13—2

196 ELEMENTARY PHYSIOLOGY. [ XVIII.

e. Examine the splenic pulp under a high power
and note that the mottled appearance is due
to the presence of red blood corpuscles scat-
tered irregularly in it,

2. Prepare a section as thin as possible of a dog’s
spleen, from which the blood has been washed
out by injection with salt solution, and which
has subsequently been injected’ with and then
preserved in ammonium bichromate 5 p.c. Stain
with dilute picrocarmine shake and mount in
glycerine. Observe under a high power.

a, There are no distinct lymph-channels.

b. The reticulum of the splenic pulp varies in
appearance in different places; in places it
appears as a network of cells having in vari-
ous directions flange-like projections which
taper off and join with the similar processes
of neighbouring cells; elsewhere the cells
may be nearly or wholly absent and a reticu-
lum of fine fibres be seen. Some leucocytes

1 As soon as possible after the animal has been killed (best by
bleeding under chloroform) all the branches of the celiac artery ex-

cept the splenic branches are tied and warm salt solution is injected.

into the artery until the spleen is quite pale; then the solution of
ammonium bichromate is injected until the spleen is yellow, the
splenic veins are then ligatured, the spleen a little distended by
further injection and the arteries tied, The spleen is removed to
5 p.c. ammonium bichromate; in two days it is cut in pieces and left
in bichromate solution for a week or longer. The pieces are then
placed in 80 p.c. aleohol which is renewed until it is no longer
coloured, sections may then be made (best with the freezing micro-
tome) or the pieces may be kept in 75 p.c. alcohol.

—_ *

xvi] THE LYMPHATIC SYSTEM. 197

and red blood corpuscles not washed out of
the reticulum will be present.

c. The reticulum of the Malpighian corpuscles
resembles that of the follicles of lymphatic
glands, in its meshes are many leucocytes,
but no red blood corpuscles.

d. The small arteries, capillaries and veins of
the pulp; the veins branch out from the
trabeculz and have sharp outlines with
nuclei at intervals (they may usually be
recognized in the dog by the spiral lines
running round them).

e. The trabeculz are chiefly composed of un-
striated muscle tissue (the amount of this
varies in different animals, in some it is very
small).

3. Prepare a section of spleen injected’ with Berlin
blue from the splenic artery under a low pressure.

1 A dog is perhaps the best for injection, but a cat or a rat answer
the purpose very well. In a dog the individual arteries and veins
which run to the spleen are large enough to be easily injected
separately, and since the fluid (especially with an arterial injection)
does not readily spread out beyond the section of the spleen directly
supplied by the vessel, a number of injections may be made in the
same animal. The whole spleen should be washed out first from the
celiac artery. To avoid clotting it is advisable to inject 10 p.c.
peptone into the jugular before bleeding. The best injection material
is a ‘2 p.c. solution of nitrate of silver, after the injection pieces of
the spleen are put in 75 p.c. aleohol for a day or two, then cut with
the freezing microtome and exposed to the light. The disappearance
of the epithelium of the capillaries and small veins of the splenic
pulp show in the clearest manner the opening of these into the spaces
of the pulp,

198

ELEMENTARY PHYSIOLOGY. [ XVIII.

Clear and mount in Canada balsam. Note under
alow power

da.

The small arteries branching off to the Mal-
pighian corpuscles; the artery penetrates the
corpuscles usually excentrically, and either

divides into a number of capillaries which

form a network in the Malpighian corpuscle,
or passes on into the pulp giving off a branch
which divides into capillaries in the corpuscle.

Capillaries in the adenoid sheath of the arte-
ries, but less numerous than in the Malpighian
corpuscle.

Small arteries dividing into capillaries in the
splenic pulp.

Small tufts of splenic pulp injected with
blue at the ends of the capillaries of the pulp.

Irregular masses of injected splenic pulp out-
side the Malpighian corpuscles and adenoid
tissue of the arteries where the capillaries
open out into the pulp.

If too great a pressure has been used in injecting,
instead of the tufts of injected pulp at the end
of the capillaries, irregular areas of the pulp or
the whole of it will be permeated with the Berlin
blue; the Malpighian corpuscles and the adenoid
tissue of the arteries will be free from injection
unless the pressure used in injecting has been

very great.

——————

cA

XVIII]

4,

THE LYMPHATIC SYSTEM. 199

Prepare sections of a spleen injected with Berlin
blue from the splenic vein under a low pressure.
Observe

a. The veins in the trabeculz filled with imjec-
tion material.

b. The veins of the pulp more or less distinctly
branching out from the trabecule.

c. The irregular masses of injected pulp at the
ends of the veins.

If the pressure used in injecting has been high
the splenic pulp will be permeated with blue as
with an arterial injection under high pressure.

Cut across a fresh spleen, preferably one from
which the blood has been removed by injecting
normal salt solution through the splenic artery,
and examine the cut surface with the naked eye.
Note the white Malpighian corpuscles. With
curved scissors cut out one including some sur-
rounding tissue, and tease it out well in normal
salt solution. Observe the large number of
leucocytes of very variable size floating about,
some not distinguishable from white blood-
corpuscles, others twice, others three times as
large; in some of the cells scattered about will
be seen brownish-red fragments, stages of the
breaking down of red blood-corpuscles. Try to
make out the constituents of the reticulum

(§ 2, b).

200

ELEMENTARY PHYSIOLOGY. [ XVII,

C. Lympmatic CAPILLARIES.

The lymphatic trunks need not be noticed, since
their structure is essentially the same as that of
the veins, (Lesson XIL)

From a frog remove the intestines (and if neces-
sary the oviducts) as in Lesson x. E. It will be
seen that the peritoneum above and to the
outside of the kidney is not adherent to the
lumbar muscles, but is separated by a space—the
cisterna magna lymphatica. Turn the frog over,
raise the vertebral column near its end with
forceps, cut it through; cut through the abdomi-
nal walls parallel to the vertebral column and
about half an inch from it, taking care not to
injure the underlying peritoneal wall of the
cisterna ; cut through the vertebral column about
an inch in front of the first cut; the dorsal sur-
face of the peritoneal cisternal wall will now be
seen attached to the kidneys in the median line
and to the abdominal walls laterally. Place a

ring of cork underneath the kidneys and attached ,
membrane and cut through the abdominal wali

close to the edge of the membrane, inserting’ a
hedgehog quill after each cut to keep the mem-
brane flat but not tense. Having thus separated
the membrane, stream it gently with salt solution
then very briefly with distilled water, imme-
diately after this pour on both sides of it a little
nitrate of silver solution 0°5 p.c. (or place it in
the solution) and leave it’ for five to ten
minutes. Wash it well with distilled water, and

XVIIL. } THE LYMPHATIC SYSTEM. 201

expose to light. After twenty-four hours, divide
it in two pieces, mount them in glycerine, one (a)
with the peritoneal surface uppermost, the other
(b) with the cisternal surface uppermost. Observe

in (a)
a. The peritoneal epithelium, consisting of large
flat cells with a slightly sinuous outline.

This is the usual character of the cells lining
serous cavities.

8. Where several more or less triangular cells
seem to radiate from a common point, note
at their apices the small granular nucleated
cells surrounding the orifice or stoma lead-
ing from the peritoneum to the cisterna.

in (0)

a. The lymphatic epithelium, consisting of flat
cells, smaller than the peritoneal epithelium,
and with a very irregular sinuous outline,

8. The stomata; very much as in (a).

2. Treat with nitrate of silver’ the peritoneal
surface of the diaphragm of a guinea-pig and
when stained mount a piece of it with the
peritoneal surface uppermost in Canada balsam.
Observe

a. The tendon bundles of the diaphragm arrang-
ed in two layers. The spaces between the

1 The Student should by this time be sufficiently familiar with
the method of using silver nitrate to require no further instructions.

202

ELEMENTARY PHYSIOLOGY. [ XVIII.

bundles mark for the most part the course of
the lymphatic capillaries of the tendon (cp.

§ 3).

b. Superficial to the tendon bundles the epithe-

lium of the peritoneum consisting of flat
polygonal cells. These are larger over the
tendinous bundles than over the intervening
spaces. Stomata similar to those of § 1 a.'8.
may be seen, situated over and communicat-
ing with the inter-tendinous spaces. Pseudo-
stomata, irregular patches of staining sub-
stance at the junctions of the cells, are
frequent.

In a guinea-pig or rabbit brush firmly the
pleural surface of the diaphragm with a camel-
hair brush to remove the surface epithelium and
treat with nitrate of silver. Mount in Canada
balsam with the pleural surface uppermost.

a. The small lymphatic vessels, running a little
above the tendinous bundles and lined with
somewhat irregular spindle-shaped epithe-
lium. In places the curved outline of a
valve may be seen,

b. The superficial lymphatic capillaries with
their characteristic sinuous epithelium con-
tinuous with the lymphatic capillaries of the
inter-tendinous spaces,

c. The origin of the lymphatics. This is best
seen in specimens deeply stained with nitrate
of silver, so as to produce the so-called

4

XVIII] THE LYMPHATIC SYSTEM. 203

negative image. Note the clear branched
spaces, whose sinuous outline resembles that
of an epithelium-cell of a lymphatic capillary.
The clear spaces, cavities containing unstain-
ed connective-tissue corpuscles, stand out in
strong contrast with the surrounding stained
matrix. The junction of these spaces with
a lymphatic capillary, may here and there be
seen,

DEMONSTRATION.

The injection of the lymphatics of the testis.
(Schiifer’s Practical Histology, p. 216.)

LESSON XIX.
STRUCTURE OF LIVER. GLYCOGEN.

A. STRUCTURE OF LIVER.

1. Prepare sections of the liver of a frog, snake or
bird, treated with 1 p.c. osmic acid and subse-
quently with alcohol. Mount a section in dilute |
glycerine. Observe with a low power that the
gland apart from the ducts and ductules consists
of anastomosing tubes between which the
blood capillaries run.

Observe with a high power

a. The tubes. In transverse section they are
seen to consist of four to six liver cells, each
cell containing a large nucleus usually inits
outer portion.

b. The bile capillaries; these are the lumina
of the tubes; in longitudinal sections of the
tubes the bile capillaries are seen to take a
zigzag course between the inner ends of the
cells.

STRUCTURE OF LIVER. GLYCOGEN. 205

c. The cell granules. According to the con-
dition of the animal from which the liver
was taken, these are seen to stretch through-
out the cell or to be grouped around the
lumen (bile capillary).

d. The fat globules, stained black with osmic
acid, they vary greatly both in number and
position according to state of the animal from
which the liver was taken.

e. The glycogen content of the cells. Mount
a section in water and run a little strong
iodine underneath the cover-slip, the parts of
the cell containing glycogen will stain a deep
brown-red (cep. B, § 2, a).

Prepare sections of a mammalian liver, prefer-
ably that of a pig, hardened in Miiller’s fluid
or potassium bichromate 2 p.c. The sections
should be made parallel to the surface of the
liver. Stain with haematoxylin and mount in
glycerine.

a. Observe with a low power
a. The division into lobules.

8. In the centre of most of the lobules the
very thin-walled hepatic or intralobu-
lar veinlet; those in which it is not
seen have been cut through near the
outer end of the lobule.

206

ELEMENTARY PHYSIOLOGY. [ XTX.

Between the lobules the thin-walled
portal or interlobular veinlets, some
of them of considerable size; and the
branches of the hepatic artery, small
but with comparatively thick walls.

Between the lobules may also be seen
small bile-ducts, with cubical or colum-
nar epithelium and distinct lumina.

The hepatic cells radiating more or
less obviously from the hepatic veinlet.

b. Observe with a high power

a.

YY:

The polygonal outline of the hepatic
cells; the cell-substance is granular
and contains one or more spheroidal
nuclei.

The capillary blood-vessels, running out
from the centre of the lobule between
the rows of cells, and joining with one
another at intervals by cross branches;
they are usually traceable by their con-
taining blood corpuscles.

The bile-ducts, their columnarepithelium
with distinct nuclei. In some of the sec-
tions the epithelium may be seen to
become shorter and more cubical as the
duct approaches a lobule. The duct it-
self often appears to end abruptly at the
margin of the lobule.

3 xIx.]

3.

STRUCTURE OF LIVER. GLYCOGEN. 207

Prepare sections of liver, the blood-vessels of
which have been injected with Berlin blue or
with carmine-gelatine. Clear and mount in
Canada balsam. Observe comparing with the
uninjected specimens.

a. The hepatic veinlet, seen according to the
plane in which the lobule is cut, either as a
more or less circular section, or as a short
veinlet passing from the centre of the lobule
to the sub-lobular vein.

b. The portal veinlet running on the petite of

the lobule.

c. The radial capillary network between the
portal and hepatic veinlets united by nu-
merous cross branches, commonly there is
only one row of cells between the radial
capillaries.

Mount in Canada balsam a section of mammalian
liver which has been injected from the bile duct.
The bile capillaries will be seen within the lobules
as a fine network of minute threads of injected
material, passing between and over the surfaces
of the cells.

Scrape a small portion of perfectly fresh liver,
and observe the pale, granular, hepatic cells,
often containing fat globules (ep. § 2, b, a.)

208

ELEMENTARY PHYSIOLOGY. [XIX,

B. GLYCOGEN,

1.

oe

Give a rabbit a full meal* and about six hours
afterwards decapitate it, cut out the liver as
rapidly as possible, disregarding the bleeding.
Remove the gall-bladder and cut up one half
of the liver into small pieces and throw them
straightway into about 200 times their bulk of
water which is already boiling. (Place the other
half in the warm, keeping it moist (cp. § 3).) In
about five minutes, when the pieces in the boiling
water are all thoroughly coagulated, and the
ferment has been destroyed, remove them and
pound them in a mortar into a paste with sand ;
mix this paste with the water previously used,
just acidulate with dilute acetic acid (to ensure
the complete coagulation of albumin) and boil
for afew minutes. Let it stand till the coagulated
proteids have settled, and filter the milky fluid
through a coarse filter. The sediment may be
squeezed in linen, and the expressed fluid thrown
on the filter. The opalescent filtrate is a crude
infusion of glycogen. The above will afford ma-
terial for several students.

If it contains much proteid material, it should be
carefully neutralized, boiled again, and filtered.

a.. Place a few c.c. of the fluid in a test-tube, and
add a drop or two of iodine solution. A port-

1 The rabbit may be fed on bran and carrots,

_

STRUCTURE OF LIVER. GLYCOGEN. 209

wine colour will result which will rapidly
disappear if much glycogen is present, if so
add more iodine until the colour is permanent.
Warm gently; the colour will disappear, but
will return on cooling (unless much proteid
matter be present).

. Test 5 cc. for sugar with Trommer’s test

(cp. Lesson xv. D § 7); a small quantity
only will be found.

Add to 10c.c. (neutralizing if acid) a little
saliva, or artificial pancreatic juice, and place
it in the warm chamber; the opalescence will
disappear, and the fluid become transparent.

a. To 5c.c. of this add iodine as before; the
port-wine colour will not appear, showing
that glycogen is no longer present.

8. . Test the other 5c.c. for sugar; much
more than before will be found.

After some hours’ warming, make a decoction of
the other half of the liver, It will probably
be acid, neutralize with sodium carbonate and
filter.

a. The decoction will be clearer, more trans-

parent, and less milky.

b. It will give less port-wine colour with iodine.

. It will contain sugar in abundance.

14

210

ELEMENTARY PHYSIOLOGY. [X1x.

By a post-mortem change, glycogen present in
the liver at the moment of death, is converted
into dextrose.

DEMONSTRATION.
Artificial Diabetes,

LESSON XX.

THE STRUCTURE OF THE LUNG. THE
MECHANICS OF RESPIRATION.

1. Curt off the head of a newt. Cut through the
skin in the median ventral line of the body, and —
expose the lungs. Cut across the aorta. Passa
silk thread under the anterior part of one lung

-and underneath the thread and lung place a
small piece of cardboard. Seize with fine
scissors a small part of the wall of the lung and
snip it with fine pointed scissors, then, pressing
a glass cannula somewhat against the underlying
cardboard introduce it into the hole made in the
lung. Tie the silk thread tightly around the

j neck of the cannula. With a pipette fill the

cannula with 0°5 p.c. gold chloride solution, and

pressing it on a small piece of india-rubber
tubing which has been previously fitted to the
cannula, distend, but not too strongly, the lung ;
on raising the cannula and alternately pressing
and leaving free the indiarubber tube, any air-
bubbles which may be in the lung will pass into
| the cannula. Clamp the tubing in such a way as

14—2

er i a i Ril le
‘

Lo

ELEMENTARY PHYSIOLOGY. pes

to leave the lung fairly distended. Pass another
thread under the lung and tie it close below the
mouth of the cannula. Holding up the thread
cut away the lung from its attachments and
place it in gold chloride 05 p.c. solution for
about fifteen minutes. Then remove it to water,
cut it open longitudinally, and get rid of the
excess of gold solution by gently shaking the
lung in water once or twice renewed. To reduce
the gold expose it to light for about 24 hours in
water acidulated with acetic acid.

Whilst cutting open the lung note that it is a
simple sac with a smooth inner surface.

Mount a piece in glycerine with the inner surface
uppermost, and observe with a high power

a. The nuclei of the epithelium cells,
occurring in the inter-capillary fosse in
groups of two to four. Sometimes a nucleus
may be seen to be surrounded by a small
amount of protoplasm coloured violet or red-
purple like the nucleus. The cell outlines
are rarely to be made out.

b. The close capillary network running between
the nuclear groups. In this will probably be
seen blood corpuscles with deeply coloured
nuclei. On focussing below the capillaries the
long stained nuclei belonging to the coat of
unstriated muscular fibres may be seen.

Distend the lungs of a frog with 30 p.c. alcohol ;
ligature each lung at its origin and place one (a)

XX.]

THE STRUCTURE OF THE LUNG. 213

in 30 p.c. alcohol for an hour or more, then in
strong spirit for about an hour; leave the other
(8) in 30 p.c. alcohol for two to three days,

Cut open (a) and observe
a. The large central space.

b. The somewhat short primary septa running
inwards from the wall of the lung, and form-
ing a number of polygonal chambers open
towards the central space.

c. Short secondary septa, projecting into the
chambers from the primary septa. <A lens
may be required to see these distinctly.

Scrape the inner surface of (8) and mount in a
drop of iodine. Observe with a high power

The isolated epithelium cells, consisting of a
thin ground plate having near one border a
nucleus with a little surrounding protoplasm
stained with iodine.

Make transverse sections from the posterior part
of a small mammalian trachea, which has been
hardened in chromic acid ‘5 p.c. The section
should be so taken as to cut through the ends of
a tracheal ring and include the membranous
part between them. Observe

a, Externally, the fibrous coat of loose connec-
tive tissue, becoming denser towards the
tube of the trachea; in this are embedded
the two ends of the ring of cartilage.

ELEMENTARY PHYSIOLOGY. ese

Stretching between the ends of the cartilage,
a transverse band of unstriped muscular
fibres; there may also be visible outside
this the cut ends of muscular fibres running
longitudinally.

The submucous coat continuous with the
fibrous coat, but of finer fibres.

The small mucous and serous glands, the
alveoli of which form a nearly continuous
layer in the submucous tissue; the ducts
will be less evident, but they may be seen
running towards the surface, and occasionally
opening on it.

The cut ends of elastic fibres, which form a
distinct longitudinal layer, on the inner side
of the submucous tissue. It will be seen
that this is most marked in the membra-
nous part of the trachea; here, too, the
fibres are frequently collected into bundles.

The mucous coat. In this may be seen

a. A number of fine elastic fibres with a
variable quantity of adenoid tissue.

The distinct basement membrane.

y. The epithelium, consisting of long colum-
nar ciliated cells, and of two, three, or
more layers of small cells lying between
the basal processes of the ciliated cells.
If the trachea has not been carefully
hardened and prepared, the cilia may

© dll de

XX.]

D.

THE STRUCTURE OF THE LUNG. 215

appear only as a granular border lying
on the columnar cells. Goblet-cells will
probably be seen amongst the ciliated
cells.

- Make longitudinal vertical sections of trachea,

taken through the tracheal rings, and compare
them with the transverse sections, noting es-
pecially the elastic and muscular elements.

Take a piece of mammalian lung which has been
distended through the trachea with chromic
acid ‘2 p.c., and hardened in the same fluid (the
lung of a foetal or new-born animal serves best).
The chromic acid having been thoroughly ex-
tracted from it by alcohol (ep. p. 69 C, footnote)
place it in dilute hematoxylin for one to two
days so that it is deeply stained. Remove it to
75 p.c. alcohol or to the fluid in which hema-
toxylin is dissolved (ep. Appendix) renewing the
alcohol as long as it becomes coloured, then place
for ten minutes or longer in strong spirit, and in
absolute alcohol. With blotting-paper remove
the excess of alcohol and transfer to bergamot
oil or oil of cloves for an hour or more. Then
remove the excess of oil and let the piece of lung
soak for two hours or more in just melted
paraffin or in a mixture of spermaceti (1 part)
and castor oil (4 parts). Imbed in the same
substance and prepare sections, using olive oil to
wet the razor; treat the sections with creosote
and turpentine and mount then in Canada
balsam. Observe

216 ELEMENTARY PHYSIOLOGY. [Xx.

a. Sections of blood-vessels (cp. Lesson X11).
b. Sections of bronchia. Note that

The sections of the larger bronchia resemble
sections of trachea, except that

a. The cartilages are irregular in form, and
irregularly scattered throughout the
fibrous coat.

8. The transverse muscular fibres now form
a complete ring in the sub-mucous coat.

From the smaller bronchia the cartilages are
absent, but the muscular fibres still con-
tinue as scattered fibres arranged trans-
versely.

c. The passage of a small bronchial tube into an
infundibulum (this may be seen occasionally
in a section), Note that

a. The columnar ciliated cells become
shorter and lose their cilia.

8. The muscular fibres apparently cease.

y. The longitudinal fibres spread out into
an irregular meshwork.

d. Sections of alveoli.

a. The outline of the alveoli: the distinct
basement membrane.

8. The flat, tesselated epithelium*. This

1 The outlines of the epithelium cells of the alveoli may be
rendered evident by treatment with silver nitrate in the following
manner. In a just killed mammal, a cannula is tied in the trachea,

xx.] THE STRUCTURE OF THE LUNG. 217

will be seen best where a portion of the
wall of the alveolus is left flat in the
section; sometimes it may be seen in

profile.

;

y. The elastic fibres of the alveolar wall,

6. The nuclei and collapsed walls of the
capillaries,

i |

Take a piece of mammalian lung the blood-
vessels of which have been injected. Treat as in
§ 5 but without staining.

Observe the close-set capillary net-work over the
alveolus, and the artery forming a circle round
the mouth of each alveolus,

DEMONSTRATIONS.

1. The circulation in the lung of toad or frog.

2. Tracing on the revolving drum of the movements
of the column of air in respiration.

the thorax punctured on each side (or the.chest walls removed) and
the lungs distended with a -2 p.c. solution of silver nitrate; by
alternately forcing the fluid in and drawing it out of the lungs the
air will be gradually displaced by the silver solution ; when this is the
case, 30 p.c. alcohol is several times injected into and drawn out of
the lung so that the nitrate of silver is displaced by 30 p.c. alcohol,
this is then displaced by 50 p.c., and so on up to strong spirit with
which the lungs are distended ; the trachea is then tied, the lungs cut
out and placed in strong spirit. Sections (best cut with a freezing
microtome) are exposed to light and treated as usual.

218

ee

aS

ELEMENTARY PHYSIOLOGY. [XX.
The movements of the diaphragm.
Apneea.
Slight dyspnea.

The action of the respiratory muscles in
dyspnea.

The collapse of the lungs on puncturing the
thorax, and the consequent futility of the respira-
tory movements.

The action of the phrenic nerve.

— + — => arr ; ,

a

—*

LESSON XXI.
THE COLOUR OF BLOOD. RESPIRATION.

1. Pour a little defibrinated blood' into several
test-tubes (a)—(d).

a. Keep for comparison with the rest.

b. Add an equal volume of water and warm to
about 50° C,

c. Add a few drops of ether or chloroform and
shake.

d. Add a little bile or solution of bile salts and
shake,

The blood in (6) (c) (d) will be laky i.e. com-
paratively transparent owing to the hemoglobin
of the corpuscles having been dissolved in the

. fluid; compare the colour with that of (a),
compare also the transparency of (a) (c) by
placing a drop of each on a glass slide and
attempting to read type through it.

1 This may be obtained from the butcher’s.

ELEMENTARY PHYSIOLOGY. [XXL

Place a rat or guinea-pig under a bell-jar with
a sponge moistened with chloroform. When it
is thoroughly under the influence of the chloro-
form, quickly open the thorax, and cut across
the heart. Collect the blood in a glass beaker,
and defibrinate it; pour the defibrinated blood
into a platinum crucible surrounded by a mixture
of ice and salt. Leave it till it is frozen. Then
remove it from the ice and salt, so that it may
thaw. By this means the blood-corpuscles will
be broken up, and the blood will become laky
(cp. § 1). If the blood does not become
thoroughly laky it should be frozen and thawed
again. ,

Place the laky blood on one side in a cool place
(it is best to surround it with ice) for a day.
A sediment will then have formed consisting
partly of hzmoglobin crystals* and partly of

1 Blood crystals in quantity may be obtained in one of the following

ways:
a.

To defibrinated blood add ether gradually, shaking continuously
until the blood becomes laky (the volume of ether required is
about 7 the vol. of the blood taken), place it then in the cool
for one to three days. Dog’s blood treated thus often yields
erystals as soon as it is cooled.

Blood is treated as in § 3, but the washing is repeated many
times; to 10 c.c. of the crude solution of hemoglobin which is
obtained add strong spirit drop by drop; shaking continually
until a precipitate is obtained, note the amount of spirit added,
add this amount of spirit minus ‘5c.c, to each 10c.c. of the
rest of the hemoglobin solution, shaking well as the spirit is
added; then place it in a mixture of ice and salt; after some
hours to a day decant as much fluid as possible and filter the
remainder; the crystals on the filter may be washed with
30 p.c. alcohol and subsequently with water both at 0°C.

XXL] THE COLOUR OF BLOOD. RESPIRATION. 221

broken-up corpuscles. Mount a little of the
sediment and examine under a high power.
Note

a. The crystals of hemoglobin (oxyhemo-
globin); those of the rat are thin rhombic
prisms; those of the guinea-pig are ap-
parently tetrahedra but in reality belong also
to the rhombic system. Look for a clump of
crystals to observe better their bright red
colour.

b. The decolorized red blood corpuscles; these
will be seen as pale rings mixed up with
a good deal of granular débris.

Sometimes defibrinated guinea-pig’s blood yields
crystals when a drop of it is simply mounted
with a drop of chloroform ; usually crystals may
be obtained without leaving the blood for a day
in the cool by placing a little of the frozen blood
on a slide, putting on a cover-slip, warming
gently over a flame for about half a minute and
then cooling; as the blood cools crystals separate
out.

3. Let blood (a large quantity is best) clot in a
beaker, leave it for a day, then pour off the
serum, mince the clot and shake the fragments
gently with an equal volume of cold water, place
a piece of muslin over the beaker and pour off
the fluid; repeat this two or three times, then
treat the residue with about three times its
volume of water (best at temperature of about

222

ELEMENTARY PHYSIOLOGY. [XXL

40°C.,) squeezing the pieces; filter through a
coarse filter. A crude solution of hemoglobin
is thus obtained.

Arrange a spectroscope so that the spectrum of
a flame and the scale are distinctly seen’, Hold
in the flame a wire having a few crystals of
common salt upon it and observe the bright
yellow sodium line (D). Shift the scale so that
the sodium line is at 58°9 of the scale and
clamp the spectroscope tubes.

The numbers on the scale indicate wave-lengths
in hundred-thousandths of a millimetre, so that
each division corresponds to a difference of a
hundred thousandth of a millimetre, and each
tenth of a division corresponds to a millionth of
a millimetre, in wave-length. The wave-length
of the line D is 589 millionths of a millimetre, so
that when this line is placed at 589 of the
scale, the wave-lengths of the parts of the
spectrum can be read off on the scale’.

The spectra described below should be carefully
drawn on a blank scale like that of the spectro-
scope, the position of Frauenhofer’s lines B, C, D,
H, F being filled in from the following table

1 Cp. Gamgee, Physiol. Chemistry 1. 93. The Demonstrator will
shew the method of using the instrument.

2 If this scale is not present in the spectroscope used, the position
of the sodium should be observed; bring then the micrometer wire
exactly over it and read off cn the vernier the position of the telescope.
In the subsequent observations when the telescope is brought into the
position read off, the micrometer wire will give the position of the D

line.

|
|
)
|

XXI.] THE COLOUR OF BLOOD. RESPIRATION. 223

of the wave-lengths of these lines expressed
(roughly) in millionths of a millimetre, B= 687,
C=657, D=589, E=527, F=486, If prac-
ticable these lines should be observed in the
solar spectrum.

Introduce between the flame and the spectro-
scope a much diluted solution of hemoglobin.
Note

a.

The two absorption bands, both between the
lines D and £; the one (a) near D being
narrower and darker than the one (8) near #
(if the solution is very dilute, (a) may be the
only band seen).

The middle of (a) is about w.L. 578’, that
of (8) about w. L. 540.

5. Increase gradually the strength of the solution.

da.

C.

The spectrum is more and more cut off both
at the blue and at the red end, but especi-
ally at the former. The absorption bands
are both blacker and broader.

As the solution becomes stronger, the two
bands run together, the ends of the spectrum
also suffering absorption, so that light passes
through only in a space in the green (middle
about w.L. 507) and a broader space in the
red (middle about w. L. 650).

With a still stronger solution, the green light

1 The mid-lines of the bands given here varies somewhat with the
strength of the solution.

224

ELEMENTARY PHYSIOLOGY. [XxXI,

also is absorbed, and only the red is visible,

and this at last disappears.

Reduce the oxyhemoglobin solution with Stokes’s
reducing” fluid in the cold, or with a few drops of
ammonium sulphide solution warming gently.

a.

Compare the claret colour of the reduced

hemoglobin solution with the bright scar-

let of the original solution.

Examine with the spectroscope. There isa
single broad band, occupying a position in-
termediate between those of the two oxy-
hemoglobin bands which have disappeared.
The band is not quite intermediate ; its mid-
line (Ww. L. about 565) lies nearer D than ZL.
This single band is much less dark than
either of the two bands produced by the
same quantity of oxyhzemoglobin.

With stronger solutions less of the blue of
the spectrum and more of the red (between
C and D) is absorbed than with a solution of
oxyhzmoglobin.

Shake well the reduced solution, pour it two or
three times from one vessel into another so as to
expose it thoroughly to air; and examine it at
once. The bright scarlet colour will be restored;
the oxyhzemoglobin spectrum will reappear.

If allowed to remain at rest, reduction, from
excess of reducing reagent present, may soon
return.

1 See Appendix.

XXI.] THE COLOUR OF BLOOD. RESPIRATION. 225

8.

10,

L.

Examine the spectrum of blood-crystals either
with the microspectroscope or by placing a thick
layer of crystals on a glass slide before the larger
spectroscope. The spectrum of oxyhzemoglobin
is seen.

Pass carbonic oxide through an oxyhzemoglobin
solution for fifteen to thirty minutes.

a.

Note the peculiar bluish tinge acquired. Ex-
amine the spectrum; two bands are seen
like those of oxyhzmoglobin, but both placed
more towards the blue end; the middle of
(a) is about Ww. L. 572, of (8) about w. L. 535.

In the absence of a wave-length spectroscope,
oxyhzemoglobin and CO-hzmoglobin may be
compared as follows. Place some of the oxy-
hemoglobin solution before the spectroscope,
bring the micrometer wire to the middle of
one of the bands, and fix the telescope in
position. Replace the oxyhzemoglobin solu-
tion by the carbonic oxide hemoglobin solu-
tion and examine; the middle of the band
will now be to the blue side of the wire.

Treat the carbonic oxide hemoglobin with
either of the reducing agents used above.
Reduction will not take place.

Take a few c.c. of a solution of crystals of hemo-
globin.

a. Boil; the proteid constituent will be co-

agulated.
15

226

xe.

ro

ELEMENTARY PHYSIOLOGY. [ XXI.

b. Add drop by drop HCl 1 p.c., the hemo-
globin will be split up and the proteid
constituent precipitated ; add an equal bulk
of ether and shake, the colouring matter
(hematin) will be largely dissolved in the
acid-ether ; with a pipette remove the lower
stratum of fluid, add a few drops more acid
and place at about 40° C., the proteid precipi-
tate will be converted into acid-albumin and
dissolved; neutralize, it will be precipitated
and may be examined for the ordinary cha-
racters of acid-albumin.

Place a drop of blood on a glass slide, and by
gently warming evaporate it to dryness: add to
it a grain of salt, and thoroughly mix it with the
blood, rubbing the whole to a fine powder.
Cover with a cover-slip, and let a little glacial
acetic acid run under it. Warm the slide, not
too rapidly, over a flame till bubbles appear
under the cover-slip; then let it cool, and ex-
amine under a microscope with a high power.
A large number of crystals of hemin as
brown-red rhombic prisms will be seen.

DEMONSTRATIONS.

The spectra of
a. Heematin in acid and alkaline solutions.
b. Heematin reduced in an alkaline solution.

The gases of the blood.

XXI.] THE COLOUR OF BLOOD. RESPIRATION. 227

3.

Or

The colour of venous and arterial blood in the
living animal, and its dependence on the pre-
sence of oxygen in the lungs.

The respiratory function of the pneumogastric.
The action of the respiratory centre,

The effect of arterial and venous blood on the
irritability of muscular tissue.

The phenomena of asphyxia.

LESSON XXII.
STRUCTURE OF THE KIDNEY.

1. a. Take a sheep’s kidney and cut it in half
longitudinally, note the ureter expanding
into the pelvis and then into several tubes,
the calices, into which the pyramids project.

b. On the cut surface of the kidney, note the
pale inner or central medulla formed by the
pyramids of Malpighi; externally the cortex
a brownish-red zone having thin pale radial
stripes (cp. § 2, c) which do not quite reach
the surface; and between the medulla and
cortex the intermediate layer forming a
dark red zone not very sharply defined, es-
pecially on the cortex side, and having pale
stripes (cp. § 2,b) continuous with those seen
in the cortex running radially through it from
the medulla.

c. Turning back to the ureter, note the connec-

| tion of its outer connective tissue coat with the
fibrous coat of the kidney, follow the renal
artery and vein running into the kidney in

XXxIL.]

STRUCTURE OF THE KIDNEY. 239

the connective tissue outside the ureter and
pelvis; both artery and vein divide into
several branches which enter the substance
of the kidney outside and between the ends
of the calices at the bases of the pyramids;
tracing them outwards they will be seen to run
to the outer portion of the intermediate-layer
and there to branch, their branches arching
through the kidney substance and so forming
a net-work (more complete in the veins
than in the arteries) stretching through the
kidney substance in the curved surface of the
outer part of the intermediate-layer.

From a mammalian kidney hardened in ammo-
nium bichromate 5 p.c. prepare radial sections
extending from the outer surface to the summit
of a papilla. The sections are best cut with a
microtome. Stain them. Observe under a low

power.

a. The medulla, with its straight tubes; some
of the numerous divisions of these as they
run outwards may be seen.

b. The intermediate layer: the straight tubes
of the Malpighian pyramids separate into
bundles the medullary rays (pyramids of
Ferrein); between the bundles are seen
numerous blood-vessels and some tubes of

Henle (ep. § 3, b).

c. The cortex: the medullary rays are seen to
run out nearly to the free surface, between

230

ELEMENTARY PHYSIOLOGY. [ XXII.

these are convoluted tubes and end-capsules
with their glomeruli arranged in double rows
between each two pyramids (the symmetrical
arrangement of the medullary rays and inter-
vening convoluted tubes may not be obvious
if the section is cut obliquely). In the outer
part of the cortex convoluted tubes only are
seen.

3. Observe under a high power.

a. In the medulla

a.

The epithelium of the straight tubes
(tubuli uriniferi recti); in the smaller
tubes, collecting tubes, this is composed
of short columnar or cubical cells with
spherical or ovoid nuclei; in the larger
tubes, outflow tubes, it is composed of
longer columnar cells with ovoid nuclei;
the lumina, distinct throughout, become
larger as the tubes increase in size.

b. In the intermediate layer

a.

B.

The continuation of the straight tubes
outwards in the medullary rays.

The loops of Henle, chiefly in the medul-
lary rays; they run down also a variable
distance into the medulla.

The ascending limbs of the loops;
these will probably be deeply stained,
they vary in size in different parts of
their course and are composed of cells

XXII]

C.

STRUCTURE OF THE KIDNEY. 231

sometimes imbricated, with striated outer
portions and containing oval nuclei; the
lumen is small.

The descending limbs of the loops;
these are much narrower, with trans-
parent flattened epithelium the nuclei
of which project into the lumen, some-
times alternately on the two sides, and
thus the tube, except for its basement
membrane, simulates a blood capillary.

The change in character of the epithelium
may take place either in the ascending or in
the descending limb of the loop.

é.

The numerous blood-vessels between the
medullary rays (cp. § 8, 6); in the outer
part of the layer rather large arteries
and veins cut transversely or obliquely
will be seen (cp. § 1, c).

In the cortex

u.

B.

The end-capsules, with the nuclei of
their epithelium.

The glomerulus in each end-capsule
(cp. § 8, c) and the nuclei of its capillaries,

The narrow neck of the capsule, this
will be obvious in those capsules only
in which the section has passed longi-
tudinally through the neck.

The coiled course of the convoluted
tubes (tubuli contorti); the outlines of

232

ELEMENTARY PHYSIOLOGY. [ XXII.

the individual cells may or may not
be distinct, they have each a spherical
nucleus and are striated in their outer
portions. Sometimes the lumen is large,
sometimes it can scarcely be made out.

e. The continuation outwards in the me-
dullary rays of the ascending loop of
Henle.

€. In the outer half of the cortex are deeply
stained short tubules running out from
the rays and sometimes seen to be con-
tinuous with the ascending loops of
Henle; their cells resemble those of the
ascending loop except that they are of
unequal size, thus giving a very zigzag
outline to the tubule; this is the ‘7r-
regular’ portion of the urinary tubule.

m. In the medullary rays will also be seen
one or two rather large tubes with con-
spicuous spherical nuclei, these are the
so-called spiral tubules. Note also in
the rays the smaller straight (collecting)
tubes.

A basement membrane may be made out in
all portions of the urinary tubule.

Cut sections at right angles to the medullary
rays through the lower part of the cortex and
observe the medullary rays surrounded by con-
voluted tubes.

- XXxir.d

D.

\

STRUCTURE OF THE KIDNEY. 233

Cut similar sections through the outer part of
the medulla and observe the cross sections of
the tubes, and their membrana propria, with a
small amount of connective tissue between them;
sections of both the ascending and descending
limb of the loops of Henle will also be seen.

Place a small piece of the cortex of a fresh
kidney in 5 p.c. neutral ammonium chromate,
tease out a fragment in the same fluid.

Observe the cells of the convoluted tubes, isolated
or in groups, shewing a very distinct striated
outer portion; in some cells the outer part may
appear as a brush of ‘rods,’

Cut as thin a section as possible of the inner
part of the cortex of a fresh kidney, tease it out
in normal salt solution and observe the appear-
ance of the fresh cells in the isolated bits of
tubules.

Take a piece of kidney which has been injected
from the renal artery, prepare sections like those
of (§ 2), clear and mount in Canada balsam.
Observe

a. The large arteries and veins in the upper part
of the intermediate layer.

b. The small arteries and veins (arterie et
vene rectz) given off from these, running
down between the medullary rays into the
medulla; they break up almost immediately
into a brush of capillaries which enter the

234

ELEMENTARY PHYSIOLOGY. [ XXII.

medulla and form a network throughout it;
the meshes are elongated in the direction of
the tubes, especially near the summit of the
papilla.

The interlobular arteries and veins run-
ning from the larger vessels outwards in the

cortex between the medullary rays; the
arteries give off on all sides (two rows will

_probably be seen in the section) small arteries

(arteriz afferentes), one to each end-capsule
where it breaks up into capillaries to form the
glomerulus.

. The small vein (vena efferens) issuing from

each end-capsule and breaking up into capil-
laries which form a network in the cortex;
the veins from the innermost capsules break
up into a brush of capillaries like the arteriz
rectz and run towards the medulla.

The small veins running from the capillary
network of the cortex to the interlobular
veins (§ c).

Here and there the small artery running to
a glomerulus may be seen to send a branch
direct to the capillary network of the cortex ;
similar direct branches will also be seen in
the outer part of the cortex running from
the ends of the interlobular arteries.

Small veins at the periphery of the cortex
(venze stellate) also arising from the capilla-
ries of the cortex.

it i i ee

XXIL.] STRUCTURE OF THE KIDNEY. 235

9. Cut vertical sections of a rabbit’s or dog’s bladder
which has been distended with and hardened in
ammonium bichromate 2 p.c. Observe

a. The thin external fibrous coat.

b. The muscular coat consisting of an outer
generally speaking longitudinal layer and an
inner generally speaking circular layer; inside
this may also be seen a third layer with fibres
running in various directions chiefly longi-
tudinally.

c. The sub-mucous coat of connective tissue.

d. The mucous coat of

a. Connective tissue rather finer but con-
tinuous with that of the sub-mucous
coat.

f8. Epithelium consisting of an inner layer
(next to the cavity of the bladder) of a
single row of roughly cubical cells, a
median layer of a single row of pear-
shaped cells, and an outer layer (next
the basement membrane) of two or
three rows of elongated cells between
and beneath the processes of the pear-
- shaped cells.

The form of the cells naturally varies with
the degree of distension of the bladder.

10. Cut transverse sections of a rabbit’s or dog’s
ureter which has distended and hardened like
the bladder (§ 9).

»

ELEMENTARY PHYSIOLOGY. [xxark 7%

Apart from the thickness of the coats, the struc-
ture is much the same as in the bladder.

DEMONSTRATION. ,
Nitrate of silver preparation to shew the epi- ©
thelium of the end-capsules.

LESSON XXIIL
URINE.

DETERMINE the specific gravity of urine by
means of the urinometer.

Test the reaction of fresh urine with litmus
paper, it will be acid; this is due to the pre-
sence of acid salts mainly of acid sodium phos-
phate, and not to free acid.

Put 200 c.c. of urine in a warm place, and ob-
serve from time to time.

a. It will, after twenty-four or more hours, lose
its acid reaction, and become alkaline.
Gently warm the litmus paper turned blue
by the urine, the blue colour will disappear,
shewing that the alkalinity is due to the
presence of ammonia or a salt of ammonium.

b. It will gradually become cloudy, and yield a
deposit of various salts.

c. Its odour will become putrefactive.

The urine has undergone alkaline fermenta-
tion, |

238

4,

ELEMENTARY PHYSIOLOGY. [ XXIII. |

A small quantity of mucus derived from the
urinary passages is occasionally present in the
form of a faint cloudy precipitate. This may
be rendered more apparent by the addition of
acetic acid. |

Urea. Place a few crystals of urea in a watch-
glass, and dissolve them in a small quantity of
water.

a. Mount a drop of the solution, and when it

b.

has partially evaporated observe under a
high power the crystals of urea, consisting
of four-sided prisms* commonly ending in
two surfaces or in a single oblique surface ;
if the evaporation is rapid the urea crystal-
lises in long spicules.

Add to another drop on a slide a drop of
pure, fairly strong, nitric acid; observe under
the microscope the rhombic and six-sided
tablets of nitrate of wrea which crystallise out.
Note the striz frequently present in these
tablets.

Repeat (b), using a concentrated solution of
oxalic acid instead of nitric acid. Oxalate of

urea will crystallise out in various forms, °

prominent among which will probably be
long thin flat crystals often in bundles;
regular rhombic prisms, or tablets resembling

1 Figures of the crystals described in this Lesson will be hung in
the laboratory.

— —

XXIII. ]

6.

URINE. 239

somewhat those of nitrate of urea, may also
be seen.

. Dilute considerably the remaining solution,

and add to a part of it a solution of mercuric
nitrate. A white precipitate of mercuric
oxide combined with urea will at once take
place.

To the rest of the urea solution add a little
sodium chloride and then mercuric nitrate
drop by drop. <A certain quantity of the
mercuric nitrate must be added before any
precipitate is formed; a double decomposi-
tion takes place between the mercuric and
sodium salts so that as long as any sodium
chloride is present, the mercuric nitrate is
not available for precipitating the urea.

Place a little urea in a test-tube and heat it
carefully over a flame, when the temperature
is somewhat over 100°C. it melts ‘and am-
monia is given off (recognized by its odour),
heat it a little longer then let cool; add a
small quantity of water to dissolve the resi-
due and pour the fluid into another test-
tube containing a strong solution of sodium
hydrate and a drop or two of cupric sulphate.
A violet colour will be produced (biuret
reaction).

Evaporate 20 c.c. of urine in a water-bath till its
volume is reduced to about 2 cc. Filter, and
examine the filtrate for urea (§ 5, D. c).

240 ELEMENTARY PHYSIOLOGY. [XXIIL

7. Uric Acid.

a. Place a few crystals of uric acid on a slide,
and add a little liquor potassze to dissolve
them; then add nitric acid just to excess.
Examine with a high power the crystals of
uric acid formed. They are generally rhombic
plates with obtuse angles, but they vary
‘greatly in form. Note the star-shaped and
dumb-bell crystals.

b. Make a saturated solution of uric acid in
strong sodium hydrate, mount a drop and
place it aside for some time. Crystals of
sodium urate will crystallise out. Note the
spheres covered with small thin projecting
prisms; hexagonal prisms and thick tablets
may also be seen,

ce. To a little uric acid in an evaporating dish
add a little fairly strong nitric acid, the uric
acid will be split up and gases given off.
Evaporate to drymess on a water-bath at
a temperature not above 40°C. A reddish
residue will be left, to this add a drop or
two of weak ammonia; a red-purple colour
will be formed; this is the murexide test
for uric acid, Add then a drop of strong
sodium or potassium hydrate, the colour will
turn to a blue-purple.

8. To 100 cc. of urine (best evaporated to a third
of its bulk) add about lec. of pure strong
hydrochloric acid. Place aside for a day or two

XXIII. ]

URINE, 241

in a tall glass jar, Brownish red crystals of uric
acid will form at the sides and bottom of the
jar. Examine one or two of these under the
microscope, to the rest apply the murexide test

(§ 7. c).

TESTS FOR SOME ABNORMAL CONSTITUENTS OF THE

9.

10,

11,

URINE,
Albumin.

a. Boil, making the fluid just acid with acetic
acid; albumin if present is coagulated (ep.
Lesson 111. § 17).

b. Apply also the tests for proteids.

‘Sugar. Coagulate any albumin that may be

present (cp. § 9) and filter; apply Trommer’s
test’ (Lesson xv. D § 7).

The colouring matter of the urine may be re-
moved by pouring it on a filter containing some
animal charcoal moistened with water; it must
be remembered however that if the quantity of
sugar present is small it may be retained by the
charcoal,

Bile Pigment. Use Gmelin’s test (Lesson
Xvil, B, § 3).

' If a faint reaction only is obtained, it cannot be relied on as
shewing the presence of sugar, since several other bodies (e.g. uric
acid) which occur in urine also reduce cupric oxide,

16

242
12.

ELEMENTARY PHYSIOLOGY. [ XXIII.

Bile-acids. Use Pettenkofer’s test (Lesson XVII.
B, § 4).

Or—Soak a small piece of white blotting paper in a
syrup of sugar, and allow it to dry. Let fall on it a
drop of the urine supposed to contain bile-acids, and
near it a drop of strong sulphuric acid, so that the
two drops run into one another. If bile-acids are
present, there will be a purple colouration at the
junction of the drops. If the quantity of bile-acids
present is small, the urine must be concentrated, but
in such cases it is best to test in the ordinary way-

QUANTITATIVE ESTIMATION OF UREA.

Knop-Hiuifner Method. (Hypobromite Method.)

When sodium hypobromite in an excess of alkali
is added to a urea solution nitrogen is given off,
thus,

CO (NH,),+3Na Br0+2NaHO=3NaBr+Na,CO,+3H,0+N,,.
According to this equation 1 gram of urea gives off
very nearly 373c.c. of nitrogen. In fact, however,
the decomposition of urea is not complete, the
amount of the deficit depends chiefly upon the
amount .and strength of the hypobromite solution,
and on the percentage of urea in the urea solution.
In making an accurate estimation of urea, the
deficit of nitrogen under the conditions of. the
observation, (determined beforehand) must be
allowed for; and as the theoretical amount of
nitrogen is estimated at a temperature of 0°C. and
a pressure of 760mm. of mercury, the volume of

XXIII] URINE. 243

the gas obtained must be corrected for this tem-
perature and pressure (cf. p. 402).

When, as in medical practice, a rough estimation
of the percentage of urea is sufficient, the tube in
which the nitrogen is collected is graduated em-
pirically, at the ordinary indoor temperature, Le. at
about 18°C. The graduation is performed by
mixing a known quantity of hypobromite solution
with 5cc. of a lp.c, solution of pure urea, and
marking on the collecting tube the volume of
nitrogen given off. Similar observations are made
with 5c.c. of a 2p.c. and of a 3p.c. solution of
urea. The tube, between each of the marks so
obtained, is divided into tenths. Hence, when
5 c.c. of a solution of urea of unknown strength are
taken, and treated as the standard solution was
treated, the percentage of urea in it is at once
roughly determined by the number of divisions of
the tube occupied by the nitrogen given off.

One form of apparatus of this kind is described in
the Appendix (cf. p. 400).

2. Liebig’s Method. (See p. 398.)

QUANTITATIVE ESTIMATION OF SUGAR.

Fehling’s fluid (cf. p. 381) contains a known amount
of cupric sulphate, and this, as has been seen in
using Trommer’s test, is reduced by sugar to cuprous
hydroxide or oxide when boiled with it in an alka-
line solution. In making an estimation, 10c.c. of

16—2

244 ELEMENTARY PHYSIOLOGY. [XXIII

Fehling’s fluid may conveniently be taken, the
cupric salt in this is just reduced by ‘05 gram of
sugar,

Preliminary estimation, Take 20c.c. of urine,
dilute it to 100c.c. with water, and put the diluted
urine in a burette. Into a small porcelain dish,
measure accurately 10c.c. of Fehling’s fluid, add
about 10 c.c. of sodium hydrate and 30 cc. of
water; place this on a piece of wire gauze over a
small Bunsen burner to boil, as soon as it boils
run in the urine from the burette in such amount
that the fluid does not cease to boil, and stir
continuously. When the fluid undergoes a distinct
change of colour, the urine should be added more
slowly, and as soon as the fluid turns bright yellow
or brick red, the amount of urine added (a c.c.)
should be noted,

Second estimation. The Fehling’s fluid is measured
out and diluted as before, a—‘5c.c. of diluted urine
run into it and the mixture boiled. It is then put
aside to allow the precipitate to partially settle;
as soon as there is a sufficient fluid free from
precipitate, the colour of the fluid should be
noted. If it is still blue, ‘lec. urine is added, the
mixture again boiled, the colour of the fluid noted,
and so on until the fluid is no longer blue, If the
fluid is colourless after adding a—5c.c. of diluted
urine; a fresh portion of Fehling’s fluid must be
measured out a—1lc.c. of diluted urine added and
the process given above repeated. This ought not

XXIII. ] URINE. 245

to be necessary for the preliminary estimation
should give the amount within ‘jcc. If b be the
number of c.c. of diluted urine required to reduce
the 10c.c. of Fehling the percentage of sugar in
05 x5x100 25
b pa

There are certain sources of error in the method given
above. (1) When the mixture is boiled, a little ammonia
is set free from some of the constituents of the urine,
this dissolves some of the precipitated cuprous oxide ;
the cuprous oxide takes up oxygen from the air and
a cupric salt is formed giving a blue tinge to the
solution. The re-oxidation of the precipitated oxide
may be lessened by boiling in a small glass flask
instead of in an open porcelain dish, but since the
former is more easily cracked during boiling, and in it
the colour of the fluid is less readily seen, it is not recom-
mended for the beginner, (2) When the diluted urine
contains more than 1 p.c. or less than °5 p.c. of sugar,
the method is inaccurate; since 5 c.c. of a 1 p.c. solution
of sugar, or 10 c.c. of a ‘5 p.c. solution contain *05 gram
of sugar, ie, will reduce 10c.c. of Fehling’s fluid,
the urine, for an accurate estimation, must be diluted
to such an extent that not more than 10 and not less
than 5c.c. of the diluted urine are required to reduce
10 c.c. of Fehling’s fluid. The Fehling’s fluid is diluted
5 times because with either a greater or a less dilution
the results are inaccurate,

the urine will be

LESSON XXIV.

SKIN AND TOUCH,

Prepare vertical sections of human scalp which
has been hardened by placing it in chromic acid
‘2p.c. for a week. Stain the sections with
picrocarmine, mount them in glycerine and
observe

\
The Epidermis: proceeding from the outside
will be seen

—

a. Non-nucleated cells, the cell substance of
which is no longer protoplasmic but has
become transformed into horny keratinous
material; they are much flattened especially
near the surface of the skin, where they may
present the appearance of a number of fibres.
This is the horny layer; in the stained
sections it will be yellow.

b. Nucleated cells, spheroidal in form: the
peripheral part of these cells may appear
striated or closely serrated (the expression
of fine projections of the surface).

- —

XXIV.]

C.

SKIN AND TOUCH. . 247

A row of columnar nucleated cells placed
perpendicularly to the tissue beneath. The
spheroidal and the columnar cells together
form the Malpighian layer.

B. The Dermis: in which may be seen

a. A thin outer layer consisting of a fine plexus

of white fibrous tissue, of connective tissue
corpuscles, and of some anastomosing elastic
fibrils, embedded in a good deal of hyaline
or faintly fibrillated matrix. The upper
portions of this form a kind of basement or
limiting membrane immediately below the
columnar epidermic cells. The lower portions
pass gradually into

A plexus of fibrous tissue with larger bands
and amore developed elastic-fibre network ;
in its deeper portion this merges into the
sub-cutaneous connective tissue in which
the spaces of the plexus are large and in
which clumps of fat cells usually occur ; there
is no definite line of demarcation between the
dermis and the sub-cutaneous tissue.

The papille: conical elevations of the der-
mis, rising up into the Malpighian layer,
which therefore appears to dip down between
them.

Indications of the blood-vessels, abundantly
distributed all over the dermis, and forming
capillary loops in the papilla. They should
be observed in injected specimens.

248

é.

ELEMENTARY PHYSIOLOGY. [ XXIV.

In the lower part of the dermis bundles of
nerves which, as they run to the upper part
of the dermis, split up into smaller bundles
and form a plexus; from this run still smaller
bundles to form a more superficial closer
meshed plexus; probably the latter will not
be very obvious in this specimen.

Hair-follicles and the roots of hairs. Note

a. The outer sheath of the follicle formed
of fibrous tissue continuous with that
of the dermis, but much denser in
arrangement.

8. The inner sheath of the follicle, con-
sisting for the most part of indistinct
transversely arranged fibres, amongst
which oblong nuclei may be discerned.

y. A hyaline limiting membrane more
marked than that of the dermis.

5. The outer root-sheath, consisting of cells
continuous with, and resembling those
of the Malpighian layer.

e. The inner root-sheath, pale and trans-
parent, its inner part consists of small
nucleated cells, near the bulb its cells
become larger. This sheath is absent
above the duct of the sebaceous gland.
At the exit of the hair from the skin
the horny layer dips down around it in
the form of a cone,

XXIV.] SKIN AND TOUCH. 249

€. The hair-bulb or the enlargement of the
hair in its deepest part, consisting of a
layer of nucleated cells continuous with
the outer root-sheath; the outer root-
sheath near this point is much diminished
in breadth.

m. The papilla of the follicle projecting into
the hair-bulb, it consists of connective
tissue like that of the outer layer of the
dermis with which it is continuous.

0. The substance of the hair consisting of
a cuticle of thin horny scales in a single
layer, and internally of long horny fibres
or in its deeper portion of spindle shaped
cells, '

h. The sebaceous gland or glands opening
into the upper part of the hair follicle; in the
body of the gland the cells are several layers
deep; they are continuous with the cells of
the outer root-sheath.

zt. A little below the opening of the gland into
the follicle may be seen a band of plain
muscular fibres, stretching obliquely from
the inner sheath of the follicle to the upper
part of the dermis.

2. Cut sections of the palm of the hand or of the
forehead", hardened in chromic acid, picric acid,

1 Instead of this a piece of the skin of the ball of the foot of a cat
may be taken, .

\,

yy

250

ELEMENTARY PHYSIOLOGY. [ XXIV.

“or alcohol, to observe the structure of the sweat-
glands. Note

a.

or)

The terminal gland-coil in the outer part of
the subcutaneous tissue, with its membrana
propria continuous with that of the dermis,
and lined by a single layer of cubical or
short columnar cells.

The gland-duct having two or three layers
of cells continuous with those of the Mal-
pighian layer of the epidermis. The lumen
is usually distinct in both duct and coil. In
the epidermis the cells proper to the duct
cease but its lumen is continued on in a
spiral course to the surface surrounded by
concentrically arranged epidermic cells.

In the specimen of the cornea of the frog pre-

pared as in Lesson v. B § 2 (cp. also Lesson x.
C § 2) observe

c

a. The primary nerve-plexus formed by the

bundles of nerves which enter the cornea at
its periphery ; this plexus has in its deeper
part rather large meshes and its fibres have
nuclei on their course; towards the surface,
and especially just beneath the anterior
basement membrane, this plexus becomes
much finer consisting chiefly of small bundles
of nerve-fibres with much less frequent nuclei;
when these occur they are usually at the
nodal points of the plexus. :

XXIV. ] SKIN AND TOUCH. 251

4, Cut out the cornea.of a recently killed cat or
rabbit, taking care not to drag it, and place it
in gold chloride *5 to 1 p.c. for an hour. Wash
well and expose to light as usual, but do not
remove its epithelium. When it is well stained,
if necessary harden with alcohol, imbed a piece
of it vertically in paraffin, and cut sections as
thin as possible. Observe

a. The bars of the primary plexus cut across,
and becoming finer and more numerous
towards the anterior membrane; from this
superficial layer of the plexus single axis
cylinders or small bundles of fibrils (rami
perforantes) run through the anterior mem-
brane and separate into a brush of fine
varicose fibrils which spread out immediately
underneath the epithelial cells forming the
sub-epithelial plexus. Probably the sec-
tion in some part will be a trifle oblique,
here a fragment of the sub-epithelial plexus
will be seen having small meshes and very
fine varicose nerve fibrils, elsewhere similar
fine fibrils will be seen running a longer or
shorter course underneath the cells. (To
see the sub-epithelial plexus well, sections
should be cut from the surface of the cornea
and a section which includes the anterior
surface of the basement membrane picked
out.)

b. Between the epithelial cells of the cornea,
the epithelial plexus of very fine varicose

252 ELEMENTARY PHYSIOLOGY. [XXIV.

nerve fibrils; in places the fibrils will be seen
to run from the sub-epithelial plexus.

The peripheral course of the nerve-fibres in
the parts of the skin where there are no
special terminal organs, is in the main like
that described above for the cornea. j

Cut sections of the skin of the tip of a finger
which has been placed fresh in osmic acid 1 p.c.
and subsequently treated with alcohol. Mount
the sections in dilute glycerine. Observe the
touch-corpuscles.

or

a. They are oval in form, and are found in the
axes of the papilla; many papilla are with-
out them, in which case a capillary may
generally be seen running up into the papilla.

b. They appear to consist of a mass of connec-
tive tissue in which nuclei are disposed trans-
versely.

c. The nerve-fibre reaches it near the base, and
may be traced coiling round it spirally for a
variable distance.

Pacinian bodies (cp. § 6) may be seen in the
subcutaneous tissue.

The touch-corpuscles are also obvious in sections
of skin hardened in potassium bichromate 1 p.c,
but the course of the nerve fibre ‘is less easily
traced,

6. Examine the mesentery in a recently killed cat ;
in it will be seen a considerable number of oval
transparent bodies the Pacinian bodies. Select

XXIV. ] SKIN AND TOUCH. 253

one which is not surrounded by fat, cut it out
and mount it in normal saline solution. With
a little care the mesentery covering it may be
removed with needles, Observe

a, The medullated nerve-fibre and the thicken-
ing of its connective-tissue sheath as it
approaches the Pacinian body.

b, The division of the connective-tissue sheath
to form the numerous concentric capsules, .
which become closer together towards the
centre. At intervals on the capsules elon-
gated nuclei will be seen.

c. The loss of medulla as the sheath splits up.
d. The hyaline core inside the innermost capsule.

e. The axis-cylinder penetrating the core, and
ending in it in a slight enlargement.
To observe the manner in which a small bundle
of nerve fibres separate and run to the Pacinian
bodies, a piece of the mesentery containing a
number of them should be stretched over a
slide, the piece cut out and the edges allowed
to dry, so that the part containing the Pacinian
bodies is kept stretched. The outlines of the
flat cells of the capsules can easily be made
obvious by placing a Pacinian body freed from
the mesentery as in § 6 in ‘5 p.c. nitrate of
silver for a quarter of an hour, then exposing
to light as usual and mounting in water. They
are not easy to preserve, the best plan is
perhaps to irrigate with dilute glycerine after
treatment with osmic acid or gold chloride,

254 ELEMENTARY PHYSIOLOGY. [XXIV.
B. Tovucu.

1. Power of localisation. a

Using a pair of small bone or wooden compasses,
determine the distance at which the points must |
be placed from each other in order that they |
may be felt as two points when applied lightly |
and with the same force to various parts of the
body. Try if the following order is correct: tip |
of tongue, tips of fingers, palm of hand, cheeks, |
back of hand, fore-head, front of fore-arm, back
of fore-arm, back of neck, &c.

ro

Sensation of temperature.

Dip a small metallic knob in hot water (e.g.
70° C.), and applying it to the same parts of the
body as before, determine roughly the sensitive-
ness to temperature of these parts.

3. Estimation of weight. The muscular sense.

a. Determine the smallest difference in weight
which can be appreciated when slightly
differing weights are held in the palm of
the hand.

b. Then laying the hand and arm supine on a
table, so that they are motionless, and placing
weights on the palm of hand, determine as
before the difference of weight which can be
appreciated. ‘This will be much less in the
case of (b) than of (a). In (a) the muscular
sense is employed ; in (0), only the sensation

XXIV.] SKIN AND TOUCH. 255

of pressure. The weight should be placed
on the same part of the palm of the hand
in each case.

4, Estimate (in either way) the difference of weight
which can be appreciated

a. When the weights are light, eg. 1, 2, 3, 4, or
5 grms.

8. When the weights are heavy, e.g. 10, 20, 30,
&e., or 100, 200, &c. grms.

Smaller differences will be appreciated in a than
in 8. In fact the difference of weight which
can be appreciated bears, roughly speaking, a
constant ratio to the whole weight.

5. Plunge the finger into mercury, and then draw
it out. The sensation will be that of a ring
passing down and up the finger, the greatest
sensation being where there is the greatest
variation of pressure,

6. Place two equal light weights, one cold the other
warm, on the corresponding fingers of the hands;
the cold one will feel the heavier.

Paint a ring of collodion on the palm of the
hand, leaving only a small central space, and
attempt to distinguish between the approach of
a hot body and gentle contact of a feather.

~I

8. Place the elbow first in warm water, and then
in a mixture of ice and water. There will be a
sensation of pain in the fingers and cold in the

ELEMENTARY PHYSIOLOGY, [XXIV.

elbow. The application of cold to the trunk of
a nerve does not cause a sensation of cold,

Tactile delusions.

Cross the second finger over the first or the
third over the second, and place between their
tips a small marble (or any solid round body a
little larger than a pea) so that it touches the
radial side of the first, and the ulnar side of the
second finger-tip, On gently rolling the body
about, a sensation as if of two distinct bodies
will be felt.

The same delusion is experienced if the tip of
the nose be gently rubbed with the tips of the
fingers so placed,

Experiments 6 and 7 will need the assistance of a _

second person,

LESSON XXV.

TASTE AND SMELL,

A. ORGANS OF TASTE AND SMELL.

tf:

Cut off the head of a newt’, remove the lower
jaw, cut off the nose by an incision carried just
in front of the eyes, with a fine pair of scissors
remove the roof of the nasal cavities, and
place the remainder of the nose in osmic acid
1 p.c. for 2 to 24 hours as convenient. Tease
out in water a small piece of the mucous mem-
brane. Or place the nose in Miiller’s fluid for a
week, and tease out the mucous membrane in
the same fluid or in water. Observe

a. The cylindrical epithelium cell, its large
oval nucleus, its broad, clear, peripheral
process, and its branching, faintly granular,
central process,

b. The rod-cell, (olfactory cell) with its spheri-

1 Instead of this the nasal mucous membrane of a mammal may
be taken, it is best to take it with the underlying bone or cartilage
attached (cp. p. 30, § 22),

17

258

ELEMENTARY PHYSIOLOGY. [XxXv.

cal nucleus, its hyaline slender peripheral, and
its still more slender varicose and branching
central process.

ce. Cylindrical cells will frequently be seen, sur-
rounded and partially hidden by three, four
or more rod-cells.

_ Take the nose of a freshly-killed newt’ as in § 1,

but instead of laying bare the olfactory chambers,
cut off the tip of the nose. Place it in chromic
acid 3 p.c. fora week. Ifthe salts of the bone
are not completely dissolved, complete the
solution with nitric acid 1 p.c. mixed with an
equal volume of spirit. Imbed it vertically, and
cut transverse sections. Stain and mount one
of the first sections (a) and one of the last
sections (b). Observe

a. In the respiratory portion of the mucous
membrane

a. That. the surface-epithelium consists of
columnar ciliated cells, interspersed with
goblet-cells. Between the deeper por-
tions of these cells and close to the base-
ment membrane, some small cells will
be seen (these with or without division
develop into the surface cells).

8. In the sub-mucous tissue, serous and
mucous racemose glands.

1 Cp. footnote on p. 257.

’

XXv.]

TASTE AND SMELL. 259

b. In the olfactory part of the mucous mem-
brane, the division of the epithelium into an
outer and an inner layer.

a.

The outer layer consisting of the peri-
pheral processes of both cylindrical and
rod-cells.

The inner or nuclear layer consisting
of several rows of nuclei belonging to
the rod-cells. The nuclei of the cylin-
drical cells form a single row in the
upper part of this layer, but these, as
well as the greater part of the processes
of the rod-cells, are generally hidden by
the nuclei of the rod-cells.

In the sub-mucous tissue the numerous
bundles of the non-medullated nerve-
fibres of the olfactory nerve. Near the
epithelium these form a close-meshed
plexus.

Serous glands, usually tubular with a
few branches; here and there a duct
may be seen running through the epi-
thelial layer as a fine tube consisting
of basement membrane and flattened

cells.

3. From a rabbit’s tongue which has been hardened
in chromic acid ‘2 p.c., or in potassium bichro-
mate 1 p.c., excise a papilla foliata (cp. p. 32, §
29) and imbed in such a way that the sections
will be at right angles to the ridges.

17—2

260

ELEMENTARY PHYSIOLOGY. [XXv.

Prepare thin vertical sections and observe

a.

The section of the ridges with the interven-
ing fosse.

In each ridge three papilliform processes of
the dermis, one median, two lateral.

The horny epidermis, consisting of cells very
flattened, but still retaining thcir nuclei,
covering the ridges and lining the fossz.

The Malpighian layer of the epidermis under-
neath the horny layer. In the fosse both
layers of the epidermis are thinner and less
marked off from one another than on the
surface.

On the outside of each lateral papilla, and
therefore lining each side of the fossa, the
flask-shaped taste-buds*. There are usually
four nearly in contact one above the other.

Each bud is in contact by its deeper part
with the dermis of the papilla, and has a short
neck running to the free surface of the
epidermis of the fossa where its circular open
mouth may often be seen.

The epidermic cells are flattened around its
median portion, forming a nest for it,
The bud itself, with

a. ‘The external cover-cells, curved and
flattened, with conspicuous oval nuclei.

1 Taste-buds in fungiform papille may be seen in sections of the
tip of the tongue of the cat.

XXvVv.]

TASTE AND SMELL. 261

8. The rod-cells, (taste-cells) with spheri-
cal or ovoid nucleus, rod-shaped hyaline
peripheral and central processes,

t. Nerve fibres running along the dermis of the
papille, and branching off to the buds. Note
also the layers of striated muscle and the
glands of the tongue, comparing the general
structure with that of the cesophagus (Lesson
XVI. B.) and of the skin (Lesson xxIv. A.).

Tease out a small piece of a papilla foliata which
has been placed in osmic acid ‘2 p.c. for one or
two days, and observe the characters of the cells
of the taste-buds, noting the branched central
process of the rod-cells,

B. SENSE oF TASTE AND SMELL.

1.

9

Pinching the nose tightly, so as to suspend the
sense of smell, attempt with the eyes shut to
distinguish by taste between pieces of an apple,
an onion, and a potato; they will taste alike.
The pieces must not be rubbed against the
mouth with the tongue, or they will be recog-
nized by their different consistency.

Wipe the tongue quite dry, and place on the tip
a crystal of sugar or on the back a crystal of
sulphate of quinine; neither will be tasted until
it is dissolved.

Apply with pointed, non-polarizable electrodes
(cp. Appendix) a constant current to the tongue ;

262

ELEMENTARY PHYSIOLOGY. [Xxv.

a, distinct sensation of taste will be felt, of acidity
at the positive pole (anode), of alkalinity at the
negative pole (kathode).

Keeping the electrodes at the same distance, say
two or three millimetres, determine the amount
of taste-sensation in different parts of the mouth.
It will be greatest at the tip and back (region of
circumvallate papillz) of the tongue, and edges
of the tongue; least or absent in the anterior
and middle surface, under surface of the tongue,
and lips. The soft palate will be found to be

highly sensitive, the hard palate little or not
at all.

Take two small pieces of sugar of equal size, and
place one on the tip, and the other on the back
of the tongue; the sensation will be most acute
at the tip. |

Place a crystal of sulphate of quinine on the tip
of the tongue; it will hardly be tasted. Place
it on the back ; it will be tasted at once.

LESSON XXVI.

THE EYE.

A. DISSECTION.

A.

2.

Take a fresh eye of an ox or sheep, and note
a. The transparent cornea.

b. Surrounding and continuous with this, the

dirty-white sclerotic which forms the outer
coat of the rest of the eye. The posterior
two-thirds will probably be covered with fat.

The conjunctiva, a continuation of the
mucous membrane of the eyelids. In taking
the eye out of the orbit this membrane is
cut through where it passes from the eyelids
to the sclerotic. Dissect it forwards in any
one place ; it will be traceable to the junction
of the sclerotic and the cornea. Its epithe-
lium is continuous with that of the cornea

(ep. B. 2).

Clear away the fat surrounding the four straight
muscles, it will be seen that their tendons form
a layer under the conjunctiva of the sclerotic.

264

ELEMENTARY PHYSIOLOGY. [XXVI.

Cut away the conjunctiva and muscles, and
remove the fat around the optic nerve; this
pierces the sclerotic on the nasal side, and not in
the axis of the eye.

Cut through the cornea close to its junction with
the sclerotic; the anterior chamber of the eye,
containing clear, limpid aqueous humour, is
thus laid bare. Observe the iris with its central
aperture through which projects the anterior
part of the lens. 3

At a little distance from the cornea, cut through
the sclerotic, being careful not to cut too deeply;
it will separate easily from the pigmented sub-
jacent choroid, except at the junction of the
sclerotic with the cornea, and at the entrance of
the optic nerve.. In other places there is only a
loose connection, largely by means of blood-
vessels. Remove a strip of the sclerotic a few
mm. breadth stretching from the optic nerve to
the cornea. Note its dark inner surface, or la-
mina fusca; note also in the front part of the
choroid, close to the cornea, the pale fibres of
the ciliary muscle spreading from the junction
of the sclerotic and cornea, backwards over the
choroid.

Carefully pinch up the choroid about half-way
between the optic nerve and the cornea with a
fine pair of forceps, and snip it through. Un-
derneath it will be seen a thin membrane, the
retina. The pigment layer of the retina will
probably be torn away with the choroid.

XXVL]

10,

THE EYE. 265

Tear away a piece of the retina to expose the
clear vitreous humour which occupies the
posterior cavity of the eye.

Tilt the eye so that the cornea is underneath ;
through the vitreous humour can be seen the
choroid coat becoming folded longitudinally as it
approaches the lens, and so forming the ciliary
processes.

The nervous elements of the retina cease at the
level of the commencement of the ciliary process.
Their termination is marked by an uneven line,
the ora serrata.

Holding up the choroid and retina, cut them
through as far forward as the ora serrata; it will
be seen that the vitreous humour separates
readily from the retina as far as that line, but
in the region beyond its thin outer coat, the
hyaloid membrane, becomes attached to the
ciliary processes. If an attempt be made to
separate them here with the handle of a scalpel,
it will be found that the pars ciliaris retin (or
the non-nervous continuation of the inner coat
of the retina) together with some of the pigment
layer of the retina (the outer coat) will come
away with the vitreous humour.

At the ora serrata the hyaloid membrane be-
comes united to the membrana limitans interna
of the retina.

Turn the eye with the cornea uppermost, and
cut away the free edge of the iris; make two

266

aa;

13.

ELEMENTARY PHYSIOLOGY. [XXVI.

incisions at right angles to one another on the
surface of the lens, it will be seen that the lens
is covered by a membrane; this is the anterior
part of the lens capsule. Carefully remove
the lens, and trace out the limits of the lens
capsule, noting that it forms a complete in-
vestment for the lens.

Gently separate with the handle of a scalpel the
lens capsule from the front part of the ciliary
processes, and observe that a membrane, the
suspensory ligament or zone of Zinn, passes
from the edge of the capsule to the ciliary pro-
cesses, of which it forms the innermost layer,
dipping. down into their depressions. Poste-
riorly it is continuous with the thin hyaloid
membrane (cp. § 9).

Looking into the eye from the front, observe
a. The entrance of the optic nerve.

b. The blood-vessels running in the retina and
radiating out from the entrance of the optic
nerve.

c. The iridescence, mainly below the entrance of ©

the optic nerve; it is caused by the irregular
reflection of light from the wavy connective-
tissue fibres of the choroid. In this region
(the tapetum) the hexagonal cells of the
retina have no pigment. =

Separate the rest of the retina from the choroid,
and observe that

——

THE EYE. 267

a. The pigment-layer generally adheres rather
to the choroid than to the retina.

b. Apart from the pigment-layer the retina ap-
pears like an expansion of the optic nerve.

c. The retina is firmly attached to the choroid
at the ora serrata,

B. HistToioey.

1,

Take an eye which has been cut open and then
hardened in potassium bicromate 1 p.c. It is
best to take the eye of a large animal, e. g. of an
ox. Cut out a piece containing the junction of
the sclerotic, cornea and iris. Stain, imbed in
spermaceti and castor oil (cp. Lesson xx. § 6),

and cut sections extending from the outer to

the inner surface. Observe

a. The irregular connective tissue of the sclerotic
passing into the lamellz of the cornea.

b. The membrane of Descemet (cp. § 2, c.)
splitting up at the junction of the cornea
and sclerotic into fine transparent bundles,
some of which curve’ round towards the iris,
the others spreading out fan-like a short
distance into the ciliary process and sclerotic.
These bundles of fibres form the ligamen-
tum pectinatium iridis.

c. The ciliary muscle; consisting of bands of
unstriped muscle, radiating from the angle
of the iris and cornea, (In the eye of man

268

ELEMENTARY PHYSIOLOGY. [XXVI.

bundles of circular fibres, the circular muscle
of Miiller, inside the longitudinal fibres will
be seen; in many lower mammals this part
of the ciliary muscle is small or absent.)

. The pigment-layer of the retina continuing

forwards to form the uvea of the ciliary pro-
cesses and of the iris. Inside this is a simple
layer of columnar cells without pigment, this
is a continuation of the inner coat of the
retina. |

The canal of Schlemm at the la: of the
cornea and sclerotic.

In sections of a cornea which has been hardened
in mercuric chloride, and stained (ep. also p. 251),
observe

a. The anterior epithelium, In this note

a. The external flattened nucleated cells.
8. The middle layers of roundish but ir-

regular cells.
y. The internal single layer of columnar

cells placed perpendicularly, cp. Lesson
XXIv, A.

b. The body of the cornea. Note

a. The narrow transparent zone immedi-
ately beneath the epithelium, the an-
terior basement membrane of Bowman.

8. The rows of connective-tissue corpuscles
seen in profile, with numerous branches
communicating not only with the cells

XXVL.]

THE EYE. 269

in the same row, but also with those
below and above,

y. The fibrous lamine between which the
cells are placed; here and there bundles
running obliquely from one lamina to
another may be seen,

c. The membrane of Descemet or posterior
basement membrane: a thick hyaline mem-
brane, with very sharp outlines.

d. The posterior epithelium, consisting of a
single layer of large nucleated tesselated
cells. In this section they are seen in pro-
file, covering the membrane of Descemet.

Lens.

Take an ox’s or sheep’s lens which has been
in potassium bichromate ‘2 p.c. for a week;
observe

a. The junctional lines of the lens fibres as a
star.

b. The fibres readily peel off in lamine.

Place the eye of a rabbit or cat, with the cornea
and sclerotic cut through, in Miiller’s fluid for
three weeks or longer; then cut out the lens
with its capsule, and cut vertical sections through
its centre and mount them in glycerine. The
sections are best cut with a freezing microtome ;
neither the lens nor the sections of it should
be touched with spirit. Observe

270

Or

ELEMENTARY PHYSIOLOGY. _[XxvI

a. The hyaline capsule of the lens.

b. The columnar epithelium of the anterior
surface of the lens ; at the sides these rapidly
but gradually elongate into fibres.

c. At the sides, the nuclei of the fibres forming
an irregular band.

Take a lens as in § 4, and cut transverse sec-
tions through its centre. Observe the cross sec-
tions of the fibres, and their junctional lines.

Place the lens of a rabbit or rat in osmic acid

‘25 p.c. for about three hours; it will swell up -

somewhat, and the outer coat becomes jelly-like
and easily torn; to obviate this, place it in gold
chloride ‘25 p.c. for a minute or less; then tear
off as long a strip as possible of the outer coat,
and tease out in water. Observe the long band-
like fibres with serrated edges usually adhering
together in layers but in some cases isolated.

In some of the fibres a nucleus will be seen;
when the fibres form a layer, the nuclei appear
as an irregular band running across it.

Iris.

Cut out a piece of an iris from an eye which
has been preserved in potassium bichromate,
stain and cut radial vertical sections. Observe

a. On the posterior surface, the uvea or thick
layer of pigment epithelium-cells (ep. § 1, d.).
The pigment will probably be too abundant
to allow the outlines of the individual cells
to be seen.

——

;
+

-

XXVI.]

THE EYE. 271

b. The cross sections of the bundles of plain
muscular fibres forming the sphincter pu-
pille near the edge of the pupil, and lying
just above the pigment-layer.

c. The body of the iris, consisting of blood-
vessels united together by a loose connective
tissue in which branched pigment-cells are
conspicuous. Near the anterior surface is a
denser layer, the so-called anterior limiting
membrane. The radial muscular fibres (di-
lator) may (with difficulty) be seen near the
posterior surface,

Take the iris from a white rat’s or an albino
rabbit's eye which has been. preserved in
Miiller’s fluid. Cut out a piece from the free
edge to the attachment, stain with hzema-
toxylin, clear, and mount in Canada balsam
with the posterior surface uppermost. Observe
the arrangement of the sphincter and dilator
muscles,

Retina.

Cut away the anterior half of the eye of a mam-
mal immediately after death: avoid dragging
the vitreous humour that the retina may not be
displaced. Place the posterior half in potassium
bichromate 2 p.c. or in Miiller’s fluid for a week,
changing the fluid once or twice. After treatment
with alcohol remove the retina carefully, carrying
with it if possible the pigment-layer. Stain a smali
piece with hematoxylin, imbed it in spermaceti

272

10.

ELEMENTARY PHYSIOLOGY. [XXVI.

and castor oil, clear the sections, and mount them

in balsam.

Remove the retina from a fresh eye to osmic acid
lp.c. After a few hours, treat with alcohol. Im-
bed and mount the sections in dilute glycerine.
In all cases take a small piece for making sec-
tions. If long vertical sections are made they
are very apt to turn over.

Examine the two preparations of § 9 and § 10,
some points will be best seen in the one, some
in the other. Observe the following structures
from within (anterior surface) to without (pos-
terior surface).

a. The inner limiting membrane; from this
the fibres of Miiller are scen to start with
a broad foot and to run vertically outwards,
becoming thinner as they go; they may -
readily be traced as far as f, and may be
followed to the outer limiting membrane (A),
here and there anastomosing branches be-
tween the neighbouring fibres of Miiller may
be seen. |

b. The layer of optic fibres. The nerve-
_ fibres here are without medullary sheaths, or
rarely possess them.

c. The ganglionic layer; this consists of a
single layer of large nerve-cells with con-
spicuous nuclei. Externally a single process
may be seen running into the optic fibre
layer, internally several processes branching
into

XXVI.]

11.

THE EYE. 273

. The molecular layer, consisting of a close

network of fine fibrils; under a moderately
high power it may appear simply granular.
The inner nuclear layer, consisting of two
to four rows of somewhat large, spherical or
oval nuclei with nucleoli.

The outer molecular layer or fenestrated
membrane of the same appearance as d.

. The outer nuclear layer. In this the

nuclei are spherical or oval, smaller but more
numerous than in c.

. The outer limiting membrane, seen as a

sharp well-defined line.

The layer of rods and cones. Both limbs
of the more numerous rods may be easily
seen; the cones are shorter than the rods
and are less obvious.

. The pigment of the choroid with the fine

pencils of processes enveloping the outer
limbs of the rods and cones.

Tear off a small piece of the retina from a
sheep’s eye preserved in Miiller’s fluid; probably
the pigment-layer of the retina will be left ad-
hering to the choroid. Take a small piece of
this pigment-layer, and mount it in glycerine.
Observe en face the single layer of hexagonal
cells with large pigment-granules (cp. § 12, d).

Remove the retina of a frog immediately after
death, and place it in osmic acid for 30 minutes

18

274

ELEMENTARY PHYSIOLOGY, [XXVI.

to 24 hours, as convenient; tease out in water
in dilute glycerine. Observe the isolated frag-
ments of the various layers, noting especially

a. The large outer limbs of the rods, their

striation and transverse lamination.

b. The pigment-cells, each consisting of a
colourless outer part, containing the nucleus, |
and a coarsely pigmented part, which splits
up into a number of fibres, amongst which
are imbedded the heads of the rods.

DEMONSTRATION.

Effect on the iris of stimulating the sympathetic
nerve.

Muscles of the eye dissected in the head of a
sheep or dog.

LESSON XXVIII.
VISION.

Accommodation.

Remove very carefully the sclerotic and choroid
from a small portion of the posterior surface of
the eye of an ox or a sheep. Place the eye
in the end of a blackened tube just large
enough to hold it, with the cornea outwards.

On looking through the tube at any object, the
inverted image will be seen shining through the

retina.
Or,

The eye of any small animal, e.g. rat, may be
used without any preparation, the sclerotic
being sufficiently thin to be transparent. It
will be found convenient to place the eye, with
the cornea downwards, on the stage of a micro-
scope, and to direct light through it by means
of the mirror.

Standing some feet before a window, close one
eye and hold up before the other, at a distance
18—2

276

ELEMENTARY PHYSIOLOGY. [XXVIL.

of about six inches, a needle, so that it is at
right angles to one of the horizontal bars of the
window.

Look at the window-bar, the needle will appear

_dim and diffuse.

Look at the needle, the window-bar will appear
dim and diffuse.

The eye can accommodate itself for either the
needle or the window-bar, but not for both at the
same time. The accommodation for the near
object is accompanied by a distinct feeling of
effort.

Facing a window, or a white surface, close one
eye and hold a fine needle vertically before the
other. At about six inches the needle will be
seen distinctly. Bring it nearer the eye, the
image will be dim and diffuse, and at the same
time larger. The dimness and apparent increase
of size are due to diffusion, resulting from
imperfect accommodation.

Prick a small clean hole in a piece of card, hold
it before the eye and again bring the needle
close to the eye. It will be seen distinctly at a
much smaller distance than before, and at the
same time will appear magnified. It will be
seen more distinctly because the diffusion circles
are cut off. It will seem larger because, as it is
nearer the eye, the rays which pass through the
hole in the card are more divergent.

For a similar reason an object at a distance
viewed through the same hole will appear

XXVIZ] VISION. 277

smaller when the eye is accommodated for a
nearer object.

4. Prick in a card two small holes close together,
so that both are within the diameter of the
pupil. Holding the card before one eye (the
other being closed) with the holes horizontal,
hold a fine needle vertically at a distance of
twelve inches. It will be seen distinctly as one
when the eye is accommodated for it, but double
when accommodation is made either for farther
or nearer objects. Both the double images will
be dimmer than the single one.

When a double image appears and

a. The eye is adjusted for far objects; stopping
the right-hand hole will make the left-hand
image disappear.

b. The eye is adjusted for near objects; stopping
the right-hand hole will make the right-hand
image disappear.

When the eye is exactly accommodated for
the object, the rays passing through the two
holes are united at the focus on the retina.
In a, the rays fall upon the retina before
they are so united. Stopping the right-hand
hole removes the image on the right-hand
side of the retina. But images on the right-
hand side of the retina are referred by us
to objects on our left-hand side, hence it is
the left-hand image which disappears.

In b, the rays are united, and cross at the

278

ELEMENTARY PHYSIOLOGY. [XXVII.

focus before they fall on the retina. Stop-
ping the right-hand hole removes the image
on the left-hand side of the retina; and the
disappearance is referred to the right-hand
side. Construct a figure to shew this.
When a single image is seen, stopping either
hole merely makes the field of vision appear

— dim.

This is called Scheiner’s experiment.

The image being one, bring the needle closer to
the eye; at a certain distance it will become
double; this marks the near limit of accommoda-
tion. ;
Fixing the needle on a sheet of paper, walk
away while looking at it through the two holes;
at a certain distance it will become double, this
marks the far limit of accommodation. This
experiment succeeds best with short-sighted
people. |

Compare the near and far limits of accommoda-
tion as fixed by looking at a vertical needle
through horizontal holes with those fixed by
looking at a horizontal needle through holes
placed vertically. The results will differ ac-
cording to the amount of astigmatism in the
eye.

Helmholtz’s Phakoscope. This should be
used in a dark room. The observed looks with
one eye through the hole opposite the needle.
The observer looks through the hole at the side.

——

XXVIl.] VISION, 279

A lamp or candle is placed at some little distance
from the prisms and shifted about until the
observer sees on the eye of the observed, when
the latter looks at a distant object, two small
bright patches of light on the cornea, two larger
but dimmer patches on the anterior surface of
the lens, and two small and very dim (not readily
seen) patches on the posterior surface of the
lens.

Let the observed now accommodate for the
needle in front of him, making every effort not
to move the eyeball. The observer will see the
two patches on the anterior surface of the lens
approach each other, while the other two pair
remain motionless, thus shewing that during
accommodation for near objects the anterior
surface of the lens becomes more convex,
Observe that in accommodating for near objects
the pupil becomes smaller, and in accommo-
dating for far objects the pupil becomes larger.

7. Astigmatism. Draw on a card a star composed
say of eight lines passing through the centre, the
angle between each two neighbouring lines
being the same and the lines of equal tint and
of equal thickness.

Place this at about the distance which has been
determined (§ 5) as the far limit of accommoda-
tion (if this distance is more than eight or ten
yards use convex spectacles).

Probably one or more of the lines will be seen
much more distinctly and with less blurring than

ELEMENTARY PHYSIOLOGY. [Xxvu. | 9

the others. The direction of the line most
distinctly seen represents the meridian of least
convexity of the cornea. Approach gradually
nearer the star and note whether the other lines
become all visible at once or in succession.
Repeat this first with one and then with the
other eye closed; the astigmatism may be different
in the two eyes.

Then holding the star at a distance a little
greater than the near limit of accommodation,
fix the centre of the star with one eye, keeping
the other closed, and bring the star gradually
nearer; the lines will probably not all become
dim at the same moment; the line which
remains distinct longest will be the meridian |
of greatest curvature of the cornea; this is
usually at right angles to that of least curvature,
so that the line last seen with near accommoda-
tion will probably be at right angles to that first
seen with far accommodation.

Instead of the star a number of parallel hori-
zontal and parallel vertical lines may be drawn.

Or,

~_ Ea —-

Fix a needle vertically on a board. Looking at
the needle with one eye, accommodate the —
eye exactly for it. Then hold another needle
horizontally before the first, and move it back-
wards and forwards until both needles are seen
distinctly atthe same time. This will be found »
to be the case when the needles are at some
distance apart.

ae

XXVIL}

VISION, 281

More exact results are however gained by
Scheiner’s method (§§ 4, 5).

Irradiation.

Cut out two patches of exactly the same size, of
white and of black paper.

Place the white on a sheet of black and the
black on a sheet of white paper.

Place them some distance off and adjust the eye
so as to throw them a little out of the range of
accommodation.

The white patch will appear larger than the
black one.

Blind Spot.

Make a bold mark on a sheet of white paper,
place the sheet on the table, and, closing the
left eye, fix the axis of vision of the other, by
steadfastly looking at the mark at a distance of
about 10 inches. Dip a new quill-pen in black
ink and place it close to the paper near the
mark. Keeping the axis of vision fixed, and the
head at the same distance from the table, move
the pen outwards (towards the right) from the
mark. At a certain distance it will become
invisible; mark this spot on the paper. Carry
the pen still farther outwards. It will again
come visible; mark this spot also. The two
spots will indicate the outer and inner limits of
the blind spot. Similarly the upper and lower
limits may be traced, and with a little practice
an outline of the blind spot, shewing even the ~

10.

1].

ELEMENTARY PHYSIOLOGY. [XXVII.

commencement of the retinal blood-vessels as
they emerge from the edge of the optic disc,
may be constructed.

The size of the blind spot may be calculated

from the formula f- >) where f is the distance

of the eye from the paper, F the distance of the
retina from the nodal point of the eye (aver-
age =15 mm.), d the diameter of the outline on
the paper, and D the outline of the blind spot.

Region of Distinct Vision.

Fix the axis of vision by steadfastly looking at
some mark. Make on a card two dots so close
that they can just be seen as two when placed
close to the mark. Keeping the axis fixed, move
the card towards the periphery of the field of
vision; the dots will soon appear as one.

Or,

Make two dots on a card ‘5 mm. from each other.
Fix the axis of vision. Place the card outside
the whole field of vision and gradually move it
inwards towards the mark; the dots will first
become visible as one, and only appear as two as
they approach the centre of the field, and enter
the region of distinct vision. |
It will be found on examination that the outline
of this region is not circular but very irregular.

Purkinje’s Figures.
Go into a dark room with a lighted candle:

—_— = - -

XXVIL] VISION. 283

looking steadfastly with one eye towards a wall’;
hold the candle to the side of that eye so that
while the eye is illuminated the image of the
candle is not seen, and gently move the candle
up and down. In a few seconds the subdued
reddish glare caused by the candle-light will be
marked by branching dark lines, which will be
seen to form an exact image of the retinal
vessels as seen with the ophthalmoscope. The
dark lines are shadows of the blood-vessels;
consequently the structures in which the physio-
logical processes which give rise to the sensation
of light begin must lie behind the retinal blood-
vessels.

A cup-shaped space, in which the blood-vessels
are absent, may with care be seen; this is the
yellow spot.

Or,

Turn the eye inwards towards the nose so as to
expose as much as possible of the thin sclerotic
behind the cornea. Let an assistant with a lens
concentrate the rays of a candle or lamp on the
sclerotic as far behind the cornea as possible, so
that the rays may pass directly through it
towards the opposite side of the eye, and
gently move the focus to and fro. The same
image is still more distinctly seen. ‘The smaller
the focus on the sclerotic, the more distinct the
image.

1 A light-coloured wall or white blind is the best. A wall, the
paper of which has any very marked pattern, should be avoided.

284

ELEMENTARY PHYSIOLOGY. [XXVIL.

If the movement of the light is stopped, the
image soon fades away.

In the first method the image moves in the same
direction as the light when the light is moved
from side to side, but in an opposite direction
when moved up and down.

In the second method the movement of the image
is in the same direction as that of the light,
whether up and down or from side to side.

2. Look through a microscope with an empty field,

illuminated, but not too brightly, by a white
cloud, and gently move the head to and fro; an
appearance of fine retinal capillaries will be seen
as a dark meshwork on a finely punctated
ground. In the centre may be seen a finely
punctated spot devoid of capillaries. If the
head be moved from side to side, a vertical
meshwork will be seen; if up and down, a
horizontal meshwork.

The Yellow Spot. Maxwell’s Method.

Place a moderately strong, but perfectly trans-
parent solution of chrome alum in a flat-sided
glass vessel. Resting the eye for a minute or
two, suddenly look through the vessel at a white
cloud. A rosy spot or cloud will appear in the
centre of vision and remain for some little time,
but will gradually become less distinct.

The pigment of the yellow spot absorbs the
blue-green rays between the lines # and F of
the spectrum, these rays removed from those

——_

XXVIL.]

14.

VISION. 285

passing through the chrome alum, viz. red and
greenish blue, leave a rose colour.

Region of normal colour-vision.

Take small pieces of paper of various colours
(about 10 mm. square). Fix the axis of vision,
on a sheet of white paper, and place each
of the coloured papers near the axis of vision,
they will all be distinct.

Place a red piece near the axis, and then
gradually move it towards the outside of the
field of vision; it will first become yellow, then
a dull white, and will finally disappear. ;
Repeat this with a green piece, similar changes
are seen, but the yellow is darker.

Repeat this with violet, this becomes blue, then
a dull white, and finally disappears.

. Positive After-Image.

When waking in the morning, close and shade
the eyes for a minute or two, then suddenly
look at the bright window for a moment or two,
and then close and shade the eyes again. The
image of the window exactly corresponding to
the natural one, i.e. with the sashes dark and
the panes bright, &c., will last for some little
time.

To succeed, the retina should be in rest before-
hand, and the exposure to the stimulus momen-
tary or nearly so.

Or, in the evening, having closed and shaded
the eyes for some time, suddenly look at a lamp

286

16.

iv é

ELEMENTARY PHYSIOLOGY. [XXVII.

and immediately close the eyes. A similar posi-
tive after-image will be seen.

This positive after-image must not be confounded
with the negative after-image which comes later.
It simply shews that the sensation is of longer
duration than the application of the stimulus.

Look for an instant at a coloured light, and
then look at a white or grey surface, a positive
image of the same colour will for a brief period
be visible. |

Negative After-Image.
Look fixedly for about twenty seconds

a, At a white patch (e.g. white wafer) on a
black ground, and then look at a white
surface (or preferably pass a white surface
over the whole, keeping the visual axis
fixed); there will be visible a corresponding
dark patch on the white ground.

b. Ata black patch on a white ground, and turn
to a grey surface; there will be visible a
white patch on a grey ground. /

c. Atared patch on a black ground, and turn
to a white surface; there will be visible a
blue-green patch.

And so with the other colours, the colour of
the negative image will be complementary to
that of the actual object.

d. At ared patch on a black ground, and turn

XXVIL]

18.

19.

~YVISION. 287

to a yellow surface; there will be visible a
green patch.

e. Look fixedly at a brightly illuminated win-
dow and then close the eye. The positive
after-image will probably not be seen; in its
place there will come the negative after-image
with the sashes as bright lines and the panes
as dark spaces. This will in turn be suc-
ceeded by coloured images.

Contrast.

Place three candles in front of a white, otherwise
un-illuminated surface; pass between them and
the surface an opaque body with a sharp clean-
cut edge, so that part of the surface is illuminated
by the three candles, part by two, part by one
and part un-illuminated; stand two or three
yards back and look fixedly at the junction lines
of the variously illuminated surfaces; it will be
seen that each area is lighter close to a darker
surface and darker close to a lighter one than it
is elsewhere.

Simultaneous Contrast.

a. Cut out a thin cross of grey paper, and place
it in the middle of a sheet of bright green
paper. Cover the whole with a sheet of thin
tissue paper. The grey patch will appear
pink. The exact tint of the patch will
depend on the tint of the green, of which
it will be the complementary colour.

288

ELEMENTARY PHYSIOLOGY. [XXVII.

Surround the grey cross with a broad dark ~
black rim. The effect of contrast will be
lost; the grey patch will appear grey.

On a red ground the grey cross will appear
green, and with the other colours similar
complementary effects will be produced; but
the results are most striking in the case of
red and green.

The effect is greatest when the patch is grey,

- not white, and is always heightened by cover-

ing with tissue paper.

. Cuta thin strip of grey paper and place it

across the junction of a red with a green
paper, and cover with tissue paper.

The grey will appear green on the red side
and pink on the green side.

Place a sheet of white paper on a table before
a window illuminated by reflection from a
white cloud, not with direct sunlight. On
the side of the paper opposite the window
place a lighted candle, and between it and
the paper place a book edge-ways, or any
object which will throw a shadow on the
paper. Between the paper and the window
place a similar object, throwing a like shadow.
The distance of the candle should be such
that the two shadows are of nearly equal
intensity.

The shadow from the candle, though illumi-
nated by the white sunlight, will appear blue,
the complement of the reddish yellow colour

20.
L.

‘

VISION. 289

of the rest of the paper illuminated by the
candle.

This effect of contrast is subjective, not
objective. To shew this, blow out or place a
screen before the candle so as to do away
with the candle-light, The place formerly
occupied by the shadow now appears white.
Through a small black tube, e.g. a piece of
black paper rolled up, with so small a bore
that the whole field of view lies within the
shadow, look at the centre of the area formerly
occupied by the shadow. It still of course
appears white. While looking, let some one
light the candle, or remove the screen. No
change will be visible to the observer looking
through the tube.

If the tube however be directed partly on
the area of the shadow, and partly outside,
the blue tint of the shadow will, on the candle
being lighted or the screen removed, become
apparent as before.

The daylight-shadow heightens the effect on
the candle shadow, but may be dispensed
with. :

In place of sunlight and candle, two coloured
lights may be used.

In the above experiments (§§ a, b, c) avoid look-
ing at the colours too fixedly and for too long a
time. Otherwise the results will be modified by
after-images,

Visual Judgements. Measure on a card two
19

290 ELEMENTARY PHYSIOLOGY. [X XVII. 9

equal squares; without putting in the outline of
the squares, fill in one with fine vertical limes
one to two millimetres apart and the other with
similar horizontal lines the same distance apart.
Place them a short distance off, they will appear
not square but oblong. The side at right angles
to the direction of the lines looking longer than
the side parallel to the direction of the lines.

DEMONSTRATIONS.
1. Kiihne’s artificial eye.

Mixing of colour sensations with the colour top.

3. Method of testing colour blindness with skeins of
coloured worsted.

LESSON XXVIII.

THE EAR.

A. SKATE.

ms

Cut through with a strong sharp scalpel or with
bone forceps the cartilaginous roof of the skull
transversely between the eyes; remove the
posterior part of the roof. Opposite the hinder
part of the brain the cartilage will be seen to be
much thicker than elsewhere; it contains the
vestibule and semicircular canals: slice it away
till one of the semicircular canals is reached;
when this is done, trace it, removing the upper
portion with a scalpel or a strong pair of scissors.
Note

a. The almost transparent membranous canal
much smaller than the cartilaginous canal
in which it lies.

b. The ampulla or spindle-shaped dilatation of
the membranous canal close to one end.

c. The opening of the semicircular canal at
either end into a large membranous bag,
the utriculus.

19—2

292

' ELEMENTARY PHYSIOLOGY. [ XXVIII.

d. The sacculus, a membranous bag not so
large as the utriculus and separated from it
only by a shallow constriction.

e. The rudiment of a cochlea appearing as a
small projection from the anterior end of the
sacculus.

Trace out the three ‘semicircular’ canals, the
horizontal canal, the anterior and posterior
vertical canals, the two latter uniting at their
non-ampullary ends. Observe that the planes
passing through these canals are at right angles
to one another. A tube may be noticed running
upwards from the utriculus, close to the point of
entrance of the two vertical canals, to open on
the surface of the body: the recessus vestibuli of
mammals is homologous with this.

Cut through a membranous semicircular canal
and pull up with forceps the part connected with
an ampulla; the canal separates from the carti-
lage readily, but the ampulla is more firmly
attached at one spot, the crista acustica,
where a branch of the auditory nerve enters;
cut through the ampulla at either end, and with
a sharp scalpel cut through the nerve close to
the cartilage ; cut open the ampulla on the side
opposite the entrance of the nerve and note the
ridge of the crista acustica running transversely
a third to a half way across the tube, where the
nerve enters it.

XXVIII] THE EAR. 293

4.

At the lower part of the utriculus and sacculus
note the white calcareous paste, lift up the
membranous bag and note that beneath the
paste, nerve fibres enter through the cartilage.
Remove the membranous vestibule and canals,
sop up any fluid with a sponge and moisten the
cartilaginous vestibule and canals with osmic
acid; in a short time the nerve fibres entering
through the cartilage become blackened and so
very distinct.

Trace towards the brain, cutting through the
cartilage, one of the nerve bundles, e.g. that
entering the utriculus.

Lay bare the semicircular canals in a fresh skate
and cut out an ampulla as in § 3; place it in
picric acid or in chromic acid ‘2 p.c. for a week,
treat with alcohol, stain with hematoxylin, and
imbed, soaking previously a short time in the
imbedding mixture, clear the sections on the
slide and mount in Canada balsam.

In sections through the crista acustica observe

a. The outer loose connective tissue, and
b. The thick connective tissue wall.

c. The numerous large nerve fibres running
from the outside towards this; the fibres
may be traced through the membrane be-
coming much smaller and some losing their
medulla,

294

ELEMENTARY PHYSIOLOGY. [ XXVIII.

d. Two or more layers of cells with large

nuclei; long processes may be seen to
proceed upwards from some of these.

e. Superficial columnar cells. |

f. Fine fibres projecting beyond the columnar

cells; if care has not been taken these will
have been broken off; in any case it will be
difficult to make out whether the processes
run between the cells or from their extremities.
In sections through any other part of the
ampulla observe,

g. The thinner connective tissue wall,

h. The single layer of short columnar or even
flattened cells,

Place another ampulla in osmic acid 1 p.c. for
half-an-hour or longer; tease out a piece of the
crista acustica in water or in dilute glycerine and
observe the nerve fibres with blackened medulla
in the wall of the tube, and the isolated cells

(§§ 0, c, d).

Scrape off a little of the epithelium from the
crista acustica of a fresh ampulla and tease out

in a drop of endolymph to observe the characters

of the fresh cells.

——————— —. ae —

dl eek ial aT ita ables Rane a

= ©

pe et ee! =

s XxvuL] | THE EAR. ux 295

B. MAMMAL.

1. Take the head of a cat’, and cut away all the
soft tissues surrounding the tympanic bulla,
having snipped off the external meatus as close
to the skull as possible, look down and observe
the tympanic membrane; it is placed obliquely
to the canal and faces forwards, outwards, and
downwards. The attached handle of the malleus
may be seen shining through it.

2. Place the head with the lower jaw uppermost,
and with a strong pair of forceps break away
piecemeal the projecting part of the bulla. The
cavity thus exposed has a floor? made irregular
by projections. Note in the centre a projection
of somewhat yellowish bone running in the long
axis of the bulla; this is the promontory of the
cochlea: at a lower level in the exterior and
posterior part of it will be seen a round depres-
sion, the foramen rotundum.

3. In front and outside the cochlea is a projection
of thin bone which prevents the membrana
tympani from being seen; it divides the tym-
panic cavity into two parts communicating by

1 The details of arrangement differ considerably in different
animals: the following description applies to the cat, the more
general features may however be made out in other mammals,

2 It must be particularly noticed that the words floor, roof,
exterior, posterior, etc. are here used with reference to the position of
the head during the dissection; the position of the parts when the
head is placed in the normal position should also be observed.

296

ELEMENTARY PHYSIOLOGY. [ XXVIII.

an aperture over the foramen rotundum; this
arrangement is a characteristic of the Felide ;
break through the bone from above and very
carefully remove it in pieces with forceps. The
rest of the cavity of the tympanum is thus ex-
posed.

Note the funnel shape of the membrana tym-

pani; it has the handle of the malleus attached

to it; this proceeds from the outside attachment
of the membrane slightly upwards to a little past
the middle point of the membrane (cp. foot-notes,
p- 295).

Running out from a bony rim behind the mem-
brana tympani in the outer part, will be seen a
band of tissue stretching to the outer, anterior
edge of the bony rim surrounding the foramen
rotundum ; from this a thin ligament proceeds
at right angles to the former band to be inserted
into the head of the malleus. This is the
posterior ligament of the malleus.

On the opposite side of the head of the malleus
and in the same straight line, note the processus
gracilis running down to the lower edge of the
membrana tympani; it is attached here to the
bony wall by the anterior ligament of the
malleus; do not attempt to trace the ligament
till the malleus is removed later on. The tissues
in the straight line thus followed down form the
axis band of the malleus, i.e. the axis about
which the ossicula auditis turn.

XXVIII] THE EAR. 297

6.

Proceeding from the head of the malleus nearly
at right angles to the axis band, inwards and
downwards is a bony process to which is attached,
by a very short tendon, the tensor tympani
muscle. Press this towards its origin with a
seeker and note that it tightens the membrane.
Press lightly.on the end of the handle of the
malleus, it has but a small excursion.

With a fine pair of scissors cut through the
attachment of the membrana tympani, except
at the handle of the malleus, and with a small
pair of bone forceps remove the upper part of
the bony ring to which it was attached. The
malleus will remain in position. The incus and
stapes may be indistinctly seen deep down on the
outer side.

In order to make out well the relation of these
great care is required. Take a fine saw and
proceeding from behind in a plane passing just
outside the incus, saw away the external piece of
bone. .

Observe then

a. The rather long slender neck, and knobbed
head of the malleus.

b. The incus with two processes, one passing
almost horizontally backward and by a liga-
ment attached to the bony tympanic wall,
the other proceeding upwards, and attached
to the head of the stapes. When the bones
are removed later, the saddle-shaped surface

298

10.

aa

12,

ELEMENTARY PHYSIOLOGY. [ XXVIII.

of articulation of the incus with the malleus
should be observed.

c. The stapes, much more transparent than
the other bones; the base fits into an oval
depression, the fenestra ovalis. Observe
the stapedius muscle passing backwards
from the head of the stapes to the aque-
ductus Fallopii at the lower, outer part of
the cochlea.

Take out now the ossicula auditus, and observe
further the shape of each.

To the inner side, rather in front of the attach-
ment of the tensor tympani muscle, observe the
opening of the Eustachian tube ; pass a probe
down it and note the pharyngeal opening.

With a small pair of bone forceps, break away
the wall of the cochlea proceeding from the
foramen rotundum towards the apex of the bony
cone. Observe the coils of the cochlea with the

_ central piece of bone or modiolus and the pro-

jecting lamina spiralis.

With a strong pair of forceps remove the periotic
bone from the skull and cut through the cochlea
down the modiolus. Observe the nerves running
up its centre.

Starting from the fenestra ovalis the vestibule
may be exposed, the openings of the semicircular
canals seen and traced out in the bone, but this

XXVILL.] THE EAR. 299

13.

is not easy, and the arrangement differs but
little from that in the skate.

Take the cochlea of a mammal, preferably of a
guinea-pig, which has been treated with picric
acid until the salts have been dissolved out, and
subsequently with alcohol. Remove all the
softened bone as close as possible to the cochlea.
Slice away the apex of the cochlea, stain the
remainder with hematoxylin and imbed in the
manner given in Lesson xx. § 6. Cut sections
through the axis of the cochlea, throw away the
first two or three and the last two or three
sections; clear the rest on a slide with creosote
and turpentine, mount in Canada balsam.
Observe
a. The division of each turn of the cochlea into
three canals by the basilar membrane
running across from the end of the lamina
spiralis, and by the membrane of Reissner
starting from the lamina farther back; the
latter will very probably have been torn
through.
b. The following modifications of the epithelium
cells of the scala media, starting from the
inner side of the basilar membrane.

a. Cells passing from cubical to columnar.

8. The single inner hair cell, columnar
with short rod-like processes, the so-
called hairs, from its free surface, its
deep pointed end is more or less hidden
by small cells with large nuclei.

300

ELEMENTARY PHYSIOLOGY. [ XXVIII.

The inner and the outer rod of Corti. _

6. The three or four outer hair cells,
long irregular cells, not perpendicular
to the membrane, but bending inwards,
with short rod-like processes projecting
from the surface, and deeply seated
nucleus; occasionally two nuclei are
seen in a cell. |

my. The rings of the reticular membrane
surrounding the tops of the inner and
outer hair cells.

6. Cells passing from columnar to cubical,
inclining inwards, like the outer hair
cells.

The membrana tectoria proceeding from a
projection of tissue on the lamina spiralis,
thence enlarging and forming a more or less
distinct pad above the organ of Corti; in the
preparation it will probably be considerably
shrunken.

. The nerves running along the lamina spiralis

towards the basilar membrane.

LESSON XXIX.
SPINAL CORD.

MAKE transverse sections of the hardened spinal
cord of a cat or dog through the region of
the cervical enlargement (say at the origin of
the 5th cervical nerve). Stain the sections with
dilute carmine or picrocarmine for a day, clear
and mount in Canada balsam.

The cord should have been placed in ammonium or
potassium bichromate 2 p.c. for three or four weeks,
then washed with water, placed in 30 p.c. alcohol
for several days changing the fluid each day, then
placed in 50 p.c. alcohol, which should be renewed
until it no longer becomes coloured. The cord may
be kept in 75 p.c. or in strong alcohol. It is best
stained by placing the piece to be cut in strong
Frey’s carmine for several days, it may then be
well washed with water, soaked in gum for a day,
and cut with the freezing microtome.

Observe under a low power the following general
features

a. The pia mater, surrounding the cord; it
consists of two coats (cp. § 3 a.); the inner

302

ELEMENTARY PHYSIOLOGY. [XXIX.

coat sends into the cord numerous septa.
Note the blood-vessels running from the pia
mater into the cord along the septa.

The anterior and posterior fissures; the
pia mater dips down into the anterior fissure;
the posterior fissure contains a prolongation
of the inner coat only of the pia mater, the
outer coat running over the fissure.

The entrance into the cord of the anterior
roots of the spinal nerves in several small
bundles.

. The entrance into the cord of the posterior

roots of the spinal nerves in a compact
mass.

The white substance forming the outer
part of the cord and divided on each side by
the entrance of the nerve roots into anterior,
lateral and posterior columns; since the
fibres of the anterior root do not enter the cord
in one bundle (cp. c.) there is no definite line
of division between the anterior and lateral
columns.

The grey matter projecting on each side
into an anterior and a posterior cornu,
but with no distinct separating line between
them.

. The central canal; it may be plugged up

with epithelial cells ;

. A ring of deeply stained neuroglia around

the central canal, and

XXIX. |

9

ae

1.

SPINAL CORD. 303

On either side of this, the anterior and pos-
terior grey commissures of deeply stained
substances in which crossing nerve fibres are
seen.

The anterior white commissure consisting
of fairly large decussating nerve fibres in

- front of the anterior grey commissure.

A round or oval deeply stained area, the
substantia gelatinosa, forming the hinder
part of the posterior cornu.

Examine the same section first under a low and
then under a high power. Observe

a. The large multipolar nerve cells of the an-

terior cornu placed chiefly in the anterior
and lateral part of the cornu; they may be
more or less distinctly arranged in two or in
three groups.

The transverse fibres (longitudinal in the
plane of the section) running from the
posterior part of the anterior cornu into
the lateral column.

The fibres of the posterior root running
(a) transversely through the substantia gela-
tinosa, (8) obliquely into the posterior
column.

Transverse and oblique fibres running from
the posterior column into the posterior
cornu.

304

ELEMENTARY PHYSIOLOGY. [XXIX.

Under a high power repeat the observations
of § F, Lesson x. and observe further

a.

The outer coat of the pia mater consists

‘chiefly of white fibrous tissue; the inner

chiefly of fine elastic fibrils.

The central canal is lined with a layer of
columnar epithelium cells.

In the grey substance, a fine small-meshed
network af fibrils, lying in a punctated or
granular-looking matrix.

The nerve fibres, many medullated, of all
sizes, throughout the grey substance (in the
ring around the central canal the nerve fibres
are very few and small, in the substantia
gelatinosa they are chiefly the fibres of the |
posterior roots). Note the fibres running
across the anterior and posterior grey com-
missures and compare the anterior grey with
the white commissure.

Prepare transverse sections of the spinal cord of
a cat or dog taken through the dorsal and lum-
bar regions (say through origin of 9th dorsal and
4th lumbar nerves) and compare them with the
section of the cervical region. Observe

a. The sections taken from the cervical and

b.

lumbar regions are larger than those taken
from the dorsal region.

The enlargements in these regions are largely
due to an increase in the quantity of grey

XXIX.]

L.

SPINAL CORD. 305

matter; note the relatively small number of
nerve-cells in the anterior cornu of the dorsal
region,

The quantity of white substance, (number of
nerve fibres) is greater in the dorsal than in
the lumbar region, and greater in the cervi-
cal than in the dorsal, i.e. the quantity of
white substance in the cord increases from
below upwards.

. Int the cervical region the posterior column

is divided into two parts, a median part, the
fasciculus of Goll, and a lateral part, the
fasciculus cuneatus, by a septum of con-
nective tissue (the fasciculus of Goll may
be traced some distance down the dorsal
region becoming smaller in its course).

In the dorsal region there is a small lateral
collection of nerve cells in the grey substance
behind and laterally of the central canal;
this is Clarke’s column, or the posterior
vesicular column. It becomes indistinct in
the upper dorsal and lower lumbar regions.

In the dorsal region there is a lateral pro-
jection of the grey substance about mid-way
between the anterior and the posterior cornu,
this is the tractus intermedio-lateralis ; in it
is a group of nerve cells; in the cervical and
lumbar regions this group of cells blends into
that of the anterior cornu.
20

306

ELEMENTARY PHYSIOLOGY. [XXIX. .

g. In the lumbar region, the nerve fibres of the
anterior root spreading out in the grey sub-
stance, some may be seen running across to
the other side of the cord and forming part
of the anterior white commissure (the course
of the fibres of the anterior root is the same
in the other regions of the cord but usually
less distinct).

The Student should be able to recognize a sec-
tion of the spinal cord as belonging to the cervi-
cal, the dorsal or lumbar region; in doing this,
the chief points to be noticed are:—the presence
or absence of Goll’s column, of the tractus inter-
medio-lateralis (lateral cornu), and of the collec-
tion of cells forming Clarke’s column; the
relative amount of white and of grey substance ;
the shape of the anterior and of the posterior
cornu ; and the position of the central canal. .

Place a short piece of the spinal cord of an ox
in potassium bichromate °2 p.c. for a few days,
and after the chromium salt has been removed
by 30 p.c. alcohol, place the piece in strong
Frey’s carmine for a week or longer. With free
hand cut a thick section from the anterior
cornu and the neighbouring part of the anterior
column and tease it out ina small quantity of
glycerine under a dissecting microscope, trying
to isolate one or more of the large nerve-cells
seen and throwing away the rest of the tissue.
In the more or less isolated cells observe

XXIX ] SPINAL CORD. 307

a. The single unbranched axis-cylinder process.

b. The numerous remaining processes (proto-
plasmic processes) frequently branching, some
may be seen to divide into very fine fibrils.

6. Cut an antero-posterior longitudinal section
passing through both the anterior and the pos-
terior nerve-roots. Note

a. The longitudinal medullated fibres of the
anterior column.

b. The anterior nerve-roots entering obliquely.

c. The nerve-cells and fibres of the anterior
cornu.

d. The substantia gelatinosa of the posterior
cornu, with vertical fibres on either side and
curved fibres running through it.

e. The longitudinal medullated fibres of the
posterior columns, in which can be seen

f. The cut ends of the posterior nerve-roots.

DEMONSTRATION.

Section of spinal cord prepared by Weigert’s
method to show medullated fibres in the grey
substance. (Cf. p. 396).

20—2

LESSON XXX,
THE BRAIN,

A. DISSECTION OF THE BRAIN‘ OF A DOG OR OF A
SHEEP.

1. Note the dura mater and pia mater (ep. Lesson
1. B§ 3, 5, c).

2. Cutting away the dura mater and carefully
handling the brain to avoid tearing the nerve-
roots, observe

a. The cerebrum overlaps the anterior part of
the cerebellum (the overlapping is less in
the sheep than in the dog),

Ss

1 The brain should be removed from the skull as carefully as
possible, especial pains being taken to cut the internal carotid arteries
and the cranial nerves close to the skull. When the brain is removed
it is best to tie the internal carotid arteries, and inject into the
basilar artery first salt solution to wash out the blood as much as
possible, then dilute and finally strong spirit; it is then placed in
strong spirit; it may be dissected in a few days, but it is better to
leave it in the spirit for three or four weeks. If the brain be not
injected, it should be placed for two or three days in weak spirit of
about 50 p.c., and then transferred to 90 p.c. spirit; in this it should
be left for a month or more to harden it thoroughly.

Xxx. }

b.

THE BRAIN. 309

The cerebellum covers the dorsal part of the
medulla oblongata.

Tearing away as much of the pia mater as may
be necessary, turn forward the cerebellum; the
posterior dorsal surface of the medulla oblongata
will be seen. Note

a.

The choroid plexuses of the fourth ventricle,
appearing on either side as a very vascular
projection of the pia mater; they lie immedi-
ately above the thin epithelial roof of the
ventricle; tear them away to expose the pos-
terior half of the fourth ventricle.

The diverging posterior columns of the spinal
cord ; the fasciculus of Goll (cp. Lesson xxrx.
§ 4, d.) is continued on as the fasciculus
gracilis which forms the lateral wall of the
posterior part of the fourth ventricle; late-
rally of this is seen the fasciculus cuneatus
continuous with the fasciculus cuneatus of
the cord (ep. Lesson XxIx. § 4, d.).

The oblique fibres running from the lateral
and anterior columns over the fasciculus
cuneatus and apparently blending with it;
the depression between the fasciculus gracilis
and the fasciculus cuneatus at the same time
disappearing, so that soon after the posterior
columns have diverged, a single rounded
eminence, the restiform body, is seen.

Anteriorly the restiform bodies run into the

310.

ELEMENTARY PHYSIOLOGY. [XXX.

cerebellum, constituting the inferior pedun-

cles of the cerebellum.

Turn back the cerebellum, tearing away the pia
mater which dips down in front of it. Observe

a.

The corpora quadrigemina, consisting of
two round and rather large anterior bodies,
and two smaller posterior bodies. The an-
terior corpora quadrigemina are partially
covered by the cortex of the cerebrum.

The superior peduncles of the cerebel-
lum, one on each side, proceeding from
the cerebellum as a roundish cord and dis-
appearing underneath the posterior corpus

quadrigeminum of the same side.

The valve of Vieussens, a thin layer of
nervous substance stretching between the
superior peduncles, and covering in the an-
terior part of the fourth ventricle. In the
anterior part of the valve the roots of the
fourth nerve may be seen rising from the
middle line, and curving round to reach the
base of the brain.

Tear away the valve of Vieussens and observe
the anterior triangular part of the fourth
ventricle, its lateral boundaries are the -
superior peduncles of the cerebellum.

Note on the under (ventral) surface of the

‘medulla oblongata, without tearing away the

pia mater,

XXX.]

THE BRAIN. 311

. The anterior pyramids, two rounded cords

one on either side of the median line.

The pons Varolii. At its lower edge are
transverse fibres forming a narrow band, the
trapezium, which dips down underneath
(dorsally of) the anterior pyramids; anteriorly
to this are transverse fibres forming a broad
band which runs over (ventrally of) the ante-
rior pyramids; the band has a median shallow
depression. In man the fibres of the trape-
zium are not seen on the surface of the pons.
Note on each side the continuation of the
transverse fibres of the pons into the middle
peduncle of the cerebellum.

The inferior olivary bodies, two slight
oval elevations one on each side, laterally of
the anterior pyramid and just below the
trapezium. These will be seen more dis-
tinctly when the pia mater has been torn

away (cp. § 10).

. The crura cerebri or peduncles of the

cerebrum, two broad roundish bands which
appear at the anterior edge of the pons, and
run forwards diverging from one another.

The posterior perforated space between
the diverging fibres of the crura cerebri.

The round projecting corpus albicans or
mammillare in front of the posterior perfo-
rated space. In the dog there is a shallow
median groove dividing it in two.

312

ELEMENTARY PHYSIOLOGY. [XXX

g. Immediately anterior to this a small area

. the tuber cinereum, from this springs »

the funnel-shaped infundibulum leading
to the round pituitary body; cut across
the infundibulum and observe the central
space, which leads into the third ventricle.
(The pituitary body may have been torn
away in removing the brain from the skull.)

h. The optic tracts, two flat bundles of fibres
coming obliquely forward over the front part
of the crura cerebri, and meeting in the
middle line to form the optic chiasma:
from this a small piece of the optic nerve
will be seen coming off on each side.

?

Observe the division of the cerebellum into a
median and two lateral lobes, cut through the
middle peduncle (§ 5, 6) on each side where it
passes from the cerebellum, and remove the cere-

- bellum; divide it into four pieces two by a longi-

tudinal and a transverse cut and note, the depth of
its fissures, the white substance radiating out-
wards in the lamine, and covered by a thin layer
of grey substance, the whole having a distinctly
arborescent appearance. In the midst of the cen-
tral mass of white substance in the lateral lobes
may be seen a greyer area representing the

' corpus dentatum of man; it is more distinct in

a brain preserved in ammonium bichromate.

Observe now more closely the exposed fourth
ventricle. It has a roughly rhomboidal shape ;

is

XXx.]

THE BRAIN. 313

its posterior triangular portion is the calamus
scriptorius; note the opening of the central
canal of the spinal cord into this; note also
at the anterior end of the fourth ventricle the
aqueduct of Sylvius or iter underneath the
corpora quadrigemina; it runs from the fourth
to the third ventricle (cp. §:19).

.Trace the chief blood-vessels running in the pia

mater of the under surface of the brain, tear-
ing away the pia mater where necessary but
being careful not to tear away at the same time
the nerve-roots. At the upper part of the
medulla will be seen two arteries. These are
the vertebral arteries, which having given
off recurrent branches along the anterior fissure
of the medulla, curve round to unite in the
median line; the vessel formed by their union
is called the basilar artery, and runs forwards
in the median line of the pons Varolii. At
the front edge of this it divides into the two
posterior cerebral arteries, each of which
running obliquely forward passes to the median
side of the third nerve as it springs from the
crus cerebri.

Just beyond this each posterior cerebral divides
into two branches. One of these runs _ back-
ward; the other proceeding forwards, is, a little
behind the optic commissure, joined by the
internal carotid artery. [Each arterial trunk
so formed passes round the optic commissure,
and divides into the middle cerebral artery

314

ELEMENTARY PHYSIOLOGY. [ XXX.

which crosses the olfactory lobe, its main branch
running in the fissure of Sylvius, ep. C, § 6,
giving off numerous branches, and the anterior
cerebral artery which passes forwards between
the front lobes of the cerebrum, having a trans-
verse communicating branch with its fellow of
the opposite side. The anastomoses between
the branches of the posterior cerebral and the
internal carotid arteries and those between the
two anterior cerebral arteries complete the
circle of Willis.

Now carefully tear away the pia mater and
observe the points of exit of the follwing nerve-
roots (cp. Lesson I. p. 12).

a. The third pair of nerves, arising from the
inner surfaces of the crura cerebri in front of
the pons Varolii (ep. § 8).

b. The fourth pair of nerves, which will be seen
curving round the frent edge of the pons;
they originate a little behind the corpora
quadrigemina (cp. § 4, c).

c. The fifth pair of nerves, large and con-
spicuous, arising from the sides of the pons.

d. The sixth pair of nerves, arising in front
of the olivary bodies and anterior pyramids
close behind the pons.

e. The seventh pair of nerves seen at the
lower edge of the pons nearly in a line with
the fifth pair, they arise from the trapezium
in a line with the outer border of the inferior

a

10.

11,

THE BRAIN. 315

olivary body. (Their superficial origin is
different in man.)

jf. The eighth pair of nerves just outside the

seventh; they cling to tke lateral surface of
the medulla, they will be seen on the dorsal
surface passing over the restiform bodies.
Note the small bundles of fibres (striz
acustice) passing over the restiform bodies
into the fourth ventricle.

g. The ninth, tenth and eleventh pairs of nerves
each arising by several roots which form a
line on the lateral part of the medulla; this
line in its lower portion is continuous with
the line of the posterior roots of the cervical
nerves; the eleventh pair has origin also
from the spinal cord, it may be traced down-
wards on the surface of the cord to about the
sixth cervical nerve.

h. The twelfth pair of nerves arising by severat
roots between the anterior pyramids and
olivary bodies.

- Observe again the ventral surface of the brain

(§ 5), removing the pia mater.

Separate the hemispheres and observe the
corpus callosum, noting its curve in front
(genu) and its curve behind (splenium).

Cut away in thin slices the dorsal surface of the
hemispheres, nearly down to the level of the
corpus callosum, noting the outer grey substance

_ and the inner white substance.

316

ELEMENTARY PHYSIOLOGY. [XXx.

Make a shallow cut along each side of the corpus
callosum and pull up the cortex on the outside
of the cut; a space, the body of the lateral ven-
tricle, will be seen; carefully cut away the roof
of this space, which will then be seen to run
forwards as the anterior cornu and backwards and
downwards as the descending cornu of the lateral
ventricle; on one side completely remove the
cortex above the cornua. Observe

a.

The nucleus caudatus of the corpus stria-
tum, seen as a large rounded projection into
the anterior cornu and continued backwards
as a tapering mass (tail of the nucleus) on the
outer side of the body of the ventricle.

The hippocampus major, a rounded projec-

tion of the floor and medial wall of the
descending cornu.

The floor of the body of the lateral ventricle
consisting of a thin lamina (cp. § 15), the

fornix; continuous posteriorly with the ~

hippocampus.

The choroid plexus of the lateral ventricle
covering the lateral edge of the fornix and of
the hippocampus. At the junction of the
body of the ventricle with the anterior cornu
the choroid plexus dips down underneath
the fornix.

Turn laterally the choroid plexus and observe
the band of white substance running from the
fornix along the edge of the hippocampus
forming the fimbria of the hippocampus,

XXX. ]

13.

14,

THE BRAIN. 317

f. Carefully pass a seeker underneath the edge of
the fornix and of the fimbria; note that they
can be raised from the subjacent parts and
that the choroid plexus dips underneath them.

Gently raise the corpus callosum,a thin membrane
the septum lucidum will be seen stretching
from its under surface to the fornix and sepa-
rating the lateral ventricles of the two sides.

Cut through the corpus callosum near its poste.
rior end and turn it forwards, cutting through
the septum lucidum ; indications of the forma-
tion of the septum from two membranes will be
seen, in its anterior portion a narrow space the
fifth ventricle may be seen. Cut through the
corpus callosum anteriorly and remove it.

The parts of the fornix previously noticed in
each lateral ventricle are now seen to be joined
in the median line forming a triangular lamina.
Anteriorly this dips down some little distance
behind the genu of the corpus callosum; as it
dips down it divides into two round cords, the
anterior pillars of the fornix (these will be.
better seen when the fornix is cut through
(§ 15)); the part of the fornix where these
pillars are joined is called the body, this is
small in lower mammals since the pillars soon
diverge posteriorly ; where they diverge they are
called the posterior pillars of the fornix, these
run on to form the fimbrize of the hippocampus
(cp. § 12, e) and also send fibres which run over
the inner surface of the hippocampus.

318

15.

ae 6.

ELEMENTARY PHYSIOLOGY. [XXxX.

Carefully cut through the anterior portion of
the fornix and turn it back, being careful not to
drag with it the choroid plexus; the choroid
plexus of each side will be seen to curve back
slightly and join with the other in the median
line; the space left on either side between
the choroid plexus and the anterior pillars of
the fornix is part of the foramen of Monro.
The remaining part of the foramen of Monro
is the space in the median line between the
anterior portions of the choroid plexuses where
they join; this leads into the third ventricle
(cp. § 19); the foramen of Monro is thus roughly
a Y-shaped space, one limb communicating with
the third ventricle and each of the other two
with a lateral ventricle at the junction of the
body with the anterior cornu.

The vascular membrane underneath the fornix
is the velum interpositum, this is seen to be
continuous anteriorly with the recurved ends of
the choroid plexuses about the foramen of Monro
and laterally with the whole length of the
choroid plexus of the lateral ventricles, in fact
the choroid plexuses are only the free borders of
the velum interpositum curving over the edge of
the fornix and over its posterior pillars. (It is to
be remembered that since there is an epithelial
membrane running from the fornix over the
choroid plexuses to the edge of the nucleus
caudatus, the lateral ventricle has no opening
except at the foramen of Monro.)

EE

XXX.]

17.

18.

19,

THE BRAIN. ’ 319

The space between the cortex and the mid-brain
in which lies the velum interpositum is the so-
called transverse fissure.

In the median line cut through the fornix, with
the posterior part of the corpus callosum which
remains attached to it;

Note that the cortex curls a short way under- —
neath the corpus callosum.

On one side pull the fornix and hippocampus
backwards; note on the outer (lower) surface of
the cortex, the dentate or hippocampal fissure;
it is shallow and is situated a short distance from
the edge of the fimbria, nearly opposite the
middle of the hippocampus; it runs from the
cortex underneath the corpus callosum (ep. § 17)
to the extremity of the cortex of the descending
cornu of the lateral ventricle; the projection of
the hippocampus seen in the descending cornu
is caused by the folding of the cortex round
this fissure; note the lower surface of the pos-
terior pillars of the fornix.

Turn back the velum interpositum; in separa-
ting the velum posteriorly note two projections
downwards into the median space, these are the
choroid plexuses of the third ventricle.

The optic thalamus will now be seen on each
side, a depression runs round its lateral boundary;
between the optic thalami is a narrow space, the
third ventricle. Note the tail of the nucleus
caudatus stretching backwards for some distance
laterally of the optic thalamus.

320

20.

21.

ELEMENTARY PHYSIOLOGY. [XXX.

Note at the posterior part of the optic thalami
in the median line the round pineal gland, and
running from this near the middle line two
thin white bands, the peduncles of the pineal
gland, one over the optic thalamus of each side.
Between the optic thalami, and running across
the third ventricle, is seen the large middle
cerebral commissure, of grey substance and
hence very readily torn through.

Cut away on one side the cortex so as to
completely expose the optic thalami and optic
tract, and trace the course of the optic tract,
carefully tearing away the pia mater. The optic
tract will be seen to curve dorsally and form an
eminence, the external corpus geniculatum,
at the posterior lateral part of the optic thala-
mus; over this may be seen fibres running to
the optic thalamus, and a rather large band of
fibres curling back and running into the anterior
corpus quadrigeminum (the so-called brachium
of the ant. corp. quad.),

Below and behind the external corpus genicula-
tum a small eminence, the internal corpus
geniculatum.

Both geniculate bodies are more marked in the dog
than in the sheep; the position of these bodies is
different in man; in man the backward projection of
the optic thalamus or pulvinar has the position here
occupied by the external geniculate body,

Disappearing underneath the posterior edge of
the internal corpus geniculatum will be seen a

-_ ee

ee

XXX.]. THE BRAIN. 321

band of white fibres, the brachium of the pos-
terior quadrigeminum.

22. Note the anterior cerebral commissure, a
small compact bundle of fibres running trans-
versely in front of the anterior pillars of the
fornix.

Cut through the anterior pillars ¢ the fornix to
expose the commissure.

23. The posterior cerebral commissure under-
neath the pineal gland.

24. Cut through the anterior and middle commis-
sures and trace the cavity of the third ventricle
into the infundibulum ; cut through the posterior
commissure and the corpora quadrigemina and
trace out the aqueduct of Sylvius.

B. PARTS OF THE BRAIN SEEN IN SECTIONS.

1. Divide a brain in half by a longitudinal sec-
tion carried carefully through the median line.
Observe the relative positions of the structures
visible on the cut surface, noting
The obliquely cut fibres in the decussation of
the pyramids; the transversely cut fibres of the
ventral part of the pons; the valve of Vieussens;
the corpora quadrigemina; the aqueduct of Syl-
vius; the posterior commissure, this will be con-
tinuous with transversely cut fibres forming a
layer in the anterior corpus quadrigeminum ; the
pineal gland, a small piece of its peduncle running
from it will be seen; the large middle commissure
occupying a considerable portion of the third

L. 21

322 ELEMENTARY PHYSIOLOGY. [Xxx.

ventricle; the corpus callosum; the septum
lucidum, deep anteriorly, between the corpus
callosum and the fornix; the pia mater entering
the transverse fissure, forming there the velum
interpositum which les underneath the fornix
and over the optic thalami; the anterior pillar
of the fornix curving downwards in the direction
of the corpus albicans; the anterior commissure
a little in front of the apparent termination of
the anterior pillar of the fornix; the posterior
perforated space behind the corpus albicans ; the
optic chiasma.

2. Cut a transverse section of the mid-brain’ passing
a little in front of the pons and through the
anterior corpora quadrigemina. Observe
a. Ventrally, and a little removed from the
median line on each side, the crusta of the
cerebral peduncle, it forms a slight project-
ing oval mass of transversely cut fibres.

b. A thin grey layer, the substantia nigra
above the crusta.

c. The central grey substance around the

- aqueduct of Sylvius.

d. The tegmentum, between the substantia
nigra and the central grey substance, else-
where it does not show distinct boundaries.

3. Cut a brain through transversely carrying the
cut a little in front of the corpus albicans and
1 The parts described in this and in the succeeding section are

much better seen in a brain which has been hardened in ammonium
bichromate than in one hardened in alcohol.

XXX.]

THE BRAIN. 323

across the Sylvian fissure (cp. C. § 6). Note on
surface exposed

a.

The band of white substance, the inner
capsule, laterally of the optic thalamus.

The bands of white substance, corona radi-
ata, continuous with the inner capsule and
spreading out into the convolutions of the
cortex,

The corpus callosum stretching between the
coron radiate of the two sides,

The nucleus caudatus of the corpus stria-
tum,a roundish grey mass, dorsal and lateral to
the optic thalamus, and partially surrounded
by the deep white band of the corona radiata;
it may be very small in this section.

The nucleus lenticularis of the corpus stria-
tum; this occupies a small triangular space
just outside the inner capsule, and has a num-
ber of fibres running transversely through it
from the inner.capsule ; on its lateral margin
is a thin white band, the outer capsule.

The anterior pillar of the fornix will also be
seen cut across, near the ventral surface of
the grey substance close to the third ventricle;
note also the position of the fornix,

C. CONVOLUTIONS AND FISSURES OF THE Doq’s BRAIN.

1,

Observe on the ventral, anterior surface of
the brain the olfactory bulb (this may have

21—2

324

~J

ELEMENTARY PHYSIOLOGY. [XxXx.

been, cut off) continuous posteriorly with the
olfactory tract; pull this a little outwards, it
is unconnected with the brain except posteriorly
where it runs into the olfactory lobe.

Behind the olfactory lobe is a somewhat pear-
shaped bulging, the lateral part of the uncinate
lobe.

Cut the brain in half through the median longi-
tudinal fissure, press down the cerebellum and
follow the course of the uncinate lobe in the
posterior part. of the cortex; it curves round as
a rather narrow -band and then runs upwards to
the end of the corpus callosum, it runs also a
little forwards underneath the corpus callosum
(cp. A. § 17).

At the end of the corpus callosum it joins the
supra-callosal convolution which runs for-
wards above the corpus callosum.

It will be seen that the above convolutions form
a nearly complete ring round the cortex, they
are all parts of the internal convolution or
limbic lobe.

Turning back now to the side of the brain, note
the deep Sylvian fissure running upwards and
backwards from about the apex of the part of
the uncinate lobe here seen (ep. § 2); around
this curves the first or Sylvian convolution.

Curving dorsally around this is the second or
inferior convolution.

XXx.]

10.

a1.

12.

13.

THE BRAIN. 325

Dorsally again of this is the third or median
convolution having a longitudinal fissure in its

posterior part.

Between the third convolution and the median
longitudinal fissure is the fourth or superior
convolution.

Looking at the median surface of the brain ex-
posed by the cut made previously (§ 3), observe
that the superior convolution stretches down to
the supra-callosal convolution (cp. § 4).

In the anterior part of the dorsal surface of
the superior convolution, note the deep crucial
fissure running transversely and a little for-
wards, follow it on the median surface of the
brain; it is seen to run into the fissure that lies
above the supra-callosal convolution.

The bend of the fourth convolution around the
crucial fissure is the sigmoid gyrus, the part
in front of it being the anterior limb, the part
behind being the posterior limb.

Anteriorly and posteriorly the four external
convolutions join.

The above general plan of the convolutions is in
special cases modified to some extent by secondary
fissures, and by fissures being absent in part of their
normal course (thus the fissure between the Ist and
2nd convolution is not infrequently incomplete) so
that the typical arrangement given above may not
be obvious at first,

526

ELEMENTARY PHYSIOLOGY. [XXX.

14, -In front of the anterior joined ends of the ex-

15.

ternal convolutions is a deep fissure (the supra-
orbital) ; the part of the brain in front of this is
called the sub-orbital lobe.

Cut away the first convolution and note the
surface of the cortex, island of Reil, lying at
the bottom of Sylvian fissure,

D. HIsToLoGy,

1.

Cut transverse sections of the spinal cord just
above the origin of the Ist cervical nerve, at the
beginning of the decussation of the pyramids.
Note :

a. The anterior and posterior cornua of the
grey substance are partially separated from
one another by bundles of medullated fibres
springing from the lateral columns and ex-
tending some little distance in the grey sub-
stance towards the median line,

b. Some oblique bundles of fibres will be seen
running through the grey matter on either
side and crossing in front (ventrally) of the
grey commissure, forming part of the decus-
sation of the pyramids.

c. Medially of the anterior column will be seen
a thin band of transversely cut small fibres ;
these are the fibres which have already
crossed, the beginnings of the anterior pyra-
mid,

XXx.] THE BRAIN. 327

d, The grey substance extending into Goll’s
fasciculus of the posterior column, and into
the fasciculus cuneatus.

The appearances vary considerably in sections taken
at different parts of the decussation.

2. Take a brain of a rabbit’ hardened in ammonium
bichromate and subsequently treated with al-
cohol. Cut out a piece from the middle of the
dorsal portion of one hemisphere, extending from
the surface to the lateral ventricle, and prepare
vertical sections (preferably with a freezing mi-
crotome), Stain then with dilute carmine or
picrocarmine, clear and mount in Canada balsam.
Observe

a. The inner layer of horizontal nerve fibres
(medullated) forming the white substance ;

1 The fresh brain should be placed in ammonium or potassium
bichromate 2 p.c., the fluid changed on the following day (when the
brain may be cut transversely in four or five pieces) and again in a
week, then left about three weeks. It should then be cut up and the
pieces washed with water for a day and with dilute spirit (first 30 p.c.
then 50 p.c.) until all the excess of the chromium salt is taken out.
The pieces may be stained with strong Frey’s carmine during a
week or more, washed well with water, soaked in gum and cut with a
freezing microtome.

Instead of a rabbit’s brain the brain of a cat or dog may be taken;
a solution of the chromium salt may advantageously be injected into
the basilar artery, but even then it is not easy to preserve the natural
form of the cells of the 3rd and 4th layers of the cortex; the structure
of the cortex in the cat and dog differs also at different points more
than in the rabbit, the chief points of difference being the occurrence
of very large cells in groups or singly in the lower part of the third
layer, and the variation in number and extent of the angular cells,
which may spread into the third layer or may be inconspicuous,

328

. Outside (oO are large pyramidal cells, the

ELEMENTARY PHYSIOLOGY. [Xxx.

(between the fibres a considerable number
of leucocytes will be seen) ; from this bundles
of fibres at fairly regular intervals run out
into the grey substance, usually ceasing to
be distinct in the third layer (cp. d).

At the outer limit of this a layer of fusiform
nerve cells lying amongst nerve fibres and
forming the 5th layer of the grey substance
of the cortex.

Outside this, a layer of small cells of various
shapes, cells with three or more obvious
though small processes predominating (angu-
lar cells); these form the 4th layer of the
cortex.

process from the apex of the cell tapering off
from it and often being traceable upwards
for a considerable distance; from the base
three, four or more processes may be seen to
proceed. The district of the large pyramidal
cells forms the 3rd layer of the cortex; it
will be seen that it is much thicker than any
of the rest, and that generally speaking its
cells diminish in size from within outwards.
Small angular cells may be seen in its deepest
part.

Outside the preceding layer is a thin layer
with numerous small pyramidal cells, the
peripheral process being usually distinct;
this is the 2nd layer.

XXX.]

THE BRAIN. 329

J. The Ist layer of the cortex, consisting of a
fine network of fibrils showing a few very
small cells.

g- Blood vessels may be made out in all por-
tions of the cortex, they are usually most
conspicuous in the outer layers of the cortex
running into it from the pia mater.

Prepare sections of a lobule of cerebellum ex-
tending from the surface to the inner white
substance and at right angles to the direction of
the folds. The hardening, staining, etc., are the
same as for the cerebral cortex. Observe

a. The inner strand of medullated nerve fibres
spreading into

b. The nuclear layer, formed mainly of small
cells closely packed together; these cells
have a very small amount of cell substance,
so that probably their deeply stained nuclei
only will be seen.

c. A single layer of large somewhat globular
cells (Purkinje’s cells); each has a large peri-
pheral process which will be seen to branch,
and the branches to branch again and so
on, eventually extending as fine branching
fibrils nearly to the surface of the cortex;
close to the surface the fibrils become lost
to view. Since the branches, especially the
larger ones, run to some extent laterally, the
processes from neighbouring cells will be

330

ELEMENTARY PHYSIOLOGY. [XXX.

seen to cross one another. In a good speci-
men a small process may be seen to run
from the deep portion of the cells towards
the nuclear layer.

. The outer layer of the cortex contains,

besides the fibres from the cells of Purkinje,
some scattered small angular cells with rela-
tively large nuclei; from these cells one or
more small branching processes may be seen
to proceed, the fibres and cells being imbedded
in a close fibrillar network; in this layer
numerous capillaries will be seen.

LESSON XXXI.
DISSECTION OF THE LARYNX,

It is preferable to obtain a fresh larynx of a
sheep or an ox from the butcher's; but the
spirit-preserved larynx of the dog (Lesson 1.)
will serve. The larynx will probably be obtained
with the upper part of the cesophagus attached,
and surrounded by a mass of muscle and con-
nective tissue.

Having slit up the esophagus lengthways, turn
back or cut away the sides and observe the
opening into the larynx bounded in front by the
epiglottis, at the sides by folds of the mucous
membrane, and behind by the large converging
yellow crests of the arytenoid cartilages. On
looking down into the larynx the opening be-
tween the vocal cords, or rima glottidis, may
be seen at some depth below. Observe that the
mucous membrane of the cesophagus is con-
tinuous with that of the larynx. Bend down
the epiglottis projecting from the front, upper,
edge of the larynx, and note that the passage to
the larynx is then quite closed,

332

ELEMENTARY PHYSIOLOGY. [XXXI.

From the posterior surface carefully remove the
cesophagus with the pharyngeal muscles. From
the sides dissect away the sterno-thyroid muscle
(Lesson I. p. 22), taking care not to injure the
muscle lying underneath it (§ 3), and clear away
the whole of the thyro-hyoid muscle which
covers the side of the thyroid. The hyoid bone
and thyro-hyoidean membrane may be left.
The outlines of the thyroid will now come into
view, clear away the connective tissue until they
are quite distinct. Note that

The thyroid cartilage consists of two lateral
lamine, which meet in front and diverge behind,
and have their upper and lower posterior angles
prolonged, forming the upper and lower cornua.
Observe in front the rounded projection or
Adam’s apple.

Observe the crico-thyroid muscle spoken of
above (§ 2); dissect it away, noting its attach- °
ments. The cricoid cartilage will come into
view, and it will be seen that the muscle in
question passes from the posterior cornu and
posterior portion of the lower margin of the
thyroid, to the front part of the cricoid. Observe
that below this muscle there is stretching be-
tween the two cartilages a membrane which
limits the movements of the thyroid.

Observe the articulations of the posterior cornua
of the thyroid to the cricoid. Disarticulate one
cornu, cut through the crico-thyroid membrane,

XXXL] DISSECTION OF THE LARYNX. 333
and remove one half of the thyroid, taking care

not to injure any of the muscles. Trace out
with the finger the outlines of the cricoid carti-
lage. Note that

It forms a complete ring, which in front is
narrow, and is covered by the edge of the
thyroid, but behind is deep and enclosed on
either side by the thyroid.

5. On the posterior surface of the cricoid note
on each side of a central ridge two lamine of
muscles, the posterior crico-arytenoids ; cut
through their attachment to the cricoid, and
reflecting them from below upwards note that
they are attached above to the external angles
of two cartilages, the arytenoid cartilages,
placed on the posterior upper edge of the cricoid
cartilage.

6. Carefully clear away the tissue from the back of
the arytenoid cartilages above the cricoid, and
observe on the posterior surface of the former
the arytenoid muscle; cutting it through the
middle, which in the sheep is frequently ten-
dinous, reflect it on either side; it will be seen
to be attached to the back of each arytenoid
cartilage ; the posterior surface of the arytenoid
cartilage will now be laid bare, and its articula-
tion with the cricoid cartilage can be made out.

7. Looking at the larynx where the thyroid has
been cut away, observe the lateral crico-

334

10.

ELEMENTARY PHYSIOLOGY. [XXXI.

arytenoid muscle. Cut away its attachment
to the lateral portion of the upper margin of the
cricoid, and, reflecting it, observe that it passes
backwards and upwards from the cricoid to be
inserted into the arytenoid cartilage just in front
of the insertion of the posterior crico-arytenoid.

Clearing away the fat and connective tissue
from the side of the larynx, observe the thyro-
arytenoid muscle stretching across from the
thyroid in front to the arytenoid behind. Cut it
through in the middle and reflect both ends.
Note its origin from the angle of the thyroid and
its insertion into the lateral surface of the
arytenoid in front of the insertion of the lateral
crico-arytenoid.

Remove the muscle altogether, and observe the
lateral surface of the arytenoid cartilage.

Cut away on the same side the underlying

mucous membrane; the interior. of the larynx
will now be laid open. On the opposite side the
indistinct vocal cord will be seen passing as a
pale band of tissue from the anterior angle of
the arytenoid cartilage to the angle of the
thyroid. The inner or median surfaces of the
arytenoid cartilages will be seen to bound a
large oval space, called the respiratory space.

Clear away on one side any muscle or connective
tissue still attached to the arytenoid cartilage,
and observe more fully its shape, noting par-
ticularly the anterior projection or processus

XXXL]

11.

DISSECTION OF THE LARYNX. 335

vocalis, the posterior lateral projection or pro-
cessus muscularis, and the articulation with the
cricoid.

Dissect from the inside the thyro-arytenoid
muscle of the opposite side, and observe more
carefully its attachments (§ 8).

The larynx of the sheep differs materially from
that of man, by the indistinctness of the vocal
cords, by the absence of the false vocal cords and
ventricles of the larynx, and by the peculiar
crested conformation of the arytenoid cartilages,

DEMONSTRATION,
The use of the laryngoscope.

LESSON XXXII.
TISSUES OF REPRODUCTION.

A. OVARY.

I. Take an ovary of a mammal, eg. cat, hardened
in a mixture of equal parts of chromic acid ‘3 p.c.
and 50 p.c. alcohol, and prepare longitudinal
sections passing through the hilus.

Observe first with a low and then with a high
power

a. The germinal epithelium, consisting of a
single layer of short columnar cells covering
the surface of the ovary except at the hilus.

b. The connective tissue radiating from the —
hilus throughout the ovary to form the
stroma; in this many blood vessels are
seen; towards the periphery the fibrous
tissue largely disappears (cp. § 2. a);immedi-
ately underneath the epithelium the stroma
forms a denser layer, the tunica albuginea.

c. Small Graafian follicles forming a zone
a short distance below the germinal epi-
thelium.

XXXII]

TISSUES OF REPRODUCTION. 337

d. Deeper (older) Graafian follicles of various

sizes scattered throughout the rest of the
stroma, in each of these there is a more
or less considerable clear space.

One or more corpora lutea (unless the
ovary has been taken from a young animal).
The corpora lutea vary greatly in size and
appearance according to their age, they
consist of radiating bands of stroma, between
which are a number of granular polygonal
cells; if the corpus luteum is young there
will be pigment (from effused blood) in its
centre.

2. Observe with a high power

a.

The peripheral parts of the stroma consist
chiefly of elongated spindle-shaped cells.

The small Graafian follicles. Note in these

a. The ovum, large and spherical, within
it lies a comparatively large spherical
nucleus, the germinal vesicle, in
which may be seen a nucleolus, the
germinal spot.

8. The membrana granulosa, a layer of
flattened epithelium cells immediately
surrounding the ovum.

y. The thin membrana propria enclosing
the membrana granulosa.

In the smallest Graafian follicles the membrana
propria will not be seen, in the larger ones the

L.

22

338

ELEMENTARY PHYSIOLOGY. [ XXXII.

cells of the membrana granulosa become cubical
or short columnar and the ovum has a membrane,
the zona pellucida.

c. The large Graafian follicles. Note in these

a.

The stroma investment of the follicles,
consisting chiefly of spindle-shaped cells,
partly of fine fibrous tissue; inside this
is the membrana propria.

The membrana granulosa, several cells
deep, those next the membrana propria
being short columnar cells, the rest
flattened polyhedral cells.

The central space of the follicle.

The cumulus proligerus projecting
into the space; it consists of a mass of
cells much like those of the membrana
granulosa and continuous with them,
and it encloses

The ovum, like the ovum of the small
Graafian follicles but with all its parts
larger and its cell-substance (beginning
vitellus) more granular; it has further ©
a distinct investing membrane which in —
some is much thickened so as to form a
zona pellucida; from this the cell-
substance frequently shrinks in the
process of hardening. The cells of the
cumulus which lie next the zona pel-
lucida are frequently arranged in a
radiating manner.

XXXIL] TISSUES OF REPRODUCTION. 339

Probably most of the stages between the small-
est and the largest follicles will be seen in the
specimen.

d. Granular polyhedral cells in the stroma some-
thing like those that occur in the interstitial
tissue of the testis. -

e. In some of the sections taken near the hilus
there may be seen lying in the stroma groups
of tubules, lined with a short cubical or
flattened epithelium, and cut at various angles.
These are the tubules of the parovarium.

3. Take the fresh ovary of a sheep or large dog;
observe the bulgings due to the more or less
ripe Graafian follicles. Holding the ovary over
a glass slide, carefully prick the most prominent
follicle and receive the contents on a glass slide.
Examine without a cover slip, with simple lens
or low objective. If the ovum is present it will
at once be recognized. When one is obtained
carefully cover with a cover slip, inserting a ring
of paper in order to avoid pressure, and examine
with a high power. Observe

a. The thick zona pellucida, with double contour
(and radiating striation).

b. The granular cell-substance (vitellus).

c. The transparent germinal vesicle, with its

germinal spot. If the follicle be quite ripe
these may have disappeared.

d. The cells of the cumulus proligerus attached
all round the zona pellucida.
22—2

340

ELEMENTARY PHYSIOLOGY. [ XXXII.

B. UTERUS.

i,

Prepare transverse sections of the fundus of a
mammalian uterus which has been hardened in
the chromic acid and spirit mixture. The uterus
may have been either distended with the fluid,
or pinned out and placed in it. Observe

a. The thin external fibrous coat.

8. The thin outer muscular coat of two layers,
an outer longitudinal and an inner circular.
Inside this and separated from it by a little
connective tissue is

y. The thick inner muscular coat of two

layers, the outer longitudinal or oblique, the

inner circular.

5. The mucous membrane, consisting of fine,
loose connective tissue into which run tubu-
lar glands from the surface; the glands are
lined by a single layer of columnar ciliated
cells. The surface of the mucous membrane
is covered by similar columnar ciliated cells
with some mucous cells.

C. TEsTISs.

1.

Take a small mammalian testis, e.g. of a guinea-
pig, which has been hardened in Miiller’s fluid,
and cut longitudinal sections through the testis
and head of the epididymis. Observe under
a low power

a. The tunica vaginalis propria, which is

XXXII]

TISSUES OF REPRODUCTION. 341

round the greater part of the testis firmly
connected with the underlying tissue, and
is hence called the tunica adnata. It sepa-
rates from the testis near the epididymis.

The tunica albuginea, the thick invest-
ment proper of the testis; near the epididy-
mis it is very much thickened, forming the
corpus Highmori; from this radiate bands
of connective tissue towards the rest of the
albuginea, forming imperfect septa which
divide the testis into lobules.

The tubuli seminiferi, contorted and ana-
stomosing tubes occupying the lobular spaces;
near the corpus Highmori the tubes unite,
and the tubes so formed, the vasa recta,
have a short, tolerably straight course; in
the corpus Highmori they anastomose with
one another, forming the rete testis ; from
the outer edge of this may be seen some
larger tubes, the vasa efferentia, which
originate from the rete and after coiling in
the coni vasculosi join to form the canal
of the epididymis.

2. Observe under a high power

a.

b.

The fine connective tissue of the tunica
adnata, indications of the flattened epithelium
covering it will be seen.

The coarser connective tissue of the tunica
albuginea, corpus Highmori and septa, con-
taining numerous blood-vessels,

ELEMENTARY PHYSIOLOGY. [XXXII

c. The bands and lamelle of fine connective
tissue running from the septa between the
tubuli seminiferi. ,

é.

Polyhedral granular cells scattered about in
the tissue between the tubuli seminiferi.
These are very numerous in some animals,
and to see them in number, a section of the
lobular part of the testis of a pig should
be cut.

The tubuli seminiferi. In these note

a.

The membrana propria; in some animals
this is thick and is formed of several
layers of flattened cells.

Tubes lined by cells lying three or four
deep and arranged as an inner single
layer of short columnar and flattened
polyhedral cells, and an outer layer of
less regular cells two to three deep.
Amongst the cells much larger ones
may sometimes be seen.

Tubes like the above but with the out-
lines of the cells bordering the lumen
less distinct and with bundles of sper-
matozoa imbedded amongst them. In
different tubes the spermatozoa are in
different stages of development; in all
cases their heads (more deeply stained)
are in the peripheral parts of the tubes

- and their tails directed to the lumen.

7
ww,

—_— =

XXXII] TISSUES OF REPRODUCTION. 343.

3.

j- The vasa recta; the tubes have a single

layer of cubical or flattened cells,

g. The rete testis ; the membrana propria of the

tubes disappears, the epithelium is rather
more flattened than in the vasa recta,

h. The vasa efferentia and canal of the epi-
didymis ;
a. The external fibrous coat.

8. Inside this a coat of circularly arranged
unstriated muscle.

y. The epithelium consisting of columnar

| ciliated cells with some small cells
between or outside their peripheral
ends near the basement membrane. In
the canal of the epididymis the ciliated
columnar cells are long and _ slender.
Masses of spermatozoa may sometimes
be seen in the lumina of the tubes,

Cut a transverse section of the vas deferens and
compare with the sections of the canal of the
epididymis.

a. The wall is very much thicker, this being
chiefly due to a great increase in the muscular
coat; there is a longitudinal muscular coat
outside the circular coat.

b. The epithelium consists of non-ciliated colum-
nar cells with one or more layers of small
peripheral cells; between the cells and the
muscular coat is a layer of fine connective
tissue (mucous coat).

344

ELEMENTARY PHYSIOLOGY. [XXXII

Tease out in dilute glycerine a fragment of a
testis which has been placed in osmic acid °5 p.c.
for a day and observe the cells and developing
spermatozoa of the tubuli seminiferi.

Cut in half the fresh testis of a rat, and gently
press the cut surface on a glass slide. Observe
the spermatozoa, each consisting of

a. An ovoid head or body.
b. A long tapering tail or proccss.
c. A short intermediate part.

Note that the spermatozoa move by a whip-like
movement of their tails.

Observe in like manner the spermatozoa of a
newt. The head is long and pointed, the inter-
mediate part small and not very distinct; from
the intermediate part starts a filament which
runs in a spiral around the long tail; the filament
is in reality the edge of a thin spiral membrane,
but this is difficult to make out.

} WH |
Hill

APPENDIX.

DESCRIPTION OF INSTRUMENTS USED IN LESSON IX.

du Bois Reymond’s Key. This is shewn at C,
Fig. 1. (See end of Appendix.) Note the arrange-
ment of the wires; when the key is closed the current
passes across the key and an infinitesimal part only
passes by the electrodes through the alternative
circuit; in the text this is called ‘arrangement of
key for short-circuiting the current.’ If two wires
only are connected with the key, one on either side,
the current cannot pass when the key is open, but
can when the key is closed; in the text this is
called ‘arrangement of key to break the current.’

Morse Key. This is shewn at 7, Fig. 1. The con-
necting wires of a Morse key are hidden by the
framework. The following diagram (Fig. 2) will
shew the way in which they are usually arranged.

It is obvious that with wires arranged as at F, Fig. 1,
the current can pass from @ to 6, except when the
end « of the lever is pressed down (ep. Fig. 2), then
the circuit is broken ; if the wires are connected with
6 and ¢c, the current cannot pass until the end & of

346 APPENDIX.

the lever is put down (arrangement of key to break

Fic. 2.

the current) ; lastly, if the wires from the battery are
connected with a and 6 respectively, and the wires
of the electrodes with a and c respectively, then the
current passes across the key until the end x of the
lever is pressed down when the current is thrown
into the electrodes (arrangement of key for short-
circuiting the current)

du Bois Reymond’s Induction Machine, This is
shewn at D, Fig. 1. The screws at the top of the
machine are connected with the ends of the wire
forming the primary coil; consequently, when the bat-
tery wires are connected with these screws, each break
and each make of the circuit produces an induction
current in the secondary coil; the make and break
can be most simply effected by means of a key
placed in the primary circuit and arranged to break
the current (cp. above); in the text this is called the
arrangement for single induction shocks.

When the wires from the battery are connected with

APPENDIX. 347

the screws of the two pillars at the front end of the
machine, cp. Fig. 3, then the current of the primary
circuit is rapidly made and broken, and a corre-
sponding number of induction shocks produced in the
secondary coil. The method in which the primary
current is made and broken will be easily understooa
from the figure. When the current is passing in the
primary coil, (b) is in contact with (c), but since the

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Fic. 3.

current also passes round the coil (m) the soft iron
core of this coil becomes magnetized, and conse-
quently attracts the plate (e); as this goes down the
contact of (b) and (c) is broken, hence the current
ceases to pass in the primary coil, the core of (m)
is no longer magnetized, the plate (e) flies up,
(b) comes again in contact with (c) and the cycle

348 APPENDIX.

of events starts again; in the text this is called the
arrangement for the interrupted current.

Moist Chamber.

One form of moist chamber is shewn at A, Fig. 1. In
arranging a nerve-muscle preparation in it the femur
(7) is clamped (c/), a hook is passed through the
lower tendon of the muscle, the hook is connected
with another hook fastened to the lever, and the
lever by means of its screw is shifted up or down
to bring it in a horizontal position. Moist blotting-
paper should be placed on the floor of the chamber,
further the nerve should be covered with blotting-
paper moistened with normal salt solution.

The nerve is placed across electrodes such as those
shewn in Fig. 4.

Fic. 4,

Non-polarizable electrodes.

One form of these is given in Fig. 5; the pointed end
(ch, c) of the glass tube is filled with china clay
worked into a stiff mass with normal salt solution ;

APPENDIX. 349

a little saturated solution of zinc sulphate (z. s) is
poured with a fine pipette into tube above the plug
of clay; into the zine sulphate solution dips the
thoroughly amalgamated end of a thin rod of zine.

Fic. 5.

Oscillating rod.

This is a thin band of steel about 9 inches long and
Zin. wide; at one end at right angles to the band
is fixed a pointer one to two inches long. The band
can be fixed at any part of its length by a wide
brass clamp; connected with the clamp is a binding
screw. A cup containing mercury is placed under-
neath the pointer and the clamp is arranged on the
stand at such a height that when the band descends
in oscillation, the pointer dips a millimetre or so into
the mercury; it is well to cover the mercury with a
thin layer of alcohol. A piece of main-spring bent
at one end will serve the purpose of the steel band.

Lever for the Frog’s Heart.
The lever used is like that given at (/) Fig. 1, but

350

APPENDIX.

a thin vertical needle is attached to it at the place
where the weight hangs in the Figure, at the end of
the needle is fixed a small piece of cork which rests
on the ventricle, the frog lying on a stage below the
lever.

Time Marker.

The two ends (w, w') of the wire wound round the

bobbins (6, b’) are connected with the binding screws
(sc, sc’). In using the instrument one binding screw
is connected with one of the poles of a galvanic
battery, the other binding screw with a clock or
metronome from which runs a wire to the other pole
of the battery. The clock or metronome is arranged
so that it makes and breaks the circuit at regular
intervals. When the circuit is made the current
passes round the bobbins, the soft core of the bobbins
becomes magnetized and the lever (/) is drawn down ;
when the circuit is broken the lever is drawn away
from the bobbins by the spring (sp). At the end of
the lever is fixed a stiff bristle (to write on smoked
paper) or a small bent glass tube filled with magenta
dissolved in dilute glycerine (to write on glazed
paper).

APPENDIX. 351

NoTEs ON THE USE OF THE MICROSCOPE.

See that the tube of the microscope moves easily, but
not too easily, up and down. It should occasion-
ally be rubbed round with a little olive-oil, and
the oil then wiped off with a dry cloth.

' With a piece of soft chamois leather or silk, remove
any dust that may be on the lenses of the eye-piece
or on the lower lens of the objective. If by accident
the objective has been smeared with glycerine,
stream it with water from a wash-bottle, and wipe it
dry with chamois leather: if with Canada balsam,
place on it a drop of chloroform or strong spirit, and
rub gently, repeating till the balsam is quite re-
moved: this must be done very cautiously, since the
lenses are sometimes fixed in with Canada balsam.

All unnecessary rubbing of lenses however should be
avoided; hence take care to put them away after
they have been used.

It is advisable for the student not to unscrew the
separate lenses of a high objective; when however
the dimness of an object under observation renders
it necessary to do so, care must be taken not to
remove the black coating on the inside of the tube
in cleaning the lenses, and when cleaned to restore
them, in proper order, to their previous positions.

With a low power (cp. foot-note, p. 35), twist down
the microscope-tube till it is rather less than a
quarter of an inch from the object; then looking
through the eye-piece, twist the tube upwards till
the specimen is in focus.

352 APPENDIX.

With a high power, lower the tube till it is + of
an inch from the object, and then slowly twist
the tube down, moving the slide about, till the
object just becomes visible, then focus by means of
the fine adjustment.

Direct sunlight should not be used to illuminate the
field ; in selecting a permanent position to work with
the microscope, it is best to face the north.

With high powers, use the corresponding small dia-
phragm; otherwise although the field may look
brighter, the outlines of the cells, etc, will not be so
well defined.

The student should accustom himself to keep both eyes
open when using the microscope, and to use either the
right or the left eye for looking at specimens. The
fatigue of microscope work is thus very much lessened.
It generally requires a little practice to keep both eyes
open, but a few minutes’ trial every time that the
microscope is used will soon overcome the difficulty.

When an object appears dim, it may be caused by some
fault in the specimen itself, or by the object-glass
not being clean. In the latter case the dimness
remains, whether the object is in focus or not. Dust
upon the lenses of the eye-piece can be recognized
by the outlines of the particles appearing well de-
fined, not dim and diffuse,

Zeiss’ camera lucida.

The ring (7) Fig. 7, is slipped over the tube of the micro-
scope (¢), and the ocular (0) inserted ; the object to be
drawn is focussed, and the prisms are brought over
the ocular in the position shewn in Fig. 7. (The
upper part of the figure represents a side view ; the

APPENDIX. 353

lower, a view from above, of the camera arranged for
use). The edge (a) of the prism of the side A of the
camera covering half of the upper lens of the ocular.

Fic. 7.

In front of the microscope, the drawing-board is placed
~ at an angle of 15—20° with the surface of the table.
On looking down the microscope both the object and
the drawing-board are seen. The illumination of
the drawing-board and of the field of the microscope
L. 23

354 APPENDIX.

should be about equal, so that the point of a pencil
on the drawing surface is distinctly seen.

Measurement of the size of microscopic objects. —
The method of doing this which has been given in
the text (p. 37, § 3) involves making a drawing of the
object ; when this is not done the size of the object —
may be measured by means of an ocular micrometer. —
This is a flat piece of glass on which a scale is
scratched (e.g. 5 mm. divided into ten parts, the actual
distances of the lines are however not important), —
this is dropped into the ocular so that it rests on a
ledge placed on a level with the focus of the upper
lens of the eye-piece. With this scale in the eye-—
piece the stage micrometer (cep. p. 37, § 3) is ob-
served, and the number of divisions of the stage
micrometer which correspond to one division of the
ocular micrometer noted. Say that one division of
the one exactly covers one division of the other,
then if (as is usually the case) one division of the
stage micrometer is ;45 mm. one division of the
ocular micrometer, with the objective, ocular and
length of draw-tube used, corresponds to >$5 mm. of
the field of the microscope. Hence any object seen
with this arrangement of the microscope can be at
once measured by the ocular micrometer, an object
which exactly occupies say 5 divisions measures in
that direction exactly ;3, mm.

If another ocular or objective be used, or if the tube be
drawn out, the divisions of the ocular micrometer
will obviously correspond to a different number of
divisions of the stage micrometer, so that the value
of the divisions of the ocular micrometer must be
determined for each arrangement of the microscope. —

APPENDIX. 355

Supposing the value of the divisions of the ocular
micrometer are known with any given arrangement
of the microscope, then the magnification of any
object drawn can easily be measured; suppose for
example that the drawing measures 5 mm. and the
object itself with the given arrangement of the
microscope occupies 5 divisions of the ocular micro-
meter, the value of each division being ;3, mm.,
then in the drawing the object is magnified
5 +755 =100 times.

The magnifying power of a microscope with any

given ocular, objective, and length of draw-tube, is
usually taken as the amount of magnification when
the image of the object is seen at a distance of
25 centimetres (10 inches) from the eye. To de-
termine this, the stage micrometer lines are drawn
at this distance and the distance between these
lines is measured ; thus if one division of the stage
micrometer equal 53, mm., and the drawing of this
measures exactly 1 millimetre, then the magnifying
power is 100. In order to compare the tables given
by different instrument makers of the magnifying
powers of their microscopes, it is necessary to know
the method used in determining it; in all cases when
an object is drawn its magnification should be
determined by the method given above or by that
given in the text.

OBSERVATION OF FREsH TISSUES.

Sections of fresh tissues may be made with the aid
of a freezing microtome (cp. p. 368); if, however,
specimens can be prepared without freezing, it is

23—2

356 APPENDIX.

generally better to do so. Such specimens may be
prepared in various ways.

A few parts of the body, e.g. the kidney, are sufficiently
thin to allow sections to be cut free-hand with a
razor ; one section should be mounted without adding
any fluid, another in a normal solution, and a third
should be teased out in a normal solution.

With structures too yielding to cut sections, e.g. the
villi of the small intestine, a piece may be snipped
off with scissors ; in the same way a good specimen
may often be obtained from the edge of a lobule of
a gland: in some cases, e.g. the coat of an artery, a
strip may be torn off with forceps; and in the case
of membranous structures, a piece may be pinned
out over a hole in a stage, the gastric glands and
pancreas of some animals for example may be thus
ubserved.

The knowledge gained by the examination of such
specimens is essential to thoroughly understanding
the appearances presented by the tissue after the
action of reagents.

There are certain fluids which, when fresh, cause very
slight changes in the tissues; they more or less
resemble the fluids with which the tissues in the —
‘body are surrounded. |

These normal fluids are

a. The aqueous humour of the eye.

b. Blood-serum.
If, then, a fresh tissue is to be observed from eg. a —

rabbit, the cornez should be punctured to obtain the
aqueous humour, or the blood should be allowed to

APPENDIX. 357

clot in a vessel, to obtain the serum. In one of
these fluids the tissue should be mounted.
c. Normal saline solution.

This is prepared by dissolving 6 grm. of sodium
chloride (pure if obtainable) in 1000 c.c, of distilled
water. . This is for some tissues rather better than
the -75 p.c. solution which is often used, but in
many cases there is nothing to choose between the
two solutions,

d. Iodized serum.

Iodine is sometimes added to serum to preserve it, so
that it may be at hand when required ;.serum thus
iodized is, however, far less a ‘normal’ fluid than
the others. It may be prepared by dropping a few
crystals of iodine into fresh serum, and shaking
occasionally. The fluid should be of a light brown
colour. Instead of serum the amniotic fluid of the
cow may be taken.

Teasing.

Be careful to take a small piece only: it should be
viewed with a low power before teasing, to ascertain
the general relation of the parts, and hence to guide
the teasing: thus, if it consists of parallel fibres,
with one needle fix the piece at one corner and draw
the other needle through it in the direction of the
fibres ; in teasing out to shew special objects, e.g.
ganglion-cells, the arrangement of the parts should
be particularly noticed under a low power, and such
pieces as do not contain the parts required should
be thrown away.

In teasing, it is important to place the slide on an

358 APPENDIX.

appropriate ground ; if the object is white or grey,
the slide should be placed on a piece of black paper ;
if it is dark, on a piece of white paper. Stained
sections should also be teased out against a white
or grey ground.

Dissociatine Fiuips.

These are fluids which, whilst preserving certain parts
of a tissue, dissolve or partially dissolve others,
principally the cementing or ground substances, so
that the former can be isolated by teasing or shaking.
As a rule the piece of tissue so treated should be
not more than one to two mm. square.

Weak iodized serum of a light brown colour is in most
cases the best. As the tissue absorbs the iodine, a
little strongly iodized serum should be added ; or
the fluid renewed. The tissue may often be teased
out after one day; but a longer time may be neces-
sary.

Osmic acid, ‘1 to 1 p.c., is a dissociating fluid of general
application; it has the advantage of altering the
normal appearances in most cases vety slightly. It
swells up some nuclei, and also the axis cylinders of
nerves.

Dilute alcohol (30 to 35 p.c.) is also good for many
tissues.

The following agents are also used, the tissues for

_ which they are most recommended are put in brack-

ets; baryta water (fibrille of white fibrous tissue);

5 p.c. neutral ammonium chromate (mucous glands);

‘02 p.c. potassium bichromate (muscle and nerve-

cells); ‘02 p.c. chromic acid (nerve-cells of spinal

cord); Muller’s fluid (olfactory cells); 5 p.c. chloral —

APPENDIX. 359

hydrate (serous glands). As a rule it is best to
tease out the tissues in the fluid in which they
have been placed, they may also be teased in water
or in dilute glycerine ; if mounted in water they may
be temporarily preserved by heating the end of a
small wax taper in a flame, and with the wick smear-
ing the wax round the cover-slip.

HARDENING AGENTS.

General Directions.

The tissues should be removed from the body to the
hardening agent as soon as possible after the death
of the animal.

Any blood which may be on the piece of tissue should
be removed by shaking gently in salt solution or by
placing on blotting-paper.

Divide the tissue (with a razor or sharp scalpel) into
small pieces before placing in the hardening fluid.
As a rule the pieces should not be more than 3 to
4mm. square. Since the brain is too soft to allow
it to be easily cut up without injury in the fresh
state, it is generally placed whole in the hardening
fluid, but in this way the deeper parts of the cortex
are in large brains rarely well preserved (cp. foot-note,
p. 327).

In all cases where a tissue is put in a fluid to harden
or have lime-salts extracted from it, the fluid in
which it is placed should be relatively abundant,
e.g. 15 to 20 times the volume of the tissue; it
should be renewed after a time varying from 12 to
24 hours.

360 APPENDIX.

When the tissue is well hardened, the hardening agent
should be extracted from it by placing it in alcohol,
first dilute 30 p.c. to 50 p.c. then stronger 50 p.c. to
75 p.c.; it should then be preserved in 70 to 95 p.c.
alcohol.

Since chromic acid, potassium bichromate, etc., are
precipitated by alcohol in the light, tissues hardened
in these reagents, after being washed with water and
placed in alcohol, should be kept in the dark, until
required for cutting, or until the alcohol, frequently
renewed, no longer becomes yellow. Many tissues
when well hardened may be left a day or more in ~

' water without injury before placing in dilute alcohol.
Tissues hardened in picric acid are best placed an
hour or two only inthe dilute alcoholic solutions,
and then transferred to'70 to 95 p.c. alcohol, which
may be renewed as often as it becomes coloured.

The tissues should be placed in a flat short bottle to
harden, rather than in a narrow high one; they should
-be kept cool during the first day or two, after this it

is of less lk didi. though usually advantageous.

actaontion of Solutions.

It is generally advisable to make of any given sub-
stance a solution of the maximum strength likely to
be required, and to dilute this when weaker solu-

- tions are needed. If-the stronger solution contains
x p.c. of the substance and it is required to make

a weaker solution of y p.c., add —lec. of water
| to each c.c. of the stronger ae

It is rather better to use distilled water in making up

- qa ee ae. hel:

APPENDIX. 361
the solutions, but in most cases this is not of much
importance.

Chromic acid. Dissolve 5 grms. in a litre of water,
from this the ‘3 and -2 p.c. solutions which are more
frequently used can readily be prepared.

A little chromic acid may often be added with advan-
tage to potassium bichromate and ammonium bichro-
mate solutions ; it may also be used mixed in various
proportions with picric acid.

Chromic acid hardens the tissues by an action analo-
gous to tanning ; but renders them, if they are left
in it too long, brittle and friable.

Chromic acid and spirit. Mix two parts of chromic
acid } p.c. with 1 part of methylated spirit (Klein),
or

Mix equal parts of chromic acid °3 p.c. and alcohol
50 p.c.

These solutions may require to be filtered before they
are used, they should be kept in the dark. It is
better to make them when required.

Erlicki’s fluid. Dissolve 2-5 grms. of potassium
bichromate and ‘5 grm. cupric sulphate in 100 c.c.
water. This is sometimes useful when a quicker
hardening agent than the chromium compounds
alone is required ; after two days the fluid with the
tissue may be kept at about 40° C. to increase the
rate of hardening.

Extraction of salts from bone.
The bone should be divided into small pieces. and

362 APPENDIX.

placed in a large quantity of chromic acid ‘1 p.c. It
is well to shake gently occasionally, in order to bring
fresh acid in contact withthe bone. The fluid should
be renewed in 24 hours, and changed for ‘25 p.c. acid
in about two days. After a week a solution of
‘5 p.c. may be used. This also may with advantage
be changed once or twice. The decalcification of
fully developed bone will take about a fortnight, but
a preliminary section should be made from one end
with a blunt razor or scalpel to see that all salts are
extracted.

To hasten the solution of the salts, 1 c.c. of nitric acid
may be added to each 100 c.c. of the chromic acid
solution, but this should only be used when the
tissue has been for 2 to 3 days in the dilute chromic
acid; as soon as the salts are dissolved the tissue
should be transferred to alcohol.

Instead of chromic acid, picric acid (either Kleinen-
berg’s or a saturated aqueous solution) may be used,
or the tissue may be placed in strong spirit for a
day or two and the salts then extracted with nitric
acid 1 to 4 p.c.; as soon as this is complete the tissue
should be replaced in spirit. A mixture of nitric
acid 1 to 3 p.c. and strong spirit is often useful to
complete the solution of salts in tissues which have
been hardened in chromic acid but from which the
salts have been only partially extracted.

Miiller’s fluid. Dissolve 25 grms. of potassium bi-
chromate and 10 grms. of sodium sulphate in 1000

c.c. of water.

Picric acid. Make a cold saturated solution of picric

APPENDIX. 363°

acid, and to 100 c.c. of this add 2 c.c. of concentrated
sulphuric acid ; filter, and to the filtrate add 300 c.c.
of distilled water (Kleinenberg).

Tissues should be left in this for a comparatively short
time, varying from three hours toa few days; the
hardening may be completed with alcohol. If the
tissue contains earthy salts to be extracted, nitric
acid should be substituted for sulphuric acid.

A simple concentrated aqueous solution is also fre-
-quently useful. |

Osmic acid. A 1 p.c. solution of this is most useful.
To avoid reduction of the osmic acid care should be
taken to obtain water free from organic matter.
The acid is obtained in tubes containing one grm.
each, the tube should be well washed and then broken
in a bottle containing 100 c.c. of distilled water and
shaken occasionally; it takes some little time to
dissolve completely. The bottle should have been
previously well washed with sodium hydrate, sul-
phuric acid and water. Ifa solution is required to
be made at once, the tube with the osmic acid may be
ground up in a mortar with a little water, but some
care is required to do this without suffering from the
hurtful vapour. If the solution begins to turn brown,
the bottle should be covered with black paper and
kept in the dark ; this however should not benecessary.
Osmic acid does not penetrate well, so that a small
piece only of the tissue should be taken; the tissue
may be left in the osmic acid any time from half-an-
hour to a day, in the latter case a ‘5 p.c. solution should
be used. Many tissues are hardened sufficiently by
a stay of 12 to 24 hours in osmic acid to allow of

364: APPENDIX.

sections being at once cut; if they are to be imbedded
they should be washed with water and placed for a
quarter of an hour or more in 30 p.c, 50 p.c. and
70 p.c. alcohol. When the tissues are to be kept
some time before cutting, they should be well washed
with water and then treated with alcohol as chromic
acid specimens are treated; of course if the tissue
has not been sufficiently hardened by the osmic acid,
its stay in dilute alcohol must be short. The tissues
become very much darker on staying in alcohol ; it is
not advisable to leave them long in this fluid before
sections are made. In osmic acid specimens the
nuclei are generally spherical and indistinct although
the nucleoli are not infrequently obvious. The
longer a tissue is left in osmic acid, the less easily —
does it stain with carmine and other reagents.

Osmic acid specimens which it is desired to preserve
should be mounted in dilute glycerine or in a con-
centrated aqueous solution of acetate of potash ; they
may, though with less satisfactory results, be cleared —
and mounted in Canada balsam.

Alcohol.

Alcohol, besides its general use of completing the
hardening begun by other fluids, is sometimes used —
alone. Jt coagulates the albumin in tissues and
thus renders them more opaque.

It is generally best to place the piece of tissue for
half-an-hour to an hour in 75 p.c. alcohol, and
then to remove it to strong spirit (90 to 95 p.c.);
in some cases it is advantageous after the tissue
has been half-an-hour in strong spirit to transfer
it to absolute alcohol, ‘Absolute alcohol. when

APPENDIX. ; 365

used alone usually causes more or less shrinking and
distortion of the outer parts of the tissue; in such
case the inner part ny should be taken in pre-
paring sections,

IMBEDDING.

Take a small piece of tissue only to imbed; of course
this cannot be done in many cases, where it is
desired to make out the general relation of the parts
of an organ; it is moreover of comparatively little
importance when the sections are to be cut with
a microtome. |

Before imbedding (cp. p. 68) press the piece of tissue
very gently between blotting-paper, to remove as
much as possible of the fluid in which it has been
placed. The fluid, if not removed, cools the imbed-
ding mixture immediately round the tissue, and not
mixing with it prevents the due adhesion between
the mixture and the tissue: with spirit specimens
the excess of fluid should not be removed till the
imbedding substance, etc., is ready, since the spirit
rapidly evaporates, and the tissue may become dry.

The imbedding substance should be heated as little as
possible above its melting point. To keep it at a
constant temperature, a water-bath heated by a

small gas jet with a regulator may with advantage
be used,

When the tissue is moved about by means of a heated
needle, to place it in position in the embedding
substance or to remove air-bubbles, bring down the
temperature of the needle to that of the substance
by moving the needle about in it before touching
the tissue.

366 | APPENDIX,

The substance in which different tissues should be
imbedded varies with the consistency of the tissue.
As a rule the imbedding substance should be of
about the same degree of hardness as the tissue.
Loose tissues, or tissues with cavities, should before
imbedding be treated in the manner given on p. 215.

A still simpler method of imbedding than that given
on p. 68 is to take a block of paraffin B, to heat a
stout wire over a Bunsen flame and with it to melt
the central portion of one end of the paraffin block,
in this the tissue is placed, it is turned into the
proper position and the bubbles are removed by ~
means of the warmed wire; since the wire is very
easily made so hot that it would injure the tissue,
it should be brought into contact with the tissue
as little as possible. This method should only be
used for dense tissues.

The paraffin ‘blocks’ can be made by taking a tube
(e.g. of copper) about a foot long and fitted with
a piston, the inside of this is oiled a little and
the melted imbedding mixture poured into it;
it is placed under a tap to solidify the mixture, the
column of substance is then pushed out and cut_
into lengths about an inch and a half long. |

The time the tissue should stay in the various agents
given on p. 215, varies with its density. With tissues
treated in this manner the following method of im-—
bedding will be found better than those previously —
given. Two L-shaped pieces of lead with sides about
half an inch high are placed on a glass slide so that the
long limb of each is in contact with the short limb of
the other; the enclosed space is about three-fourths —
filled with the imbedding mixture, a little water _

APPENDIX. 367

is then poured on the glass to solidify the lower
portion of the mixture, and just when a thin crust
is forming on the upper surface, the space is filled
up with the mixture and the tissue placed in it.
The layer next the glass is by this time solid, and on
this the tissue can be arranged with a warm needle
in any position required.

An oblong box made out of not too thin paper may be
used instead of the pieces of lead.

The sections, in this case, are best cut with a razor
moistened with olive oil. The imbedding mixture
must be dissolved out, e.g. with creosote and turpen-
tine; if it is desired to mount the sections in
glycerine, they should be placed in absolute alcohol
to remove the creosote and turpentine, they can then
be mounted at once or after having been placed in
more dilute spirit,

Before cutting sections, cut away the imbedding sub-
stance close around the tissue. It is easier to cut
thin sections when the surface to be cut is small
than when it is large.

In preparing the following mixtures, the constituents
should be placed in a capsule on a water-bath and
kept at a temperature just above melting point for
an hour or more, the liquid being occasionally
stirred,

Paraffin A.
Solid paraffin 15 grms,
Liquid paraffin 15 c.c.
This is used to surround the cover-slip in observing
the ameboid movements of white blood-corpuscles,
on the warm stage. It begins to melt at about

368 APPENDIX.

37° C., and thus serves as an indication of the
limit of temperature that can be used in the
observation without injury.

Paraflin B.
Paraffin 2 parts.
Vaseline 1 part.

This is one of the best imbedding mixtures for general
purposes, it can be made harder by adding a little
more parafiin.

Seepeenaness and Castor oil.

Spermaceti 40 grms.
Castor oil 10 grms.

SECTION CUTTING.

In order to make thin sections it is absolutely necessary
to have a sharp razor. It sometimes happens that a
student who has resigned himself to the belief that
he cannot cut thin sections finds no difficulty. in
doing so on buying a new razor.

In carrying on a class, where it is required to cut a
considerable number of equally good sections, a
microtome is almost indispensable; perhaps the
most useful for a Demonstrator’s purpose is the
freezing microtome made by Swift, Optician, Totten-
ham Court Road, London. The one in which
ice and salt is used, is recommended in preference.
to the one in which ether is used, since it is less
liable to get out of order, and when once started
will continue freezing for two to three and a half
hours. Equal parts of ice and salt are taken, and
pounded up together in a mortar, then packed
tightly in the box of the microtome.

a

APPENDIX. 369

Tissues to be cut with the freezing microtome are soak-
ed in gum; if they have been kept in strong spirit,
they should be placed for a short time in weak spirit,
and for an hour or more in water, before being placed
in the gum; it is best to let them soak in the gum for
a day; but sections may be made, though probably
less good, after a very short period of soaking; in
this case the crystals of ice, which form, rapidly
blunt the razor.

The gum solution is prepared by dissolving gum arabic
in warm water, and filtering through linen ; a rather
thin solution may be used to soak the tissue in,
a thicker one to surround the tissue on the micro-
tome. Since gum alone freezes into a hard mass,
a little sugar solution may with advantage be added
to it.

The plate of the microtome, and the grooves in it, must
be quite clean; a layer of gum is then spread over
it, care being taken that the gum fills up the grooves.
When the tissue has become frozen on the plate of
the microtome, the gum around it should be bevelled
off so that the section to be cut is not much larger
than the piece of tissue.

A hollow ground razor should not be used, since it
bends, and so makes a section, which, under the
microscope will be found to consist of bands of
unequal thickness.

The proper rate to carry the razor through the tissue
varies with the temperature of the frozen mass ;
when it is a few degrees only below zero, the move-
ment may be the quickest possible ; when it is frozen
hard, it should be carried slowly through the tissue,
otherwise the sections are apt to curl or break up.

L. 24

370 APPENDIX.

In cutting not very large sections, a dozen or more
sections may be cut in rapid succession; the sec-
tions heap up on the razor, and may be removed
to water with a camel-hair brush; three or more
pieces of a well hardened tissue may thus be cut
simultaneously. In cutting large sections, it is best
to place on the razor a number of small drops
of water and to cut slowly; the section folds up on
the razor. The razor (with the frame) should then be
dipped under water and the section floated off; it
should be taken out on a glass slide and treated —
on the slide with 30, 50, 75 p.c. alcohol, ete. ; care
should be taken to remove as much as possible of
the clearing agent, otherwise the Canada balsam in
which the section is mounted may remain a long
time fluid. Smaller sections should be treated in
the glass dish or watch glass-with the various —
reagents.

Sections of imbedded tissues may also be cut with the
microtome, but as a rule this is more trouble than,
and has no advantage over, the freezing method.

Fresh tissues should be frozen as little as possible below
0°C., they are apt to shew crossing bands brought
about by the crystallization of the water in them ; they
should be transferred with a brush to salt solution.
If it is required to stain them they should be
transferred to 30 p.c. alcohol and treated with
Spiller’s purple, methylene blue or methyl-violet or
removed from 30 p.c. to stronger alcohol and stained
with carmine, hematoxylin, etc.

When one or two tissues only are to be cut, it is
simpler to use a microtome with ether spray as the

—oe lm re ee

OL <-— =

Se eee ee

APPENDIX. 371

freezing agent; Swift’s ether freezing microtome
may be used.

STAINING.
Hematoxylin.

a. 1. Kleinenberg’s hematoxylin. Take crystallized cal-
cium chloride, and add it in excess to 70 per cent.
alcohol; shake well and let it stand. Draw off the
saturated solution and add alum to excess, shake up
and let stand fora day. Filter.

2. To one volume of the filtrate from (1) add six to
eight volumes of 70 p.c. alcohol.

3. To the mixture thus obtained, add drop by drop a
saturated solution of hematoxylin in absolute alco-
hol, till it is a moderately dark purple.

The solution will become considerably darker after
some days. It is better to make it some wecks,
or even months, before it is required for use.

It may be diluted to any extent required with the
mixture (2). Ifa section is placed in hematoxylin
for some time it must of course be covered up, or
the spirit will evaporate, and the solution become
concentrated. Im many cases it is well to stain
a piece of tissue as a whole in hematoxylin before
imbedding, in this case the tissue should be left in
the diluted fluid one to two days; a tolerably porous
tissue naturally serves best for this method, A
small piece only should be taken,

24—2

APPENDIX.

It is perhaps best to wash tissues or sections stained
with hematoxylin in the diluting fluid (2); they
may however be washed with alcohol (best about
70 p.c.); if it is desired to stain more particularly
the nuclei, the stained tissue may be placed for a
short time in a mixture of 2 parts alcohol (70 p. c.)
and 1 part 1 p. ec. hydrochloric or nitric acid.

If a section, which it is wished to preserve, has been
stained too deeply, the colour may be partially
extracted by placing the section in dilute acetic
acid. If the sections are washed with water,
distilled water must be used, otherwise alumina
may be precipitated.

The hematoxylin after it has been used should be put
aside in a separate bottle, since after filtration it
may be used again. If it has been mixed with acid,
it will have a reddish tint and will be useless; the
acid may be neutralized with sodium carbonate, but

the mixture rarely stains so well as the original
solution.

Alum-hematoxylin. To a *3 to ‘5 p.c. solution of ©
alum, add a few drops of a saturated solution of
hematoxylin in absolute alcohol.

Alum-logwood. Place in a mortar 5 to 10 grms. of
alum, 5 grms. extract of logwood, and 100 c.c. of
water, grind well, leave for a day and filter.

To both (6) and (c) a little thymol or salicylic acid
should be added, both should be filtered before being
used; these solutions stain well the nuclei of sections
of most tissues, but they cannot be satisfactorily
used for staining pieces of tissue before sections are
made,

am

APPENDIX. 373

Carmine. (Frey’s method.)

a.

Carmine 0:3 grm.
Glycerine 30°0 ,,
Alcohol 40 ,,
Distilled water 30-0 _,,
Ammonia q. 5.

Add the distilled water to the carmine, and then add

dilute ammonia drop by drop until the carmine is
dissolved, taking care that the least possible excess
is added. If the solution smells strongly of ammonia
expose it in a flat vessel to the sunlight for a day
or two. Finally add the glycerine and alcohol, and
shake. It should be kept in a stoppered bottle,
since otherwise the ammonia evaporates and the
carmine is precipitated.

It may be diluted with water to any extent required.

In the strong solution tissues hardened in chromic
acid or chromium salts may be left for days or weeks
without being over-stained.

As a rule sections stain better with carmine when they

are left in a dilute solution for 12 to 24 hours.

In another glass, under the same bell-jar as the sections,

a little dilute solution of ammonium carbonate may
be placed.

If sections are too deeply stained with carmine, or if

there is a precipitate of carmine upon them, they
should be placed in very dilute ammonia and gently
moved about in it; as soon as the excess of car-
mine is removed, they should be poured into a large
quantity of water and all trace of ammonia washed
away.

374 APPENDIX.

Most preparations stained with carmine are best pre-

served in glycerine containing 1 p.c. formic acid
1:16 Sp. Gr.

Picrocarminate of Ammonia or Picrocarmine.

Prepare a saturated solution of picric acid, and to it
add a saturated ammoniacal solution of carmine till
a precipitate occurs. Evaporate in a water-bath till
the volume is reduced by four-fifths. Filter, and
evaporate the filtrate to dryness. A crystalline
mass of picrocarmine is thus obtained. It dissolves
readily in distilled water; a 1 to 5 p.c. solution
should be made, and further diluted as occasion
requires,

If the picrocarmine is found to stain sections too
yellow, a little Frey’s carmine may be added to it,
and the mixture filtered.

Most of the yellow colour may however be removed
from tissues stained with picrocarmine by repeated
washing with water or alcohol.

Picrocarmine after being used may be filtered and used
again.

Borax carmine.

Borax 4 parts
Carmine 2°5 ,,
Water 100 *

Warm until the carmine is dissolved, being careful not
to let the mixture boil, When the fluid is cold, add
an equal volume of 70 p. c. alcohol and filter.

APPENDIX. 375

The proportions of borax and carmine given here are
; not very important; less carmine and more borax
may be used. The fluid may be used without the
; addition of alcohol, or glycerine may also be added
to it; further the precipitate from the alcohol may
; also be dissolved in water and used.

Borax carmine is principally used when it is desired
to stain nuclei deeply; the staining is usually im-
proved by placing the sections for a few minutes in

a mixture of 2 parts alcohol and 1 part hydrochloric

acid 1 p. c.

Spiller’s purple and Methylene blue.

—

These are used in a strong aqueous solution so that in
a minute or two the sections are sufficiently stained,
they are then washed with water, and mounted in
water; if the staining is too deep, the sections are
placed for a short time in alcohol; the sections
cannot be mounted satisfactorily in glycerine, since
the glycerine dissolves the colouring matter and
renders the staining diffuse. To preserve the sec-
tions permanently they should be over-stained, then
passed rapidly through 30, 50 and 70 p. c. alcohol ;

they should then be washed with strong spirit

(which dissolves the colouring matter very rapidly)

until nearly the proper tint is obtained, transferred

to absolute alcohol, and in a minute or two to cedar
wood oil; when they have become transparent, they

;

are mounted in Canada balsam. Cedar wood oil
instead of clove oil is used to clear the sections, since
it does not dissolve the colouring matter; water

376 APPENDIX.

must be entirely removed from the sections by abso-
lute alcohol.

Todine.
Dissolve 2 grms. of iodide of potassium in 100 c.c. of

distilled water, and add flakes of iodide to slight
- excess,

Gold chloride.

The method of using this for ordinary cases is suffi-
ciently described in the Text, Less. 1v. $3; Less.
xxiv. A, § 4. The tissue should be touched with
metal as little as possible, it may be removed from
one fluid to another with a small brush. In cold
weather it is well to place the acidulated water with
the tissue in a glass chamber kept at 20—30° C.

The following methods are advantageous in particular
cases,

a. Place the piece of tissue, which should be small, in
rather strong formic acid (1 vol. formic acid 1-16
Sp. Gr. and 3 or 4 vols. water) for a few minutes
until it is fairly transparent throughout, then place
it for about 20 minutes in 1 p. c. gold chloride, wash
it well with water; then place it in formic acid
diluted with three vols. water and leave it in the
dark fora day. The formic acid should be renewed
once or twice in the first two or three hours. After
the tissue has been placed in gold chloride it should
be exposed to light as little as possible. When the
tissue is stained it should be well washed with water
and mounted in formic glycerine. (Léwit.)

APPENDIX. 377

6. Filter fresh lemon-juice through flannel and place
the tissue in the fi'trate for about 5 minutes before
placing it in gold chloride. After treatment with
gold chloride, wash the tissue well with water and
expose to light in the ordinary way in acidulated
water, or place in formic acid in the dark as in (a).
(Ranvier. )

c. The tissue after treatment with gold chloride is
washed and placed in distilled water for 6 to 20
hours until it is of a steel-grey colour ; it is removed
to a saturated solution of tartaric acid for about 10
minutes and then warmed in the solution up to
40°—50° C. until it becomes dark; this generally
takes about ten minutes (Klein). If it is desired to
stain the tissue more quickly, it may be left in the
gold chloride solution for about three-quarters of an
hour, then well washed and at once treated with
tartaric acid as above.

Silver Nitrate.

For the method of using this ep. Less. x11. A, § 2, C, § 4,
and Less. xvii. C,§ 1. It serves mainly to trace out
flat tesselated epithelium, such as that of arteries,
veins, or lymphatics. By exposure to light, the silver
is reduced ; in a fresh tissue that has been placed in
nitrate of silver, the reduction on exposure takes place
more readily in the homogeneous substance between
the cells or ‘cement substance’ than in the cells.

As a consequence, where there is a single layer of cells,
with a small amount of cement between them, the
reduced silver in this substance marks very distinctly
the outline of the cells.

378: APPENDIX.

The success of silver preparations largely depends upon
not dragging the tissue, on washing it well, and on
exposing it to a bright light. The reduction of the
silver may be hastened by placing it in dilute alcohol
instead of in water.

The reduction of silver goes on though slowly in the
other parts of the tissue, so that it becomes darker
and darker, until it is useless; this is rendered less
rapid by mounting.

MountTInea.

The fiuids in which tissues are mounted serve to make —
them more transparent and to preserve them.

Glycerine.

It is best to allow sections to soak in glycerine for
5—10 minutes before mounting so that the glycerine
may thoroughly penetrate them.

Some unstained tissues are made too transparent by
strong glycerine, these should be mounted in gly-
cerine diluted with one or two vols. of water.

Many gold chloride and most carmine-stained specimens
are best mounted in glycerine containing | p. ec, of -
formic acid 1:16 Sp. Gr. (formic glycerine).

Some hardened tissues, stained or unstained, may with
advantage be kept in glycerine instead of in alcohol;
before sections are made the glycerine must be
extracted from the tissue with water or alcohol.

Creosote and Turpentine. ,
Add 1 part of creosote to 4 of turpentine, shake well

APPENDIX. 379

and set aside until the cloudiness which forms on
mixing the fluids disappears.

Sections may be transferred to this from strong spirit,
they may however have to be gently warmed to
render them clear throughout. Before mounting in
Canada balsam, the sections should be examined
under a low power in creosote and turpentine to see
that the clearing process is complete; if they are
not, fresh creosote and turpentine should be added
and the sections warmed init. Cp. also p. 75, foot-
note.

Before mounting the sections in Canada balsam as
much of the clearing fluid as possible should be
removed by means of a piece of blotting-paper with
a clean cut edge.

Canada Balsam.

Put some Canada balsam into a capsule, and place it in
the warm chamber at about 65°C. for twenty-four
hours to drive off all water. Let it cool and dissolve
it in a sufficient quantity of chloroform or benzole to
make a fairly fluid solution; it should be kept in
a bottle with a ground glass cap fitting over the
neck of the bottle, instead of a stopper, as the
stopper is apt to become fixed in the bottle; if any
balsam is allowed to get on the neck of the bottle
wet it with spirit and rub it off with a cloth.
Dammar may be used instead of Canada balsam.

Canada balsam renders tissues much more transparent
than glycerine.

Tissues stained in carmine or any watery solution
should be placed in alcohol of increasing strengths

SO APPENDIX.

up to 95 p.c. or absolute and then cleared previously
to being mounted in Canada balsam.

Injection mass.

a. Make a 2 p.c. aqueous solution of Berlin blue (this
may be bought at a chemist’s); it may be injected
either cold or warm. The tissues after injection
should be placed in alcohol. The sections should be
cleared, and mounted in Canada balsam ; if the blue
has become a lighter colour during the stay of the
tissue in alcohol, the sections should be placed for
a day in turpentine in an uncovered watch-glass
or bell-jar. Injections made with this solution are
rarely so good as those made with the following

() (¢).

b. Place 20 grms. of gelatine in cold water until it is
well swollen, then pour off the water, and place the
gelatine in a water-bath at about 40° C. (covering it
up to prevent evaporation) until it forms a fluid
mass.

Rub 8 grms. of carmine into a paste with water, add
about 10 grms. of strong ammonia and mix well,
then add about 100 cc. of water, shake well and
filter ; if a suction-pump is not used it will probably

- take 10 to 20 hours to filter. Warm the filtrate to
about 40°C. Pour it then slowly into the gelatine
kept warm over a water-bath, stirring continuously ;
when the fluids are well mixed, add gradually strong
acetic acid, stirring as before; when the smell of
ammonia becomes faint, use acetic acid diluted
5 to 10 times, adding it until the mixture smells of
acetic acid. The mixture previously a lake red

APPENDIX. 3S1

(carmine dissolved) will become an opaque carmine |
red (carmine in suspension). Add a few drops of
thymol, remove from the water-bath and let cool ;
if it does not smell of acetic acid when again melted,
a little more of the acid should be added.

The gelatine mixtures must of course be injected warm;
the blood should be washed out of the organ to be
injected with warm salt solution; during the in-
jection warm salt solution should be poured over the
organ or the whole animal should be immersed in
warm salt solution. The injected tissue should be
placed in alcohol.

c. Prepare gelatine as in (b). Take 100 c.c. of a 2 p.c.
solution of Berlin blue warmed to 40°C, and pour it
slowly into the gelatine kept warm on a water-bath,
stirring continuously.

CuemiIcaAL REAGENTS,
Millon’s reagent.

Weigh out 50 grms. of purified mercury and an equal
weight of pure strong nitric acid. Place the mercury
in a flask in the closed chamber, add the nitric acid
to it, and leave the mixture as long as any chemical
action continues. If all the mercury is not dissolved
warm it gently to complete the solution.

Add then to it twice its volume of water, and place

aside for some hours ; a white crystalline precipitate
will fall; the supernatant fluid is Millon’s reagent.

Fehling’s fluid.
(1) Dissolve 103-92 grams of pure cupric sulphate in
warm water and add water to make up exactly a
litre.

382 APPENDIX.

(2) Dissolve 320 grams of the double tartrate of sodium
and potassium in warm water, add a little carbolic
acid to prevent the growth of fungi, fill up with
water to exactly a litre and filter.

(3) Dissolve 150 grams of caustic sodium hydrate in
water dilute to a litre with water. If the fluid is
cloudy, filter through asbestos, or let stand and
decant the clear fluid.

From the above, Fehling’s fluid should be made at the
time it is required for use. It will not keep. To
make it, equal quantities of (1), (2) and (3) are
mixed together. The cupric sulphate should be
shaken up and a given quantity, say 100 cc,
accurately measured, to this, 100 c.c. of (2) is added,
and then (3) to make up the volume to exactly
300cc. From 10cc. of this mixture, the cupric
salt is reduced by ‘05 gram of dextrose.

Stokes’ fluid.

This should be made afresh when required ; it consists
of a solution of ferrous sulphate, to which ammonia
has been added after the previous addition of
sufficient tartaric acid to prevent precipitation.
An excess of tartaric acid is not of consequence;
roughly speaking, three parts by weight may be
added to two parts of ferrous sulphate.

Mason ARTERIAL SCHEME,
This (see Fig. 8) is a modification of Weber’s original

scheme of the circulation. It is not intended to

imitate the details of the circulation of the blood,

but simply to illustrate the points to which attention

is called in Lesson xin. It can without much

APPENDIX, — $88

difficulty be made by joining together india-rubber
tubing by means of three-way glass tubes.
Clamps are placed upon the straight tubes ¢, c’, c”,
: and the dilated tubes along the same line are stuffed
with sponge, until on closing the clamps and pump-
) ing steadily, a mean pressure of two or three inches
| of mercury is obtained in the arterial tube,

Fic. 8.

The arterial tube P is connected with an india-rubber
bag; from the opposite end of which a tube passes
to a vessel full of water. The bag has two valves,

i one at each end, opening in the same direction,

so that when it is compressed with the hand the

water it contains is driven onwards through the

; arterial tube, and on its dilating water is drawn

t up from the vessel,

384 APPENDIX.

Any other form of pump can be used.

At S,, S,', S,’ are placed vertically above one another
either the usual sphygmograph levers or light levers
such as are used to take tracings of the movements ©
of the frog’s heart (cp. p. 349). The tubing can
be supported on pieces of lead bent to fit its course —
and fastened to a stand. Each of the tubes A and
V communicates with a mercurial manometer.

The rest of the scheme requires no explanation further —
than that which is given in the text.

Minor ARTERIAL SCHEME.

This consists of an india-rubber bag like that used in ©
the major arterial scheme. By means of a three-way —
tube, the end of this, towards which the valves open, —
is connected both with a piece of glass tubing about —
a yard and a half in length, and with a piece of —
india-rubber tubing of similar length and bore.

There are clamps upon the long india-rubber tube close
to its junction with the three-way tube and upon ~
the small piece of india-rubber which connects the
three-way tube with the glass tube, so that the flow —
of water may be through either the glass or the —
india-rubber tube.

A small piece of india-rubber tubing is also placed on ,
the end of the glass tube, into which a tube finely
drawn out can be inserted.

To DESTROY THE BRAIN OF A FROG.

Place the frog under a small bell-jar with a sponge

moistened with ether; when the animal is motion-

APPENDIX. 385

less and under the influence of the anesthetic, take
it up in a cloth, hold the hind legs between the
third and fourth fingers, and with the fore-finger
press down the head. Cut transversely through the
skin a little behind the skull with a pair of scissors,
carrying the incision only a short way on either side
of the median line; make a slight incision also
through the skin in the median line. The end of
the occipital bone can then be easily made out; it is
just above the level of the anterior edges of the
scapule. With a sharp scalpel cut through the
muscles just behind the occipital bone and as nearly
as possible in the median line, so as not to cut the
vertebral arteries; the spinal cord will be exposed.
Thrust then a blunt stout needle into the brain and
move it about until the brain is quite destroyed,

The following rough table may be useful to the student.

1 litre =1 cubic decimetre=1¢ pints nearly.
1 minim = ‘059 cubic centimetres,
1 pint = 0°57 litres.

1 gallon = 4°54 litres.

1 fluid ounce =z/5 pint.

1 sq. inch = 6°45 sq. centimetres,

1 cubic inch = 16°39 cubic centimetres.
1 gram = 15°43 grains.

1 kilogram = 85°27 oz. Avoird. =2°2 Ibs
1 grain =0°065 gram.

1 drachm = 3°88 gram.

1 oz. Troy =31-1 gram.

1 oz. Avoird. = 28°35 gram.

1 Ib. Avoird. = 0°45 kilogram,

1 centimetre = inch.

1 metre =1 yard 3} inches.

1 inch = 2°54 centimetres.

1 foot = 3°05 decimetres.

1 yard =0°91 metre.

xtsv inch = 7, millimetre about.
a0s7 inch =12°7 u.

avon inch = 10°16 Me

i. 25

FAHRENH

APPENDIX.

386

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ADDITIONS.

Platelets of Blood. (Blutplittchen.) The platelets

are colourless bodies about } the size of the red
corpuscles ; in shape fairly like the red corpuscles,
and nucleated when the red corpuscles are nucleated.
They very readily break down into formless granular
masses. In the frog or newt they are readily seen
in the capillary circulation of the mesentery or
other part of the animal. During inflammation (cp.
p- 133) they will be seen clinging in numbers to the
inner walls of: the small vessels. They may also be
seen in a drop of blood if it is examined immediately
after being drawn from the body; certain fluids
retard the rate of disintegration of these corpuscles,
the corpuscles are longer preserved if a little blood
be allowed to run from the body into 3 to 5 times
its volume of one of the following: sodium chloride
°6 p.c. with a little methyl-violet (about 3 c.c. sodium
chloride solution and one drop of a strong aqueous
solution of methyl-violet); 20 p.c. magnesium sul-
phate; 1 p.c. osmic acid.

HARDENING AGENTS,
Mercuric chloride. A saturated aqueous solution is a

good general hardening agent for glands and epi-
thelium of skin and cornea. The tissue is left in
the mercuric chloride solution for several days,

25—2

888 ADDITIONS.

placed in a stream of water for some hours to re-
move the mercuric salt, and further hardened in
alcohol. As the mercuric salt is soluble in alcohol,
the tissue may be removed from mercuric chloride
to alcohol. In this case, however, the alcohol should
be renewed several times.

Flemming’s mixture is used chiefly to harden tissues
in which it is desired to make dividing nuclei
obvious, and consists of |

- Chromic acid 1 p.c.—15 parts.
Osmic acid 2 p.c. —4 parts.
Glacial acetic acid —43 to 1 part.

In this the tissue should be left two or three days,
washed with water, and treated in the usual way
with alcohol,

ImBeppinG, Section Crttine, Mountine.

When a number of sections have to be prepared for
distribution amongst students, the freezing method
given in the Appendix is only satisfactory with
tissues that are firm and hold well together. Other —
tissues should be either imbedded in celloidin, or —
imbedded in hard paraffin with a view to preparing
ribbons of sections. .

Paraffin. Ribbons of Sections.

Imbedding and cutting. The piece of hardened and —
stained tissue is placed in absolute alcohol for 1 to
24 hours. From this it is removed to either chloro-
form, turpentine, or oil of cloves. It is then placed
in hard paraffin (melting-point about 58°C.), kept at
a constant temperature, not greater than 60°C., for —
two or more hours, and imbedded in the manner

i

Tare Eo

_——— a -

—— =

EE ee ——_s ae

ADDITIONS, 389

given on p. 366, § 4. The paraffin is sliced away
until the surface is very nearly reached, then the
parafiin at the sides of the tissues is sliced away in
such a manner that the surface to be cut is rect-
angular. The shaped end of the paraffin block is
dipped for a moment in melted soft paraffin, i.e.
paraffin melting at about 40°C, The block is fixed
in a microtome, and arranged so that the edge which
meets the razor is parallel to the edge of the razor,
the soft paraffin may be cut away from the two
sides of the block which are at right angles to the
razor edge. The soft paraffin causes each section as
it is cut to stick to the preceding one, and thus a
ribbon of sections is obtained. Of the less expensive
forms of microtome adapted for cutting ribbons of
sections the most convenient is the Rocking Micro-
tome of the Cambridge Scientific Instrument Com-
pany. In this a short tube is filled with hard
paraffin, and the base of the block of paraffin con-
taining the tissue to be cut is sliced away within a

few millimetres of the tissue, this is held down on

the paraffin mass of the microtome tube and a hot
knife passed between the two surfaces and around
the junction of the block with the paraffin mass.
Thus the tissue to be cut is firmly fixed when the
paraffin solidifies. It is best to do this before coat-
ing with soft parafiin.

Mounting. Take a thin solution of white shellac in

kreosote and paint a thin film of it over a clean
dry glass slide. On this place the section or sec-
tions it is desired to mount, press each gently with
a camel hair brush to make it lie flat, Place the
slide in a warm chamber until the paraflin is melted;

390

ADDITIONS.

remove then the slide to bottle of turpentine for a
few minutes until the paraffin is dissolved. Let the
turpentine drain from the slide. . Place a streak of
rather thin balsam (dissolved in a mixture of chloro-
form and benzol) near one edge of the cover-slip,
and lower this gently on the sections. By this
method the tissue sticks to the shellac and the parts
of the section are not displaced on dissolving out
the paraffin.

Instead of the shellac solution, equal parts of filtered white of
egg and glycerine to which a little thymol has been added,
may be used. The treatment is the same as that given above,
except that after the paraffin has been dissolved by turpentine,
the slide should be placed for a minute or so in absolute

alcohol, If required the sections can then be stained on the
slide.

Celloidin.

Imbedding in celloidin has the advantage that it is
not necessary to warm the tissue, and thus there is
little risk of the tissue shrinking. By the simple
celloidin method, however, such thin sections can-
not be obtained as by the paraffin method. But
when the tissue imbedded in celloidin is soaked in
gum, frozen and cut, this objection to the method
no longer holds.

Celloidin solution. The celloidin solution is cut up
into small pieces and dissolved in a mixture of equal
parts of absolute alcohol and ether to make a fairly
fluid solution. It is sometimes advisable to use a
thicker solution after this.

Treatment of tissue. (a) The tissue, dehydrated with
absolute alcohol—after which it may be placed in a
mixture of absolute alcohol and ether or in ether
alone—is placed in celloidin solution and left in it

aa

— CT

SS

ADDITIONS. 391

for one day to a week or more. (+) Whilst the
tissue is in absolute alcohol } vol. ether is added,
and then celloidin in small fragments from day to
day, so that solution slowly goes on and a fairly
thick solution is obtained in 8 to 10 days.

Celloidin hardened in 80 p.c. alcohol. The tissue is

removed from the celloidin solution, placed on a
cork and a little of the solution poured over it;
when a thin film has formed on the surface of the
celloidin the whole is put in alcohol 80 p.c. In a
day or more the tissue can be cut. During cutting,
the razor should be kept wet with 70 to 80 p.c.
alcohol. Instead of fixing the tissue to cork it may
be placed in a small paper case, celloidin poured
over it and treated as above. When the outer part
is firm the paper should be removed.

Celloidin hardened in chloroform. Chloroform is used

instead of the 80 p.c. alcohol of the previous section ;
or a little celloidin solution is gently poured on chlo-
roform, when the lower surface is firm the tissue
is placed on the celloidin and more of the solution
poured over it. The celloidin shrinks much less
and remains more transparent with chloroform than
with alcohol.

Celloidin hardened by evaporation. The tissue is re-

moved from celloidin solution to a small watch glass
with a thin layer of celloidin over it, the small watch
glass covered with a larger one for two or three hours
till the celloidin is fairly firm, it is then coated with
fresh celloidin, allowed to dry partially, and so on
until (in 1 to 3 days) a sufficient coating of firm
celloidin is obtained, Or the tissue with celloidin
may be poured into a small glass dish, this covered

392 ADDITIONS.

with a flat piece of glass, allowing of a slow evapora-
tion of the alcohol and ether, so that the celloidin
becomes fairly hard in two or three days,

Preparation for freezing. The celloidin around the
specimen, imbedded in one of the above ways, is
pared away and the block placed in water for an
hour or two, then transferred to gum for one or two
days or for any longer time. The block can then be
frozen and cut. Thinner sections can thus be ob-
tained than from celloidin alone.

Staining sections. The sections still permeated by
celloidin may be stained with carmine or with hema-
toxylin. Most aniline dyes stain the celloidin also.
If required, the celloidin may be dissolved out by
absolute alcohol, or by a mixture of absolute alcohol
and ether.
Mounting. The celloidin need not be dissolved before
- mounting. If sections are to be mounted in Canada
balsam and other substances which render it neces-
sary to dehydrate the tissue, this should be done
with 90 to 95 p.c. alcohol and origanum or bergamot
oil, instead of with absolute alcohol and clove oil,
since the latter dissolve the celloidin. Clove oil does
not however dissolve the celloidin very rapidly, so
. that, if the sections are large, clove oil may be used ;
in this case the sections should be removed and
mounted as soon as they are clear.

Mounting i in glycerine-jelly.
_ The glycerine-jelly may be made as follows:
Glycerine 70 c.c.
Water 60 c.¢.
Gelatine 10 grams.
A little thymol or camphor, or carbolic acid.

ADDITIONS. 393

Place the gelatine with the water in a porcelain cap-
| sule, heat the mixture in a small water-bath over a
Bunsen burner, until the gelatine is dissolved, stirring
the while and taking care that the gelatine does not
stick to the capsule. The water should not be
: allowed to boil, or at any rate for a short time
only. To the hot solution add the glycerine and a
drop or two of a strong alcoholic solution of thymol.
If necessary cool the mixture to about 40°C., add
} the beaten up white of an egg and well mix. Then
heat as before to about 90° C, stirring continuously.

Filter through a hot water filter.
The gelatine may be left for a day in the cold water to swell up,
then dissolved by warming to about 40°C. The glycerine,

warmed to about the same war psec cst is added. Then
q proceed as above.

To mount a section in glycerine jelly, place a small

bottle containing glycerine-jelly in a warm chamber.
Remove the section from water to a slide, sop up
the excess of water, and let fall on it a drop of the
melted glycerine-jelly.. Place the slide and a cover-
slip in the warm chamber for a minute or two, if the
section rises to the top of the fluid press it down
with a needle so that it lies on the glass. Then
cover with the warm cover-slip.

If a warm chamber is not at hand melt the glycerine-
jelly in the flame of a Bunsen burner, warm a slide
in the flame and place a drop of the melted jelly on
it. With a needle transfer a section to the jelly,
and spread it out flat on the glass, Pass a cover-

: slip through the flame, and cover the section.

Before surrounding with Canada balsam or other
cement the specimen should be examined with a

394 ADDITIONS.

high power to see that the layer of jelly is not too
thick. If it is, warm the specimen and gently press
the cover-slip, let cool, scrap away the pressed out
jelly, and surround the edges of the cover-slip with
cement,

Mounting in Farrant’s solution.
Pure gum arabic 40 grams,
Water 40 c.c.
Glycerine 20 c.c.
A little thymol or other preservative.

The mixture may be made thus:—Take lumps of gum
arabic which are nearly free from colour, weigh
out 40 grams, grind to powder, place in about
150 c.c. water, warm or boil to dissolve. Add a
few drops of a strong alcoholic solution of thymol.
Filter through a hot filter, changing the filter when
clogged. Evaporate the filtrate until it is about
80 c.c., then add the glycerine. The mixture is best
kept in a bottle with a glass cap.

When mounting in this, let a section soak for a minute
in glycerine, then in a drop of Farrant’s solution on
the slide. Before cementing, the solution should be
allowed to become firm at the edges.

STAINING. .
After treatment with osmic acid. When it is re-
quired to harden a tissue in osmic acid and after-
wards to stain it, the tissue should be left for half-

an-hour to two hours only in the acid ; then washed
well with water and kept in alcohol for a few days.

Nitrate of silver preparations. The reduction of
silver in these may be greatly retarded by placing
them for a few minutes in a 2 to 10 p.c. solution of

a

ADDITIONS. 395°

sodium hyposulphite, after the required degree of
reduction has taken place. The sodium hyposulphite
must be thoroughly washed out with water.

Delafield’s hematoxylin.
Make (1) A saturated solution of hematoxylin in
absolute alcohol.

(2) A saturated solution of ammonia alum in
the cold.

Add 4 c.c. of (1) to 150 ec. of (2), leave for a week,
filter, to the filtrate add 25 c.c. glycerine and 25 c.c.
of methyl-aleohol.

This is often called Grenacher’s hematoxylin. It stains
nuclei well and quickly. To dilute it add some of
the mixture minus the hematoxylin.

Staining for indirect nuclear division. The tissue
should be hardened in Flemming’s mixture (cp. p.
388). The sections are placed in a strong alcoholic
solution of safranin, e.g. a saturated solution of
safranin in absolute alcohol diluted with an equal
quantity of water, and left for a day. They are
treated with dilute alcohol and then placed in abso-
lute alcohol containing ‘3 to ‘5 p.c. hydrochloric
acid. In this fluid they are left until colour ceases
or nearly ceases to be extracted from them. It is
best to examine a section from time to time, when
in such a section the cell-substance is decolourized
the remaining sections should be removed (twice) to
absolute alcohol free from acid. They are then placed
in clove oil, and as soon as they become transparent
mounted in Canada balsam. Sections may also be
stained with Delafield’s hematoxylin.

396 ADDITIONS.

To observe the typical changes in indirect nuclear —
division, a larval salamander 2 to 3 c.m. long, ~
hardened in Flemming’s mixture, may be conveni-
ently taken, divided into two or three pieces, and
then cut longitudinally. .

Staining medullated fibres in grey substance.
Weigert’s method.

Prepare

(1) Mordant solution. An aqueous solution of normal
cupric acetate saturated in the cold, and add an —
equal volume of water.

(2) Staining solution. Dissolve 1 gram of hema-
toxylin in 10 c.c. of absolute alcohol, add 90c¢.c. —
of water and 1 c.c. of a cold saturated aqueous
solution of lithium carbonate (or sodium car- —
bonate), aie + tues

(3) Decolourizing solution. Dissolve 2} grams of —
potassium ferricyanide and 2 grams of borax
in 200 c.c. of water,

Sections are placed in the cupric acetate in the warm —
for a day, washed in water, and transferred to hema- —
toxylin for two hours toa day. The sections, now
quite black, are washed with water and placed in the —
potassium ferricyanide solution for a few minutes to —
two hours, until the grey substance becomes of a
light tint ; if the decolourization is rapid, it is better
to dilute with water the ferricyanide solution; the —
sections are then well washed with water, treated
with alcohol, and mounted in Canada balsam.

Another method of decolourizing is given by Pal; the sections

after having been stained, are washed with water containing
a little lithium carbonate, placed in ‘25 p.c, potassium per- —

Se ee ar may

_ADDITIONS. 397

manganate for 15 to 20 secs., washed with water, placed for
a few seconds to a few minutes inamixtureof

1 gram pure oxalic acid

1 gram potassium sulphite

200 c.c. water,
and again washed well with water. The parts of the tissue
other than medullated fibres are thus more completely de-
colourized than by Weigert’s method. The sections may
finally be stained with alum-carmine.

Solutions for Liebig’s method of estimating urea.
(a) Standard mercuric nitrate solution, Take 71°48

grams of pwre mercury, add 5 vols. nitric acid Sp.
Gr. 1:425, and warm on water-bath until the mercury
is completely dissolved; evaporate the solution to
a syrupy consistence, until the addition of a few
drops of nitric no longer causes red nitrous fumes
to come off, evaporate further until the fluid acquires
a faint yellowish tinge, Stir and add about 10 vols,
of water; if a precipitate is formed let stand, pour
the clear fluid into a litre flask, to the residue add
the minimal quantity of strong nitric acid necessary
to dissolve the precipitate, add this to the fluid in
the flask, wash the vessel out with water, add this
to the previous fluid and fill up to exactly a litre.

One c.c. of this solution precipitates 10 mgs. of urea
and leaves just enough mercuric nitrate over to be
detected by sodium carbonate.

The method given above can only be used when the
mercury is pure, if there is any doubt about this, the
mercuric nitrate solution diluted to somewhat less
than a litre should be titrated with a solution of pure
urea and then further diluted to the proper amount.

(6) Baryta mixture, Add two volumes of barium
nitrate, saturated in the cold, to one volume of
barium hydrate, also saturated in the cold,

398 ADDITIONS.

A standard urea solution for titrating the mercuric
nitrate solution may be made by drying urea over
sulphuric acid, weighing out 4 grams and adding
water to make up 200 cc. 10 cc. of this just
give the end-reaction with 20 ¢.c. of standard mer-
curic nitrate solution. This however is only the
case when the mercuric nitrate solution is neut-
ralized by sodium carbonate immediately after
being added to the urea solution. As this point is
neglected in the method given in the text, it may —
be neglected in preparing the standard solution.

Liebig’s method.

Take 40c.c. of urine and add 20c.c. of the baryta
mixture a copious precipitate of barium phosphate,
sulphate, carbonate will be formed. Shake to mix
well the fluids and filter through a dry filter. The
object of this is to remove the phosphates, these give
a precipitate with mercuric nitrate.

Fill a burette with the standard solution of mercuric
nitrate. Ona glass plate lying on a dull black surface, —
place a number of drops of a saturated solution of
normal sodium carbonate.

Preliminary estimation. In a beaker place exactly
1l5c¢c.c. of the filtered urine-mixture, into this run
slowly the mercuric nitrate solution from the burette.
Stir with a glass rod, and from time to time, add a
drop of the mixture to one of the drops of sodium
carbonate ; as soon as a yellow colour is thus produced,
note the amount of mercuric nitrate which has been
added. The yellow colour is produced when all the
urea has been precipitated and there is an excess of
mercuric nitrate ; this with sodium carbonate gives a

ADDITIONS. 399

yellow precipitate of oxide of mercury or of a basic
mercuric carbonate.

Second estimation. In the preliminary estimation too
niuch mercuric nitrate will probably have been added ;
so the process should be repeated, adding at once the
amount of mercuric solution found in the preliminary
estimation less le.c. After well mixing, a drop is
added to a drop of sodium carbonate; if a yellow
colour results, the process must be repeated adding
less mercuric nitrate; probably however the mixed
drops will be colourless, if so add ‘le.c. mercuric
nitrate, mix and test again, and so on adding ‘lc.c. at
a time until a yellow colour is obtained.

Correction. Since the sodium chloride in the urine
prevents the precipitation of urea by mercuric nitrate
(cf. § 5e) more mercuric nitrate is added to the urine
than is required to precipitate the urea in it. The
excess thus added is found experimentally to be about
2c.c. for 10c.c. of urine. Hence 2 ¢.c. must be deducted
Srom the number of c.c. of the standard solution actually
used.

If v be the corrected number of c.c. of mercuric nitrate
used, the 10c.c. of urine taken, contain ‘Olv grams of
urea, i.e. the percentage of urea in the urine is the O-v.

Note. The method as above given is very rough. For
an accurate estimation, the sodium chloride as well as
the phosphates must be separated from the urine, the
filtered mixture must be neutralized, and again neu-
tralized with sodium carbonate after each addition of
mercuric nitrate. It must also be remembered that
the amount of urea found by this method is too high,

400 ADDITIONS.
since ammoniacal salts, and all the nitrogenous bodies
in the urine give a precipitate with mercuric nitrate.

Hypobromite method of estimating urea. (Cf —
p. 242). q
Hypobromite solution. Dissolve 100 grams of —
sodium hydrate in 250c.c. water, cool, and add
gradually 25c.c. of bromine, cooling the mixture

in astream of water as the bromine is added. The

&

, a
/ Sa
’
4

| Wil

~—

EEE EEO —— <=

" ADDITIONS, | 401

sodium hydrate solution should be kept ready, and
bromine added just before the mixture is required.
It is convenient to have thin glass capsules, each
containing 2°3c.c. of bromine, if one of these be
placed in 25c.c. of the hypobromite mixture
broken by a sharp shake, the proper amount of
hypobromite solution for a single observation is
obtained,

Method of using Gerrard's apparatus. In this, as
in many other forms of apparatus, the collecting
tube is graduated so as to show percentages of urea
when 5c.c. of a urea solution are taken.

Raise the tube (a) to about the height shown in
the figure, pour water into it until the tube (0) is
filled with water to the zero mark. Pour 25c.c, of
hypobromite solution into the bottle (c), measure
from a burette 5c.c. of urine into the tube (d).
With the aid of a forceps, place the tube (d) in the
bottle, being careful not to spill any of the urine.
Fit the cork, with the tube attached to it, tightly
into the bottle, and clamp the tube (e). Fill up
and gently shake the bottle so that the urine and
the hypobromite solution mix. Place the botile
in a basin of water. Lower (a) until the level of
the water in it is the same as that in (b). Leave
for about five minutes, re-adjust the level of the
water in the two tubes, and note how much
nitrogen has been given off,

An estimation of urea should also be made, using

two burettes instead of the tubes (a) and (0).
The volume of nitrogen evolved is measured in

L, 26

. Deficit of Nitrogen. The chief causes of variation

instead of giving off 373 vols. of nitrogen gives off

_ volume of the gas is measured at a temperature

ADDITIONS.

cubic centimetres, and from this the percentage i
urea is calculated in the manner given below.

in the amount of nitrogen given off, have been
mentioned in the text. Hiifner, Pfliiger and
Schenk find with the apparatus used by them, and
with a 1 p.c. solution of urea, that the deficit of
nitrogen is very nearly 44 p.c., ie. 1 gram of urea

357.
Russell and West using a 2p.c. solution of urea
find with their apparatus a deficit of nearly 8 p.c.,
i.e. 1 gram of urea, instead of giving off 373 vols,
of nitrogen gives off about 343 vols. When the

of 18°C., no correction being made for the tension
of the aqueous vapour, the deficit of 8 p.c. is nearly
corrected by the expansion of the gas, so that
according to Russell and West, in making an
approximate estimate of urea with their apparatus, —
no account need be taken of. a deficit of nitroge
nor any correction be made for temperature and
pressure. |

Correction for temperature and pressure. Ifv' be the |
volume in cubic centimetres of the nitrogen obtained, —
at temperature ¢’ C. and pressure B in mm. of mer-

cury, and 7’ be the tension of aqueous vapour at CO,
the volume v at 0°C. and 760 mm. pressure will be

v B-T |

1+ 003665 ¢ ~~ 760 ’
‘003665 being the coefficient of expansion of gases, —

ADDITIONS. 403

Calculation of percentage of urea. Theoretically
373 c.c. of nitrogen are given off by 1 gram of
urea, if then vec. of nitrogen, after correction
for temperature and pressure, are obtained in
an experiment, this will have been given off by
Ps grams of urea, that is there are 573 grams of
urea in the 5c.c. of urea solution taken, so that

the percentage of urea in the solution is ie

But since in practice the whole of the nitrogen is
not given off, the deficit must be allowed for, if we
take Hiifner’s estimate of the deficit of the volume
of the nitrogen, viz. 44 p.c., the percentage will be
20 v

357 .

Sources of error in estimating urea in urine. Nitro-
genous bodies other than urea are partially decom-
posed. Further certain substances increase and
others decrease the completeness of the decom-
position.

26—2

INDEX.

A.

Abdominal aorta, 23

Abdominal muscles, of dog and
rabbit, 1

Absorption of fat in small intes-
tine, 183

Accommodation of the eye, 275

Acetic acid, effect on blood cor-
puscles, 38, 39, 42; effect on
cartilage cells, 53; on elastic
fibres, 58; on white fibrous
tissue, 59, 62; on striated
muscle, 83

Acid-albumin, 45, 101, 102, 173

Adenoid tissue, 179, 191

After-image, positive, 285; nega-
tive, 286, 287

Albumin, 51; in milk, 175; in
urine, 241

Alcohol, for dissociating tissues,
358 ; for hardening tissues, 364

Alkali-albuminate, 102, 103

Alkalinity of blood, 45

Alum-logwood, 372

Ampulla, 291

Anterior nares, 30

Aorta, in heart of sheep, 136, 137

Appendix of large intestine of
rabbit, 3

Aqueous humour, 264

Arterial scheme, minor,
major, 148

Arteries, caeliac, 4; superior me-
senteric, 4; renal, 4; splenic,
6; hepatic, 6, 7; lieno-gastric,
6; pancreatic, 6; common
carotid, 18, 20; innominate,
20, 137; pulmonary, 21; fe-
moral, 23, 91; coronary, 140;

147;

interlobular, 234; vertebral,
313; basilar, 313; internal
carotid, 313; posterior cere-
bral, 313 ; middle cerebral, 313;
anterior cerebral, 314; struc-
ture of, 124

Artery, epithelium of, 126

Arytenoid cartilage, histology, 70;
dissection, 333

Arytenoid muscle, 333

Astigmatism, 279

Auricular nerve of rabbit, 25

Automatic action, 120

Axial zone of blood current, 131

Axis band, of malleus, 296

Axis-cylinder, 106

Azygos vein, in heart of sheep,
138

B.

Basilar membrane, 299; artery,
313

Bidder’s ganglia, 145

Bile, reaction of, 184; effect on
gastric digestion, 187; capil-
laries, 204; ducts, 206; -pig-
ment, 241; -acids, 242

Bile-salts, preparation of, 185

Biuret reaction, 239

Bladder, 10; structure of, 235

Blind spot, 281

Blood pressure, 147

Blood, structure of, 35 ff; of frog
or newt, 35; of man, 41; co-
agulation of, 44 ff; circulation
of, 131, 134; colour of, 219

Bone, 30; structure of, 72; ex-
traction of salts from, 359, 361

Boracic acid, effect on red corpus-
cles, 40

— _—

INDEX, 405

Brain, of dog or sheep, dissection,
308; method of hardening,
308, 327; parts seen in sec-
tions, 321

Brunner, glands of, 181

Bulb, olfactory, 323

Bulbus arteriosus, in heart of
frog, 141, 142

C.

Caecum, 2

Caeliac artery, 4

Calamus scriptorius, 313

Calices, of kidney, 228

Camera lucida, 352

Canada balsam, 379

Canaliculi, 73

Capillaries, structure of, 127

Cardiac muscle, 88

Carmine, 373; picro-, 374; borax-,
374

Carotid artery, common, 18, 20;
internal, 313

Cartilage, hyaline, 52; costal, 33,
54, 55; of cuttle fish, 56; paren-
chymatous, 56, 57; interverte-
bral, 68 ; fibro-, 70; elastic, 71;
arytenoid, of sheep, 70

Casein, in milk, 175, 176

Cedar wood oil, 375 ~

Central canal, 302

Cerebellum, of dog or sheep, 308;
inferior peduncles of, 310; su-
perior peduncles of, 310; mid-
dle peduncle of, 311; structure
of, 329

Cerebral arteries, posterior, 313;

_ middle, 313 ; commissures, 320,
321; anterior and posterior,
321

Cerebral hemispheres, of rabbit,
11; of dog, 12

Cerebrum, of dog or sheep, 308

Cervical ganglion, inferior, 20;
superior, 28

Chemical stimulation of muscle,

93
Cholesterin, 187
Chorda tympani fibres, 26, 27
Chordae tendineae, 139

Choroid, 264; pigment of, 273;
plexus, 309, 316, 319

Chromic acid, staining after, 69 ;
for hardening, 361

Ciliary action, 81, 122; muscle,
264, 267 ; processes, 265

Ciliated cells, of frog, 80

Circulation of blood (frog) in
web, 131; in tongue, 134; in
mesentery, 135

Clarke, column of, 305

Cochlea, promontory of, 295;
structure of, 299, 300

Commissure, of spinal cord, 303;
of brain, 320, 321

Common iliac, 23

Condyloid foramen, 29

Conjunctiva, 263

Connective tissue, 58; sub-cuta-
neous, 247

Constant current, effect on nerve,
90, 91

Contrast, simultaneous, 287

Convolutions, of dog, supra-callo-
sal, 324; internal, 324; Sylvian,
324; inferior, 324; median,
325; superior, 325

Cornea, of frog, 61, 109, 250, 268;
of cat, 251, 263, 264

Cornu, anterior and posterior, 302

Corona radiata, 323

Sey vein, in heart of sheep,

8

Corpora, quadrigemina, 310;
lutea, 337

Corpus, Arantii, 139; albicans,
311; callosum, 315, 322; ge-
niculatum, 320; Highmori, 341

Corpuscles, in blood of frog, 36,
38, 39, 133; in blood of man,
41, 42; connective-tissue, 60;
bone,74; Malpighian, 195, 197;
touch-, 252

Corpus dentatum, of cerebellum,
312

Cortex of cerebrum, structure of,
327

Corti, rods of, 300

Cover cells, 260

Cranial nerves, 11; superficial
origin, 314°

406

‘Creosote, 378

Crico-arytenoids, posterior and
lateral, 331

Cricoid cartilage, 332

Crico-thyroid muscle, 332

Crista acustica, 292

Crucial fissure, 325

Crura cerebri, 311

Crural vein, 23

Crusta of brain, 322

Crusta petrosa, 77, 78

Cumulus proligerus, 338

D.

Decussating fibres of bone, 75

Decussation of the anterior pyra-

mids, 326
Demilune cells, 156
Dental nerve, inferior, 28
Dentate fissure, 319
Dentine, 77, 78
Depressor nerye, 19, 21
Dermis, 247
Descemet, membrane of, 209
Dextrin, 162
Dextrose, 162
Diaphragm, of dog and rabbit,
7,11; structure of, 201, 202
Diffusion circles, 276
Digastric muscle, 26, 27
Dissection, of rabbit and dog, 1
Dissociating fluids, 358
Dorsal aorta, 4
Dorso-lumbar fascia, 2
Duodenum, 3, 4, 9
Dura mater, of dog, 11, 129, 308
Dialyser, 165

E.

Elasticity of red corpuscles, 40
Electrodes, non-polarizable, 348
Enamel of teeth, 77, 78
Epidermis, 24

Epididymis, 10, 341

Epiglottis, 30

Epithelial nerve plexus, 251
Erlicki’s fluid, 361

Eustachian tube, 31, 298

INDEX.

Eustachian valve, in heart of ©
sheep, 138 ‘

Eye of sheep, dissection of, 263 ;
retina of, 264

F,

Facial nerve, 24

Fallopian tube, 10

Falx cerebri, 11

Fasciculus, of Goll, 305; cuneatus, —
305, 309; gracilis, 309

Fat-cells, 65

Fat-ferment of pancreatic juice,
190

Fehling’s fluid, 381

Femoral artery, 23, 91

Fenestra ovalis, 298

Fenestrated membraneof arteries,
126 ry

Fibres, elastic, 58, 214; white,
58; perforating, 74; of Muller, —
272

Fibrin ferment, 49

Fibrin, 44; for digestion experi- —
ments, 172

Fibrin fibrils, under microscope,
41 /

Fibrinogen, 48

Fimbria, 316

Fissures of spinal cord, anterior
and posterior, 302 ; (of brain of
dog,) crucial, 325; Sylvian,
324; dental or hippocampal,
319

Foramen, jugulare, 19, 29;
condyloid, 29; rotundum, 295;
of Monro, 318

Fornix, 316; anterior and pos-
terior pillars of, 317

be ovalis, in heart of sheep,
I *

Freezing microtome, 369

Fresh tissues, methods of ob-
serving, 355

G.

Gall-bladder, 7

Ganglion, spinal, 14, inferior
cervical, 20; first thoracic, 20;
of the vagus, 28; superior

INDEX.

cervical, 23;
112; celis, 111

Gasserian ganglion, 32, 112

Gastric juice, artificial, ;
action of, 172, 173; action of,
on milk, 176

Gastric veins, 6

Germinal epithelium, 336;
vesicles, 337 ; spot, 337

Gland, thyroid, 23; parotid, 24;
submaxillary, 26 ; Harderian,
31; orbital, 31; lachrymal,
31; intestinal, 179; of Lieber-
kiihn, 179; of Brunner, 181 ;
serous and mucous, 214; seba-
ceous, 249; pineal, 320

Globulin, 51, 101

Glomeruli, 231

Glosso-pharyngeal nerve, 29, 141

Glycerine, 378

Glycogen, 205, 208

Gmelin’s test, for bile pigments,
185, 241

Gold chloride, to stain cartilage,
53; to stain cornea, 61; sub-
cutaneous connective tissue, 63 ;
septum auricularum, frog, 144;
lung of newt, 211, 376

Goll, fasciculus of, 305

Graafian follicles, 336, 337

Gum arabic for imbedding, 369

Gasserian, 32,

H.

Haematoxylin, to stain cartilage,
56; Kleinenberg’s, 371; alum,
372

Haemin, crystals of, 226

Haemoglobin, 40; to prepare,
220, 221; crystals of, 221;
reduced, 224

Hair-follicles, 248; -cells of organ
of Corti, 299, 300

Hardening agents, method of
using, 359; to make solutions,
360

Harderian gland, 31

Haversian canal, 72

Heart, of frog, 120, 140; of sheep,
136, 137 —

Helmholtz, phakoscope of, 278

407

Hepatic artery, 6, 7, 206; duct
7; veins, 8; cells, 206
Hippocampal fissure, 319
Hippocampus major, 316
Hyaloid membrane, 265
Hydrophilus, muscle of, 84
Hypoglossal nerve, 28, 29

Z.
Tleum, 3
Imbedding, 365 ff
Tneus, 297

Induction machines, 346
Induction shocks, effect on nerve,
92

Inferior cervical ganglion, dog, 23

Inflammation, 133

Infundibulum, 312

Inhibition of heart by vagus, 142

Injection mass, 380

Inner capsule, 323

Innominate artery, 20; in heart
of sheep, 137

Innominate veins, dog, 22

Intermediate layer of sheep’s
kidney, 228, 229

Intermedio-lateral tract of nerve
cells in spinal cord, 305

Internus obliquus abdominis, 2

Interrupted current, effect on
nerve, 93

Intercostal muscle, srioron’ 15;
internal, 16

Intestinal glands, 179, 182

Intestine, small, of dog and
rabbit, 2; large, 3, 9; method
of hardening, 33; structure of,
178

Intralobular veinlet, 205, 206,
234

Iodine, 376
Iodized serum, 357, 353
Irradiation, 281
Iris, 270
Iter, 313

J,
Jacobson’s nerve, 29
Jejunum, 3

Jugular veins, 18, 19

408

K.

Keys of du Bois-Reymond and
Morse, 345

Kidneys of dog and rabbit, 9, 10 ;
of sheep, 228

L.
Lachrymal gland, 21
Lactic acid, 176
Laky blood, 219
Lamina fusca, 264
Lamina spiralis, 298
Laryngeal branches of vagus, 18,
20

Larynx, dissection of, 331
Lateral, columns of spinal cord,
302; ventricle of brain, 316

Lens of ox or sheep, 269

Levatores costarum, 15

Lever for frog’s heart, 349

Liebig’s method of estimating
urea, 398

Lieno-gastric veins, 5; artery, 6

Lieberkiihn, glands of, 179

Ligamentum pectinalium iridis,
267

Ligamentum nuchae, 58

Limbic lobe, 324

Linea alba, of dog and rabbit, 1

Lingual nerves, 26

Liver, dissection of, 7; histology,
204

Longissimus dorsi, 15

Lumbar fascia, 2

Lung, structure of, 211 ; injection
with silver nitrate, 216, 217

Lymph-channels, 193

Lymph follicles of intestine, 180

Lymph-hearts of frog, 120

Lymph-sinus, 191

Lymphatic capillaries, structure
of, 200

Lymphatic-glands, structure of,
191; method of hardening, 192

M.

Malleus, posterior ligament of,
296; anterior ligament of, 296;
axis band of, 296

INDEX.

Malpighian corpuscles, 195, 197,
199

Malpighian layer of epidermis,
247

Mammary glands, dissection in
rabbit, 1

Masseter muscle, 25

Maxillary nerve, inferior, 28;
superior, 31

-Maxwell’s method of detecting

yellow spot, 284

Measurement of microscopic ob- —

jects, 37, 354

Mechanical stimulation of nerve,
94

Mediastinum, 16

Meduila oblongata, 11, 309

Medullary sheath of nerves, 105; —

-cords of lymphatic glands, 193;

-rays of kidney, 229
Medullated nerve-fibres, 105
Membrana granulosa, 337
Membrane,mucous(Schneiderian),

30; hyaloid, 265; of Descemet,

269; inner limiting, 271; outer —

limiting, 273; tympanic, 295;
basilar, 299; of Reissner, 299
Membranous semicircular canal,

291
Mercurial manometer, the, 148

Mesenteric lymphatic glands, 43

veins, 5

Mesentery, of dog and rabbit, 3;
nerves in, 109

Methylene blue, 375

Micrometer, ocular, 354

Microscope, magnifying power of,

35, 355; notes on use of, 351, —

352
Microtome, 368 389
Millon’s reagent, 381
Milk, fat globules in, 174; alka-

linity of, 175; albumin in, 175; —
casein in, 175, 176; fermenta- —

tion of, 176
Mitral valve, 140
Modiolus, 298
Moist chamber, 348
Molecular layers, retina, 273
Monro, foramen of, 318
Morse key, 345

*:

INDEX.

Mounting, 378, in glycerine, 55;
method for thin membrane, 59;
in Canada balsam, 66

Mucin, 161; of bile, 184

Mucous cells, of salivary glands,
156; of stomach, 168; of in-
testine, 179

Mucous coat of stomach, 168, 214

Mucous membrane, stomach of
rabbit, 9

Miller, fibres of, 272

Miiller’s fluid, 362

Murexide test, for uric acid, 249

Muscle, pectoral, 14; external
intercostal, 15; internal inter-
costal, 16; masseter, 25; digas-
tric, 26, 27; niylohyoid, 26, 27;
structure of, 82, 85, 88; reaction
of, 98, 99; ciliary, 264; tensor
tympani, 297; stapedius, 298;
arytenoid, 334

Muscular coat of stomach, 168

Muscular contraction, in frog, 9,
95

Mylohyoid muscle, 26, 27

Myosin, 100, 101

N.

Nasal bones of rabbit, 30

Negative after-image, 286

Nerve fibres, method of harden-
ing, 34; medullated, 105; non-
medullated, 108

Nerve-muscle preparation, 95

Nervoussystem. Splanchnicnerve,
4, 5, 21; pneumogastric nerves,
5, 18, 27, 29, 142; cranial, 11;
optic nerve, of rabbit, 12; of
sheep, 264; fourth and fifth
nerve, 12; ninth, tenth, eleventh
and twelfth,13; phrenic nerve,
17, 19; sympathetic, 18; superi-
or cardiac, 18; crural, 23; facial,
24; sciatic, 24,91, 95; auricular,
of rabbit, 25; lingual, 26; pha-
ryngeal, 27; hypoglossal, 28,
29; inferior maxillary, 28; in-
ferior dental, 28; glossopharyn-
geal, 29, 142; spinal accessory,
29; Jacobson’s, 29; olfactory,

409

30; ophthalmic, 31; superior
maxillary, 31; trigeminal, 32;
superficial origin of, 314
Nodes of Ranvier, 106, 107
Normal fluids, 52, 356
Nuclear layers, retina, 273
Nucleus, of cells (hyaline carti-
lage) ,52,53; of primitivesheath,
107 ; of capsule of nerve cells,
111; of capillaries, 128; of ad-
ventitia, 128; of salivary glands,
158, 159; of alveolar cells of
lung, 212; caudatus, 316, 323;
lenticularis, 323

O.

Obliquus externus abdominis, 2

Oleic acid, action of bile on, 186;
effect on pancreatic digestion,
189

Oesophagus, of dog and rabbit, 2,
7; structure of, 170, 171

Olfactory, lobes, 11, 324; nerves,
30; mucous membrane, struc-
ture of, 257 ff; bulb, 323; tract,
324

Olivary bodies, inferior, $11

Omentum, 3

Optic, fibres, layer of, 272; tracts,
312; chiasma, 312; thalamus,
319

Optic nerve of rabbit, 12

Ora serrata, 265

Orbital gland, 31; lobe, 326

Oscillating rod, 349

Osmic acid, for dissociating tis-
sues, 358; general directions
for use, 363

Ossification, 75

Osteoblasts, 76, 77

Osteoclasts, 76

Ovary, dissection, 10; histology,
336

Ovoid cells of stomach, 168

Ovum, 337

P,

Pacinian bodies, 252
Pancreas, dissection, 3, 6; histo-
logy, 159, 160

410

Pancreatic duct, 8, 4; arteries, 6;
_ veins, 63; jetice, artificial, 188

Papilla foliata, of rabbit, 32;
structure of, 259 ff

Papillae, of skin, 247

Parotid gland, 24; duct of, 25

Paraglobulin, 48

Parovarium, 339

Paraffin, 367, 368

Pectoral muscle, 14

Pelvis of sheep’s kidney, 228

Pepsin, preparation of, 171; de-

. struction of by acid, 173

Peptone, characters of, 174

Perforated space, posterior, 311

Pericardium, 17

Perichondrium, 64

Periosteum, 77

Peripheral zone of blood current,
131

Peritoneum, 3

Pettenkofer’s test, for bile acids,
185, 242

Peyer’s patches, 8, 180, 191

Pharyngeal nerve, 27

Phrenic nerve, 17, 19.

Pia mater, 11, 129, 301, 308

Picric acid, 362

Pigment-cells, of frog’s web, 65;
of retina, 274

Pigment of choroid, 273

Pineal gland, 320; peduncles of,
320

Pituitary body, 312
Plasma of blood, 45, 46
Plasmine, 47
Platysma, 18
Pleura, 16, 17
Pneumogastric trunks, 20;

nerves, 5, 18, 22, 23, 27, 28,
142 ; superior Jaryngeal branch
of, 18; pharyngeal branch of,
(ioe

Pons Varolii, 311

Portal vein, 5, 7

Positive after-image, 285

Posterior root of spinal nerve of
rabbit, 13, 302; columns, 302

Preserving agents, method of
using, 359; to make nenreons,
360

INDEX,

Primary nerve-plexus, 250 ©

Primitive sheath, 105

Processus gracilis of malleus, 296

Proteids, general reactions of,
49; in saliva, 161

Piyalin, to prepare extract of,
165; destruction of by acid,
165

Pulmonary arteries and veins, 21,
137

Pulse waves, progression of, 151

Purkinje’s figures, 282

Pyloric glands, 170

Pyramids, anterior, 511

Q.
Quantitative estimation of urea,
hypobromite method, 242, 460

R.

Rabbit, dissection of, 1
communicantes, 21

Ranvier, nodes of, 106, 107

Recti abdominis, 2

Rectum, 3, 7

Red corpuscles, method of count-
ing, 42

Reflex action, 116

Reil, island of, 326

Reissner’s membrane, 299

Renal, artery, 4; vein, 5

Rennet-ferment, 176

Restiform body, 309

Rete testis, 341 :

Retina, dissection of, 264; histo-
logy of, 271

Rigor mortis, 99

Rima glottidis, 331

Rod cells of olfactory mucous
membrane, 257; of taste-buds,
261

Roots of spinal nerves, 13

s.
Sacculus of semicircular canals,
292

Saline solution, normal, 357
Saliva, 161

INDEX.

Salivary glands, structure of, mu-
cous, 155; serous, 158

Sartorius muscle, rabbit, 23.

Scalenus medius, 15; — anticus,
15; — posticus, 15

Scheiner’s experiment, 278

Schneiderian membrane, 30

Sciatic nerve, 24, 91

Sebaceous gland, 249

Section cutting, 368

Semicircular canals of ear, 290 ff

Septum auricularum, in heart of
sheep, 138; in heart of frog,
145; structure of in frog, 144

Septum lucidum, 317, 322

Serratus anticus, major, 14;
minor, 15; serratus posticus, 15

Serum, iodized, 357, 358

Sigmoid gyrus, 325

Silver nitrate, 125, 130, 197,217,

Solar plexus, 4, 5

Solutions, preparation of, 360

Spectrum of haemoglobin, 222

Spermaceti and castor oil, 363

Spermatozoa, 344

Sphincter pupillae, 271

Sphygmograph, 150 _

Spiller’s purple, to stain red cor-
puscles with, 39; to stain con-
nective-tissue cells, 60, 375

Spinal accessory nerve, 29

Spinal cord, 114, 301; structure
of, 110; method of hardening,
301

. Spinal ganglion, dissection of, 14;
structure of, 110

Spiral process of nerve cells, 145

Splanchnic nerves, 5, 21

Spleen, dissection, 3; histology,
194, 195; method of hardening,
196; method of injecting, 197;
Malpighian corpuscles of, 197,
199

Splenic artery, 6; pulp, 195, 196

Stapedius muscle, 298

Stapes, 297, 298

Starch mucilage, preparation of,
162

411

Sterno-cleido-mastoid musele, 22
Sterno-hyoid muscle, 21
Sterno-mastoid muscle, 18
Sterno-thyroid muscle, 22
Stokes’ fluid, 382, 383
Stomach, of dog and rabbit, 2, 8;
method of hardening, 32;
structure of, 167, 168
Stomata, 201
Stylo-mastoid foramen, 25
Subclavian veins, 19, 20
Sub-epithelial nerve plexus, 251
se gland, 26; — duct, 26,

Submaxillary gland, disseetion,
26, 27; duct, 26, 275 histology,
155

Substamtia gelatinosa, 303

Substantia nigra, 322

Sugar, quantitative estimation in
urine, 243

Superior mesenteric artery, 4

Superior vena cava, dog, 22

Suprarenal body, 4

Sweat-glands, 250

Sylvius, aqueduct of, 313; fissure
of, 324

Sympathetic ganglia, dissection
of, 20, 21, 28; structure of, 113

Sympathetic nerve in neck of dog,
22, 23, 28; in rabbit, 18, 20,

28
Syntonin, 101, 102

T.

Tactile delusions, 256
Tail, of tadpole, 60; of mouse, 58,
62

Taste, 261

Taste-buds, 260

Teasing, 357

Teeth, structure of, 77
Tegmentum, 322
Temperature, sensation of, 254
Tendon, 62

Tensor tympani muscle, 297
Tentorium, 11

Testis, 10, 340

Tetanus, 97

Thalamus, optic, 319

412

Thoracic duct of dog, 17
Thoracic ganglion, 20

Thymus, 19

Thyro-arytenoid muscle, 334, 335
Thyro-hyoid muscle, 22

Thyroid, gland, 23; cartilage, 332
Time marker, 350

Touch, 254

Touch-corpuscles, 252

Trachea, histology of, 215
Transversalis abdominis, 2
Tricuspid valve, 138

Trigeminal nerve, 32

Trommer’s test, 162, 163

Tuber cinereum, 311

Tubuli seminiferi, 341, 342
Tunica albuginea, 340

Tunica vaginalis, 10; propria, 310
Turpentine, 378

Tympanic membrane, 295

U.

Uncinate lobe, 324

Unstriated muscle, structure of,
86, 87

Urea, 237; crystals of, 237; ni-
trate of, 237; oxalate of, 237

Ureter, dissection, 9; histology,
228; structure of, 235

Uric acid, 240

Urinary bladder, 10

Urine, acid in, 237; alkaline fer-
mentation in, 237; mucus in,
237; albumin in, 241; sugar in,
241

Uterus, 10, 340

Utriculus, 291

Uvea, 270

V.

Valsalva, sinuses of, 139
Vas deferens, 10, 343
Vasa efferentia, 341, 343

INDEX.

Veins, renal, 4; portal, 5; lieno-
gastric and mesenteric, 5; por-
tal, 5; gastric and pancreatic, —
6; external jugular, 19; sub-
clavian, 19, 20; pulmonary, 21,
137; innominate, 22; femoral, —
23; structure of, 127; azygos
in heart of sheep, 138; coro- —
nary, 138; interlobular, 234

Velum interpositum, 318

Vena cava, inferior, 4, 19, 121,
137, 142; superior, 19, 121, 137,
142

Ventricle, fourth, 309, 310; late-

. el, 316; fifth, 317; third, 319;

- 320

Vertebral artery, 20, 313

Vieussens, valve of, 310

Villi, 9; structure of, 178, 181

Vision, region of distinct, 282;
region of normal colour, 285

Visual judgments, 289

Vitreous humour, 265

W.

Weight, estimation of, 254, 255.
Willis, circle of, 314

te
Xanthoproteic reaction, 49, 50,
174

Xe
Yellow spot, 284

Z.

Zeiss, camera-lucida of, 352, 353

Zinn, zone of, 266

Zona pellucida, 338, 339

— of pancreatic gland-cells,
6

ADDITIONS TO INDEX.

Blood corpuscles, white, of frog,
method of obtaining, 43

Celloidin, 390

Farrant’s solution, 394
Flemming’s mixture, 388

Glycerine jelly, 392
Hematoxylin, Delafield’s, 395
Imbedding in paraffin, 388
Mercurie chloride, 387

Mounting ribbons of sections, 389

Nerve-fibres, staining by Weigert’s
method, 396

Nitrate of silver, treatment of
preparations, 394

Nuclear division, methods of
staining for, 395

Osmic acid, staining after, 304

Platelets of blood, 387

Ribbons of sections, 388

Sympathetic ganglia, dissection
of 23, 113; isolation of cells,
112, 115

Vienssens, annulus of, 23

Weigert’s method of staining
nerve-fibres, 396

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CONTENTS.

CLASSICS— ; PAGE
EvLementary Crassics ‘ = ° ote. 3
CLASSICAL SERIES ° ° . 7
CriassiIcaL LIBRARY, (x) Text, ®) Translations ~ e ° 12
GRAMMAR, COMPOSITION, AND PHILOLOGY ° e é 17
ANTIQUITIES, ANCIENT HISTORY, AND PHILOSOPHY . e 22

MATHEMATICS—

ARITHMETIC AND MENSURATION .¢ rs ‘* S é 24
ALGEBRA . ° “ > . 26
EUCLID, AND ELEMENTARY GroMETRY > é 3 ; 28
TRIGONOMETRY . ‘ 3 e ° > ° 29
HIGHER MATHEMATICS . e ° . e . . . 30
SCIENCE— :
NATURAL PHILOSOPHY . ” ¢ “ > ‘ ; > 37
ASTRONOMY. . . . . e - ° . 43
CHEMISTRY . e e e e : ° ° « 43
BioLocy . ° a © . . e ° , 45
MEDICINE . . se . . ° 7 . tal 49
ANTHROPOLOGY . Bs ° e e . 50
Puysicat GEOGRAPHY AND Giiotos¥ = ‘ * s 50
AGRICULTURE . > e e . ° > 52
PouiticaAL Economy §. Fy e . . ° 52
MENTAL AND Morat Putrosopxy > . ° . > 53

HISTORY AND GEOGRAPHY . oly jsf i Se 9955-99
Macmillan’s Geographical Series.

MODERN LANGUAGES AND LITERATURE—

ENGLISH . . - . . . . . ° 59
FRENCH = . - a . * e ° ° 65
GERMAN . e . e . e . e . 68
Mopvern GREEK . ‘ : . e e ° e 7°
ITALIAN @ ‘ e ° . ° e e qo
SPANISH . . € e ° . ° ° 7o

DOMESTIC ECONOMY e ° ‘ ‘ ° : 7
ART AND KINDRED SUBJECTS . ‘ ° ° 7%
WORKS ON TEACHING . * ° : > ‘ 72

DIVINITY a ¢ . 2 > e e . 73

29 AND 30, BEDFoRD STREET, CoveNT GARDEN,
Lonpon, W.C., Afri, 1888.

CLASSICS.

ELEMENTARY CLASSICS.
18mo, Eighteenpence each.

Tuis SERIES FALLS INTO TWO CLASSES—

(1) First Reading Books for Beginners, provided not
only with Introductions and Notes, but with
Vocabularies, and in some cases with Exercises
based upon the Text.

(z) Stepping-stones to the study of particular authors,
intended for more advanced students who are beginning
to read such authors as Terence, Plato, the Attic Dramatists,
and the harder parts of Cicero, Horace, Virgil, and
Thucydides,

These are provided with Introductions.and Notes, but
no Vocabulary. The Publishers have been led to pro-
vide the more strictly Elementary Books with Vocabularies
by the representations of mnany teachers, who hold that be-
ginners do not understand the use of a Dictionary, and of
others who, in the case of middle-class. schools where the
cost of books is a serious consideration, advocate the
Vocabulary system on grounds of economy. It is hoped
that the two parts of the Series, fitting into one another,
may together fulfil’all the requirements of Elementary and
Preparatory Schools, and the Lower Forms of Publie
Schools. are j

2

4 MACMILLAN’S EDUCATIONAL CATALOGUE.

The following Elementary Books, with Introductions,
Notes, and Vocabularies, and in some cases with
Exercises, are either ready or in preparation :—

Aeschylus.—pROMETHEUS VINCTUS. Edited by Rev. H.
M. STEPHENSON, M.A.

Arrian.—SELECTIONS. Edited for the use of Schools, with
Introduction, Notes, Vocabulary, and Exercises, by JOHN Bonp,
M.A., and A. S. WALPOLE; M.A.

Aulus Gellius, Stories from. Edited, with Notes and
Vocabulary, by Rev. G. H. NALL, M.A., Assistant Master in
Westminster School. . [Zn the Press.

Czesar._THE HELVETIAN WAR. Being Selections from
Book I. of the ‘‘ De Bello Gallico.” Adapted for the use of
Beginners. With Notes, Exercises, and Vocabulary, by W.
Wetcu, M.A., and C. G. DUFFIELD, M.A,

THE INVASION OF BRITAIN. Being Selections from Books
IV. and V. of the “De Bello Gallico.” ‘Adapted for the use ‘of
Beginners. With Notes, Vocabulary, and Exercises, by W.
Wetcu, M.A., and C. G, DUFFIELD, M.A.

THE GALLIC WAR. BOOK I, _ Edited by A. S. WALPOLE,

M.A.
THE GALLIC WAR. BOOKS II. anp III. Edited by the

Rev. W. G. RUTHERFORD, M.A., LL.D., Head-Master of West-
minster School.

THE GALLIC WAR. BOOK IV,. Edited by CLEMENT BRYANs,
M.A., Assistant-Master at Dulwich College.

THE GALLIC WAR. SCENES FROM BOOKS V. anv VI.

Edited by C. CoLBEck, M.A., Assistant-Master at Harrow;
formerly Fellow of Trinity College, Cambridge. ,
THE GALLIC WAR. BOOKS V. anp VI. (separately)... By
the same Editor. Book V. ready. Book VI. in preparation.
THE GALLIC WAR. BOOK VII. Edited by JoHN Bonn,
_ M.A,, and A. S. WALPOLE, M.A.
Cicero.—DE SENECTUTE. Edited by E. S. SHuCKBURGH,
M.A., late Fellow of Emmanuel. College, Cambridge.
DE AMICITIA. By the same Editor.
STORIES OF ROMAN HISTORY. Adapted for the Use of
Beginners. With Notes, Vocabulary, and Exercises, by the Rev.
G. E. Jeans, M.A., Fellow of Hertford College, Oxford, and
A. V. Jonzs, M.A. ; Assistant-Masters at Haileybury College.»
Eutropius.— adapted for the Use of Beginners. With Notes,
Vocabulary, and Exercises, by WILLIAM WELCH, M.Az, and C,
G. DUFFIELD, M.A., Assistant-Masters at Surrey County School,
Cranleigh,
Homer.—ILIAD. BOOK I. Edited by Rev. JoHN BOND, M.A.,
and A. S, WALPOLE, M.A. |

-. a ee

a

—— ee

ELEMENTARY CLASSICS. 5

Homer.—ILIAD, BOOK XVIII. THE ARMS OF ACHILLES,

Edited by S. R. James, M.A., Assistant-Master at Eton College.

ODYSSEY. BOOK I, Edited by Rey. JOHN Bonp, M.A. and
A, S. WALPOLE, M.A,

Horace.—ODES. BOOKSI.—IV. Edited by T. E. Pace, M.A.
late Fellow of St. John’s no Cambridge ; Assistant-Master
at the Charterhouse, Each Is,

Latin Accidence and Wicreised Arranged for Be-
GINNERS. By WILLIAM WE cH, M.A., and C. G. DuFFIELp,
M.A., Assistant Masters at Surrey County School, Cranleigh.

Livy. —BOOK I, Edited ‘by H. M. STEPHENSON, M.A., late
Head Master of St. Peter’s School, York.

THE HANNIBALIAN WAR, Being part of the XXIJ. AND
XXII. BOOKS. OF LIVY, adapted for the use of beginners,
by G. C. Macautay, M, A. late Fellow of Trinity College,
Cambridge.

THE SIEGE OF SYRACUSE. Being part of the XXIV. AND
XXV. BOOKS OF LIVY, adapted for the use of beginners.
With Notes, Vocabulary, and Exercises, by GEORGE RICHARDS,
M.A., and A. S. WALPOLE, M.A.

LEGENDS OF EARLY ROME. Adapted for the use of begin-
ners, With Notes, ‘Exercises, and Vocabulary, by HERBERT
WILKINSON, M.A. [nu preparation.

Lucian.—EXTRACTS FROM LUCIAN. Edited, with Notes,
Exercises, and. Vocabulary, by Rev. JOHN Bonp, M.A., and
A. S. WALPOLE, M.A.

Nepos.—SELECTIONS ILLUSTRATIVE OF GREEK AND
ROMAN HISTORY. Edited for the use of beginners with
Notes, Vocabulary and Exercises, by G. S, FARNELL, M.A.

Ovid.— SELECTIONS. Edited by E. S. SHuUcKBURGH, M.A.
late Fellow and Assistant-Tutor of Emmanuel College, Cambri

EASY SELECTIONS FROM OVID IN ELEGIAC VERSE.
Arranged for the use of Beginners with Notes, Vocabulary, and
Exercises, by HERBERT WILKINSON, M.A.

STORIES FROM THE METAMORPHOSES. Edited for the
Use of Schools. With Notes, Exercises, and Vocabulary. By
J. Bonn, M.A., and A. S. WALPOLE, M.A.

Phzedrus.—SELECT FABLES. Adapted for the Use of Be-
ginners. With Notes, Exercises, and Vocabularies, by A. S.

WALPOLE, M.A.

Thucydides.—THE RISE OF THE ATHENIAN EMPIRE.
BOOK I. cc. LXXXIX. — CXVIL anp CXXVIII. —
CXXXVIII. Edited with Notes, Vocabulary and Exercises, by F.
H. Coison, M:A., Senior Classical Master at Bradford Grammar
School ; Fellow of St. John’s College, Cambridge.

Virgil.— 2NEID. BOOK I. Edited by A. S. WALPoLE, M.A,

AENEID. BOOK IV. Edited by Rev. H. M. STEPHENSON,
M.A. [Zn the press.

6 MACMILLAN’S EDUCATIONAL CATALOGUE,

pi eee BOOK V. Edited by Rev. A. CALVERT,
, late Fellow of St. John’s College, Cambridge.
ENED. BOOK VI. Edited by T. E. Pacg, M.A.
ZENEID. SOUs IX. Edited by Rev. H. M. STEPHENSON,

M.A
GEORGICS. BOOKI. Edited by C. Bryans, M.A.

[ln preparation.
SELECTIONS. Edited by E. S. Souckspuren, M.A.

Xenophon.—ANABASIS. BOOK I. Edited by A. S.
WALPOLE, M.A.

ANABASIS. BOOK I. Chaps. I.—VIII. for the use of Beginners,
with Titles to the Sections, Notes, Vocabulary, and Exercises, by
E. A. WELLS, M.A., Assistant Master in Durham School.

ANABASIS. BOOK II, Edited by A. S. WALPOLE. M.A.

[Ln the press.

ANABASIS. BOOK 1V. THE RETREAT OF THE TEN
THOUSAND. . Edited for the use of Beginners, with Notes,
Vocabulary, and Exercises, by Rev. £. D. Stone, M.A., formerly
Assistant-Master at Eton.

[/n preparation,

SELECTIONS FROM THE CYROPA:DIA. Edited, with
Notes, Vocabulary, and Exercises, by A. H. CooKE, M.A., Fellow
and Lecturer of King’s College, Cambridge.

The following more advanced Books, with Introductions
and Notes, but no Vocabulary, are either ready, or in
preparation :—

Cicero.—SELECT LETTERS. Edited by Rev. G. E. JEANs,
M.A., Fellow of Hertford College, Oxford, and Assistant-Master
at Haileybury College.

Euripides.—HECUBA. Edited by Rev. JoHN Bonp, M.A.
and A. S. WALPOLE, M.A,

Herodotus.—SELECTIONS FROM BOOKS VIL anp VIIL.,
THE EXPEDITION OF XERXES. Edited by A. H. Cooke,
M.A., Fellow and Lecturer of King’s College, Cambridge.

Horace. — SELECTIONS FROM THE SATIRES AND
EPISTLES. Edited by Rev. W. J. V. BAKER, M.A., Fellow of
St. John’s College, Cambridge.
SELECT EPODES AND ARS POETICA. Edited by H. A.
DALTON, M.A., formerly Senior Student of Christchurch ; Assistant-
Master in Winchester College.

Plato.—EUTHYPHRO AND MENEXENUS. Edited by C. E.
Graves, M.A., Classical Lecturer and late Fellow of St. John’s
College, Cambridge.

Terence.—SCENES FROM THE ANDRIA. Edited by F. W.
CoRNISH, M.A., Assistant-Master at Eton College,

CLASSICAL SERIES. 7

The Greek Elegiac Poets.— FROM CALLINUS TO
CALLIMACHUS. _ Selected and Edited by Rev. HERBERT
Kywnaston, D.D., Principal of Cheltenham College, and formerly
Fellow of St, John’s College, Cambridge.

Thucydides.—BOOK IV. Cus. L—XLI. THE CAPTURE
OF SPHACTERIA, Editee by C. E, Graves, M.A.

Virgil.—GEORGICS. BOOK II. Edited by Rev. J. H. Skring,
M.A., late Fellow of Merton College, Oxford ; Warden of Trinity
College, Glenalmond.

*,* Other Volumes to follow,

CLASSICAL SERIES
FOR COLLEGES AND SCHOOLS.
Feap. 8vo,

Being select portions of Greek and Latin authors, edited
with Introductions and Notes, for the use of Middle and
Upper forms of Schools, or of candidates for Public
Examinations at the Universities and elsewhere.

Attic Orators.—Selections from ANTIPHON, ANDOKIDES,
LYSIAS, ISOKRATES, AND ISAEOS. Edited by R. C;
Jess, M.A., LL.D., Litt.D., Professor of Greek in the University
of Glasgow. [New Edition in the press.

ZEschines.— IN CTESIPHONTEM. Edited by Rev. T.
GwarTkin, M.A., late Fellow of St. John’s College, Cambridge.

[ln the press.
fEschylus.— PERS. Edited by A. O. Prickarp, M.A.
Fellow and Tutor of New College, Oxford. With Map. 3s. 6d.
SEVEN AGAINST THEBES. Edited by A. W. VERRALL,
M.A. School Edition prepared by Rev. M. A. BAYFIELD, M.A.
[J the press.

Andocides.—pE MYSTERIIS. Edited by W. J. Hicxiz, M.A.,
formerly Assistant-Master in Denstone College. 2s. 6d.

Czsar.—THE GALLIC WAR. Edited, after Kraner, by Rev.
Joun Bonn, M.A., and A. S, WaALPoLe, M.A. With Maps. 6s.

Catullus.—SELECT POEMS. Edited by F. P. Smmpson, B.A.,
late Scholar of Balliol College, Oxford. New and Revised
Edfttion. 5s, “The Text of this Edition is carefully adapted to
School use.

Cicero.—THE CATILINE ORATIONS. From the German
of Kart Haim. Edited, with Additions, by A. S. WILKINS,
M.A., LL.D., Professor of Latin at the Owens College, Manchester,
Examiner of Classi¢s to the University of London, New Edition,
35. 6d,

8 MACMILLAN’S EDUCATIONAL CATALOGUE.

Cicero.—PRO LEGE MANILIA. Edited, after HaLM, by Pro-
fessor A. S. WILKINS, M.A., LL.D. 2s, 6d.

- THE SECOND PHILIPPIC ORATION. — From the German
of Kart HAtm. Edited, with Corrections and Additions,
by Joun E, B. Mayor, . Professor of Latin in the University of
Cambridge, and Fellow of St. John’s College. New Edition,
revised. 55.

PRO ROSCIO. AMERINO. Edited, after HALM, by E. H. Don-
KIN, M.A., late Scholar of Lincoln College, Oxford ; Assistant-
Master at Sherborne School. 45. 6d.

PRO P. SESTIO. Edited by Rev. H. A. HotpEn, M.A., LL.D.,
late Fellow of Trinity College, Cambridge ; and late Classical
Examiner to the University of London. §s.

Demosthenes.—DE CORONA. Edited by B. Drake, M.A.,
late Fellow of King’s College, Cambridge... New and revised
Edition. . 6d.

ADVERSUS LEPTINEM. Edited by Rev. J. R. Kinc, M.A.
Fellow and Tutor of Oriel College, Oxford. 4s. 6d.

THE FIRST PHILIPPIC, Edited, after C. REHDANTZ, by Rev.
T. GwaTkIn, M.A., late Fellow of St. John’s College, Cambridge.

2s. 6d,
IN MIDIAM. Edited by Prof. A. S. WiLkins, LL.D., and
HERMAN HAGER, Ph.D., of the Owens College, Manchester.
In preparation.
Euripides.—HIPPOLYTUS. Edited by J. P. MAHAFFY, M.A.,
Fellow and Professor of Ancient History in Trinity College, Dub-
lin, and J. B. Bury, Fellow of Trinity College, Dublin. 3s. 6d.
MEDEA. _ Edited by A.W. VERRALL, M.A., Fellow and
Lecturer of Trinity College, Cambridge. 3s. 6d.
IPHIGENIA IN TAURIS. Edited by E. B. ENGLAND, M.A.,
Lecturer at the Owens College, Manchester. 4s. 6d.

Herodotus.—pooksS V. anp VI. Edited by J. STRACHAN,
M.A., Professor of Greek in the Owens College, Manchester.
[Jn preparation.
BOOKS VII. anp VIIL Edited by Miss A, RAMSAY. — >:
: : [Zu the press.
Hesiod.—_THE WORKS AND DAYS. Edited by W. T.
LENDRUM, Assistant Master in Dulwich College. [J preparation.
Homer.—iLIAD. BOOKS I., IX., XL, XVL—XXIV.. THE
STORY OF ACHILLES. Edited by the late J. H. Pratt,
M.A., and WALTER Lear, M.A., Fellows of. Trinity College,
Cambridge. 6s.
ODYSSEY. BOOK IX. Edited’ by Prof. Joun E. B. Mayor.
2s. 6d.
ODYSSEY. BOOKS XXI.—xXxXIV. THE TRIUMPH OF
ODYSSEUS. Edited by S. G. Hamitton, B.A., Fellow of
Hertford College, Oxford. 35. 6d.

er es
> ene

hy ot ee Mei ere

CLASSICAL SERIES. 9

_ Horace.—THE ODES. Edited by T. E. Pace, M.A., formerly
Fellow of St. John’s College, Cambridge ; Assistant-Master at
the Charterhouse, 6s. (BOOKS L, IL, III., and IV. separately,

2s. each.) .

THE SATIRES. _ Edited by ARTHUR PALMER, M.A., Fellow of
Trinity College, Dublin; Professor of Latin in the University of
Dublin. 6s..,

THE EPISTLES anp ARS POETICA. Edited by A. S.
WILKINS, M.A., LL.D,, Professor of Latin in Owens College,
Manchester; Examiner in Classics to the University of
London. 6s. 3 Oa }

Isaeos.—THE ORATIONS.; Edited: by WimLiam RipcEway,
M.A., Fellow of Caius College, Cambridge; and Professor of
Greek in the University of Cork. [Jn preparation.

_Juvenal. THIRTEEN SATIRES, Edited, for the Use of
Schools, by E.G. Harpy, M.A., late Fellow of Jesus College,
Oxford, 55... aulita ttc

The Text of this Edition is carefully adapted to School use.

SELECT SATIRES. Edited by Professor JoHn E,. B, Mayor.

_ X. AND XI... 3s. 6d. XIL.—XVI. . 4s. 60,

Livy.—BOOKS II. anp III. Edited by Rev. Hi M! STEPHENSON,
M.A. 55. 3 )

BOOKS XXI. AND XXII. Edited by the Rev. W. W. Capes,
M.A. Maps. 55. |

BOOKS XXIII. anp XXIV. Edited by G. C. Macautay, M.A.
With Maps. 5s.

- THE LAST TWO KINGS OF MACEDON. EXTRACTS
FROM THE FOURTH AND FIFTH DECADES OF
LIVY. Selected and Edited, with Introduction and Notes, by
F. H. RAW.LInNs, M.A., Fellow of King’s College, Cambridge; and
Assistant-Master at Eton. With Maps. 35.

THE SUBJUGATION OF ITALY. SELECTIONS FROM
THE FIRST DECADE. Edited by G. E. Marinpin, M.A.,
formerly Assistant Master at Eton. [/n preparation.

Lucretius. BOOKS L+III. Edited by J. H. WARBURTON
Leg, M.A.; late Scholar of Corpus Christi College, Oxford, and
Assistant-Master at Rossall, 4s.6¢. °

Lysias.—SELECT ORATIONS. Edited by E. S. SuucKBuRGH,
M.A., late Assistant-Master at Eton College, formerly Fellow and
Assistant-Tutor of Emmanuel College, Cambridge. New Edition,
revised. j

Martial. — SELECT EPIGRAMS. Edited by Rev. H. M.
STEPHENSON, M.A. New Edition, Revised and Enlarged. 6s. 6¢.

Ovid.—FASTI. Edited by G. H. Haviam, M.A., Fellow of St.
John’s Collége, Cambridge; and Assistant-Master at Harrow.
With Maps. 5S.

HEROIDUM EPISTULZ XIII. Edited by E. S, SHucksurcGu,
M.A. 45. 6d.

Io MACMILLAN’S EDUCATIONAL CATALOGUE,

Ovid.—METAMORPHOSES. BOOKS I.—III. Edited by €.
Simmons, M.A. [/n preparation.
METAMORPHOSES. BOOKS XIII. Ann XIV. Edited by
C. Simmons, M.A. 4s. 6d.
Plato.—MENO. Edited by E. S. THompson, M.A., Fellow of
Christ’s College, Cambridge. [ln preparation.
APOLOGY AND CRITO._ Edited by F. J. H. JENKINSON,
M.A., Fellow of Trinity College, Cambridge. [/u preparation.
LACHES. Edited, with Introduction and Notes, by M. T. TATHAM,
M.A., Balliol College, Oxford, formerly Assistant Master at
Westminster School. [Zn the press.
THE REPUBLIC. BOOKS I.—V. Edited by T. H. WARREN.
M.A., President of Magdalen College, Oxford. [ln the press.
Plautus.—MILES GLORIOSUS. Edited by R. Y. TyRRELL.
M.A., Fellow of Trinity College, and Regius Professor cf Greek in
the University of Dublin. Second Edition Revised. 5s.
AMPHITRUO. Edited by ARTHUR PALMER, M.A., Fellow of
- Trinity College and Regius Professor of Latin in the University

of Dublin. [Jn preparation.
CAPTIVI. Edited by A. Ruys Smirn, late Junior Student of
Christ Church, Oxford. [Zu preparation.

Pliny.—LETTERS. BOOK III. Edited by Professor Joun E. B.
Mayor. With Life of Pliny by G. H. RENDALL, M.A. © 55.
LETTERS. BOOKS I. and If. Edited by J. Cowan, B.A.,
Assistant-Master m the Grammar School, Manchester.
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Plutarch.—LirE OF THEMISTOKLES. Edited by Rev.
H. A. Hotpen, M.A., LL.D. 5s.

Polybius.—THE HISTORY OF THE ACHAZAN LEAGUE
AS CONTAINED IN THE REMAINS OF POLYBIUS.
Edited by W. W. Capes, M.A. 6s. 6d.

Propertius.—SELECT POEMS. Edited by Professor J. P.
PostcaTr, M.A., Fellow of Trinity College, Cambridge. Second
Edition, revised. 6s.

Sallust.—CATILINA anp JUGURTHA. Edited by C. Mert-
VALE, D.D., Dean of Ely. New Edition, carefully revised and
enlarged, 4s. 6d. Or separately, 2s. 6d. each.

BELLUM CATULINAE. Edited by A. M. Cook, M.A,, Assist-
ant Master at St. Paul’s School. 45. 6a.

JUGURTHA. . By the same Editor. [Zz preparation.
Sophocles.—ANTIGONE. Edited by Rev. Jonn Bonn, M.A.,
and A. S. WALPOLE, M.A. [Jn preparation,

Tacitus.—AGRICOLA anp GERMANIA. Edited by A. J.
Cuurcu, M.A., and W. J. Bropriss, M.A., Translators of
Tacitus. New Edition, 3s. 6¢. Or separately, 2s. each.

THE ANNALS. BOOK VI. By the same Editors. 2s. 6d.
THE HISTORIES. BOOKS I. anp LI, Edited by A, D,
Goprey, M.A. 55,

CLASSICAL LIBRARY. ir
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tor. [fx alion,
THE ANNALS, BOOKS I. anp II. Edited by J. S. Rerp,
M. L,, Litt. D. [Ln preparation.

Terence.—HAUTON TIMORUMENOS. Edited by E. S.

SHUCKBURGH, M.A. 3s. With Translation, 4s. 6d.
PHORMIO. Edited by Rev, JoHN Bonp, M.A., and A. S
WALPOLE, M.A. 4s. 6d.

Thucydides. BOOK IV. Edited: by C. E. Graves, M.A.,
Classical Lecturer, and late Fellow of St.. John’s College,
Cambridge. 5s. i

BOOKS II. anp V._ By the same Editor. To be published
separately. [ln preparation. (Book V. in the press.)
BOOKS I. anp II. Edited by C. Bryans, M.A. [Jn preparation.
BOOKS VI. anpD VII. THE SICILIAN EXPEDITION. Edited
by the Rev. PERcIVAL Frost, M.A., late Fellow of St. John’s
College, Cambridge. New Edition, revised and enlarged, with

__Map. 55. .

Tibullus.—sELECT POEMS. . _Edited by Professor J. P.
PosTGATe, M.A. , [Ze preparation.

Virgil—ZNEID. BOOKS IL ann III. THE NARRATIVE
OF AZENEAS. Edited by E. W. Howson, M.A., Fellow of King’s
College, Cambridge, and Assistant-Master at Harrow. 3s.

Xenophon.—HELLENICA, BOOKS I. anv II. Edited by
H. HartstToneE, B.A., late Scholar of Peterhouse, Cambridge.
With Map. . Od.

CYROPZEDIA. BOOKS VII. anp VIII. Edited by ALFRED
Goopwin, M.A., Professor of Greek in University College,
London. 5s.

MEMORABILIA SOCRATIS. Edited by A. R. Curr, B.A.,
Balliol College, Oxford. 6s.

THE ANABASIS. BOOKS I.—IV. Edited by Professors W. W.
GoopwIN and J. W. WuitTEe. Adapted to Goodwin’s Greek
Grammar. Witha Map. 5s.

HIERO. | Edited by Rev. H. A. HotpEn, M.A., LL.D. 35. 6d.

OECONOMICUS. By the same Editor. With Introduction,
Explanatory Notes, Critical Appendix, and Lexicon, 65,

** Other Volumes will follow.

CLASSICAL LIBRARY.
(1) Texts, Edited with Introductions and Notes,
forthe use of Advanced Students, (2) Commentaries
and Translations.

4Eschylus.—THE EUMENIDES. The Greek Text, with
Introduction, English Notes, and Verse Translation. By BERNARD
DRAKE, M.A., late Fellow of King’s College, Cambridze.
$vo, 45,

12 MACMILLAN’S EDUCATIONAL CATALOGUE.

ZEschylus.—AGAMEMNON. © Edited, with Introduction and
Notes, by A. W. VERRALL, M.A. 8vo. [Jn preparation.
AGAMEMNON, CHOEPHOR@, AND EUMENIDES.

. Edited, with Introduction and Notes, by A. O. PRicKARD, M.A.,

Fellow and Tutor of New College, Oxford. 8vo. — -
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AGAMEMNO. Emendavit DAvip S, MARGOLIOUTH, Coll. Nov.
Oxon. Soc. Demy 8vo. 25. 6d.

THE “SEVEN AGAINST THEBES,” Edited, with Introduc-
tion, Commentary, and Translation, by A. W. VERRALL, M.A.,
Fellow of Trinity College, Cambridge. 8vo. 7s. 6d.

SUPPLICES. Edited, with Introduction and Notes, by T, G.
TUCKER, M.A., Professor of Classics in the University of
Melbourne. 8vo. ; [Zn preparation.

Antoninus, Marcus Aurelius.—BOOK IV. OF THE

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6 ORR On ee

. MATHEMATICS. | 27

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MATHEMATICS. 35

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Tait—HEAT. By P. G. Tart, M.A,, Sec. R.S.E., formerly
Fellow of St. Peter’s College, Cambridge, Professor of Natural
Philosophy in the University of Edinburgh. Crown 8vo. 6s.

Thompson,—ELEMENTARY LESSONS IN ELECTRICITY

* AND MAGNETISM, By Sitvanus P. THompson, Principal
and Professor of Physics in the Technical College, Finsbury. With
Illustrations, New Edition, Revised. Twenty-Eighth Thousand.
Feap. 8vo. 45. 6d. ‘

SCIENCE. 41

Thomson, Sir W.—ELECTROSTATICS AND MAG-
NETISM, REPRINTS OF PAPERS ON. By Sir Wii1am
TuHomson, D.C.L., LL.D., F.R.S., F.R.S.E., Fellow of St.
Peter’s College, Cambridge, and Professor of Natural Philosophy
a the University of Glasgow. Second Edition. Medium 8vo.

Thomson, J. J.—THE MOTION OF VORTEX RINGS, A
TREATISE ON, An Essay to which the Adams Prize was
adjudged in 1882 in the University of Cambridge. By J. J.
THOMSON, Fellow of Trinity College, Cambridge, and Professor of
Experimental Physics in the University. With Diagrams. 8vo. 6s.

APPLICATIONS OF DYNAMICS TO PHYSICS AND
CHEMISTRY. By the same Author. Crown 8vo.
[/n the press.

Todhunter.—NATURAL PHILOSOPHY FOR BEGINNERS.
By I. TopHUNTER, M.A., F.R.S., D.Sc.

Part I, The Properties of Solid and Fluid Bodies. 18mo. 35. 6d.
Part II. Sound, Light, and Heat. 18mo. 3s. 6d.

Turner.—HEAT AND ELECTRICITY, A COLLECTION OF
EXAMPLES ON. By H. H. TURNER, B.A., Fellow of Trinity
College, Cambridge. Crown 8vo. 2:5. 6d.

Wright (Lewis).— LIGHT; A COURSE OF EXPERI-
MENTAL OPTICS, CHIEFLY WITH THE LANTERN.
By Lewis WRIGHT. With nearly 200 Engravings and Coloured
Plates. Crown 8vo. 75. 6d.

ASTRONOMY.

Airy.—POPULAR ASTRONOMY. With Illustrations by Sit
G. B. Arry, K.C.B., formerly Astronomer-Royal. New Edition.
18mo, 45. 6d.

Forbes.—TRANSIT OF VENUS. By G. Forses, M.A.,
Professor of Natural Philosophy in the Andersonian University,
Glasgow. [Illustrated. Crown 8vo. 35. 6d. (Mature Series.)

Godfray.—wWorks by HucH Goprray, M.A., Mathematical
Lecturer at Pembroke College, Cambridge.

A TREATISE ON ASTRONOMY, for the Use of Colleges and
Schools. Fourth Edition. 8vo. 12s. 6d.

AN ELEMENTARY TREATISE ON THE LUNAR THEORY,

with a Brief Sketch of the Problem up to Sy time of Newton.
Second Edition, revised. Crown 8vo. 55. 6

Lockyer.—wWorks by J. NorMAN LockvEr, . R.S.
PRIMER OF ASTRONOMY. | With numerous Illustrations.
New Edition. 18mo. 15s. (Scéence Primers.)
ELEMENTARY LESSONS IN ASTRONOMY. With Coloured
Diagram of the Spectra of the Sun, Stars, and Nebule, and
numerous Illustrations. New Edition. Feap. 8vo. §s. 6d.

42 MACMILLAN’S EDUCATIONAL CATALOGUE,

Lockyer—continued. .
QUESTIONS ON LOCKYER’S ELEMENTARY LESSONS IN
ASTRONOMY. For the Use of Schools. By JoHN ForRBEs-
RoBerRTSON. 18mo, cloth limp. Is. 6d.
THE CHEMISTRY OF THE SUN. With Illustrations. 8vo. 14s.
Newcomb.—ProPpuLAR ASTRONOMY. By S. Newco,
LL.D., Professor U.S. Naval Observatory. With 112 Illustrations
and 5 Maps of the Stars. Second Edition, revised. 8yvo. 18s.
‘It is unlike anything else of its kind, and will be of more use in circulating a

knowledge of Astronomy than nine-tenths of the books which have appeared on the
subject of late years.””—SaTurRDAY REVIEW,

CHEMISTRY.

Armstrong.—A MANUAL OF INORGANIC CHEMISTRY.
By HENRY ARMSTRONG, Ph.D., F.R.S., Professor of Chemistry
in the City and Guilds of London Technical Institute. Crown 8vo.

[lu preparation,

Cohen.—THE OWENS COLLEGE COURSE OF PRAC-
TICAL ORGANIC CHEMISTRY. By JuLius B. CoHEN,
Ph.D., F.C.S., Assistant Lecturer on Chemistry in the Owens
College, Manchester. With a Preface by SIR HENRY ROSCOE,
F.R.S., and C, SCHORLEMMER, F.R.S. Feap. 8vo. 2s. 6d.

Cooke.—ELEMENTS OF CHEMICAL PHYSICS. By JosIAH
P. CooKE, Junr., Erving Professor of Chemistry and Mineralogy
in Harvard University. Fourth Edition, Royal 8vo, 2is.

Fleischer.—A SYSTEM OF VOLUMETRIC ANALYSIS.
By EMIL FLEISCHER. Translated, with Notes and Additions,
from the Second German Edition by M. M. PATTISON Murr,
F,.R.S.E. With Illustrations. Crown 8vo. 7s. 6d.

Frankland.—AGRICULTURAL CHEMICAL ANALYSIS,
A Handbook of. By PERCY FARADAY FRANKLAND, Ph.D.,
B.Sc., F.C.S. Associate of the Royal School of Mines, and
Demonstrator of Practical and Agricultural Chemistry in the
Normal School of Science and Royal School of Mines, South
Kensington Museum. Founded upon Leit/aden fiir die Agriculture
Chemiche Analyse, von Dr. F, KROCKER. Crown 8vo. 7s. 6d.

Hartley.—A COURSE OF QUANTITATIVE ANALYSIS
FOR STUDENTS. By W. Nort Hart ey, F.R.S., Professor
of Chemistry, and of Applied Chemistry, Science and Art Depart-

_ ment, Royal College of Science, Dublin. Globe 8yo. 5s.

Hiorns. — A TEXT-BOOK OF METALLURGY AND

ASSAYING, By A. H. Hiorns. Illustrated. Globe 8vo.
[Ln the press,

Jones.—Works by Francis Jones, F.R.S.E., F.C.S., Chemical
Master in the Grammar School, Manchester.

THE OWENS COLLEGE JUNIOR COURSE OF PRAC-
TICAL CHEMISTRY. With Preface by Sir HENRY Roscor,
F.R.S., and Illustrations. New Edition. 18mo. 2s. 6d.

’
:

SCIENCE, 43

Jones— continued.

QUESTIONS ON CHEMISTRY, _ A Series of Problems and
Exercises in Inorganic and Organic Chemistry. Feap. 8vo. 3s.

Landauer.—BLOWPIPE ANALYSIS. By J. LANDAUER.
Authorised English Edition by J. TAyLor and W. E. Kay, of
Owens College, Manchester. Extra feap. 8vo. 45. 6d.

Lupton.—CHEMICAL ARITHMETIC, With 1,200 Problems.
By Sypney Lupton, M.A., F.C.S., F.1.C., formerly Assistant-
Master at Harrow. Second Edition, Revised and Abridged.
Feap. $vo. 45. 6d.

Meldola.—PHOTOGRAPHIC CHEMISTRY. By RArHAkgL
MELDOLA, Professor of Chemistry in the Technical College,
Finsbury. Crown 8vo. [/n preparation.

Muir.— PRACTICAL CHEMISTRY FOR MEDICAL STU-
DENTS. Specially arranged for the first M.B. Course. By
M. M. Pattison MuIR, F.R.S.E. Feap. 8vo. 1s. 6d.

Muir and:Wilson.—THE ELEMENTS OF THERMAL
CHEMISTRY. By M. M. Pattison Muir, M.A., F.R.S.E.,
Fellow and Przlector of Chemistry in Gonville and Caius College,
Cambridge; Assisted by DAVID Murr Witson. 8vo. 12s. 6d.

Remsen.—Works by IRA REMSEN, Professor of Chemistry in the
Johns Hopkins University.

COMPOUNDS OF CARBON ; or, Organic Chemistry, an Intro-
duction to the Study of. Crown 8vo. 6s. 6d.

AN INTRODUCTION TO THE STUDY OF CHEMISTRY
(INORGANIC CHEMISTRY). Crown 8vo. 6s. 6d.

THE ELEMENTS OF CHEMISTRY. A Text Book for
Beginners. Fcap. 8vo. 2s. 62.
Roscoe.—wWorks by Sir Henry E. Roscor, F.R.S., formerly

Professor of Chemistry in the Victoria University the Owens College,
Manchester.

PRIMER OF CHEMISTRY. With numerous Illustrations. New
Edition. With Questions. 18mo. 1s. (Science Primers.)

LESSONS IN ELEMENTARY CHEMISTRY, INORGANIC
AND ORGANIC. With numerous Illustrations and Chromolitho
of the Solar Spectrum, and of the Alkalies and Alkaline Earths.
New Edition. Feap. 8vo. 4s. 6d. (See under THORPE.)

Roscoe and Schorlemmer.—INORGANIC AND OR-
GANIC CHEMISTRY. A Complete Treatise on Inorganic and
Organic Chemistry. By Sir Henry E. Roscor, F.R.S., and Prof.
C. SCHORLEMMER, F.R.S. With Illustrations. Medium 8vo,

Vols. I. and Il.—INORGANIC CHEMISTRY.

Vol. I.—The Non-Metallic Elements. 21s. Vol. II. Part L—
Metals. 18s. Vol. II. Part II.—Metals. 18s.

Vol. III.—ORGANIC CHEMISTRY.
THE CHEMISTRY OF THE HYDROCARBONS and their
Derivatives, or ORGANIC CHEMISTRY. With numerous
Illustrations. Four Parts. Parts I., Il., and IV. 21s. each.
Part III. 18s.

44 MACMILLAN’S EDUCATIONAL CATALOGUE,

Thorpe.—a SERIES OF CHEMICAL PROBLEMS, prepared
with Special Reference to Sir H. E. Roscoe’s Lessons in Elemen-
tary Chemistry, by T. E. THorprr, Ph.D., F.R.S., Professor of
Chemistry in the Normal School of Science, South Kensington,
adapted for the Preparation of Students for the Government,
Science, and Society of Arts Examinations. Witha Preface by Sir
Henry E. Rosco, F.R.S. New Edition, with Key. 18mo. 2s,

Thorpe and Riicker—A TREATISE ON CHEMICAL
PHYSICS. By T. E. THorreE, Ph.D., F.R.S. Professor of
Chemistry in the Normal School of Science, and Professor A. W.
RUCKER. Illustrated. 8vo, [Jn preparation.

Wright.—METALS AND THEIR CHIEF INDUSTRIAL
APPLICATIONS. By C. ALDER Wricut, D.Sc.,. &c.,
Lecturer on Chemistry in St. Mary’s Hospital Medical School,
Extra feap. 8vo. 35. 6d.

BIOLOGY.

Allen.—ON THE COLOUR OF FLOWERS, as Illustrated in
the British Flora. By GRANT ALLEN, With Illustrations.
Crown 8vo. 3s. 6d. (Nature Series.)

Balfour.— A TREATISE ON COMPARATIVE. EMBRY:
OLOGY. By F. M. Batrour, M.A., F.R.S., Fellow and
Lecturer of Trinity College, Cambridge. With Illustrations.
Second Edition, reprinted without alteration from the First
Edition, In 2vols. 8vo. Vol. I. 18s, . Vol. II. 21s.

Balfour and Ward.—a GENERAL TEXT BOOK OF
BOTANY. By Isaac BAyLEy BatFour, F.R.S., Professor of
Botany in the University of Edinburgh, and H. MARSHALL WARD,
Fellow of Christ’s College, Cambridge, and Professor of Botany
in the Royal Indian Engineering College, Cooper’s Hill. 8vo.

[/n preparation,

Bettany.— FIRST LESSONS IN PRACTICAL BOTANY,
By G. T. BeTTANY, M.A., F.L.S., formerly Lecturer in Botany
at Guy’s Hospital Medical School. 18mo. Is.

Bower—Vines.—A COURSE OF PRACTICAL INSTRUC-
ION. IN BOTANY. By F. O. Bower, M.A., F.L.S.,
Professor of Botany in the University of Glasgow, and SYDNEY
H. Vines, M.A., D.Sc., F.R.S., Fellow and Lecturer, Christ’s
College, Cambridge. With a Preface by W. T. -THISELTON
Dyer, M.A., C.M.G., F.R.S., F.L.S., Director of the Royal
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Part L—PHANEROGAMAI—PTERIDOPHYTA. 6s. Part
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Darwin (Charles).—MEMORIAL NOTICES OF CHARLES
DARWIN, F.R.S., &. By THomas HENRY HUXLEY, F.RS.,
G. J. RomAnes, F.R.S., ARCHIBALD GEIKIE, F.R.S., and
W. T. THISELTON Dyer, F.R.S, Reprinted from ature.
With a Portrait, engraved by C. H. JEENs. Crown $8vo.
2s. 6d. (Nature Series.)

SCIENCE. 45

Fearnley.—A MANUAL OF ELEMENTARY PRACTICAL
HISTOLOGY. By WILLIAM FEARNLEY. With Illustrations.
Crown 8vo. 7s. 6d.

Flower and Gadow.—aAN INTRODUCTION TO THE
OSTEOLOGY OF THE MAMMALIA. By Wi LLiAmM HENRY
Flower, LL.D., F.R.S., Director of the Natural History De-
lee of re British cate, late Hunterian Professor of

omparative Anatomy an ysiology in the Royal Co of
Surgeons of England. With phiileroes Theat Seem "Briel
Edition. Revised with the assistance of HANs Gapow, Ph.D.,
M.A., Lecturer on the Advanced Morphology ef Vertebrates and
sone Curator in the University of Cambridge. Crown 8vo.
10s. 6d. x

Foster.—Works by Micuazt Foster, M.D., Sec. R.S., Professor
of Physiology in the University of Cambridge.
PRIMER OF PHYSIOLOGY, With numerous Illustrations.
New Edition. 18mo. Is.

A TEXT-BOOK OF PHYSIOLOGY. With Illustrations. Fourth
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Foster and Balfour.—THE ELEMENTS OF EMBRY-
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Professor of Physiology in the University of Cambridge, Fellow
of Trinity College, Cambridge, and the late Francis M. BALFour,
M.A,, LL.D., F.R.S., Fellow of Trinity College, Cambridge,
and Professor of Animal Morphology in the University: Second
Edition, revised. Edited by: ADAM SEDGwIcK, M.A., Fellow
and Assistant Lecturer of Trinity College, Cambridge, and WALTER
HlzAPE, Demonstrator in. the Morphological Laboratory of the
University of Cambridge, With Illustrations, Crown 8vo. Ios. 6¢.

Foster and Langley.—A COURSE OF ELEMENTARY
PRACTICAL PHYSIOLOGY. By Prof. MicHArL Foster,
M.D., Sec. R.S., &c., and J. N. LANGLEY, M.A., F.R.S., Fellow
of Trinity College, Cambridge. Fifth Edition. Crown 8vo. 7s. 6d.

Gamgee.—A TEXT-BOOK OF THE PHYSIOLOGICAL
CHEMISTRY OF THE ANIMAL BODY, | Including an
Account of the Chemical Changes oceurring in Disease. By A.
GaMGEE, M.D., F.R.S., formerly Professor of Physiology in the
Victoria University the Owens College, Manchester. 2 Vols. 8vo.
With Illustrations, Vol. I. 18s. [Vol. LL, in the press.

Gray.—STRUCTURAL BOTANY, OR ORGANOGRAPHY

ON THE BASIS OF reba pn tn Me To wiles =

the principles of Taxonomy and Phytography, and a Glossary o

Botanical Terms. By Professor ASAGRAY, LL.D, 8vo. 10s. 6d.

Hamilton,—A PRACTICAL TEXT-BOOK OF PATHO.

LOGY. By D. J. HAMILTON, Professor of Pathological Anatomy
(Sir Erasmus Wilson Chair), University of Aberdeen. 8vo.

[ln the press.

46 MACMILLAN’S EDUCATIONAL CATALOGUE,

Hooker.—Works by Sir J. D. Hooker, K.C.S.I., C.B., M.D.,
F.R.S., D.C.L.

PRIMER OF BOTANY. With numerous Illustrations. New
Edition. 18mo. Is. (Science Primers.)

THE STUDENT’S FLORA OF THE BRITISH ISLANDS.
Third Edition, revised. Globe 8vo. Ios. 6d.

Howes.—AN ATLAS OF PRACTICAL ELEMENTARY
BIOLOGY. By G. B. Howes, Assistant Professor of Zoology,”
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Preface by THoMAS HENRY HUXLEY, F.R.S. Royal 4to. 14s,

Huxley.— Works by THomas Henry Huxtey, F.R.S.
INTRODUCTORY PRIMER OF SCIENCE. 18mo. 1s.
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QUESTIONS ON HUXLEY’S PHYSIOLOGY FOR SCHOOLS.
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Huxley and Martin.—a COURSE OF PRACTICAL IN-
STRUCTION IN ELEMENTARY BIOLOGY. By T. H.
Hux ey, F.R.S., LL.D., assisted by H. N. Martin, M.A.,
M.B., D.Sc., F.R.S., Fellow of Christ’s College, Cambridge.
New Edition, revised and extended by G. B. Howes, Assistant
Professor of Zoology, Normal School of Science, and Royal School
of Mines, and D. H. Scott, M.A., PH.D., Assistant Professor of
Botany, Normal School of Science, and Royal School of Mines.
New Edition, thoroughly revised. With a Preface by T. H.
HuXLEY, F.R.S. Crown 8vo. Ios. 6d.

Kane.—EUROPEAN BUTTERFLIES, A HANDBOOK OF.
By W. F. DE Vismres KANE, M.A., M.R.I.A., Member of the
Entomological Society of London, &c. With Copper Plate Illustra-
tions. Crown 8vo. Ios. 6d.

A LIST OF EUROPEAN RHOPALOCERA WITH THEIR
VARIETIES AND PRINCIPAL SYNONYMS. _ Reprinted
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Klein.—MICRO-ORGANISMS AND DISEASE. An Intro-
duction into the Study of Specific Micro-Organisms. By E.
KLEIN, M.D., F.R.S,, Lecturer on General Anatomy and Physio-
logy in the Medical School of St. Bartholomew’s Hospital, London.
With 121 Illustrations. Third Edition, Revised. Crown8vo. 6s.

THE BACTERIA IN ASIATIC CHOLERA. By the Same.

Crown 8vo. [ln preparation.
Lankester.—Works by Professor E. RAY LANKESTER, F.R.S. |
A TEXT BOOK OF ZOOLOGY. §8vo. [Jn preparation.

DEGENERATION: A CHAPTER IN DARWINISM. LIllus-
trated. Crown 8vo. 2s. 6d. (Mature Series.)
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THE ORIGIN AND METAMORPHOSES OF INSECTS.
With numerous Illustrations, New Edition. Crown 8yo, 3s. 6d.
(Nature Series.)

SCIENCE. 47

Lubbock—cntinued.

ON BRITISH WILD FLOWERS CONSIDERED IN RE-
LATION TO INSECTS. With numerous Illustrations. New
Edition. Crown 8vo. 4s: 6d. (Nature Series).

FLOWERS, FRUITS, AND LEAVES. With Illustrations
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Martin and Moale.—ON THE DISSECTION OF VERTE.
BRATE ANIMALS, _ By Professor H. N. MarTIN and W. A,
_MOoALE. Crown 8vo. [Zn preparation,
Mivart.—wWorks by St. GzorcE Mivart, F.R.S., Lecturer on
Comparative Anatomy at St. Mary’s Hospital.

LESSONS IN ELEMENTARY ANATOMY. With upwards of
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THE COMMON FROG. Illustrated. Cr. 8vo. 3s.6d. (Mature Series.)

Muller.—THE FERTILISATION OF FLOWERS. By Pro-
fessor HERMANN MULLER. Translated and Edited by D’Arcy
W. THOMPSON, B.A., Professor of Biology in University College,
Dundee. With a Preface by CHARLES DARWIN, F.R.S. With
_numerous Illustrations. Medium 8vo. 2Is.
Oliver.—Works by DAnrex Oxiver, F.R.S., &c., Professor of
Botany in University College, London, &c.
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trations. Extra fcap. Svo. 6s. 6d.
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Parker.—A COURSE OF INSTRUCTION IN ZOOTOMY
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With Illustrations. Crown 8vo. 8s. 6d.
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Parker and Bettany.—THE MORPHOLOGY OF THE
SKULL. By Professor W. K. PARKER, F.R.S., and G. T.
BeTrany. Illustrated. Crown 8vo. tos, 6d.

Romanes.—THE SCIENTIFIC EVIDENCES OF ORGANIC
EVOLUTION. By Gerorce J. Romanes, M.A., LL.D.
F.R.S., Zoological Secretary of the Linnean Society. Crown
8vo, 2s, 6d. (Nature Series.)

Sedgwick.—A SUPPLEMENT TO F..M. BALFOUR’S
TREATISE ON EMBRYOLOGY. By ADAM SEDGWICK,
M.A., F.R.S., Fellow and Lecturer of Trinity College, Cambridge.
8vo. Illustrated. [/n preparation.

Smith (W. G.)—DISEASES OF FIELD AND GARDEN
CROPS, CHIEFLY SUCH AS ARE CAUSED BY FUNGI.
By WortTuincton G. Smiru, F.L.S., M.A.I., Member of the
Scientific Committee R.H.S. With 143 New Illustrations drawn
and engraved from Nature by the Author. Feap. 8vo. 45. 6d.

Ward.—TIMBER AND ITS DISEASES. By H. MARSHALL
Warp, Professor of Botany in the Royal Indian Engineering
College, Cooper’s Hill. Crown 8vo, Illustrated. [7m preparation.

48 MACMILLAN’S EDUCATIONAL CATALOGUE,

Wiedersheim (Prof.).—ELEMENTS OF THE COM-
PARATIVE ANATOMY OF VERTEBRATES, Adapted
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tomy, and Director of the Institute of Human and me sep
Anatomy in the University of Freiburg-in-Baden, by W.
NEWTON PARKER, Professor of Biology in the University College
of South Wales and Monmouthshire. With Additions _ by the
Author and Translator. With Two Hundred and Seventy Wood-
cuts. Medium 8vo. 12s. 6d.

MEDICINE,

Brunton.—wWorks by T. Lauper BruNnToN, M.D., D.Sc.,
F.R.C.P., F.R.S., Assistant Physician and Lecturer on Materia
Medica at St. Bartholomew’s Hospital; Examiner in Materia
Medica in the University of London, in the Victoria University,
and in the Royal College of Physicians, London; late Examiner
in the University of Edinburgh. Sy ear 8

A TEXT-BOOK OF PHARMACOLOGY, THERAPEUTICS,
AND MATERIA MEDICA, Adapted to the United States
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Third Edition, Adapted to the New British Pharmacopeeia, 1885.
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TABLES OF MATERIA MEDICA: A Companion to the Materia
risa . With Illustrations. New£Edition Enlarged.

vo. 10s. 6d.

Griffiths.—LESSONS ON PRESCRIPTIONS AND THE
ART OF PRESCRIBING. By W. HanpDsEL GRIFFITHS,
Pu.D,, L.R.C.P.E. New Edition, Adapted to the Pharmacopeeia,

ne 18mo. 35. 6d.

Hamilton.—A TEXT-BOOK OF PATHOLOGY. By D. J.
HAMILTON, Professor of Pathological Anatomy University of
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Klein.— MICRO-ORGANISMS AND DISEASE, An Intro-
duction into the Study of Specific Micro-Organisms. By E.
KEIN, M.D., F.R.S., turer on General Anatomy and Physio-
logy in the Medical School of St. Bartholomew’s Hospital, London.
With 121 Illustrations. Third Edition, Revised. Crown 8vo 6s.

THE BACTERIA IN ASIATIC CHOLERA. By the Same
_ Author. Crown 8vo. [Jn preparation.

Ziegler-Macalister.—TEXT-BOOK OF PATHOLOGICAL
ANATOMY AND PATHOGENESIS. By Professor ERNST
ZIEGLER of Tiibingen. Translated and Edited for English
Students by DONALD MACALISTER, M.A., M.D., B.Se., F.R.C.P.,
Fellow and Medical Lecturer of St. John’s College, Cambridge,
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the University. With numerous Illustrations. Medium 8vo.

Part I—GENERAL PATHOLOGICAL ANATOMY. Second
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Part IL—SPECIAL PATHOLOGICAL ANATOMY. . Sections
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SCIENCE, 49

ANTHROPOLOGY.

Flower.—FASHION IN DEFORMITY, as Illustrated in the
Customs of Barbarous and Civilised Races. By Professor
FLowWER, F.R.S., F.R.C.S. With Illustrations. Crown 8vo.
2s. 6d. (Nature Series.)

Tylor.—ANTHROPOLOGY. An Introduction to the Study ot
Man and Civilisation: By E. B. Tytor, D.C.L., F.R.S. With
numerous Illustrations. Crown 8vo. 7s. 6d.

PHYSICAL GEOGRAPHY & GEOLOGY.

Blanford.—_THE RUDIMENTS OF PHYSICAL GEOGRA:
PHY FOR THE USE OF INDIAN SCHOOLS; with a
Glossary of Technical Terms employed. By H. F. BLANForD,
F.R.S. New Edition, with Illustrations. Globe 8vo. 25. 6d.

Geikie.— Works by ARCHIBALD GzIkiE, LL.D., F.R.S., Director
General of the Geological Survey of Great Britain and Ireland, and
Director of the Museum of Practical Geology, London, formerly
Murchison Professor of Geology and Mineralogy in the University
of Edinburgh, &c.

PRIMER OF PHYSICAL GEOGRAPHY. With numerous
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ELEMENTARY LESSONS IN PHYSICAL GEOGRAPHY.
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THE SCENERY AND GEOLOGY OF SCOTLAND,
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(See also under History and Geography.)

Huxley.—PHYSIOGRAPHY. An Introduction to the Study
of Nature. By Tuomas Henry Huxtey, F.R.S. With
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Lockyer.—OUTLINES OF PHYSIOGRAPHY—THE MOVE-
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HISTORY AND GEOGRAPHY, 53

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HISTORY AND GEOGRAPHY. 55

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56 MACMILLAN’S EDUCATIONAL CATALOGUE.

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58 MACMILLAN’S EDUCATIONAL CATALOGUE,

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MODERN LANGUAGES AND LITERATURE. 59

“ENGLISH CLASSICS FOR INDIAN
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[Zn preparation.
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Cowper.—THE TASK. Edited by W. T. WexB, M.A., Pro-
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BUSINESS. Edited by F. J. Rowe, M.A., Professor of English
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Milton.—PARADISE LOST, Books I. and II. Edited by
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[Ready.

Scott.—THE LADY OF THE LAKE. Edited by G. H. Stuart,

Professor of English Literature, Presidency College, Madras.

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MARMION. Edited by MicHAEL MACMILLAN, B.A, Globe 8yo.
; ai 6d. [Ready.
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Shakespeare.—MUCH ADO ABOUTNOTHING. Edited by
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25,
HENRY V. By the same Editor. [Jn preparation,
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60 MACMILLAN’S EDUCATIONAL CATALOGUE,

Tennyson.—SELECTIONS. Edited by F. J. Rowe, M.A.,
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This Volume contains:—Recollections of the Arabian Nights—The Lady of

. Shalott—CEnone—The Lotos-Eaters—A Dream of Fair Women—Morte
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ey san on a ny aie

eMODERN LANGUAGES AND LITERATURE, 61

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MODERN LANGUAGES AND LITFRATURE. 63

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MODERN LANGUAGES AND LITERATURE. ~ 65

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f

66 MACMILLAN’S EDUCATIONAL CATALOGUE,

FRENCH.

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MODERN LANGUAGES AND LITERATURE. 67

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68 MACMILLAN’S EDUCATIONAL CATALOGUE,

Macmillan’s Foreign School Classics (continued)—
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DIVINITY.

*,* For other Works by these Authors, see THEOLOGICAL
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DIVINITY. 73

Abbott—Rushbrooke.—THE COMMON TRADITION OF
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The Acts of the Apostles. — Being the Greek Text as
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Arnold. —A BIBLE-READING FOR SCHOOLS. — THE
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Calvert.—GREEK TESTAMENT, School Readings in the. A
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John’s College, Cambridge. Feap. 8vo. 45. 6d,

Cassel. MANUAL OF JEWISH HISTORY AND. LITERA-
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Cheetham,—a CHURCH HISTORY OF THE FIRST SIX
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Cross.—BIBLE READINGS SELECTED FROM THE
PENTATEUCH AND THE BOOK OF JOSHUA. By
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Curtéis.—MANUAL OF THE THIRTY-NINE ARTICLES.
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74 MACMILLAN’S EDUCATIONAL CATALOGUE,

Davies.—THE EPISTLES OF ST, PAUL TO THE EPHE-
SIANS, THE COLOSSIANS, AND PHILEMON; with
Introductions and Notes, and. an Essay on the Traces of Foreign
Elements in the Theology ot these Epistles. By the Rey. J.
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Drummond:—THE STUDY OF THEOLOGY, INTRO.
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Gaskoin.—THE CHILDREN’S TREASURY OF BIBLE
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Golden Treasury Psalter.—students’ Edition. Being. an
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Greek Testament.—Edited, with Introduction and Appen-
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GREEK TESTAMENT, SCHOOL READINGS IN THE. Being
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Hardwick.—Works by Archdeacon HARDWICK :—

A HISTORY OF THE CHRISTIAN CHURCH. Middle
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of Modern History in the University of Oxford. With Four
Maps, New Edition, Crown 8vo. 10s. 6d,

DIVINITY, a

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Jennings and Lowe.—THE PSALMS, WITH INTRO-
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Kay.—T. PAUL’S TWO EPISTLES TO THE CORIN-
THIANS, A COMMENTARY ON. By the late Rev. W.
Kay, D.D., Rector of Great Leghs, Essex, and Hon. Canon of
St. Albans ; formerly Principal of Bishop’s College, Calcutta ; and
Fellow and Tutor of Lincoln College. Demy 8vo. 9s.

Kuenen.—PENTATEUCH AND BOOK OF JOSHUA: an
Historico-Critical Inquiry into the Origin and Composition of the
Hexateuch. By A. KUENEN, Professor of Theology at Leiden.
Translated from the Dutch, with the assistance of the Author, by
PuiLure H. WicksTeep, M.A. 8vo. 14s.

The Oxrorp MaGaAzinE says:—‘‘ The work is absolutely indispensable to all
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Lightfoot.—Works by the Right Rev. J. B. Licutroor, D.D.,
D.C.L., LL.D., Lord Bishop of Durham,

ST. PAUL’S EPISTLE TO THE GALATIANS. A Revised
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ST. PAUL’S EPISTLE TO THE PHILIPPIANS. A Revised
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Edition, revised. 8vo. 12s.

ST. CLEMENT OF ROME—THE TWO EPISTLES TO
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ST. PAUL’S EPISTLES TO THE COLOSSIANS AND TO
PHILEMON. A Revised Text, with Introductions, Notes, -
and Dissertations, Eighth Edition, revised. 8vo. 12s.

THE APOSTOLIC FATHERS. Part II. S. IGNATIUS—
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Maclear.—works by the Rev. G. F. MAcLEAr, D.D., Canon of
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A CLASS-BOOK OF OLD TESTAMENT HISTORY, New
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A CLASS-BOOK OF NEW TESTAMENT HISTORY,
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A SHILLING BOOK OF OLD TESTAMENT HISTORY,
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New Edition,

' 96 MACMILLAN’S EDUCATIONAL CATALOGUE,
Maclear.—wWworks by the Rev. G. F., continued.

inued.

A SHILLING BOOK OF NEW TESTAMENT HISTORY,
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New Edition.

These works have been carefully abridged from the Author’s
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A FIRST CLASS-BOOK OF THE CATECHISM OF THE
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Maurice.—THE LORD’S PRAYER, THE CREED, AND
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Pentateuch and Book of Joshua: an Historico-Critical
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A. KUENEN, Professor of Theology at Leiden. Translated from
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WICKSTEED, M.A: 8vo. 145.

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Procter and Maclear.—AN ELEMENTARY INTRO-
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The Psalms, with Introductions and Critical

Notes.—By A, C. JenninGs, M.A.,, Jesus College, Cambridge,
Tyrwhitt Scholar, Crosse Scholar, Hebrew University Prizeman,
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Ramsay.—THE CATECHISER’S MANUALS; or, the Church
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Ryle.—AN INTRODUCTION TO THE CANON OF THE
OLD TESTAMENT. By Rev. H. E. Ry et, M.A., Fellow
of King’s College, and Hulsean Professor of Divinity in the
University of Cambridge. Crown 8yo. [Zn preparation.

DIVINITY, 77

St. James’ Epistle.—The Greek Text with Introduction and
Notes. By Rev. JosePpH Mayor, M.A., Professor of Moral
Philosophy in King’s College, London. 8vo. [/# preparation.

St. John’s Epistles.—The Greek Text with Notes and Essays,
by Brooxe Foss Werstcott?, D.D., Regius Professor of Divinity
and Fellow of King’s College, Cambridge, Canon of Westminster,
&c. Second Edition Revised. 8vo. 12s. 6d,

St. Paul’s Epistles.—Greek Text, with Introduction and Notes.

THE EPISTLE TO THE GALATIANS. Edited by the Right

Rev. J. B. Licurroot, D.D., Bishop of Durham, Ninth
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THE EPISTLE TO THE PHILIPPIANS. By the same Editor.
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THE EPISTLE TO THE COLOSSIANS AND TO PHI-
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THE EPISTLE TO THE ROMANS, Maed by the Very Rev.
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THE EPISTLE TO THE THESSALONIANS, COMMENT-
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CAIRNS. $8yo. . 12s,

THE EPISTLES TO THE EPHESIANS, THE COLOSSIANS,
ray PHILEMON; with Introductions and Notes, and an

ssay on the Traces of Foreign Elements in the Theology of these
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Christ Church, St, Marylebone; late Fellow of Trinity College,
Cambridge. Second Edition, revised. Demy 8vo. 75. 6d.

THE TWO EPISTLES TO THE CORINTHIANS, A COM-
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Westcott.—Works by Brookg Foss Westcort, D.D., Canon of
Westminster, Regius Professor of Diyinity, and Fellow of King’s
College, Cambridge,

78 MACMILLAN’S EDUCATIONAL CATALOGUE,

Westcott.— Works by Brooxe Foss, continued. ~~ .

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80 MACMILLAN’S EDUCATIONAL CATALOGUE,

MAGMILLAN’S GEOGRAPHICAL SERIES,

Edited by ARCHIBALD GEIKIE, F.R.S., Director-General of the
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The following List of Volumes is contemplated :—

THE TEACHING OF GEOGRAPHY, A Practical Handbook for
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GEOGRAPHY oF INDIA,

GEOGRAPHY OF THE UNITED STATES.

ADVANCED CLAss-BooK OF THE GEOGRAPHY OF EuROPE.

RICHARD CLAY AND SONS, LONDON AND BUNGAYs

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