Histological Atlas of the

Laboratory Mouse
Histological Atlas of the
Laboratory Mouse
William D. Gude
Oak Ridge National Laboratory
Oak Ridge, Tennessee

Gerald E. Cosgrove
San Diego Zoological Society
San Diego, California

and

Gerald P. Hirsch
Veterans Administration Wadsworth Hospital Center
Los Angeles, California

SPRINGER SCIENCE+BUSINESS MEDIA, LLC

 Library of Congress CatalogIng In Pub!tcation Data

Cude WIllIam O
HistologlCal atlas of the laboratory mouse
BIbltography p
Includes Index
1 MICe~Cytology~Atlases 2 Histology~Atlases 1 Cosgrove Cerald E
II Hirsch Cerald P 1939~ III TItl"
QLS 13 M55CS 599 3233 81 8708

ISBN 978-1-4613-5701-8 ISBN 978-1-4615-1743-6 (eBook) AACR2

DOI 10.1007/978-1-4615-1743-6

© 1982 Springer Science+Business Media New York

Originally published by Plenum Press, New York in 1982

AII rights reserved

No part of this book may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without wriHen permission from the Publisher
 Foreword

The Biology Division of the Oak Ridge National Laboratory ducted with very large numbers of mice, and mice proved to
was organized in 1946 for the purpose of studying the imme- be especially suitable for cancer induction studies.
diate and long-term implications of man's exposure to ioniz- As this work progressed, we became convinced that a
ing radiation. The program that developed concentrated on strong histology department was needed to prepare the tis-
the basic mechanism of the effects in biological organisms sues in a uniform manner and also to examine and interpret
from the genetic, biochemical, biophysical, and molecular bio- them. With the support of Dr. Furth at that time, we secured
physical points of view. the services of William D. Gude, who organized this section
Most of its activities at the beginning concentrated on of the Biology Division and whose dedicated management
nonmammalian work (bacteria, fungi, Drosophzla, plants, etc.) developed it into a central information source for histology
since no facilities to perform mammalian studies were availa- work, not only for our Biology Division but also for this area
ble at that time. It became most obvious that specimens more of Tennessee, thus establishing its excellent reputation.
closely related to mammalian tissue would likely yield more I am most pleased to see that Mr. Gude has assembled this
conclusive data to extrapolate these effects upon man. work into a detailed atlas of the laboratory mouse. As essential
At Oak Ridge, the first study was the Mammalian as such information is to basic mammalian research, such a
Genetics Study under William L. Russell designed to investi- volume of complete data and illustrations is not always avail-
gate the genetic changes produced by exposure to ionizing able in a single source, and I believe this volume will serve as
radiation and the implication for man. A couple of years later, an important handbook for researchers utilizing these species.
this was followed by the study of somatic cell effects through
the efforts of Jacob Furth, later by Arthur Upton, and finally Alexander Hollaender
by John Storer. In all of these assays, we concentrated on work AssoClated UnlversltIes, Inc
Council for Research Planmng In BlOloglcal SClences
with the mouse since more was known about its genetics than 1717 Massachusetts Avenue, N W
about the genetics of any other mammal. Studies could be con- Washmgton, 0 C 20036

v
 Acknowledgments

We are most appreciative of the encouragement by Dr. Alex- Professional Ektachrome Film 6118. The color film was pro-
ander Hollaender, Associated Universities, Washington, D.C., cessed by Color Technique, Inc., Chicago.
to undertake this work and of his efforts in helping to arrange We want to express our gratitude to Mrs. Charlotte Rains
the publication of this book. Dr. Wanda Haschek of the Biol- for her diligence and patience in preparing the many type-
ogy Division, Oak Ridge National Laboratory, has been most written copies of all sections of the manuscript.
helpful in offering valuable suggestions concerning the selec- We are grateful to Katharine Hummel for granting per-
tion of chapter headings and the clarification of histological mission to copy descriptive material from the microscopic
descriptions. Our thanks go also to other members of the anatomy sections of Chapter 13, "Anatomy," of Biology of the
Biology Division for their suggestions and help, including Dr. Laboratory Mouse, 2nd ed., Blakiston Division, McGraw-Hill
Neal Clapp, Dr. E. F. Oak berg, Dr. Ray Popp, Dr. Kowetha Book Co., New York, 1966.
Davidson, and Dr. J. M. Holland. Dr. Conrad Richter, Medical For assistance with proofreading we are indebted to
Division, Oak Ridge Associated Universities, kindly partici- David A. Hambright and Craig Whitmire of the Technical
pated in the critical selection of photomicrographs, offering Publications Department, Information Division, Oak Ridge
many suggestions for improvements. Dr. Desmond Doyle, National Laboratory.
University of Tennessee Veterinary School, helped in identi- The cover of the book was designed by Nancy Harrell
fying specific structures of the male reproductive system. Lockwood, Graphic Arts Department, Oak Ridge National
Gene Watkin of the Information Section, Fusion Energy Divi- Laboratory.
sion, provided us with several ideas for improving the orga- Research reported herein was sponsored by the Office of
nization of the book and for planning the arrangement of col- Health and Environmental Research, U.s. Department of
ored prints. Energy, under contract W-7405-eng-26 with the Union Car-
We owe many thanks to Joyce Feezell and Jimmy Wesley bide Corporation, and supported in part by U.s. Department
for preparing excellent paraffin sections of the tissues and of Energy contract EY-76-C-02-0016 with Associated Univer-
organs. Jimmy also prepared high-quality thin sections of sities, Inc., Brookhaven National Laboratory. W. D. G.
methacrylate-embedded tissues. The photomicrographs were G. E. C.
taken through a Zeiss Ultraphot II microscope using Kodak G. P. H.

vii
 Contents

Introduction 1
Nervous System 3
Special Sense Organs 5
Endocrine System 9
Reproductive System 13
Digestive System 17
Urinary System 21
Skin 23
Respiratory System 25
Lymphatic System 27
Bone, Cartilage, Fat 29
Muscular System 31
Circulatory System 33
Color Figures 35
Fixatives 131
Staining Methods 133
References 145
Index 147

ix
Histological Atlas of the
Laboratory Mouse
 Introduction

The purpose of this book is to provide investigators and stu- example, mucus present in goblet cells of the intestine was
dents with an atlas of tissues of mice, commonly used exper- stained brilliant red with periodic acid-Schiff reagent (PAS).
imental animals in laboratory studies. Many strains of mice Neurons and nerve ganglia stained black or dark brown with
are available for research and clinical laboratory studies, but silver, as in the Bodian stain, are outstanding against a color-
histological differences between strains are usually minor. We less background. Procedures for obtaining the color combi-
have taken samples from six strains presently used in the nations of many other tissues are given in the Appendix for
Biology Division, Oak Ridge National Laboratory: BALB/c, those who wish to reproduce them for their own investigative
RF, C3H, C57BL, BC3F 1 /Cum, and B6D2F 1 /Jax. Both males and research.
females between the ages of 3 and 10 months were used. They With the development of embedding materials other than
were killed with ether and the dissected tissues were placed paraffin (methacrylate and epon) and the availability of micro-
in an appropriate fixative immediately upon removal from the tomes that section tissues 1-2 JLm thick, it is now practical to
mouse. The choice of fixative depended on the stain to follow obtain very thin sections of cells in which fine cytoplasmic
and included 10% neutral buffered formalin, Bouin's fixative, and nuclear structures can be identified that would be very
Zenker-formol (Helly's fluid), formal-mercuric chloride, and difficult to see in a 5-JLm-thick section.
Williams's fixative (formulas given in the Fixatives section). We believe this atlas may also be used as a supplement to
Many of the sections pictured in this book were stained with standard histology texts. For more detailed descriptions of tis-
hematoxylin and eosin (H & E). In addition, various special sues and organs characteristic of the mouse see Biology of the
stains were used to illustrate specific structures or features. For Laboratory Mouse (see Recommended Reading).

1
 Nervous System
(Figures 1-19)

Irritability and conductivity are two characteristics of.the ner- body at the axon hillock, the axon or axis cylinder. Impulses
vous system that require specialized cells and tissues. Anatom- travel away from the cell body down the axons, which
ically, the nervous system is divided into the central nervous decrease in diameter as the distance from the cell body
system (CNS), including the brain and spinal cord, and the increases. Collateral branches may be given off at intervals.
peripheral nervous system (PNS), which includes all other Other processes, dendrites, are direct extensions of the cell
nerve elements in the organism (peripheral nerves, ganglia, body and they connect with axons of functionally related neu-
etc.). Specialized cells that do not conduct nerve impulses but rons at synapses or junctions. Impulses travel along dendrites
contribute supporting tissue to the nervous system comprise toward the cell body from the synapsis.
the neuroglia. Nervous tissue also may be divided into gray The axis cylinder in peripheral nerves is covered by a
and white matter. Nerve cell bodies, dendrites, and unmyeli- neurolemma or Schwann's sheath. In nerves having a myelin
nated portions of axis cylinders comprise gray matter. White sheath covering the axon the neurolemma forms the outer-
matter consists primarily of myelinated nerve fibers, which most covering. Nuclei of Schwann cells may be identified
give it a glistening white appearance. Neuroglial cells are alongside the axis cylinder. A node of Ranvier occurs at the
present in both gray and white matter. junction of two merging segments of myelin sheath.
The functional, impulse-conducting unit of nervous tis- Neurons may be further categorized by the number of
sue is the neuron. Its basic structure consists of a large glob- processes extending from the cell body. Those having only
ular nucleus within which are a few chromatin granules and one axon are unipolar neurons. Bipolar neurons have an axon
usually one large nucleolus. The cytoplasm or perikaryon sur- and one dendrite extending from the cell body at opposite
rounding the nucleus contains abundant, irregular masses of poles; multipolar neurons are determined by the number and
basophilic material rich in ribonucleic acid also known as arrangement of dendrites. Ganglia are the chief paths of com-
Nissl or tigroid substance. Following silver staining numerous munication between the CNS and outer ganglionic plexuses.
fine threads of neurofibrils may be identified in the Their nerve cell bodies are usually large and spherical with a
cytoplasm. predominant nucleolus and abundant chromatin in the
All neurons have one process extending from the cell cytoplasm.

3
4 Nervous System

Neuroglia or specialized interstitial tissue of the nervous nucleus and a small amount of cytoplasm. They are scattered
system includes ependyma, a type of cuboidal or low colum- throughout the brain and spinal cord.
nar epithelium that lines the ventricles of the brain and spinal The meninges, connective tissue coverings of the brain
canal, and satellite cells of peripheral ganglia. Three types of and spinal cord, include (1) the outermost dura mater, a dense
neuroglia are identified: astrocytes (fibrous and protoplasmic), and tough material next to the bone, (2) the arachnoid, a thin
oliogodendroglia, and microglia. Protoplasmic astrocytes, network of loose connective tissue devoid of blood vessels
which are found chiefly in gray matter, exhibit processes, and closely adherent to (3) the pia mater, which is the inner-
many of which are attached to blood vessel walls and to the most thin membrane that contains blood vessels. The dura
pia mater. Fibrous astrocytes, more commonly found in white mater, arachnoid, and pia mater together form the lepto-
matter of the brain between nerve fibers, have a larger elon- meninges.
gated nucleus than the protoplasmic type, but they also send The choroid plexuses found in the roof of the third and
out processes that attach to blood vessel walls. fourth ventricles and in part of the walls of the lateral ventri-
Oliogodendroglia resemble astrocytes but are much cles are lined by specialized epithelial cells that differ from
smaller cells and have only a few processes with few ependymal cells. Cuboidal cells of the choroid plexus are
branches. They are associated with supporting nerve fibers arranged in a single layer and many blood vessels occupy its
rather than blood vessels, and are called "satellite" cells when folds. The choroid plexus is the source of cerebrospinal fluid.
found adjacent to nerve cell bodies. Microglia have a small
 Special Sense Organs
(Figures 20-31)

EAR thelium becomes pseudostratified with tall columnar cells,

many bearing cilia.
The ear may be divided into three parts: external ear, middle The inner ear is divided into a bony labyrinth containing
ear, and inner ear. The pinna and external auditory meatus a clear fluid, perilymph, lined by endothelium, and a mem-
comprise the external ear. The middle ear consists of the tym- branous labyrinth that contains endolymph. Within the bony
panic membrane, bony ossicles, and the auditory or eusta- portion are the vestibule, three semicircular canals filled with
chian tube, connecting the ear with the oropharynx. Included perilymph, and the cochlea. Enclosed in the membranous lab-
in the inner ear are the bony labyrinth and spiral cochlea, yrinth are the utricle, utriculosaccular duct, the saccule, endo-
which contains the organ of Corti. lymphatic sac and duct, three membranous semicircular canals
The outer third of the external ear is composed of elastic containing endolymph, and the cochlear duct (scala media).
cartilage covered by hairy skin in which sebaceous and ceru- In each semicircular canal are cristae-ridges lined by an
minous glands are distributed. epithelium of tall columnar cells of two types, hair cells and
Simple squamous epithelium lines the tympanic cavity of sustentacular cells. In each utricle and saccule are maculae
the middle ear, which contains the three bony ossicles: stapes, similar to the cristae with both hair and supporting cell types.
malleus, and incus. The tympanic membrane is an oval, In each macula are tiny crystals of a mixture of calcium car-
semitransparent membrane that is composed of two layers of bonate and protein that are called otoliths. The cristae and
collagenous fibers and fibroblasts similar to a tendon plus maculae are predominantly involved with maintaining
epithelium on the inner and outer surfaces. It divides the mid- equilibrium.
dle ear from the inner ear.
Both elastic and hyaline cartilage are found in the audi-
tory tube from its beginning to its terminus at the oropharynx. ORGAN OF CORTI
It is lined by mucous membrane with many folds. At the bony
tympanic portion the epithelium is low, ciliated columnar but On the floor of the cochlear duct is the organ of Corti,
as the tube approaches the cartilaginous oropharynx the epi- resting on a basement membrane of low cuboidal cells. This

5
6 Special Sense Organs

structure extends from the bony spiral lamina to the spirallig- body, and the lens, which is characteristically spherical in the
ament, and consists primarily of hair cells and supporting rodent, filling almost all of the eyeball.
cells. One particular structure is the tectorial membrane, a thin The retinal layer is divided into two layers; the outermost
gelatinous membrane in intimate contact with the cilia of hair is pigmented and the inner or nervous portion contains pho-
cells. Sound waves transmitted to the organ of Corti by the toreceptors, cones, and, in the mouse, a few rods.
endolymph of the cochlear duct from the perilymph of the The jellylike vitreous humor fills the interior of the eye
scala vestibuli activate the hair cells. Nerve fibers around the between the lens and retina. The anterior and posterior cham-
bases of the hair cells receive the stimulus and transfer it to bers are filled with aqueous humor, secreted by the ciliary
nerves in the spinal ganglion and finally, by way of bipolar processes.
cells, to the cochlear division of the acoustic nerve.
Many strains of mice are affected by mutants involving
the ear structures. Malformed cristae and absence of otoliths CORNEA
produce conditions resulting in imbalance. Other strains are
born with defects that eventually produce deafness owing to The cornea is a transparent avascular structure consisting
degenerating hair cells and a malformed organ of Corti. of four identifiable strata. A stratified squamous epithelium
Waltzing mice, twirlers, and shaker mice fit into this category. covers the cornea, resting upon a basement membrane. Below
is a substantia propria of specialized, dense connective tissue
containing a few flattened fibroblasts dispersed among colla-
EYE gen fibers bound together by an amorphous cement sub-
stance. Behind this layer is Descemet's membrane, a band of
The principal structures of the eye consist of three layers refractile, homogeneous, elastic substance adjacent to a very
constituting the wall of the eye: the supporting layer, middle thin squamous epithelium.
layer, and retinal layer. The sclera or "white" of the eye is the The choroid lying internal to the sclera is heavily pig-
primary supporting structure of dense connective tissue. The mented and contains elastic fibers in addition to a single layer
cornea, forming the bulging anterior portion of the eye, func- of capillaries.
tions primarily as a transparent tissue transmitting light rays The nervous portion of the retina consists of nine layers
through the lens to the retina. However, in the rodent it also made up of cell bodies of many neurons and ganglia in addi-
provides support, covering about half of the surface of the tion to the rod and cone special photoreceptors.
eyeball, and is considered a part of this layer. The spherical lens lies just behind the pupil between the
The vascular middle layer or uvea provides nourishment anterior and posterior chambers of the eye. This transparent
to ocular structures and contains smooth muscle. It includes structure contains a highly refractive capsule that coats the
the pigmented iris and choroid, the ciliary process and ciliary outer surface of the lens epithelium of flat cuboidal cells. Lens
 Special Sense Organs 7

fibers constitute the substance of the lens, forming elongated pared sections, appears vacuolated. The lamina propria of
prisms. There is no epithelium in the capsule covering the fibrous connective tissue contains melanocytes filled with pig-
posterior portion. ment. A single excretory duct is present. The gland produces
Mutations resulting in a wide variety of eye defects- an oily secretion for lubricating the surface of the eye.
such as microophthalmia, cataracts, and abnormal lens for-
mation and development-are common findings in several
strains of mice. Incomplete retinal development has resulted LACRIMAL GLAND
in either abnormally short rods or absence of rods.
Paired tubuloalveolar lacrimal glands lie in two areas,
extraorbital and intraorbital. The extraorbital gland is below
HARDERIAN GLAND and in front of the ear. The intraorbital gland is located where
the excretory duct opens into the conjunctival sac. The pyr-
Partially surrounding the eyeball posteriorly is this amidal secretory cells have spherical nuclei lying near the
tubuloalveolar gland. It is covered by a thin capsule whose base and intensely basophilic cytoplasm. Myoepithelial cells
strands divide the gland into lobules. The pyramidal epithe- are present between the epithelial cells and the basement
lial cells contain a nucleus with two or more nucleoli lying membrane. The serous secretion moistens and lubricates the
along the base and lipid-containing cytoplasm, which, in pre- surface of the eyeball and the eyelids.
 Endocrine System
(Figures 32-43)

The endocrine glands include the pituitary, thyroid, parathy- acid-Schiff (red) and produce gonadotrophins, follicle-stimu-
roid, adrenal, pineal, ovary, placenta, testis, and islets of Lan- lating hormone (FSH), and luteinizing hormone (LH).
gerhans in the pancreas. Endocrine features of the ovary and The intermediate lobe is separated from the anterior lobe
testis will be discussed under the reproductive systems of the by a cleft lined by cuboidal epithelium. Cells in this lobe have
male and female. intensely staining oval nuclei and light basophilic cytoplasm.
The hormone elaborated by this lobe is melanin-stimulating
hormone (MSH).
PITUITARY GLAND The posterior lobe is filled with nonmyelinated nerve
fibers, neuroglial cells (pituicytes), and connective tissue sup-
The pituitary gland (hypophysis) lies within the bony porting capillaries. Two hormones stored and released from
sella turcica of the floor of the skull, the dorsal basisphenoid the pars nervosa are oxytocin and vasopressin.
bone. It consists of a pars distalis or anterior lobe, pars inter-
media or intermediate lobe, and pars nervosa (neurohypoph-
ysis) or posterior lobe. THYROID GLAND
The anterior lobe is very vascular with many capillaries.
Histologically, three types of cells may be identified: (1) The characteristic arrangement of various-sized spherical
Acidophils contain small, round nuclei and eosinophilic cyto- follicles containing eosinophilic colloid identifies the thyroid
plasm and are characterized by elaborate growth and the pro- gland. The follicles are lined by simple cuboidal epithelium.
duction of lactogenic hormones. (2) Agranular chromophobes Increases in the height of these cells as well as changes in the
whose cytoplasm does not stain may be stem cells. (3) Baso- staining quality and amount of colloid materials indicate lev-
phils may be divided into two classes: beta basophils, whose els of secretory activity. Such changes occur as an aging pro-
granules are aldehyde-fuchsin-positive and secrete thyro- cess in some strains of mice in which tall columnar follicle
trophic hormone, and delta basophils, whose granules are cells decrease in cell size in young mice, and increase in fol-
aldehyde-fuchsin-negative but stain positive with periodic licle size in mature mice. Senile changes include increased

9
10 Endocrine System

fibrous connective tissue between follicles and merging of fol- from neuroectodermal cells and form parenchymal cells of the
licles to form very large structures with flattened epithelium. body of the gland, and neuroglial cells. Parenchymal cells are
The follicular cells produce the colloid substance, which con- spherical with slightly basophilic cytoplasm. Neuroglial cells
tains thyroxin and triiodothyronine. support the parenchymal cells. Mesenchymal cells in the pia
"Ultimobranchial bodies," "parafollicular cells," and C mater covering give rise to connective tissue of the capsule,
cells are synonymous terms for lightly staining cells distrib- which provides incomplete partitions that divide the body
uted throughout the thyroid connective tissue adjacent to but into lobules.
outside the follicular cells. These cells are producers of thy-
rocalcitonin, a hormone that acts in collaboration with the
parathyroid gland in maintaining calcium homeostasis in ADRENAL GLAND
body tissues and blood. Parafollicular cells have been
described in most mammals, including the mouse. The adrenal glands are paired, each gland located adja-
cent to the anterior pole of the kidney. The adrenals ar-e larger
in the female and occasionally show strain differences. A
PARATHYROID GLANDS fibrous connective tissue capsule surrounds the gland, which
is divided into a cortex and medulla. In most mammals the
The parathyroid gland, consisting of two or more lobes, cortical division can be subdivided into three zones of epithe-
is embedded within the thyroid gland but separated from it lial cells, but in mice only two zones are clearly seen. The sub-
by a capsule of connective tissue. The cells form masses of capsular zona glomerulosa consists of small cells with large
densely packed groups interspersed with capillaries or sinus- nuclei arranged in the form of arches, with capillaries
oids. At least two cell types can be identified. These chief cells between the cells. Beneath the glomerulosa is the wide zona
consist of the dark, actively secreting cells and the light, inac- fasciculata, which consists of columns of cells separated by
tive cells. A third type (similar to the oxyphil cell) has been connective tissue. These cells are large with vesicular nuclei
described by some investigators. and contain lipid material, appearing foamy in sections when
lipid is removed in processing. The zona reticularis identified
in most mammals is rarely seen in the mouse. However, a
PINEAL GLAND zone between the cortex and medulla is identified as the X
zone in males and females before sexual maturity. The zone
The pineal gland lies on the dorsal surface of the brain at disappears in sexually mature males and with the first preg-
the junction of the cerebral cortex and cerebellum. A very thin nancy in the female. It has been observed to persist in virgin
capsule surrounds the gland and merges with the choroid females for varying lengths of time and in castrate males. In
plexus. Two types of cells can be identified: cells that originate aging mice of certain strains (RFM, BALB/c, etc.) large foamy
 Endocrine System 11

cells with brown pigment form in the cortical zone adjacent noradrenaline form the medulla. Two cell types, the sites of
to the medulla. The pigment is similar to ceroid in the ovary origin of these hormones, have an affinity for potassium
and is referred to as "brown degeneration." In some strains of bichromate, and brown cytoplasmic granules can be seen fol-
mice the change is more pronounced. lowing treatment with this chemical-the result of the chro-
Clusters of polyhedral cells that produce adrenaline and maffin reaction.
 Reproductive System
(Figures 44-76)

FEMALE REPRODUCTIVE SYSTEM become quite numerous in some strains of mice, often occur
in the hilus of the vascular medulla.
Ovary
Uterus
The paired ovaries are enclosed by thin connective tissue cap-
sules. Although in most mammalian ovaries a cortex and The uterus is a bicornuate tubular structure. The endo-
medulla are clearly identified, in the mouse these areas are not metrium or mucosa of the uterus is composed of a simple col-
easily differentiated. The surface of the ovary is covered by a umnar epithelium that lines the uterine lumen, branched
single layer of cuboidal cells called the "germinal" epithe- tubular glands, and vascular connective tissue. The myome-
lium. Beneath the germinal epithelium are developing folli- trium consists of an inner circular and outer longitudinal
cles and corpora lutea, interspersed with interstitial cells and layer of smooth muscle separated by very vascular, loose con-
blood vessels. The follicle is composed of an oocyte enclosed nective tissue. An outer thin serosa covers the uterus. At the
by a jellylike substance called the zona pellucida and multiple junction of the uterine horns with the body, a partition is
layers of granulosa cells. During follicular maturation the present in the midline that extends through the lumen to the
oocyte enlarges, the granulosa cells proliferate, and a fluid- vagina. As the uterine lumen approaches the vaginal terminal,
filled space (antrum) appears as the follicle continues to the epithelium changes from columnar epithelium to strati-
enlarge. fied squamous epithelium, which is continuous with the
At ovulation the secondary oocyte breaks through the vagina.
ovarian surface and moves into the periovarian space. The
remaining follicular cells (granulosa cells) undergo various Vagina
changes to form a corpus luteum.
In addition to indistinct areas in the ovary, tne mouse The vagina, a nonglandular tubular organ, consists of a
organ is unique in that corpora lutea may persist through sev- stratified squamous epithelium, fibrous lamina propria, and
eral estrous cycles. In addition, cysts, which enlarge and thin muscular layer. During the estrous cycle the vaginal

13
14 Reproductive System

epithelium also undergoes cyclic changes, characterized by Placenta

proliferation of the basal cells, cornification, leukocytic infil-
tration, and variation in the number of epithelial cell layers. The placenta is a flat, spherical structure that is smooth
As the phases of estrus progress, histological examination on the fetal side where it is covered by cuboidal amitotic epi-
may indicate overlapping of one phase with another. The pre- thelium. This rests on the chorion, a layer of embryonic con-
dominating phase may be determined by the abundance or nective tissue. Most of the placenta is composed of chorionic
scarcity of cornified cells or mitotic figures, the presence or villi bathed in a pool of maternal blood. The trophoblast cov-
absence of leukocytes, and the thickness of the epithelial cell ers the villi, the cores of which contain fetal blood vessels. The
layers. Estrus consists of five phases, two of which-proestrus trophoblast is divided into a syncytial and deeper cellular lay-
and estrus-represent growth, and two-metestrus I and 11- ers. Giant cells are easily identified in the syncytiotrophoblast
in which degenerative changes in the epithelium occur. Dies- of the mature placenta. Thick portions of this region are the
trus represents a transition between degeneration and sources of placental hormones: estrogen, progesterone, lacto-
renewed growth, i.e., rebuilding the epithelial lining. Proes- gen, and chorionic gonadotrophin. The maternal portion of
trus is characterized by an increase in cell layers, cornifying the placenta is the decidua basalis. The decidual cells are large
granulosa layers, active cell division, and a scarcity of leuko- polygonal cells and are rich in glycogen and lipids. Irregular
cytes. As the estrus phase predominates, the cornified layer masses of an eosinophilic, homogeneous substance called
becomes superficial prior to desquamation, the number of fibrinoid are found in young animals and increase with the
mitoses are reduced, and leukocytes disappear. Metestrus I is age of placenta. Endometrial glands, fibrinoid, and some tro-
identified by desquamation of cornified cells into the vaginal phoblast cells are also present.
lumen and the appearance of leukocytes in the outer layer of
the epithelium. Metestrus II includes a marked increase in Mammary Glands
leukocyte numbers, reduction in epithelial cell layers, and the
presence of an occasional mitotic figure. The onset of diestrus Mammary tissue develops in both the male and female
marks a return to active growth of the now thin epithelium embryos. In the male, because of the inhibitory effect of tes-
and is characterized by an increase in numbers of cell layers, tosterone secreted by the fetal testis, the mammary tissue does
reduced numbers of leukocytes, and the appearance of a few not mature into nipples and ducts.
dividing cells. Large, mucoid, PAS-positive cells may appear In the female embryo, mammary buds appear by the fif-
between the proestrus and estrus phases, probably stimulated teenth day within the epidermis and gradually lengthen into
by progesterone. The mucous cells rest superficially on the epithelial cords. A few days later, tubules form within these
outer cornified layer, desquamating just prior to that layer. cords. The epidermal cells surrounding the cords develop into
 Reproductive System 15

nipples. At birth the female gland consists of a nipple opening are believed to nourish the germ cells.
into a primary duct. This duct branches into secondary ducts, Spermatogenic cells occur in layers. The first and most
which lie within adipose tissue and are lined by low cuboidal primitive are types A and B spermatogonia, which are dis-
epithelium. A few myoepithelial cells lie between the epithe- persed among the Sertoli cells lying against the basement
lium and basement membrane. Increasing ductal branching membrane. Upon mitotic division the spermatogonia form
occurs by the fourth week as the glands rapidly grow. By 6 primary spermatocytes, which undergo meiosis to form sec-
months alveoli develop, although the degree of development ondary spermatocytes. These cells, after dividing mitotically,
varies with mouse strain. form spermatids, small cells with eccentric nuclei and a tail,
During pregnancy and lactation the number of alveoli many of which are in direct contact with Sertoli (nurse) cells
increases, with hypertrophy of epithelial cells accompanying while transforming into immature spermatozoa. Each mature
widening of the lumen. As milk is secreted in the alveoli, the sperm contains a sickle-shaped head (though width of the
cuboidal cells are flattened into a squamous pattern. At cessa- head varies considerably in several strains of mice), midpiece,
tion of suckling, alveolar epithelial cells resume cuboidal and tail. These cells now lie next to the lumen.
shape. The interstitial tissue of the testis is the endocrine por-
tion, lying between the seminiferous tubules. The loose con-
nective tissue contains blood and lymphatic vessels, nerves,
MALE REPRODUCTIVE SYSTEM
and groups of epithelial cells, the cells of Leydig, which are
Testes the source of testosterone. The oval cells contain spherical to
oval nuclei and cytoplasm, which is often vacuolated.
A thick capsule, the tunica albuginea, covers each testis. Removal of lipid material during preparation of the tissue
The internal structure consists of tubules separated by thin results in the formation of cytoplasmic vacuoles.
septa. Inside are the highly convoluted seminiferous tubules,
which are separated by loose connective tissue with some elas- Epididymis
tic fibers and the cells of Leydig. The tubules are lined by a
highly specialized stratified epithelium, seminiferous or ger- The epididymis is covered by a capsule of fibrous connec-
minal epithelium, which rests on a basement membrane. The tive tissue. It is divided into a head, body, and tail. Most of the
cells touching on the basement membrane are of two types, efferent ductules are located in the head. The greatly coiled
spermatogonia and Sertoli or sustentacular cells. The Sertoli duct of the epididymis occupies the body and tail. The effer-
cells are large pale cells with vesicular nuclei and a prominent ent ductules are lined by low to tall columnar epithelial cells
nucleolus. These cells lie against the basement membrane and with stereocilia projecting into the lumen.
16 Reproductive System

Vesicular Glands (Seminal Vesicles) prostate glands. Both glands have several ducts that enter the
urethra. Ducts from the dorsal gland enter laterally; the ven-
These elongated and curved paired glands lie immedi- tral ducts enter ventrally. The glands are surrounded by
ately above the bladder. Internally the alveolar mucosa in smooth muscle and a fibromuscular stroma. The secretion
multiple folds is lined by tall columnar epithelium whose often condenses, forming globules that give a granular, aci-
cells shorten as the gland becomes distended with acidophilic dophilic appearance.
secretion.
Bulbourethral Glands (Cowper's Glands)
Vas Deferens
These paired glands lie approximately at the junction of
This thick-walled tube is lined by pseudostratified colum- the membranous urethra and penis on the lateral sides. Each
nar epithelium. Loose connective tissue with many elastic gland is divided into a body and a tail, both of which are
fibers surrounds the star-shaped lumen. The muscularis is embedded in skeletal muscle. The glands consist of tubular
thick with smooth muscle, composed of inner longitudinal, alveoli, which are lined by tall columnar cells of variable
middle circular, and outer longitudinal layers. height. The small, dark nuclei lie flattened in the cell with
basophilic cytoplasm and rest on a well-developed basement
Coagulating and Ampullary Glands membrane.

Immediately below and attached to the vesicular glands Preputial Glands

are the paired coagulating glands whose folded mucosa is
lined by columnar cells with centrally located spherical nuclei The flat broad cells of this gland are identical in appear-
and acidophilic cytoplasm. ance and function to sebaceous glands. Characteristically, the
Lateral to the vesicular glands is a group of small glands nucleus degenerates and becomes part of the oily secretion,
with many short ducts forming the ampullary glands. Their which is then excreted. The duct of the gland is lined with
epithelium is low columnar with large oval nuclei, and their stratified squamous epithelium; it opens into the preputial
secretion is eosinophilic. space. Occasionally, pigment is observed in the surrounding
connective tissue.
Dorsal and Ventral Prostate

Immediately below the bladder are the dorsal and ventral

 Digestive System
(Figures 77-104)

TONGUE neuroepithelial cells. Sustentacular cells are elongate and

somewhat banana-shaped and follow a curved course in the
The tongue consists principally of striated muscle covered by taste bud. They are arranged so that they are pointed toward
a mucous membrane. Muscle fibers run longitudinally and a small opening in the bud, the pore, which empties into the
horizontally. In the fibroelastic tissue between muscle bun- lumen of the papilla, which in turn opens on the dorsal sur-
dles, blood vessels and nerves are found. The epithelium of face of the tongue. In the central part of the taste bud, susten-
the mucosa is thick, stratified squamous and is cornified on tacular cells mix with neuroepithelial cells. These cells are
the dorsal surface. long and narrow, with darkly staining nuclei that end in tiny
On the rough dorsal surface of the tongue are areas of hairs projecting from the pore.
various papillae projecting from the surface. Four types may
be identified: foliate, vallate, filiform, and fungiform. Filiform
(cone-shaped) papillae have a broad base and pointed tips, SALlY ARY GLANDS
which project posteriorly. A few fungiform papillae, some-
what square in shape with a connective tissue core, are among There are three salivary glands in the mouse: parotid,
the filiform papillae. Several rows of foliate papillae are found submandibular, and sublingual. They are compound tubu-
along the epithelial surface of the sides of the tongue opposite loalveolar glands divided into lobules by connective tissue
molar teeth. Behind the molars, a single, roughly square val- septa and surrounded by a capsule. Tall pyramidal cells line
late papilla occurs, bounded by deep furrows on either side in the secretory alveoli and lie on a basement membrane. Myo-
the tongue surface. epithelial or basket cells lie between glandular cells and the
Taste buds are located on both sides of the dorsal surface basement membrane.
of the vallate papilla and on the surface of the foliate papilla. The parotid gland produces a serous fluid. The cells have
Oval taste buds are arranged perpendicular to the surface of a spherical, centrally located nucleus with basophilic cyto-
the mucous membrane epithelium of the mouth. Taste buds plasm. The short, narrow ducts extending from the gland are
contain two types of cells, sustentacular or gustatory cells and lined by low cuboidal epithelium.

17
18 Digestive System

The secretory alveoli of the submandibular gland are SMALL INTESTINE

lined by tall pyramidal cells that produce mucus and serous
fluid. In male mice the intercalated tubules and some alveoli Three regions of the small intestine may be identified:
have tall columnar cells whose nuclei lie at the base of the duodenum, jejunum, and ileum. All three have characteristic
cell. Abundant eosinophilic granules fill the cytoplasm. villi, fingerlike projections that increase the surface area. Col-
Nuclei in cells of the female are centrally located and only a umnar epithelial cells cover the villi. Their nuclei are located
few cytoplasmic granules are seen. They increase in number at the base, while a striated border is seen along the lumen.
as young females as well as young males mature. Goblet cells filled with mucus are distributed among the epi-
In the sublingual gland, tall pyramidal cells whose nuclei thelial cells, increasing in number in the ileum. The villi
lie basally have pale-staining cytoplasm. Compared with the decrease in size from the duodenum to the ileum. Between
other two glands these mucous-secreting cells appear pale, adjacent villi are the crypts of Lieberkuhn. Glands of Brunner
bluish white in hematoxylin-stained preparations. The short are present only in the duodenum, extending below the bases
intercalated ducts are lined by low cuboidal epithelium. Strat- of the villi into the submucosa. Paneth cells with prominent
ified squamous epithelium lines the chief excretory duct. eosinophilic granules are found at the bases of the crypts,
especially in the jejunum. Lymphoid nodules (peyer's
patches) are distributed in the submucosa throughout the
ESOPHAGUS small intestine.

The esophagus has a folded mucosa lined by cornified,

stratified squamous epithelium. No glands are present sub- LARGE INTESTINE
mucosally. The muscle layer consists of striated muscle
throughout the length of the esophagus. The large intestine is characterized by columnar epithe-
lium with many goblet cells and a lack of villi. In the anal
region the crypts of Lieberkuhn disappear and a transitional
STOMACH zone between columnar and stratified squamous epithelium
occurs between mucous membrane and skin. The opening is
The stomach is divided into two approximately equal surrounded by the external sphincter (striated) muscle and
nonglandular and glandular portions by a ridge. Cornified, sebaceous glands.
stratified squamous epithelium lines the nonglandular stom-
ach. Mucus-secreting cells, zymogenic or chief cells, and acid-
secreting parietal cells comprise the glandular portion.
 Digestive System 19

ENTEROCHROMAFFIN SYSTEM with a prominent central nucleus and eosinophilic cytoplasm.

Occasionally two nuclei and one or more nucleoli may be
Cells of the enterochromaffin system have been described seen.
throughout the gastrointestinal system. They are character-
ized by the presence of cytoplasmic granules that react to
treatment with silver salts and with bichromate. These cells GALLBLADDER
are known to contain serotonin (5-hydroxytryptamine).
The gallbladder, a sac for bile storage, consists of a folded
mucosa lined by simple columnar epithelium, beneath which
LIVER lies the vascular lamina propria. This is surrounded by smooth
muscle and an outer serosa. The common bile duct leading
The liver is divided into indistinct lobules having in their from the gallbladder to the intestine is lined by cuboidal
center hepatic (central) veins from which cords of hepatic epithelium, which changes to columnar as it approaches the
cells, separated by sinusoids, radiate out to the periphery. The duodenum.
outer boundaries of the lobules are poorly demarcated by the
portal triads (hepatic artery, portal vein, bile duct). Reticular
fibers, originating from the fibrous capsule (Glisson's capsule) PANCREAS
covering the liver, penetrate the organ supporting the
parenchyma. The pancreas is both an exocrine and an endocrine gland.
Interlobular branches of the hepatic artery and portal The exocrine pancreas consists of cells that form tightly
vein provide the afferent blood supply to the liver sinusoids. packed acini. These acinar cells are pyramidal in shape, with
The sinusoids in turn drain into the central veins of the lob- their apical ends bordering the lumen and broad, intensively
ule, which are branches of the hepatic vein. The sinusoids are basophilic bases resting on a basement membrane. The apical
lined by endothelium and specialized reticuloendothelial cytoplasm of these cells contains eosinophilic zymogen
cells, the Kupffer cells, that function as phagocytes. granules.
Between adjacent hepatocytes, on the opposite side of the The endocrine pancreas consists of clusters of endocrine
sinusoid, are bile canaliculi. These extend from the central cells with their capillary network, the islets of Langerhans,
vein outward to the periphery of the lobule, where they open which are embedded in acinar tissue. Alpha cells that secrete
into bile ducts lined by cuboidal epithelium. These bile ducts glucagon occupy the periphery of the islet while beta cells
carry bile to the gallbladder. that secrete insulin are more numerous and fill the central
The hepatic cells or hepatocytes are large polygonal cells area of the islet.
 Urinary System
(Figures 105-111)

KIDNEY convoluted tubule empties into the collecting tubules located

within the medulla. Although cuboidal epithelium lines the
The paired, bean-shaped kidneys are covered by a capsule of distal tubules, the brush border is absent. The collecting
connective tissue. The shorter concave border is called the tubules with columnar epithelial cells connect with the ureter
hilus; from it extend the renal artery and vein, nerves, and the and the epithelium becomes transitional (urothelium).
ureter on its way to the bladder.
The kidney is divided into two regions, an outer cortex
containing tubules and glomeruli and an inner medulla com- BLADDER
posed primarily of blood vessels and collecting tubules. The
functional unit of this organ is the nephron, consisting of a The pear-shaped bladder is lined with transitional epithe-
glomerulus and associated urinary tubules lined with cuboi- lium (urothelium) of two to four cell layers, depending upon
dal epithelium. The glomerulus, composed of a tuft of capil- the degree of distention. The lamina propria of the bladder is
laries surrounded by Bowman's capsule, is supplied by an very vascular and is surrounded by a thick muscularis com-
afferent arteriole and drained by an efferent arteriole. In the posed of several layers of smooth muscle. This muscle forms
mouse a marked sex difference is noted in the type of epithe- a sphincter at the bladder neck, where it joins the membra-
lium of Bowman's capsule: cuboidal epithelium for most glo- nous portion of the urethra.
meruli of the male, squamous epithelium in the female. The
segment of nephron that emerges from a glomerulus and
courses through the cortex is the proximal convoluted tubule, URETHRA AND PENIS
lined by cuboidal epithelium with a pronounced brush bor-
der. The tubule continues into the medulla as the descending The membranous urethra extends from the neck of the
portion of the loop of Henle and returns to the cortex as the bladder to the pelvic girdle, where it enters the penis. It is
ascending portion of this loop. From this structure the distal lined by transitional epithelium, and its vascular lamina pro-

21
22 Urinary System

pria forms a sheath around the urethra. Embedded in this fills these spaces, producing erection of the penis. Most of the
structure are the urethral glands of Littre, clusters of secretory urethra from the bladder to the tip of the penis is lined by
alveolar cells with oval nuclei and basophilic cytoplasmic transitional epithelium (urothelium), which changes abruptly
secretion. In the penis, the corpus cavernosus urethrae is at the orifice into stratified squamous. Dorsally, in the septum
formed by a sheath of smooth muscle and an outer fibrous between the paired corpora cavernosa, is a small compact
connective tissue sheath. Cavernous spaces lined by endothe- bone, the os penis.
lium form from tissues of the inner and outer sheaths. Blood
 Skin
(Figures 112-122)

The skin consists of two distinct components: the epidermis nosus) are also found in the dermis. Pigmented areas contain
of keratinizing, stratified squamous epithelium, and the der- melanocytes. Dermal papillae containing blood vessels and
mis. Thin skin occurs over most of the hairy regions while nerves project up into the epidermis. Hair projects from the
thick skin appears over the hairless areas, i.e., feet, tail, nip- skin surface; the follicle containing the shaft, bulb, and matrix
ples, and genital and anal areas. is located in the dermis. Actively dividing epidermal cells in
Thick skin consists of four layers, the deepest consisting the bulb produce growth of the shaft. Sebaceous glands
of a single layer of cells with oval nuclei and indistinct cell accompany each hair follicle, as does an arrector pili. Pig-
membranes resting on a basement membrane immediately mented hair results from melanocytes in basal epidermal cells.
adjacent to the dermis. This is the stratum germinativum, in Specialized tactile hairs are found abundantly distributed
which mitotic figures are often seen and whose cells replace about the lips, cheeks, and mouth. These elongated whiskers
cells of the outer layers. The stratum spinosum, four to five are called vibrissae. Although their structure is similar to that
cells thick, is the next layer. These cells are connected across of hair follicles, the vibrissae are much larger and are sur-
intercellular spaces by tonofibrils. The stratum granulosum is rounded by venous blood sinuses and nerve fibers. Striated
identified by cells containing keratohyalin granules easily muscle fibers replace smooth muscle, and a sebaceous gland
stained with hematoxylin. The outermost layer of desquamat- accompanies each vibrissa.
ing dead cells is the stratum corneum. Sebaceous glands have ducts that open along the hair
The dermis consists primarily of connective tissue as well shaft. The gland cells are large, oval cells with prominent
as many blood vessels, nerves, collagen and elastic fibers, and nuclei in the basal layer. As the secretion accumulates in the
fat cells. Thin strands of smooth muscle (arrector pili) associ- gland, the cells lose their nuclei and become part of the secre-
ated with hair follicles and striated muscle (panniculus car- tion, which is called sebum.

23
 Respiratory System
(Figures 123-132)

The respiratory system includes the nasal passages, pharynx, dostratified, ciliated columnar epithelium with numerous
larynx, trachea, and lung. The epithelium of the nasal pas- mixed glands and goblet cells. The tube is prevented from col-
sages is of the pseudostratified, ciliated columnar type with lapsing by the presence of ten to twelve C-shaped hyaline car-
numerous goblet cells. The nasal passages also contain olfac- tilage rings. The trachea bifurcates into bronchi, which branch
tory epithelium, including speCialized bipolar nerve cells sup- successively into bronchioles. In the mouse there are no bron-
ported by two cell types, basal and supporting cells. The sup- chial glands, nor is there cartilage beyond the bifurcation of
porting cells form tall columns of cells with oval nuclei near the trachea.
the surface; many olfactory nerve cells are subjacent. The In the lungs the bronchioles are lined by epithelium con-
olfactory cells are bipolar ganglion cells, the apical portion sisting of ciliated and nonciliated cells, the latter known as
forming a dendrite that extends to the surface as a base for Clara cells. Clara cells are identified in thin methacrylate sec-
nonmotile cilia. Below the nucleus the cytoplasm extends to tions by characteristic bulging into the lumen and absence of
a fine thread to form an unmyelinated axon that eventually cilia. The bronchioles branch successively to form alveolar
constitutes, along with others, the olfactory nerve bundles. ducts, alveolar sacs, and finally alveoli.
Basal cells are roughly triangular and are seen between other The alveoli are thin-walled tubes lined by epithelial cells
cells along the basement membrane. Glands in the interstitial and supported by interstitial tissue of the alveolar wall (septa),
tissue membrane are the tubuloalveolar type (mucous and which consists of a few reticular fibers, septal cells, and many
serous). capillaries. The alveolar epi.thelial cells are of two types, the
The pseudostratified, ciliated columnar epithelium and squamous or type I epithelial cell and the great alveolar or
glands of the nasal mucosa continue into the nasopharynx, type II epithelial cell. The type II cell can be recognized in
interrupted by stratified squamous epithelium as the orophar- thin methacrylate sections by its location at the junction of
ynx and larynx are crossed, and continue down the major por- alveolar septa, its cuboidal shape, and its vacuolated cyto-
tions of the respiratory tract. plasm (loss of lamellar bodies during tissue preparation).
The trachea is lined by a mucous membrane with pseu-

25
 Lymphatic System
(Figures 133-140)

The lymphatic system includes lymph nodes and nodules stained medulla with far fewer cells and containing aggrega-
present in areas of the digestive tract as well as lymph and tions of a few large epithelial cells forming Hassall's corpus-
lymphatic vessels that carry the fluid from tissue spaces to the cle. In other animals keratin is often observed within the
circulatory system. Tonsils are absent in the mouse. corpuscle but is absent in the mouse. The medulla is well
Lymphatic vessels are lined by a single layer of endothe- supplied by blood vessels.
lial cells. Collagen and elastic fibers as well as smooth muscle
fibers envelop larger vessels. In the hilus of lymph nodes one
can identify cross sections of efferent vessels, while afferent SPLEEN
lymphatics are found entering the node at the peripheral mar-
ginal sinuses. The spleen is a very vascular organ covered with a cap-
Lymph nodes are surrounded by a fibrous capsule and are sule of dense connective tissue from which trabeculae project
divided into a marginal subcapsular sinus, the cortex contain- into the spleen pulp. Branches of the splenic artery and vein,
ing dense masses of lymphocytes and the medulla composed nerves, smooth muscle fibers, and reticular fibers accompany
of lymphoid elements with large sinusoids lined by littoral the trabeculae to form supporting connective tissue for red
cells. Supporting tissue in the node consists of reticular cells and white pulp. White pulp is lymphoid tissue and forms
and fibers. Germinal centers containing precursors of mature characteristic nodules, also called Malpighian corpuscles.
lymphocytes are occasionally seen in cortical lymphatic cen- Occasionally, germinal centers are seen, surrounded by a
ters. spherical mass of lymphocytes. A central arteriole occupies
the center of the nodule.
Red pulp consists primarily of erythropoietic cells, lym-
THYMUS phocytes, granulocytes, plasma cells, mast cells, and mega-
karyocytes. In the mouse, extramedullary hemopoiesis is com-
The thymus, a lymphoid organ, is composed of a cortex monly found in red pulp. Apparently, there are no true
packed with small lymphocytes, giving a very dark blue sinuses in mouse spleen. Some strain differences in patterns
appearance when stained with hematoxylin, and a lighter- of reticulum have been described.
27
 Bone, Cartilage, Fat
(Figures 141-149)

BONE tilage grows in two directions. Growth toward the epiphysis

mimics the growth of the epiphyseal plate, while growth
There are two kinds of ossification, endochondral and mem- toward the surface aims at replacement.
branous, implying the presence or absence of a prior cartila-
ginous model. In both processes bone is formed by osteoblasts,
which originate from mesenchyme. Osteoblasts are spindle-
CARTILAGE
shaped to columnar cells with an eccentric nucleus and baso-
philic cytoplasm. They congregate on surfaces and lay down
the organic matrix (osteoid), which then becomes mineral- Cartilage is a supporting connective tissue consisting of
ized. When the osteoblasts become trapped in small spaces three types: hyaline cartilage, elastic cartilage, and fibrocarti-
(lacunae) within this osteoid, they are called osteocytes. lage. Hyaline cartilage has a white and glossy appearance in
Bone is resorbed by two different mechanisms: osteoclasia the living state and is composed of cells (chondrocytes)
and osteolysis. Osteoclasia is the surface resorption of bony enclosed in lacunae. The cells are found individually or in
tissue by multinucleated, giant cells (osteoclasts) typically groups within a hyaline matrix of intercellular substance. It is
located in Howship's lacunae during the active phase. Osteo- from this type of cartilage that some areas of bone formation
lysis refers to a deep-seated resorption centered around the are derived.
activity of old osteocytes. Elastic cartilage, as the name implies, contains abundant
Long bones and vertebrae grow from epiphyseal plates elastic and some collagen fibers within intercellular sub-
and articular cartilage. In the growing animal there are three stance. Examples are the pinna of the ear, nasal cartilage, and
zones in the epiphyseal plate: zone of resting cartilage (pro- epiglottis.
liferation), zone of columnar cartilage (maturation), and zone Fibrocartilage is a tough substance containing an excess
of vesicular cartilage (hypertrophy). Cessation of longitudinal amount of collagen fibers; its lacunae are arranged in sym-
growth coincides with sexual maturity, and the plate gradu- metrical rows. It is found closely associated with tendons
ally decreases in size. In the growing individual, articular car- attached to bone.

29
30 Bone, Cartilage, Fat

FAT Brown fat consists of groups of polygonal cells with lipid

droplets in granular cytoplasm. The nucleus is centrally
Adipose tissue in the mouse is divided into two types: located. Numerous capillaries and reticular fibers support the
white and brown fat. Most white fat is abundantly found in fat cells. Brown fat is also found in specific locations, such as
several specific areas, i.e., in the mesenteries around blood in lobes between scapular bones, adjacent to the thymus, and
vessels, kidneys, adrenals, ovaries, and testes and in the axil- at the hilus of the kidney. The fat lobes between scapulae are
lary and inguinal regions. In H & E-stained sections a white called hibernating glands, although there is no evidence that
fat cell is identified as a round or polyhedral cell whose fat these structures secrete hormones.
has been removed during processing and whose nucleus lies
flat against the cell membrane. Some reticular fibers surround
and support the capillaries among the fat cells.
 Muscular System
(Figures 150-153)

Two characteristics of muscle tissue are contractility and, to a STRIATED MUSCLE

lesser extent, conductivity. The muscle fibers are the units of
contraction. Abundant connective tissue is found among mus- Striated (voluntary or skeletal) muscles are capable of
cle fibers as a supporting substance and also as a medium in powerful contraction or maintaining tonus (e.g., holding the
which blood vessels and nerves are placed in intimate contact head erect). Connective tissue also accompanies this type of
with muscle fibers. muscle, bearing blood vessels and nerves as in smooth muscle.
Morphologically, muscle tissue may be classified into However, skeletal muscle cells are larger and multinucleate
three types: smooth involuntary, striated voluntary (skeletal), (even syncytial), ranging in length from a few millimeters to
and striated involuntary (cardiac). a few centimeters. The nuclei are elongated and oval and
appear on the outside of the muscle fiber bundle in longitu-
dinal section.
SMOOTH MUSCLE Each fiber is enclosed by a very thin covering, the sarco-
lemma, which adheres very closely to the sides of the fiber
Smooth muscle is found in the walls of most tubular and is not easily recognized in stained preparations. When
structures and organs (gastrointestinal tract, arteries and viewed in longitudinal section the striations, for which this
veins, most hollow viscera, uterus, bladder, etc.). Smooth mus- type of muscle is named, are striking. Vertical light (I) bands
cle fibers are tapered and have a wide center around the and dark (A) bands alternate with each other. A dark (Z) line
eccentrically placed nucleus and pointed or blunt ends; the bisects each light (I) band.
cells are roughly cigar-shaped. The muscle fibers are arranged All skeletal muscle fibers have an associate nerve fiber
in sheets or bundles surrounded by connective tissue. In a whose cell body is located in the anterior horn of the spinal
longitudinal view the nuclei appear to lie on top of the sheet cord. Its axon extends to a terminal point on the muscle, the
of fibers. Blood vessels and nerves accompany the muscle motor end plate. Some of these structures in the photomIcro-
fibers in their connective tissue covering. Generally all muscle graph section have been demonstrated by silver staining.
fibers in a specific area contract as a unit, as in the intestinal
tract during peristalsis.
31
32 Muscular System

CARDIAC MUSCLE muscle fibers. Cross striations are seen in the cytoplasm,
though not stained as intensely as those in skeletal muscle. In
Cardiac (involuntary) striated muscle appears as a syncy- addition to cross striations, transverse dark lines are often
tium of fibers branching and anastomosing; between the fiber identified at various intervals in a steplike pattern. These are
bundles is connective tissue bearing blood and lymph capil- intercalated disks, and they mark the boundary between adja-
laries and nerves. Nuclei of cardiac muscle are ovoid with cent muscle fibers.
blunt ends and appear to be on top and in the middle of the
 Circulatory System
(Figures 154-168)

HEART ARTERIES AND VEINS

The heart is a hollow muscular organ enclosed in a fibroelastic The walls of arteries consist of three layers: tunica intima,
pericardial sac. In the wall of the heart, three distinct layers tunica media, and tunica adventia. The intima consists of an
can be identified. The inner layer lining the chambers and endothelial lining of the lumen with subendothelial connec-
covering the valves is the endocardium. The thick cardiac tive tissue and abundant elastic fibers. A thick wall of smooth
muscle of the heart is the myocardium. The epicardium is the muscle and elastic fibers constitutes the media, which varies
thin layer of mesothelium and connective tissue that covers in thickness with the caliber of the artery. The outermost layer
the surface of the heart as the visceral pericardium. The peri- or adventia contains connective tissue and collagen, which
cardial cavity, filled with fluid, separates the pericardium and gradually merges into the surrounding tissues. Valves are
epicardium. absent in arteries.
The impulse-conducting system of the heart consists of Although similar to arteries, veins are generally thinner
specialized cardiac muscle fibers in large amounts of fibro- and have fewer elastic fibers in the intima and less smooth
elastic tissue abundantly supplied by nerves from the auto- muscle in the media, although the adventia is very similar to
nomic nervous system. Among the constituents of the system that of arteries. In mice the smooth muscle layer of the pul-
are the sinoatrial (SA) node (the pacemaker of the heart), the monary vein is replaced by cardiac muscle. Paired valves are
atrioventricular (A V) node, and Purkinje fibers. The latter are present throughout the venous system.
very wide fibers having two nuclei in a clear mass of sarco-
plasm rich in mitochondria and glycogen, with cross striations
but no intercalated disks. The myofibrils are peripherally CAPILLARIES
located, leaving a wide central space, seen in prepared sec-
tions. The Purkinje fibers conduct impulses for ventricular The capillary wall is composed of a single layer of endo-
contraction, reacting much more rapidly than ordinary cardiac thelial cells on an underlying basement membrane with no
muscle. smooth muscle but many reticular fibers.

33
34 Circulatory System

PERIPHERAL BLOOD CELLS- WRIGHT'S STAIN of cytoplasmic staining. In Figures 165-168 several of these
variants can be compared.
Blood smears made from tail vein preparations include Erythrocytes appear in the background of Figures 161-
monocytes, small and large lymphocytes of the agranulocytic 168 as gray, biconcave disks of uniform size.
series, and neutrophils and eosinophils of the granulocytic Platelets appear in clusters with red granules and blue
series. Basophils are very rarely seen. Eosinophils vary consid- cytoplasm.
erably in nuclear morphology, size of granules, and intensity
Col or Fig ure s
36 Nervous System

1. Head SagIttal sectIOn H & E lOX

The upper half of thIs mIdsagIttal sectIOn through the head extends from
2. Cerebellum and medulla Masson staIn 25X
the lIps and InCIsor tooth on the rIght through the nasal cavIty and Its
SInuses to the trachea, ImmedIately above whIch IS the braIn The homoge-
In thIS cross sectIOn of braIn the patches of green are the granular layer of
neous pInk cerebral cortex contrasts wIth the arbor vItae or "tree of lIfe"
the cerebellum ImmedIately adjacent are areas of reddIsh brown, the molec-
appearance of the dark blue cerebellum at the posten or braIn, at left The
ular layer on the edge of whIch consIsts of cell bodIes of PurkInJe cells not
small tnangular structure ImmedIately below the posten or cerebral cortex IS
seen at thIS low magmfIcatlOn The lower half of the sectIOn IS occupIed by
the pItuItary gland The pInk trIangle Includes the IntermedIate lobe and
the medulla, whose nerve cell bodIes are located In the dark regIOns, whIle
neurohypophysIs, whIch lIe on top of the honzontal dark, antenor lobe The
the nerve fIbers course through the alternate pInk areas
lower Jaw Includes a tooth and overhangIng tongue
1 Cerebellum
Skull 7 Mouth
2 Nerve fIbers and neuroglIa
2 SkIn 8 Tooth
3 Medulla
3 Cerebellum 9 Tongue
4 ChorOId plexus In ventncle
4 Cerebral cortex 10 Esophagus
5 Nasal sInuses 11 PItUItary gland
6 Trachea

3. Cerebral cortex Luxol Fast Blue 500X 4. ChorOId plexus H&E 250X
Nuclear Fast Red
The choroId plexus of capIllarIes lIes In the ventncle, lIned by a speCIalIzed
The blue-staIned fIbers croSSIng the fIeld are myelInated fIbers Densely red- epIthelIum of ependymal cells Blood In a venous SInUSOId appears at center
staIned nucleI of neuroglIal cells appear between fIbers The group of left
nucleated nerve cells at lower left forms a ganglIon, many of whose cells
have bnght red nucleolI ChorOId plexus
2 Ependymal cell layer
MyelInated (blue) fIbers 3 Ventncle
2 GanglIon cells 4 Cerebral cortex
3 NeuroglIal cell nucleI 5 Erythrocytes In venous SInUSOId
 .10

Figure 1 Figure 2
38 Nervous System

5. Spmal cord Antenor horn Bodlan sliver 500X 6. GanglIon Masson staIn 500X

The entIre fteld IS ftlled WIth varIOUS nerve ftbers Several large motor cells, CharactenstIcally large round ganglIon cell bodIes ftll most of the fteld, wIth
two of whIch exhIbIt pronounced axons, are seen at lower left and upper a veIn at lower nght and green bone at lower left Two nerve trunks m cross
rIght sectIOn are present at upper nght wIth a few Schwann cell nucleI m focus

Motor cell 1 GanglIon cell bodIes

2 NeuroglIal cell nucleus 2 Nerve trunk
3 Nerve ftbers 3 Vem
4 Axon of nerve cell 4 Bone
5 Neuroftbnllae (ftne brown lmes) 5 Fat cells

8. Motor neurons m cerebral cortex Bodlan SIlver 500X

7. Cerebral cortex Bodlan Sliver 500X
The axons and dendntes of each cell are easIly IdentIfted Wlthm the cyto-
Several large motor neurons exhIbIt axons, and many neuroftbnllae can be plasm are ftne, dark lInes, the neuroftbnllae Neurogbal cells and nerve
seen wlthm theIr cytoplasm In the background are nucleI of neurogbal cells ftbers completely ftll the background
and nerve ftbers
1 Nucleus and nucleolus of neuron
Motor cell nucleus and nucleolus 2 Axon
2 Dendnte 3 Dendnte
3 Nerve ftber 4 Neuroftbnllary meshwork
4 Neurogbal cell nucleus 5 Nucleus of neurogbal cell
6 Nerve ftber
Figure 5 Figure 6

Figure 8
40 Nervous System

9. Cerebral cortex Luxol Fast Blue 500X

Cresylecht vlOlet
10. Cerebellum Luxol Fast Blue 250X
A large motor cell body m the center foreground exhibits a nucleus with Cresylecht violet
nucleolus and abundant Nissl substance m the cytoplasm Purple myeh-
nated nerve fIbers extend across the fIeld, among which are several darkly The molecular layer sparsely occupied by neuroghal cells and the cellular
stamed nuclei of neuroghal cells granular layer fIlled with the same type of supportmg cells form two marked
dlvlslOns The PurkmJe cell layer hes between the two layers
Motor neuron cell body
2 Nucleus Molecular layer
3 Nucleolus 2 Granular layer
4 Nissl substance 3 PurkInJe cell neurons
5 Myehnated fIbers (purple)
6 Nucleus of neuroghal cell

11. Cerebellum H&E 75X 12. Cerebellum Bodlan Silver 500X

The characteristic appearance of hght areas (molecular layer) and dark areas The left half of the fIeld IS fIlled with nucleated cells of the granular layer
(granular layer) has earned the name arbor vitae ("tree of hfe") The pmk Along the Inner border of thiS layer are large PurkmJe cells exhibiting axons
cerebral cortex hes below extendIng mto the molecular layer occupymg the right fIeld Interspersed
among the fIbers are neuroghal cells
1 Cerebellum
2 Cerebral cortex 1 PurkInJe cells
3 ChorOid plexus 2 Axon from PurkmJe cell
4 Bone (skull) 3 Granular layer
5 Granular layer 4 Molecular layer
6 Molecular layer 5 Nucleus of neuroghal cell
 .. " .
" •

• "0
.-
••• .
"

...
•0 •

•
•
Figure 9 Figure 10
42 Nervous System

13. Spmal cord Bodlan sliver 75X 14. SpInal cord AnterIor horn cells Luxol Fast Blue 500X
Nuclear Fast Red
In thIS cross sectIon of the thoracIc spInal cord, the posterIor horn IS m the
upper half of the cord, wIth the anterIor horn m the lower half near the At thIS magnI£IcatlOn the red nucleolI and chromatIn materIal In the nucleus
vertebral body A large ganglIon from whIch a nerve trunk extends IS seen of large anterIor horn motor cells are easIly dIstIngUIshed A blue-staIned
at lower left capillary IS In the rIght center One large motor neuron In upper center
exhIbIts an axon and dendrItes
1 AnterIOr horn
2 PosterIor horn 1 Motor cell neuron
3 GanglIon 2 NeuroglIal cell
4 Vertebral column 3 Myehnated nerve £Ibers
5 Nerve trunk 4 NonmyelInated nerve £Ibers
6 Central canal 5 Blood vessel

15. SpInal cord Luxol Fast Blue lOOX

Nuclear Fast Red
16. SpInal ganglIon Bodlan Silver 250X
In thIS cross sectIon of lumbar spInal cord, the "butterfly" pattern of anterIor
and posterIOr horns IS apparent, partIcularly the large red motor cells In the HIgher magnI£IcatIon enables one to IdentIfy the large round ganglIOn cells
anterIor horn Blue myelInated £Ibers £Ill out the surroundIng spInal cord A and nerve £Ibers extendIng toward the lower left
thIn menIngeal coverIng IS seen In the space between cord and vertebra
1 GanglIon nerve cell body
AnterIor horn 2 Nerve £Ibers
2 PosterIOr horn
3 Nerve trunk
3 Cerebrospmal canal
4 Skeletal muscle
4 MyelInated nerve £Ibers
5 MenInges
6 PIa mater
5

Figure 15
44 Nervous System

18. Motor end plates Pestronk and Drachman 500X

17. Motor end plate Bodian silver 500X stain

Between the bundles of striated muscle fibers are several black nerve fibers Acetylcholinesterase appears as an abundant blue mass superimposed on
terminating on oval, dark brown motor end plates. motor end plates. Black nerve fibers enter the end plates from a branch
between striated muscle fibers.
1. Nucleus striated muscle
2. Striated (skeletal) muscle 1. Nerve fiber
3. Nerve 2. Motor end plate
4. Motor end plate nucleus 3. Acetylcholinesterase (blue)
4. Striated muscle

19. Pineal gland H&E lOOX

The pineal gland lies embedded in brain tissue at the junction of the cerebral
cortex and cerebellum. Surrounded by blood vessels, it is a highly vascular
member of the endocrine system.

1. Pineal gland
2. Cerebral cortex
3. Cerebellum
4. Blood vessel
Figure 17 Figure 18

Figure 19
46 Special Sense Organs

20. Eye H&E sox

The pInk epIthehal-hned cornea IS the curved tIssue on the nght extendIng
around half the CIrcumference of the eye The red central sphere IS the lens
The pIgmented trIS above and below the lens merges Into the chorOId Just
postenor to the CIhary body, the small proJechng hssue above and below the
Ins The rehna IS the pInk band of hssue In the posten or porhon of the eye
bounded by the CIhary body The glandular tIssue surroundIng the eye
above postenorly and below on the left IS the Hardenan gland (pInk)

1 Lens S ChOroId (pIgmented)

2 Cornea 6 VItreous humor
3 Ins (pIgmented) 7 Hardenan gland
4 RetIna 8 Lacnmal gland
 8

Figure 20
48 Special Sense Organs

21. Cornea 1 5 f.Lm Methacrylate soox

Methylene Blue
BasIc fuchsm
22. Eye H&E soox
Four distinct layers may be Identified In the outer layer of stratified squa-
mous eplthehum, two mitotic figures may be seen, one cell in telophase The large optic nerve fills the central area as It enters the postenor part of
Below the eplthehum the wide substantia propna layer extends down to a the eye ending m Immediate contact With the retina Below, brown fibers of
very thin, homogeneous, pink Descemet's membrane, which m turn IS the chorOid layer blend With pmk fibers of the sclera
Immediately adjacent to a posterIOr endothehum
1 Optic nerve
1 Stratified squamous eplthehum 2 Retina
2 Basal eplthehal cell in diVISIOn 3 ChorOid
3 Substanha propna 4 Blood vessel
4 Fibroblast 5 Sclera
5 Descemet's membrane
6 Endothehum
7 Conjunctiva

24. Eye Lens, UIS, cornea H&E 2S0X

23. Eye Lens, retina, Hardenan H&E 2S0X Across the top of the sectIOn, the bow of the lens, covered by a Single cell
gland layer of epithelium, contams many nuclei of lens fibers The wide band of
the mne-Iayered retma extends from the left of the clhary body m the Imd-
Immediately below the red-stamed lens are the deeply pigmented Ins on die two-thuds of the sectIOn The space below the lens and to the left con-
the nght and nme-Iayered retma on the left Separation of the retina from tams vitreous humor m the livmg ammal The antenor chamber IS the space
the pigmented chorOid IS an artifact The Hardenan gland occupies the to the nght of the ciliary body, whose pigmented structure IS in the center
lower thud of the section Immediately below the lens ContinuatIOn of the heavily pigmented Ins
extends laterally to the nght The pigmented layer extending to the left IS
Hardenan gland the chorOid The lower thud of the sectIOn mcludes the pmk sclera Imme-
2 Conjunctiva diately subjacent to the chorOid, below which IS the mucocutaneous eplthe-
3 Clhary body hum of the conJunctiva, whose upper layer blends With the cornea and
4 Ins (pigmented) whose lower layer contmues as the conjunctiva
5 Retina
6 Lens 1 Lens 5 Sclera 9 ChorOid
7 ChorOid (black) 2 Clhary body 6 Conjunctiva
3 Ins 7 AnterIOr chamber
4 Retma 8 Posten or chamber
Figure 21 Figure 22

Figure 23 Figure 24
50 Special Sense Organs

25. Eye: Conjunctiva and cornea PAS 250X

26. Eye: Conjunctiva and cornea PAS 275X
In this section PAS-positive material occurs in the posterior layer (endothe-
lium) of the cornea and at the base of the stratified squamous epithelium. The mucus-laden cells of the conjunctiva appear brilliant red, and the endo-
PAS-positive mucus is also seen in cells of the conjunctiva. The sebaceous thelium at the posterior cornea as well as some areas of the iris in the bottom
glands of Meibom are present at lower left. of the section are also PAS-positive.

1. Substantia propria-cornea 1. Epithelia of cornea

2. Epithelium-cornea 2. Substantia propria
3. Conjunctiva-mucous cells (red) 3. Iris
4. Sebaceous gland (Meibomian gland) 4. Conjunctiva mucous cells (red)

27. Eye: Conjunctiva Heidenhain's Azan 250X

28. Eye: Lacrimal and Harderian Masson stain 250X
glands
B~own pigment fills the iris at the top of the section. The substantia propria
wlth packed blue collagen fibers dominates the cornea. Below is the con-
In the left lower third of the section is a portion of the lacrimal gland. Its
junctiva with blue-staining mucoid cells in its epithelium. Striated muscle
basophilic glandular appearance resembles the serous-type salivary gland.
fibers appear pink at lower right and Meibomian sebaceous glands appear at
lower left. The pink glandular tissue on the right is part of the Harderian gland.

1. Striated muscle 6. Choroid (black)

1. Conjunctiva mucous cells (light blue)
2. Harderian gland 7. Lens
2. Meibomian (sebaceous) gland
3. Lacrimal gland 8. Iris (pigmented)
3. Striated muscle fibers
4. Ciliary body 9. Conjunctiva (eyelid)
4. Cornea
5. Retina 10. Vitreous humor space
5. Pigmented iris
 Figure 26
Figure 25

Figure 28
Figure 27
52 Special Sense Organs

30. Ear Organ of CortI HeidenhaIn's Azan 2S0X

29. Ear Organ of CortI H&E 7SX At each end of the center structure the blue tectonal membrane overhangs
the tIny haIr cells restIng on the basIlar membrane The thIn structure sep-
Th,s IS a sectIon cut through the bony cochlea to Illustrate the delIcate struc- aratIng the cochlear duct from the scala vestIbulI IS the vestIbular or
tures IdentIfied In the captIons assocIated wIth the photomIcrograph Relssner's membrane The scala tympam IS the space below the basIlar
membrane
1 LImbus lamIna spIralIs 7 SpIral lIgament
2 VestIbular membrane (Relssner's) 8 Tectonal membrane 1 Tectonal membrane
3 Scala vestIbulI 9 Scala tympam 2 VestIbular membrane (Relssner's)
4 Cochlear duct 10 Stna vasculans 3 LImbus spiralIs
S BasIlar membrane 11 HaIr cells 4 Scala tym pam
6 Internal spIral sulcus S HaIr cells
6 Cochlear duct
7 Scala vestIbulI

31. External ear (pInna) H&E sox

StratIfied squamous epIthelIum of epIdermIs forms the outer border The
pmk area enclosmg spaces m the center IS elastIc cartIlage The pInk con-
nectIve tIssue between outer epIdermIs and layer of cartIlage IS dermIS,
mterspersed wIth groups of large, pale sebaceous gland cells A few spher-
Ical cross sectIOns of haIr follIcles are found m the dermIS

1 StratIfied squamous epIthelIum

2 DermIS
3 Sebaceous gland
4 HaIr follIcle
S Fat cells
6 ElastIc cartIlage
7 Stnated muscle
1 2 7

Figure 29 Figure 30

'-4
2
5
6

Figure 31
54 Endocrine System

33. PItUItary gland AnterIor lobe Aldehyde-fuchsIn 250X

32. PItUItary gland PerformIc aCId 200X MartIn-Mallory staIn
Aldehyde-fuchsIn
Red aCIdophIls predomInate In thIS sectIOn, wIth an occasIOnal purple beta
The three lobes of the pItUItary are clearly seen The posterIor pItUItary has cell (gonadotroph) and many chromophobes (gray) Two arterIes wIth dIs-
reacted posItIvely to the staIn, whIch demonstrates the presence of neuro- tInctIve mternal elastIc membranes (purple) are present In upper rIght
secretory substance
1 Aldehyde-pOSItIve basophIl
AnterIor lobe 2 ACIdophIl
2 IntermedIate lobe 3 Chromophobe
3 PosterIOr lobe 4 Artery
5 Collagen fibers (blue)

34. PItUItary gland HypophYSIS Masson staIn 75X

35. PItUItary gland AnterIor lobe Aldehyde-fuchsIn 500X
The pItUItary gland lIes anatomIcally WIthIn the bony sella turCIca (green), ACId fuchSIn
WIth marrow cells wIthIn ItS bony dIVIsIOns The red-staInIng anterIor lobe Orange G
IS separated by a cleft from the narrow IntermedIate lobe and the lIghter
pInk posterIor lobe (neurohypophysIs) Aldehyde-fuchsIn-posItIve gonadotrophs (purple) are eaSIly dIfferentIated
among red aCldophIls and gray chromophobes
AnterIor lobe
2 IntermedIate lobe 1 Aldehyde-fuchsIn-posItIve beta cells
3 PosterIOr lobe 2 ACIdophIls
4 Bone marrow 3 Chromophobes
5 Bone
6 StrIated muscle
Figure 32 Figure 33

Figure 35
56 Endocrine System

36. Adrenal gland H&E lOOX

DIvIsIOn between cortex and medulla IS apparent In the cortex the outer
zona glomerulosa lIes ImmedIately below the capsule and IS adjacent to the 37. Adrenal gland H&E 200X
zona fasClculata The zona fasClculata IS WIde and extends to the next zone,
whIch IS the X zone, also known as the "Juvemle zone" The zona retIculans The three zones of the cortex and the medulla can eaSIly be IdentIfIed
does not appear In thIS sectIon The medulla (center) contaInS several blood
vessels 1 Capsule
2 Zona glomerulosa
1 Zona glomerulosa 3 Zona fasClculata
2 Zona fasClculata 4 X zone
3 X zone S Medulla
4 Medulla
S Blood vessel
6 KIdney

38. Adrenal gland PAS sox

At low magnIfIcatIon the extent of "brown degeneratIon" can easIly be seen
39. Adrenal gland H&E lOOX
The presence of lIpId In thIS substance results In a posItIve reactIon (red)
wIth perIOdIC aCId-SchIff reagent concentrated between the zona fasClculata
A large amount of cortIcal "brown degeneratIOn" pigment fIlls the central
(cortex) and margIn of the medulla
and nght fIeld A small area of medulla IS seen at lower nght
1 Zona fasClculata (cortex)
Zona glomerulosa
2 "Brown degeneratIon" pIgment
2 Zona fasClculata
3 Medulla
3 "Brown degeneratIOn" pigment
4 KIdney cortex
4 Medulla
S Glomerulus

Sample from H-yr-old mouse Brown degeneratIon Increases as mouse ages

Figure 36 Figure 37

Figure 38 Figure 39
58 Endocrine System

40. Thyroid and parathyroid glands PAS sox

The anatomical relationships of the thyroid gland, trachea, parathyrOid
41. ThyrOid and parathyroid glands H&E SOOX
gland, and esophagus are seen In thiS low-power section The cartilaginous,
rInged trachea IS bound by the lateral lobes of the thyrOid gland, whose
CubOidal epithelium lines the collOid-filled follicles of the thyrOid gland
brIghtly staIned red collOid IS apparent The Isthmus lobe, which lies on the
Chief and oxyphil cells of the parathyroid gland appear as light and dark
anterIor surface of the trachea, IS not present In thiS section PosterIOr to the
blue-staIned cells
trachea IS the esophagus
1 CollOid WithIn follicle-thyroid
1 Lumen of trachea 6 StrIated muscle 2 Follicular cells-thyrOid
2 Lumen of esophagus 7 ThyrOid gland 3 ParathyrOid gland
3 Hyaline cartilage 8 ParathyrOid gland
4 Mucous glands of trachea 9 StrIated muscle
S Stratified squamous epithelium-esophagus 10 Epithelium-trachea

42. ThyrOid and parathyroid glands H&E 2S0X

43. ParathyrOid gland H&E 2S0X
CubOidal epithelium lines the thyrOid follicles, which are filled With pInk
Two cell types can be Identified chief cells (OVOId, very dark nuclei, eOSIn-
collOid The dark-purple-staIned parathyroid gland IS penetrated by several
ophilic cytoplasm) and pnnClpal cells (large veSicular nucleus and smaller
blood vessels StrIated muscle of the esophagus appears at upper left
amounts of basophilic cytoplasm)
1 ThyrOid follicles
2 Parathyroid gland 1 Chief cells
3 StrIated muscle 2 Oxyphil cells
Figure 40 Figure 41

Figure 42 Figure 43
60 Reproductive System

44. Ovary H&E sox 45. Ovary H&E soox

The ovary IS completely surrounded by a capsule of loose connective tissue
The "germinal" epithelium forms the upper boundary of the ovary, beneath
Beneath the capsule IS the "germinal" epithelium of cubOidal cells resting
which are several primordial cells Two oocytes are below the primordial
on a basement membrane Adipose tissue around the ovary IS seen at upper
cells, left and right A larger growing follicle occupies the center The
right A small porhon of uterine tube appears at upper left In the ovary
mature ovum at lower center consists of the nucleus and cytoplasm of the
several small primordial follicles can be seen adjacent to the capsule at lower
ovum, and the antrum (space), containing follicular flUid, IS surrounded by
left, while a growing follicle With follicular flUid In the antrum IS at upper
a ring of follicular cells
right Immediately below this follicle are two corpora lutea With theu char-
acteristic pink luteal cells and capillaries
1 Germinal epithelium
2 Primordial follicle
1 Growing follicle
3 Growing follicle
2 Corpus luteum
4 Nucleus of ripening follicle } Ovum
3 Primordial follicle
S Cytoplasm of npenlng follicle
4 Germinal epithelium
6 Follicular cells
S Fat cells
7 Interstitial cells
6 Capsule

46. Ovary H&E soox

47. Vagina H&E sox
A large matunng follicle With ovum In the center IS seen In the middle of
The external vaginal opening IS formed between layers of stratified squa-
thiS section Surrounding the ovum IS a pink, oval ring, the zona pelluCl-
mous epithelium Sebaceous glands lie on both Sides, surrounded by smooth
da, outSide of which are several rows of follicular cells The space at left
muscle Skin With hair follicles and sebaceous glands merges With the vagi-
center IS fdled With follicular flUid One follicular cell IS diViding OutSide
nal epithelium
the ring of follicular cells are cells forming the theca OccaSIOnally one can
differentiate a theca Interna separated from theca externa by capillanes Part
1 Vaginal opening
of a growing follicle IS seen above the mature ovum
2 Stratified squamous epithelium
3 Urethra
Mature follicle 6 Blood vessel
4 Skin
2 Ovum nucleus 7 Pnmordlal follicle
S Clitoral gland
3 Follicle cell In mitosIs 8 Theca
6 Sebaceous gland
4 Follicular flUid 9 Zona pelluCida
7 Fat cells
S Growing follicl
Figure 44 Figure 45

Figure 46 Figure 47
62 Reproductive System

48. Uterus PAS 150X

Uterine glands with PAS-positive (red) glycogen are easily recognizable in

49. Fimbria of oviduct Heidenhain's Azan 500X
the endometrium occupying the right half of the section. The myometrium
of circular and longitudinal smooth muscle is seen at center and to the left
Ciliated cuboidal epithelium lines this structure, with a pronounced red
bounded by the serosa.
striated border appearing as a base from which cilia extend. Blue collagen
fibers are part of the connective tissue supporting the epithelium.
1. Uterine glands (glycogen, red)
2. Lumen of uterus
1. Ciliated columnar epithelium
3. Simple columnar epithelium
2. Lumen of oviduct
4. Endometrium
3. Striated border (red)
5. Myometrium
6. Blood vessels
7. Serosa

51. Placenta H&E 200X

50. Placenta and embryo H&E sox
The upper two-thirds of the section is the decidua basalis with maternal
In this section an embryo is located in the center, surrounded by a very thin blood cells within the blood vessels. The middle section contains a few giant
amnion, outside of which is the visceral yolk sac. The fetal placenta is seen cells separated by spaces (labyrinths). Below is the fetal placenta, whose
at the bottom of the photo while the maternal placenta (decidua basalis) blood vessels are filled with embryonic blood cells.
occupies the upper third of the section.
l. Giant-cell area
l. Embryo 5. Blood island 2. Fetal placenta
2. Visceral yolk sac 6. Amnion (very thin) 3. Decidua basalis
3. Decidua basalis 7. Decidua capsularis 4. Labyrinth
4. Endometrium 8. Metrial gland 5. Maternal blood cells
6. Embryonic blood cells
Figure 48 Figure 49

Figure 50 Figure 51
64 Reproductive System

53. Vagma Diestrus H&E 2S0X

52. Placenta Maternal side H&E sox
The diestrus phase IS one m which the eplthehum begms to rebUild, the
The maternal structures are Identified as the myometnum on the nght
stratum germmatlvum mcreasmg to a thickness of several cell layers There
adJolmng the endometrlUm to the left, which m turn IS adjacent to the very
IS also an abundance of polymorphonuclear leukocytes movmg from the
vascular decidua basahs, below which IS the fetal placenta
eplthehum out mto the lumen
Myometnum
Stratified squamous eplthehum (mcludes prohferatmg stratum germl-
2 EndometrlUm
natlvum)
3 Decidua basahs
2 Polymorphonuclear leukocytes
4 Fetal placenta
3 Connective tissue and smooth muscle

54. Vagma Proestrus 55. Vagma Proestrus/estrus H&E 2S0X

H&E 4S0X

OccasIOnally, under the mfluence of progesterone, a layer of mucous cells

A thm, red cormfied layer appears Just below the stratum granulosum dur-
mg the begmmng of proestrus A few mitotic figures appear forms on the outer stratum granulosum They slough mto the lumen, fol-
lowed by mcreased thlckenmg of the cormfied layer
1 Granulosa layer of eplthehum begmnmg cormficatlOn
2 Thlckenmg stratified squamous eplthehum 1 Mucous cells
3 Mitotic figures 2 Thm red hne of cormficatlOn
3 Stratlfied squamous eplthehum
Figure 52 Figure 53

Figure 54 Figure 55
66 Reproductive System

57. VagIna Estrus H&E 250X

56. Vagma Proestrus / estrus PAS 250X
Dunng estrus the cornIfIed granulosum layer thIckens and appears super-
Same fIeld as prevIOus H & E section Mucous cells stimulated by progester- fICially No leukocytes or mItotic fIgures are seen Many dead cells and cel-
one appear PAS-posItive (red) lular debns are sloughed mto the lumen

1 Mucous cells (red) 1 CornIfIed layer

2 StratifIed squamous epIthelIum 2 Cells and cellular debns In lumen
3 StratifIed squamous epIthelIum

58. Vagma Metestrus I H&E 250X

59. VagIna Metestrus II H&E 250X
DurIng metestrus I cornIfIed cells appear In the lumen, wIth polymorpho-
nuclear leukocytes appeanng In the epIthelIum The cornIfIed layer desqua-
An abundance of polymorphonuclear cells are seen m the squamous epIthe-
mates, and many cornIfIed cells appear In the lumen Leukocytes are present
lIum before they move out mto the lumen WIth other cellular debns
In the epIthelIum
Polymorphonuclear leukocytes and cormfIed cells m lumen
CornIfIed layer desquamatIng Into lumen
2 Leukocytes m epIthelIum
2 Polymorphonuclear leukocytes
3 StratifIed squamous epIthelIum
3 StratifIed squamous epIthelIum
4 Smooth muscle and connective tissue
Figure 56 Figure 57

Figure 58 Figure 59
68 Reproductive System

61. Mammary gland Nonlactatmg H&E 50X

60. NIpple Lactating breast Heldenham's Azan 150X
Mammary tissue m the mouse IS found on both SIdes of the ventral abdom-
The outer skin surrounds the mpple on the left The large, oval space center Inal surface extendmg from the neck to the grom ThIs section was made
nght IS the lactiferous duct, below whIch are lobules Subcutaneous fat through the outer skm layer through fat and stnated muscle to the thIck
occurs between collagen fibers fatty tissue, m whIch lobes of mammary tissue and mtralobular ducts are
located A large vem IS at lower nght
Skm
2 Lactiferous duct EpIdermIs
3 Lobules 2 DermIs
4 HalT folhcles 3 HaIr folhcle
5 Collagen fibers (blue) 4 Fat cells
6 Sebaceous gland 5 Stnated muscle
7 Fat cells 6 Intralobular ducts
7 Blood-filled vem

62. Mammary gland Lactating H&E 100X

Thm skm wIth haIr folhcles forms the upper border, beneath whIch are a
layer of fat cells and a few strands of skeletal muscle Most of the gland
consIsts of lobules of secretmg alveoh, wIth two lactiferous ducts m the
center

Skm
2 Fat cells
3 Lactiferous duct
4 Lobules of alveoh
5 Stnated muscle
Figure 60
Figure 61

Figure 62
70 Reproductive System

A. Male Urogenital System

I. Left kidney 10. Urinary bladder

2. Ureter II. Ventral prostate
3. Left testis 12. Dorsal prostate
4. Epididymis caput 13. Ischiocavernosus muscle
5. Epididymis cauda 14. Bulbourethral gland
6. Vas deferens 15. Bulbocavernosus muscle
7. Ampullary gland 16. Preputial gland
8. Vesicular gland (seminal vesicle) 17. Penis (within sheath)
9. Coagulating gland
 2

Figure A
72 Reproductive System

63. Pems H&E 50X 64. Pems Heldenham's Azan lOOX

In the center of the sechon lS the os pems, below whlch lS the urethra Sur- Starhng at the center of thlS cross sechon of pems, the os pems hes duectly
roundmg these structures are the corpora cavernosa of connechve hssue above the urethra, whlch lS hned by transltlOnal eplthehum The vascular
Eplthehum of the glans completely surrounds the organ, separatmg lt from erechle hssue (deep blue) surroundmg the urethra lS the corpora caver-
the prepuhal cavlty The prepuhal glands he on each slde of the organ The nosum Folded strahhed squamous eplthehum (bnlhant red) forms the pre-
skm, wlth halr folhcles and fat cells, forms the outer border puce whlle the enhre organ hes m a space lmed by speClahzed eplthehum
Prepuhal glands he on elther slde of the pems near the external openmg,
1 Os pems (bone) whlch lS surrounded by the strahhed squamous eplthehum of the skm
2 Urethra Strands of smooth muscle and fat cells appear wlthln connechve hssue
3 Erechle hssue
4 Eplthehum of glans 1 Os pems (bone) 6 Skm
5 Eplthehum of prepuce 2 Urethral lumen 7 Blood vessel
6 Prepuhal cavlty 3 Eplthehum of glands 8 Fat cells
7 Prepuhal gland 4 Corpora cavernosum 9 Eplthehum of prepuce
8 Skm 5 Prepuhal gland

66. Teshs H&E 250X

Thls cross sechon of a semlmferous tubule reveals three types of cells The
65. Teshs H&E 250X
supporhng, nonprohferatmg Sertoh cells are located lmmedlately adjacent
to the basement membrane and are dlstnbuted among spermatogoma They
The teshs lS covered by a thlck, hbrous capsule (tumca albugmea) Flbers
are the most pnmlhve sperm cells The next layer of cells lS composed of
from the capsule form thlck septa These dlvlde the teshs mto semlmferous
pnmary spermatocytes, whlch undergo melOS1S and form secondary sper-
tubules, whlch he on a basement membrane contammg supporhng cells
matocytes, located m the thlrd layer of cells Spermahds are now produced,
(Sertoh cells) and spermatogemc cells
whlch form mature spermatozoa In the lntershhal hssue, amld connechve
hssue and blood vessels, are cells of Leydlg, the slte of ongm of testosterone
1 Semlmferous tubule
2 Intershhal cells of Leydlg
Sertoh cell
3 Developmg sperm
2 Intershhal cells of Leydlg
4 Capsule-tumca albugmea
3 Spermatogomum
4 Pnmary and secondary spermatocytes
5 Spermahds and mature sperm
Figure 63 Figure 64

Figure 65 Figure 66
74 Reproductive System

67. Testis Heldenhalfi's Azan 500X

In thIS sectIOn, bnlliant red mature spermatozoa occupy the lumen of a sem-
68. Ductus deferens H&E 200X
InIferous tubule DevelopIng stages can be IdentIfled from spermatogoma
lImng the penphery through cells movlfig toward the lumen, where sper-
The duct IS fllled WIth spermatozoa and llfied by sImple columnar epIthe-
matIds are formIng mature sperm
lIum Smooth muscle and connective tissue surround the ducts
1 SertolI cell
Columnar epIthelIum
2 Spermatogomum
2 Smooth muscle flbers
3 Pnmary spermatocyte
3 Mass of spermatozoa lfi lumen
4 Secondary spermatocyte
5 Spermatozoa
6 Interstitial cells of LeydIg

70. Efferent duct H&E 500X

69. EpIdIdymIs Caput H&E 250X
CoIled segments are lIned by tall columnar epIthelIum WIth stereoCllIa pro-
The head of the epIdIdymIs contalfis many ducts lIned by columnar epIthe-
jectlfig from the free surface of the cells Several dlVldlfig cells, as well as
lIum wIth stereOCIlIa on theIr free surfaces The lumlfia of many tubules con-
spermatozoa, are present In the lumen
taIn spermatozoa FIbrous connective tissue separates the tubules
1 Duct
1 Lumen wIth sl'ermatozoa
2 StereOCIlIa
2 Interstitial tissue
3 Columnar epIthelIum
3 Capsule
4, Cells lfi mItosIs
,
Figure 67 Figure 68
76 Reproductive System

72. Male reproductive system H&E lOOX

71. Bulbourethral gland H&E lOOX The vas deferens, lined by columnar epithelium and surrounded by smooth
muscle, occupies the upper left. Connective tissue with fat cells separates
This gland is characterized by alveoli of tall columnar cells, dark nuclei, and
the vas from the ventral prostate in the lower right corner of the field.
very light cytoplasm. A rim of striated muscle partially surrounds the gland.
1. Vas deferens
1. Striated muscle
2. Smooth muscle and connective tissue
2. Alveoli-tall columnar cells
3. Ventral prostate
4. Adipose tissue

73. Male reproductive system H&E sox

A section of rectum containing a lymphoid nodule lies at the extreme right.
Left of the rectum is the urethra, surrounded by smooth muscle and con-
taining pink residue in the lumen. The ventral prostate occupies the left
center of the field next to the urethra. At upper left is a cross section of vas
deferens; the urinary bladder lies at the lower left.

1. Rectum
2. Urethra
3. Ventral prostate
4. Urinary bladder
5. Pelvic bone
6. Vas deferens
Figure 71 Figure 72
78 Reproductive System

74. Male reproductive system H&E sox 75. Male reproductive system H&E 100 X

Red secretion and large alveoli with infolding of mucosa characterize the At a higher magnification the differences in epithelial linings of the coagu-
vesicular gland; the coagulating gland immediately adjacent is filled with lating gland (above) and vesicular gland (below) are clearly seen. The char-
pink secretory material and lightly stained glandular tissue. acteristic red secretion of the vesicular gland is apparent.

1. Vesicular gland 1. Vesicular gland

2. Coagulating gland 2. Coagulating gland

76. Male reproductive system Heidenhain's Azan 500X

Columnar epithelium and red-stained secretion of the vesicular gland may

be compared with cuboidal epithelium and pink secretion of the coagulating
gland.

1. Vesicular gland
2. Coagulating gland
Figure 74 Figure 75

Figure 76
80 Digestive System

77. Tongue: Posterior dorsal H&E lOOX

surface
78. Taste bud H&E 500X
The surface of the tongue in this region is covered with stratified squamous
epithelium. Two taste buds are present in the center. Above the epithelium A taste bud on the dorsal surface of the tongue in which the principal inter-
are bundles of striated muscle fibers. At upper left and in the center of the nal structures are easily seen.
muscle are two groups of serous glands; mucous glands are seen at right.
1. Stratified squamous epithelium
1. Mucous glands 2. Gustatory cell
2. Serous glands 3. Sustentacular cell
3. Stratified squamous epithelium 4. Location of pore
4. Taste buds
5. Striated voluntary muscle

79. Filiform papillae: Tongue Masson stain 500X

Pointed filiform papillae extend from a modified stratified squamous epithe-

lium. The other four layers (comparable to thick skin) are easily identified. 80. Taste buds Heidenhain's Azan 500X
Green collagen fibers and cross sections of striated muscle are immediately
adjacent to and below the basal layer. A dermal papilla projects up into the A group of taste buds in the tongue illustrate internal structures. The pores
epithelium on the left. empty into a lumen exiting on the dorsal surface of the tongue.

1. Filiform papilla 1. Stratified squamous epithelium

2. Stratum granulosum 2. Taste bud pore
3. Stratum germinativum 3. Taste bud
4. Basal layer of germinativum 4. Collagen fibers (blue)
5. Dermal papilla-collagen fibers (green)
6. Striated muscle
7. Nerve trunk
Figure 77 Figure 78

Figure 79 Figure 80
82 Digestive System

82. Esophagus H&E SOOX

81. Esophagus H&E 100X
Several layers of keratinized, stratified squamous epithelium can be identi-
In this section the esophagus consists of folds surrounding a space, the fied. One cell in the basal layer is undergoing division. Striated muscle
lumen. The epithelial lining is stratified squamous; surrounding it is subep- fibers in a longitudinal plane mixed with connective tissue and blood vessels
ithelial connective tissue. Striated muscle completely surrounds the entire occupy the area below the epithelium.
structure.
1. Stratified squamous epithelium
1. Stratified squamous epithelium 2. Dividing cell in basal layer of epithelium
2. Lamina propria-connective tissue 3. Striated muscle
3. Striated muscle 4. Lamina propria-connective tissue

83. Salivary glands H&E sox

Lymph node

Three types of salivary glands can be identified. Immediately adjacent to and 84. Salivary glands PAS lS0X
above the lymph node in the upper center is the serous parotid gland. The
submandibular gland to the left of the parotid is of the mixed type, serous The three types of salivary glands are easily differentiated: sublingual, cen-
and mucous. The mucus-secreting sublingual gland is the lightly stained ter to left (mucus-secreting) (brilliant red); parotid (serous), lower right; and
gland at center right. submandibular (mixed serous and mucous), upper left and center.

1. Parotid gland (serous) 1. Mucous type (sublingual gland)

2. Lymphatic nodule 2. Serous type (parotid gland)
3. Lymphatic vessels 3. Mixed type (submandibular)
4. Sublingual gland (mucous)
S. Submandibular gland (mixed)
6. Blood vessel
Figure 81 Figure 82

Figure 83 Figure 84
84 Digestive System

85. Submandibular gland, male Methylene Blue 500X

BaSiC fuchsm 86. Submandibular gland, female Methylene Blue 500X
BaSiC fuchsm
Histological structures charactenshc of the adult male mouse are apparent
m thiS 1 5-/Lm-thlck methacrylate sectlOn The nuclei of the columnar mter- In adult females and young males the nuclei of tubular cells are located m
calated tubular cells are located at the base of the cell Bnght red secretory the center of the cell Secretory granules are absent or are very sparse m the
granules fiJI the cytoplasmic area between the nucleus and lumen of the cytoplasm of thiS methacrylate sechon
duct
1 Centrally located nucleus
1 Basally located nucleus 2 Alveolus
2 Secretory granules 3 Plnk-stammg mltochondna
3 Alveolus

87. Stomach, glandular and H&E lOOX

non glandular 88. Stomach glands Alclan Blue 250X
Tartrazlne
Kerahmzed strahhed squamous eplthehum forms the non glandular porhon
of the stomach There is an abrupt change to a modlhed columnar eplthe- In the glandular porhon of the stomach three types of cells can be ldenhhed
hum, forming the glandular porhon Submucosa of loose connechve hssue by speClal stams The submucosa hes immediately beneath the glandular
is seen between the mucosa and circular smooth muscle layer, next to which mucosa At the base of the mucosa are chief or zymogemc cells, stamed
is the layer of longltudmal muscle The thm outer covermg is serosa brown The sphencal cells With small round nuclei and blue cytoplasm
occupymg most of the middle porhon are pane tal cells The cells located
1 Strahhed squamous eplthehum (kerahmzed) near the top, colored green, are mucous neck cells
2 Glands
3 Smooth muscle-Circular 1 Chief cell layer (dark brown)
4 Smooth muscle-Iongltudmal 2 Panetal cell layer (blue) (tan sphencal central nucleus)
5 Submucosa-connechve hssue 3 Mucous neck cells (hght green)
6 Serosa
 !5
5 5 3
4
4
'6/
Figure 87
86 Digestive System

90. Stomach Duodenum Hematoxyhn-PAS 75X

The glandular stomach with PAS-positive mucOid secretlOn occupies the left
89. Stomach mucosa H&E 500X
half and center of the sectlOn At the Junction of the pylonc end of the stom-
ach with the duodenum, the lumen Widens as the epithehum becomes Sim-
Sphencal cells with round, centrally located nuclei and pmk cytoplasm are
ple columnar Brunner's glands m the submucosa of the duodenum react
parietal cells Chief or zymogemc cells are grouped near the bottom of the
positively (red) to penodic aCid-Schiff reagent
mucosa and contain dark blue nuclei and gray cytoplasm
1 Smooth muscle-stomach wall
Chief cells
2 Stomach glands, mucous (red)
2 Pane tal cells
3 Pylonc openmg
3 Submucosa
4 Brunner's glands of duodenum
5 Villi-duodenum
6 Pancreas

92. Small mtestme Ileum H&E 150X

91. Stomach Duodenum Hematoxylm-PAS 200X
Peyer's patch is a collection of lymphocytes grouped mto a lymphOid nod-
At higher magnificatlOn the structural differences between mucous glands ule Aggregates of hghtly stamed cells occupy germmal centers, from which
of the stomach and Brunner's glands and columnar epithehum of the duo- mature lymphocytes prohferate Cross sections of vllh of varymg lengths
denum become apparent occupy the upper portlOn of the reglOn

Stomach glands, mucous (red) 1 LymphOid nodule (Peyer's patch)

2 Villi-duodenum 2 Germmal center
3 Brunner's glands-duodenum 3 Serosa
4 Fat cells 4 Vlli/
5 Smooth muscle-stomach wall 5 Lumen With fibrous debns
Figure 89 Figure 90

Figure 92
88 Digestive System

94. VIlh Jejunum Methacrylate IOOX

93. Small mtestme Ileum H&E SOOX Methylene Blue
BasIc fuchsm
Lymphocytes formmg part of Peyer's patch appear m the submucosa below
the connectIve tIssue Extendmg to the left are four crypts, at the base of This low-power view shows the relatIonship between the fmgerhke villi
each are brick-red granules of Paneth cells and crypts of Lieberkuhn Cross sectIons of crypts are seen on the right
adjacent to smooth muscle
1 Paneth cell granules (orange-red)
2 Submucosa 1 Vtllus
3 Crypt of Lieberkuhn 2 Lamma propria
4 Goblet cell 3 Crypt of Lieberkuhn
5 LymphOid nodule (peyer's patch) 4 Goblet cell
5 Smooth muscle

95. Vtllus Methacrylate SOOX

Methylene Blue
BasIc fuchSin 96. Cecum H&E 60X

In thiS 1 S-~m thm sectIon, mternal structures of the villus are apparent A The extensIOn of mtestme from the Ileum to the begmmng of the large
promment striated border outlines the lumen side of the simple columnar Intestine IS the cecum The mucosa, occurring m many folds, IS lined by
epithelium Mitochondria, darkly stamed, occupy the distal portion of each columnar epithelium Beneath the submucosa IS a layer of smooth muscle
epithelial cell Goblet cells appear as spaces m the epithelium covered by serosa

1 Simple columnar eplthehum 1 Mucosa

2 Mitochondria 2 Smooth muscle
3 Striated border 3 Intestmal contents
4 Goblet cell
5 Lamina propria
6 Blood vessel
Figure 93 Figure 94

Figure 95 Figure 96
90 Digestive System

97. Large IntestIne PAS 500X 98. Anus H&E 50X

Many mucus-hlled goblet cells appear bnght red The absence of v1llI and The anal opemng 1S bounded by strahhed squamous ep1thelIum of the skm
Paneth cells also a1ds In 1denhfyIng the large IntestIne Layers of smooth Columnar ep1thelIum of the rectum abruptly changes to strahhed squamous
cucular and long1tud1nal muscle are covered by the outermost thIn layer, Stnated muscle forms the anal sphIncter
the serosa
1 Skm
1 Mucus In ep1thelIai cells (red) 2 Stratlhed squamous ep1thelIum
2 Submucosa 3 Slmple columnar ep1thelIum
3 Smooth muscle 4 Stnated muscle
4 Serosa 5 Sebaceous glands

100. Pancreas Mod1hed aldehyde- 250X

99. Pancreas H&E 250X fuchs1n

Surrounded by dark red aClnar cells, three lIghtly staIned 1slets of Langer- In th1S sectlOn the alpha and beta cells of the 1slets of Langerhans are d1ffer-
hans appear at left, nght, and lower center An ep1thelIal-IIned pancreahc enhated beta cells are purple and alpha cells are red
duct 1S adjacent to the left 1slet, and portlOns of an artery and veIn appear at
upper left and nght Islets of Langerhans
2 Purple beta cells
1 Pancreahc duct 3 Red alpha cells
2 Artery (w1th veIn at upper nght) 4 AClnar cells
3 Islet of Langerhans 5 Pancreahc duct
4 AClnar cells 6 Blood vessel
Figure 97 Figure 98

Figure 99 Figure 100

92 Digestive System

102. LIver H&E 250X

101. LIver Heldenham's Azan 500X
The portal tnad IS surrounded by hepatocytes In the center IS the portal
Blue-staIned rehculum forms the walls of slnusOlds, spaces that con tam vem fIlled wIth blood In the upper left corner adjacent to the vem IS a
erythrocytes FIxed macrophages (Kupffer cells) are attached to the smusOld branch of the hepahc artery One branch of a bIle duct hes at the upper and
wall Hepahc cell nucleI are stamed deep blue and cytoplasm IS tan lower nght areas

Hepahc cell 1 Hepatocytes

2 SmusOld 2 Branch portal vem }
3 Kupffer cell 3 BIle duct Portal tnad
4 Hepatic artery

103. LIver H&E 800X 104. Gallbladder H&E 500X

The cytoplasm of these hepahc cells IS fIlled wIth stored matenal nch m The mner lmmg of the gallbladder IS SImple columnar eplthehum Beneath
RNA Each nucleus contaIns one or more large nucleoh m addlhon to chro- IS a layer of loose connective tissue and blood vessels A very thm layer of
mahn granules The large spaces between cords of hepahc cells are smus- smooth muscle separates thIS area from serosa CharactenstIc hepatic cells
Olds, one Kupffer cell (fIxed macrophage) IS seen attached to the wall (upper occupy the lower thIrd of the sectlOn
nght) and hes m the smusOld In the center between two hepahc cells IS
seen a small openmg, a bIle canahculus SImple columnar epIthelium
2 Lamma propna
1 Hepatocyte 3 Blood vessel
2 SmusOld 4 Smooth muscle
3 Nucleus-Kupffer cell 5 Serosa
4 Openmg-blle canahculus 6 Hepatic cells
Figure 101
94 Urinary System

105. KIdney and adrenal H&E SOX

106. KIdney, male H&E SOOX
In thIS sectIOn the anatomIcal relatIOnshIp between kIdney and adrenal IS
clear The cortex and medulla of the adrenal on the rIght are IdentIfIed In Pronounced cubOIdal epIthelIum of Bowman's capsule characterIzes thIS tIs-
the kIdney the cortex, wIth ItS proxlInal and dIstal convoluted tubules, may sue m the male In the female the epIthelIal cells are much smaller
be compared wIth the longltudmal sectIons of collectmg tubules m the
medulla 1 Glomerulus
2 CubOIdal eplthelIum-Bowman's capsule
Adrenal medulla 3 ProXImal convoluted tubule
2 Adrenal cortex 4 DIstal convoluted tubule
3 KIdney cortex 5 Artery
4 KIdney medulla

108. KIdney tubules Methacry la te 800X

107. KIdney tubules Heldenhaln's Azan SOOX Methylene Blue
BaSIC fuchsm
ProxImal convoluted tubules WIth a blue brush border may be contrasted
WIth dIstal convoluted tubules havmg no brush border MItochondrIa In thIS thm sectIOn the red mItochondrIa m the cytoplasm of tubular cells
wlthm the cytoplasm of tubular cells are stamed brIllIant red A few tubules are clearly IdentIfIed Between the tubules m the lower rIght and central
have dark blue casts WIthIn them, whIch may IndIcate mmor damage to portIons of the sectIOn are sectIOns of blood vessels WIth erythrocytes Endo-
some glomerulI thelIal cells lme the blood vessels

1 ProxImal convoluted tubule (brush border) (blue) 1 Nucleus of tubular cell

2 MItochondrIa (red) m tubular cells 2 MItochondrIa (red)
3 DIstal convoluted tubules 3 EndothelIal cell
4 Cast m tubule (blue) 4 Erythrocytes
5 Nucleolus of tubular cell nucleus
Figure 105 Figure 106

1 1

Figure 107 Figure 108

96 Urinary System

109. Bladder H&E 200X 110. Bladder Masson stain 250X

Transitional epithelium (urothelium) lines the inside bladder wall, connec- Green collagen fibers form most of the lamina propria underlying the tran-
tive tissue fibers form the lamina propria. A thick wall of smooth muscle sitional epithelium lining the bladder cavity. A lymphatic nodule is seen at
surrounds the bladder. upper left.

1. Transitional epithelium 1. Transitional epithelium

2. Lamina propria 2. Collagen-connective tissue
3. Blood vessel 3. Lymphatic nodule
4. Smooth muscle

111. Urinary bladder H&E 250X

At higher magnification the characteristic morphology of transitional epi-

thelium is apparent.

1. Transitional epithelium
2. Lamina propria
3. Smooth muscle
Figure 109 Figure 110

Figure 111
98 Skin

113. Connective tissue HematoxylIn 500X

Mast cells Aldehyde fuchsm
112. Connective tissue Heldenham's Azan 500X Fast Green

A clump of mast cells, each wIth blue cytoplasm and a red nucleus, IS nestled A group of mast cells WIth green nucleI and purple granules lIes among thm,
among elastic and collagen &bers Both types of &bers have taken the blue dark elastic &bers and green collagen &bers
stam, elastic &bers are very thm and wavy
1 Mast cell
Mast cell 2 Thm, purple elastIC &ber
2 Thm green elastic &ber 3 Green collagen &bers
3 Collagen &bers (wavy green) 4 Macrophage
5 FIbroblast

115. Mast cells Methacrylate 250X

114. Connective tissue HematoxylIn 500X Methylene Blue
Aldehyde fuchsm BaSIC fuchSIn
Fast Green
In the connective tissue of the dermiS of the skm are several elongated &bro-
In thiS sectIOn of abdommal connective tissue faSCia, thm, purple elastic blasts and a row of tissue mast cells, below which are two empty fat cells
&bers, green collagen &bers, elongated nucleI of &broblasts, and oval nuclei One medIUm artery IS m upper left, the endothelIal cells and smooth muscle
of macrophages are clearly IdentI&ed Green-stammg erythrocytes are m cells are easily IdentI&ed
lower left
1 Mast cell (tissue basophil) blue granules
1 Fibroblast 2 Fat cell
2 Elastic &ber (purple) 3 Artery
3 Wavy green collagen &bers 4 EndothelIal cells
4 Erythrocytes 5 Fibroblast
6 Two smooth muscle cell nuclei
Figure 112 Figure 113

2 2

Figure 114 Figure 115

100 Skin

117. HaIr follIcle H&E soox

116. HaIr bulb H&E soox
ThIS portlOn of the half projects from the dermIS through the epIdermIs to
ThIS portion of the haIr IS 10 the dermIS Many fat cells surround the half
the sk10 surface Keratin appears 10 the center of the haIr Sebaceous glands
One cell In the bulb reglOn IS undergOlng dlvlslOn
lIe along the half shaft
1 Connective tissue root sheath S Blood vessel
EpIdermIs
2 Outer eplthehal root sheath 6 Fat cell
2 Half shaft
3 Root of haIr 7 Nucleus of fat cell
3 HaIr follIcle
4 UndIfferentiated bulb cells 8 Cell In dlvlslOn
4 Sebaceous gland

118. Vlbnssae follIcles H&E sox 119. Vlbnssa follIcle Masson staIn sox
Vlbnssae are very senSItive tactile haIrs located on the lIps and other areas SImIlar structures Identified 10 the H & E sectlOn are seen WIth the Masson
of the face They are larger than normal half follIcles and the root and bulb stam The half shaft IS bnght red Green collagen fibers occur 10 the con-
of the vlbnssa are surrounded by nerve fibers and a nch blood supply nective tissue sheath and hyalIne membrane Blood IS present 10 the nng
SInus
Sk10
2 Half root HaIr bulb 7 HyalIne membrane
3 Nerve fibers 2 PapIlla 8 Sebaceous gland
4 PUIVlnUS 3 Nerve fibers 9 EpIdermIs
S R10g smus WIth few erythrocytes 4 Pulv10us 10 Half shaft
6 Half follIcle S Connective tissue sheath 11 Fat cells
7 Stnated muscle 6 RIng smus 12 Stnated muscle
Figure 116 Figure 117

Figure 118 Figure 119

102 Skin

120. Mucous gland and vessels Aldehyde-fuchsm lOOX 121. ThICk skm H&E SOOX
Masson staln Sole of foot

The mucous gland shown centrally stams purple One large and two The outer layer of flattened dead cells represents the stratum corneum Ker-
medlum-slzed arterIes wlth promlnent lnternal elastic membranes are seen atohyahne granules are present m the underlymg stratum granulosum The
at rIght and at center left Green collagen &bers are present m the tUnIca next layer of four to &ve cells forms the stratum germlnatIvum Connective
medla and advenha Brown strIated muscle &bers are at lower rIght Two hssue of collagen and elastlc &bers plus blood and lymphahc vessels and
nerve trunks In cross sectIOn are located between the mucous gland and nerves comprIse the dermls Two dermal papIllae contammg blood vessels
large artery and nerves extend up mto the epldermls at center and left

1 Purple mucous secrehon 1 Dermls connective tissue

2 Internal elastic membrane (artery) 2 Dermal paplllus
3 Erythrocytes m artery 3 Stratum germmatIvum
4 Nerve trunk 4 Stratum granulosum
5 Strlated muscle 5 Stratum corneum
6 Desquamatmg keratm

122. Fat Brown and whIte H&E 2S0X

Brown fat lS easlly dlfferentIated fr m whlte by the central posltlon of the

nucleus m the cell WhIte fat cell, 'pear as spaces whose nucleI are located
at the perIphery of the cell

Brown fat cells

2 Whlte fat cells
3 Erythrocytes m vem
4 Small artenole
Figure 120 Figure 121

Figure 122
104 Respiratory System

123. Junction: Nasal passage, H&E 50X

trachea, esophagus
124. Nasal mucosa Masson stain 100X
At low magnification the nasal passage curves into the trachea at top center.
The tracheal lumen curves to the left. Hyaline cartilage in the tracheal wall Ciliated columnar epithelium with goblet cells lines the sinuses of the nose
supports it and keeps it from collapsing. The epiglottis is seen covering the as it passes around an island of hyaline cartilage and forms the characteristic
esophageal opening at center right. Bone and striated muscle are seen in the specialized olfactory epithelium with serous and mucous glands.
lower third. Bony vertebrae are located at the top.
1. Hyaline cartilage
1. Nasal passage 6. Soft palate 2. Ciliated columnar epithelium
2. Mouth 7. Vertebra 3. Serous and mucous glands with goblet cells
3. Epiglottis 8. Tracheal cartilage 4. Sinuses
4. Trachea 9. Pharynx
5. Esophagus 10. Striated muscle

125. Nasal mucosa H&E 500X

The nasal mucosa is lined by pseudostratified ciliated columnar epithelium 126. Trachea Masson stain 250X
with goblet cells. Tall sustentacular cells with oval nuclei lie just beneath
the epithelium. Olfactory cells (bipolar ganglion cells) with dark spherical Ciliated columnar epithelium with goblet cells lines the trachea. Beneath
nuclei lie in the deepest part of the mucosa and send axons into the under- the epithelium are serous and mucous glands with blood vessels and nerves
lying connective tissue to form olfactory nerve trunks. Bone forms a bound- lying immediately adjacent to hyaline cartilage.
ary through which a blood vessel passes.
1. Ciliated columnar epithelium
1. Pseudostratified ciliated columnar epithelium 2. Serous gland
2. Sustentacular cells 3. Mucous glands
3. Mucous glands 4. Hyaline cartilage
4. Bone 5. Goblet cell
5. Erythrocytes in blood vessels
6. Olfactory cells
 3
4
9

Figure 123 Figure 124

Figure 125 Figure 126

106 Respiratory System

128. Lung Methacrylate 250X

127. Lung H&E TolUIdme Blue
lOOX

A termlnal bronchIOle endlng m an alveolar duct m upper center Small cav- The lumen of a resplratory bronchIOle lS hned by low columnar eplthehum
lhes opemng on thlS duct are alveolar sacs These are surrounded by alveoh The spongy appearance anses from alveolar ducts and sacs m the resplratory
hssue
1 Termmal bronchIOle
2 Alveolar duct Alveolus
3 Alveolar sac 2 Alveolar sac
4 Alveolus 3 BronchIOle
4 Blood vessel
5 Alveolar duct

129. Lung Snook's Sliver Stam 250X

Nuclear Fast Red

Black rehcular fibers occur wlthm connechve hssue of respuatory tract

Rehcular fibers (black)

2 Alveolar duct
3 Alveolar sac
Figure 127 Figure 128

Figure 129
108 Respiratory System

130. Lung Aldehyde-fuchsin 500X 131. Bronchiole Methacrylate 500X

Hematoxylin Methylene Blue
Fast Green Basic fuchsin

Abundant purple elastic fibers are seen throughout the lung tissue. In this 1.5-l'm methacrylate section of bronchiole lining, ciliated cells may
be distinguished from nonciliated Clara cells.
1. Alveolus
2. Alveolar sac 1. Ciliated cell
3. Bronchiole 2. Clara cell
4. Elastic fiber (purple) 3. Connective tissue
5. Blood vessel 4. Alveolus

132. Lung Methacrylate 500X

Toluidine Blue

Type I and II alveolar cells can be distinguished dearly in this 1.5-l'm

section.

1. Type II (great alveolar) cell

2. Type I cell
3. Endothelial cell
4. Capillary
5. Alveolus
6. Septum
Figure 130 Figure 131

Figure 132
110 Lymphatic System

133. Thymus H&E sox

The very dark cortex consIsts of masses of small lymphocytes Bands of pmk
trabeculae ongmate m the capsule and penetrate the cortex The lIghter
134. Thymus H&E soox
medulla IS located centrally Large blood vessels as well as masses of fat cells
are located In the margms DIfferentIatIOn of dark-stammg cortex and lIghter medulla can be made eas-
Ily, WIth several groups of large, pale epIthelIal cells formmg Hassall's cor-
1 Cortex puscle In the medulla
2 Medulla
3 Fat cells 1 Cortex
4 Artery 2 Medulla
Artenal wall 3 Hassall's corpuscle
S
6 Stnated muscle
7 Trabeculae

135. Spleen H&E sox

Dense connectIve tIssue forms a tough capsule surroundIng the spleen 136. Spleen H&E lOOX
Extendmg mward from the capsule are bands of trabeculae, composed of
retIcular fIbers and smooth muscle, whIch form a framework for whIte and At a hIgher magmfIcatlOn, structures In the red and whIte pulp are more
red pulp Nodules of lymphocytes form whIte pulp, whIch contams a central eaSIly seen
artenole The red pulp IS composed of erythrocytes, granular leukocytes,
lymphocytes, and megakaryocytes 1 LymphOId nodule
2 Central arterIOle
Capsule 3 Trabeculum
2 Germmal center 4 Megakaryocyte
3 Trabeculum S Red pulp
4 Red pulp
S WhIte pulp
Figure 133 Figure 134

Figure 135 Figure 136

112 Lymphatic System

137. Spleen H&E 500X

138. Lymph node H&E 50X
In thIS magmfIed sechon fIve megakaryocytes m the red pulp are present m
the left and center areas Aggregahons of erythrocytes m va no us stages of The lymph node, WIth ItS nodules, smuses, and blood vessels, IS surrounded
matunty surround them At lower nght lymphocytes form a porhon of a by porhons of the sahvary glands
nodule m whIte pulp
1 LymphOId folhcle 5 Intercommumcahng smus
Capsule 2 HIlus 6 Blood vessel
2 Trabeculum 3 Subhngual gland 7 Paracorhcal area
3 Megakaryocyte 4 SubmandIbular gland 8 Penpheral and margmal smuses
4 Red pulp

140. Spleen Snook's Sliver Stam 100X

Nuclear Fast Red
139. Lymph node H&E 250X
A well-defIned capsule surrounds the spleen Trabeculae of connechve tis-
A germ mal center IS seen m the upper left area of the node The dark para-
sue, blood vessels, and black rehcular fIbers extend from the capsule mto the
corhcal area on the nght contams lymphocytes Fat cells he between the
spleen proper One large folhcle of whIte pulp IS m rIght center, surrounded
lymph node and blood vessel on the left
by red pulp Two megakaryocytes are m the red pulp at upper left
1 Germmal center m lymphOId folhcle
1 Capsule
2 Lymphocytes m node
2 Trabeculum-black reticular fIbers
3 Artery
3 Maipighian corpuscle-whIte pulp
4 AdIpose hssue
4 Red pulp
5 Megakaryocytes
Figure 137 Figure 138

Figure 139 Figure 140

114 Bone, Cartilage, Fat

141. CartIlage H&E SOOX 142. Bone H&E SOOX

Rows of oval cells arranged 10 lmes on the nght are those of fIbrocartIlage Two osteocytes are present m the trabeculae In the center of the fIeld On
Large oval cells not so arranged but occupymg the lower center are cells of the nght IS an mdentatlOn, Howshlp'S lacuna, with an osteoclast Immedi-
hyalme cartIlage Elongated cells adjacent to the edges of bone are osteo- atelyadJacent Bone marrow cells fIll the area on the nght
blasts A few erythrocytes are present In center
1 Osteocyte
1 Osteoblasts 2 Osteoblasts
2 FibrocartIlage 3 Osteoclast
3 Hyahne cartIlage 4 Bone marrow cells
4 Bone 5 Howshlp'S lacuna
6 Bone

143. FibrocartIlage H&E lS0X

FibrocartIlage 10 the vlcmlty of a Jomt IS attached to bone and tendon

1 Flbrocartliage
2 Bone
3 Tendon attached to hyalme cartilage
4 Stnated muscle
4

Figure 141 Figure 142

Figure 143
116 Bone, Cartilage, Fat

144. Femur EpIphyseal plate H&E 200X 145. Knee JOInt H&E sox

Th,S sectIOn from a 1-month-old mouse shows the developIng zones of bone The artIcular surfaces of bone formIng the JOInt are of hyalIne cartIlage,
formatIOn from cartIlage at rIght to bone marrow cells at left of the held below whIch IS bone Marrow cells are seen as dark masses between bone
spIcules Above, below, and In between the artIcular surfaces are trIangular
1 Zone of prolIferatIOn structures of bone, hbrocartIlage, and blood vessels-the menISCI
2 Zone of maturatIOn
3 Zone of hypertrophy 1 ArtIcular cartIlage (hyalIne)
4 Trabeculae 2 Memscus
S Bone marrow cells 3 Bone marrow
4 Bone
TermInology from Bloom and Fawcett (1968) S Trabeculae (bone lIght, cartIlage dark)
6 StrIated muscle

146. Tall Cross sectIon H&E lS0X

StratIhed squamous epIthelIum formIng the epIdermal coverIng completely

surrounds the tall The four large pInk oval structures located In each of the
four quadrants are tendons The central homogeneous red structure sur-
roundIng spaces IS bone Very few marrow cells are found In the spaces
between the bone

1 StratIhed squamous epIthelIum

2 Tendon
3 Marrow caVIty WIth very few cells
4 HaIr follIcle
Figure 144 Figure 145

Figure 146
118 Bone, Cartilage, Fat

148. Tooth H&E 250X

147. InCisor tooth H&E 50X In thIs sectIon the enamel has been removed and the outer layer IS dentIne
Extendmg perpendIcularly through thIS layer are dentInal tubules, mInute
The mCIsor tooth projects from the lower pw, showmg the structures above canals that orIgInate from the pulp cavIty and penetrate all parts of the den-
and below the gum lIne tIne A row of odontoblasts IS promInent between the dentIne and the pulp
cavIty Macrophages, lymphocytes, blood vessels, and connectIve tIssue
1 Enamel fIbers comprIse the pulp An acellular cementum covers the dentIne below
2 Dentme the gum lIne
3 GmgIva (gum)
4 Alveolar bone 1 DentIne WIth dentmal tubules (strIatIons)
5 Pulp caVIty 2 Odontoblasts
3 Dental pulp
4 Acellular cementum

149. Tooth H&E 250X

The outermost layer of tIssue IS the stellate retIcular layer of enamel pulp
Subjacent are the ameloblast cells, whIch aId ,n enamel formatIon Below IS
a WIde band of enamel that covers the dentIne layer A row of odontoblasts
IS seen between the dentIne and dental pulp The latter IS a connectIve tIssue
layer WIth macrophages and lymphocytes scattered throughout

1 Enamel
2 DentIne
3 Odontoblasts
4 Dental pulp
5 Ameloblast layer
6 Enamel pulp
Figure 147 Figure 148

Figure 149
120 Muscular System

151. Skeletal muscle Heldenham's Azan 500X

150. Stnated muscle Masson stain 500X
CharactenstIc vertical cross lme~ along the muscle fibers readily Identify this
The vertical stnatlOns and locatlOn of muscle nuclei at the side of the bundle type of muscle Oval nuclei between muscle bundles are muscle nuclei
of fibers charactenze this type of muscle Because this sectlOn IS 5 ~m thick, Other round nuclei are those of wandenng leukocytes Abundant collagen,
muscle fibers overlap each other and their nuclei do not always he at the elastic, and reticular fibers form a connective tissue, conveymg blood, lymph
edge of the muscle bundle vessels, and nerves and formmg a boundary between muscle bundles

1 Artery 1 Vertical stnatlons along muscle bundle

2 Collagen fibers (green) 2 Muscle fiber nucleus
3 Muscle fiber nucleus 3 Leukocytes m connective tissue
4 Muscle fibers with stnatlOns 4 Blood vessel
5 Blue collagen fibers

153. Cardiac muscle Heldenham's Azan 500X

152. Smooth muscle H&E 500X
The presence of nuclei m the center of muscle bundles and cross stnatlOns
charactenze this type of muscle Another Identlfymg feature IS the dlvlslOn
Long spmdle-shaped fibers are arranged m parallel The oval, elongated
between sarcolemma Junctions forming Intercalated disks, stamed blue with
nuclei he m the middle of each fiber
this stam Blood and lymph vessels and nerves accompany blue collagen
fibers between muscle bundles
Muscle fiber nucleus
2 Muscle fiber
1 Muscle fiber nucleus
3 Blood vessel
2 Intercalated disk
3 Blood vessel
 4

Figure 150 Figure 151

Figure 152 Figure 153

122 Circulatory System

155. CardIac muscle In pulmonary HeIdenhaIn's Azan 500X

vem

154. Pulmonary veIn Phosphotungstic aCld- sox The spherIcal structure at upper nght IS a cross sectIOn of a nerve trunk
hematoxylIn CardIac muscle of the pulmonary veIn forms an arc at the bottom of the
field, below whIch are blood cells In the lumen Broad elastic fibers staIned
Part of a valve IS seen extendIng across the lumen of the veIn lIght blue form part of the tUnIca medIa of a large artery at upper left, blood
cells filhng the arterIal lumen
1 Valve leaflet
2 Alveolus of lung 1 CardIac muscle
3 Blood In lumen of veIn 2 ElastIc fibers-smooth muscle
3 Blood In artery and veIn
4 Nerve trunk
5 Blue collagen fibers

156. Pulmonary veIn Phosphotungstic aCld- 500X 157. Heart valves H&E 75X
hematoxylIn
The valve and valve leaflets are located In the upper left of the field, below
Elastic fibers In the Internal elastic membrane and tUnIca medIa charactenze whIch are blood cells wIthIn the aorta MyocardIUm occupIes the central
the large artenal wall In the upper field Just below are promInent cross field, and another valve lIes In the lumen of the pulmonary veIn at upper
stnatIOns of cardIac muscle, formIng the wall of the pulmonary veIn Blood rIght
cells fill the lumIna of both artery and veIn
1 Valve
1 Blood cells In lumen of artery 2 Valve leaflet
2 Elastic fibers and smooth muscle In artenal wall 3 CardIac muscle
3 Cross stnatIons of cardIac muscle In wall of pulmonary veIn 4 Blood m heart chamber
4 Blood cells In lumen of veIn 5 Aorta
1
3

Figure 154 Figure 155

Figure 156 Figure 157

124 Circulatory System

158. Bone marrow Giemsa stain 500X

159. Bone marrow Giemsa stain 500X
Many primitive, maturing, and mature cells are found in bone marrow. Cells
of the myelocyte and erythrocyte series are present, as are plasma cells and Two megakaryocytes are seen in the lower right of the field.
megakaryocytes.
1. Megakaryocyte
1. Plasma cell 2. Plasma cell
2. Monocyte 3. Normoblast
3. Neutrophil 4. Neutrophil
4. Normoblast
5. Neutrophil myelocyte

160. Peripheral blood New Methylene Blue 500X

Reticulocytes with a pronounced reticular network are scattered among

erythrocytes.

1. Reticulocyte (blue reticular net)

2. Erythrocyte
Figure 158 Figure 159

1
I
2

Figure 160
126 Circulatory System

161. Monocyte Wright's stam 950X 162. NeutrophIls Wright's stam 950X

ThIS cell IS eaSIly IdentIfIed by the mdented, horseshoe-shaped, reddIsh pur- Both cells exhIbIt small, blUlsh cytoplasmIC granules, but the nuclear shapes
ple nucleus wIth lIght blue cytoplasm Concave dIsks of umform SIze are dIffer The "doughnut"-shaped cell suggests a nucleus that has not yet
erythrocytes formed three lobes, as are seen m the other cell

164. Lymphocytes Wright's stam 950X

163. Platelets Wright's stam 950X
Companson between large and small lymphocytes can be made, although
Red granules and blue cytoplasm are readIly dIfferentIated m thIS cluster of each cell has a large, reddIsh purple nucleus occupymg most of the cell, wIth
platelets a mmImum of blue cytoplasm A clump of platelets wIth red granules and
blue cytoplasm lIes between the two cells
Figure 161 Figure 162

Figure 163 Figure 164

128 Circulatory System

165-168. Eosinophils Wright's stain 950X

These four cells illustrate variations of the nuclear shape and the size and
staining intensity of large cytoplasmic granules in eosmophils.
Figure 165 Figure 166

Figure 167 Figure 168

 Fixatives

10% NEUTRAL BUFFERED FORMALIN (pH 7.0) Store in 70% ethanol. Mercuric chloride must be removed
from tissue on slides by alcohol-iodine when staining. For
Formaldehyde (37 -40%) 100 ml pieces of liver and spleen 2-4 mm thick, 8- to lO-hr fixation is
Distilled water 900 ml sufficient.
Dibasic sodium phosphate (Na 2 HP0 4 ) 6.5 g
Excellent for H & E and connective tissue stains. Presence of
Monobasic sodium phosphate (NaH 2P0 4 ) 4g
mercuric chloride may interfere with silver staining. Recom-
mend 10% formalin.
BOUIN'S FIXATIVE
CARNOY'S FIXATIVE
Picric acid, saturated aqueous solution 750 ml
Formaldehyde (37-40%) 250 ml
Absolute ethanol 60 ml
Glacial acetic acid 50 ml
Chloroform 30 ml
Fix 8-24 hr. Store tissues in 70% ethanol. Picric acid is usually Glacial acetic acid 10 ml
removed when sections pass through 70% and 50% ethanol, 10
Very fast in penetrating tissues. Excellent for preserving
min each.
nucleic acids (DNA and RNA).

ZENKER-FORMOL (HELLY'S FIXATIVE)

WILLIAMS'S FIXATIVE FOR BODIAN STAIN
Distilled water 1000 ml
Formaldehyde (37 -40%) 40 ml
Mercuric chloride 50 g
Glacial acetic acid 10 ml
Dissolve with heat
80% Ethanol 100 ml
Add potassium dichromate 25 g
Picric acid 2g
Just before adding tissues add 5 ml 35-40% formaldehyde per
100 ml fixative. Fix 8-24 hr. Wash 24 hr in running water.
131
132 Fixatives

FORMOL-SALINE FIXATIVE FORMOL-ALCOHOL-ACETIC (FAA)

Formaldehyde (37-40%) 100 ml Formaldehyde (37 -40%) 10 ml

Sodium chloride 9g 80% Ethanol 90 ml
Distilled water 900 ml Glacial acetic acid 5 ml
Used as a rapid fixer.
FORMALIN -MERCURIC CHLORIDE
VANDERGRIFF'S FIXATIVE
10% Neutral buffered formalin 100 ml
Mercuric chloride 2g 95% Ethanol (or 99% isopropyl) 80 ml
Dissolve with aid of heat
Formaldehyde (37 -40%) 12 ml
Glacial acetic acid 4.5 ml
Picric acid 4.0 g
Mercuric chloride 0.2 g
Urea 0.5 g
 Staining Methods

There are as many ways of preparing tissues for microscopic 70% Ethanol * 5 min
examination as there are technicians preparing them. The Tap water Rinse
methods we have used and from which our photomicrographs
Desired specific staining method may now be applied.
were made are slight modifications from standard procedures
given in the references. We have included them in detail in
this book to aid the investigator or technician. HEMATOXYLIN AND EOSIN

Fixative: Any general fixative. Zenker-formol recommended.

DEP ARAFFINIZING SECTIONS Harris's hematoxylin 3-5 min

Hematoxylin crystals 5g
The standard procedure for deparaffinizing sections in Absolute ethanol 50 ml
our laboratory is as follows: Ammonium or potassium alum 100 g
(aluminum ammonium sulfate or aluminum potassium
Paraway (Technicon Instruments Corp., Tarrytown, 5 min sulfate)
New York 10591) or Xylene Distilled water 1000 ml
Xylene 5 min Mercuric oxide (red) 2.5 g
Drain excess xylene on paper towel before going into abso- Dissolve hematoxylin in absolute ethanol.
lute ethanol.
Heat approximately 900 ml of water and dissolve alum.
Absolute ethanol Two changes: First rinse 8-10 times;
Bring to boil rapidly. Remove from heat; place flask in sink
second, 2-3 min
in case contents overflow when adding mercuric oxide.
95% Ethanol Rinse 8-10 times
Reheat briefly to bring to purple color; cool in running cold
80% Ethanol plus iodine 5 min
water. For use add 1 ml glacial acetic acid for each 100 ml of
4% Tincture of iodine 10 ml
stain. Filter before use if film forms on surface. Rinse in
80% Ethanol 95 ml
water 8-10 times.
(Used only for tissues fixed in solutions containing mercuric
chloride) 'Tissues may be left in this solution overnight.

133
134 Staining Methods

Lithium carbonate, saturated aqueous solution 5-10 sec STAIN FOR MOTOR END PLATES 2
(Used to "blue" hematoxylin in nuclei; pH of LiC0 3 solu-
tion basic to counteract slightly acid hematoxylin) A. Place frozen sections 20-25 /lm thick of skeletal muscle
Rinse in water. (gastrocnemius, solenus, or tibialis anterior recommended)
Rinse in 70% ethanol. on a slide in a drop of 3% disodium EDTA (used to prevent
Eosin,l stock solution contraction of muscle fibers). Let air dry at room
Eosin Y, alcohol and water solution 5g temperature.
Distilled water 100 ml B. Immerse slides in 20% aqueous solution of sodium sulfate
Add 95% ethanol. 400 ml for 3 min to prevent shrinkage of sections in cholinester-
Working solution 2-3 min ase stain. Rinse in distilled water.
Eosin stock solution, 1 part C. Stain sections for acetylcholinesterase by incubating at
80% ethanol, 3 parts 37°C for 10-25 min in the following solution:
Add 0.5 glacial acetic acid per 100 ml stain. 5-Bromoindoxyl acetate* (dissolve first in 4mg
Rinse in 95% ethanol. 2 changes absolute ethanol)
Formalin-fixed tissues may lose eosin more readily than Absolute ethanol 0.3 ml
other fixatives. Potassium ferrocyanide 63 mg
Suggest only one change. Potassium ferricyanide 50mg
Rinse in absolute ethanol. Tris-HCl (Sigma Chemicals) 42 mg
Carbol-xylol 1-2 min Tris-base (Sigma Chemicals) 4mg
Phenol, fused, crystals melted (J. T. Baker, Calcium chloride 33 mg
USP grade) 20 ml Deionized water 30 ml
Xylene 80 ml
Continue staining until microscopy shows end plates to be
(Removes any residual moisture present in alcohols)
clearly defined transparent blue. The Tris (mixed to pH 7.2),
Xylene 2 changes, 5 min each
calcium chloride, and water can be premixed and kept as stock
Mount in Permount (Fisher), Coverbond (Harleco, Gibbs-
solution.
town, New Jersey) or other suitable medium.
Results: Nuclei: Dark blue * Available from leN Pharmaceuticals, Inc., Life Sciences Group, Plainview,
Cytoplasm: Orange red New York.
 Staining Methods 135

Nerve-Staining Procedure ml. This solution may be used several times if the edges
and backs of slides are cleaned before immersion. How-
1. To prevent loss of the sections from slides, dehydrate in ever, it must be discarded if a precipitate forms.
70% and 100% ethanol for 1-2 min each and then immerse 8. Optional: To darken axons and endings, immerse in 1%
in 0.5-1.0% (w Iv) celloidin in 1:1 ether-ethanol solution oxalic acid for 20-120 sec.
for 20 sec. Wipe excess from back of slide and harden in 9. Immediately fix again for 5 min in 5% sodium thiosulfate.
70% ethanol for 30 sec. 10. Dehydrate through 70% and 100% ethanol. Remove cel-
2. Fix sections for 30 min at room temperature in buffered loidin by immersion in 1:1 ether-ethanol solution for 5
formol-saline (37 -40% formaldehyde, 20 ml; sodium chlo- min. Rinse in xylene and mount.
ride, 4.25 g; acid sodium phosphate monohydrate, 0.80 g;
anhydrous disodium phosphate, 1.30 g; deionized water, The reaction may be inhibited by placing the slides
180 ml) at pH 7.0. The fixing solution must be less than immediately after step B in DFP (diisopropyl flurophosphate).
one week old. CAUTION: DFP is an extremely poisonous compound. It should
3. Pretreat for 30 min at 37°C in 10% chloral hydrate (w Iv) be handled only by a person wearing surgical gloves working
with 1% pyridine (v Iv). Chloral hydrate alone results in under a well-ventilated hood.
distinct staining of terminal nerve branches, but it has the For use: approximately 0.1 g DFP in 500 ml distilled
unwanted effect of darkening muscle nuclei and connec- water. Treat sections for 30 min at room temperature. Then
tive tissue. The addition of pyridine suppresses nuclear rinse with two changes of distilled water and proceed with
staining and increases staining of the more proximal step C.
axons.
4. Incubate for 40 min at 37°C in 20% silver nitrate contain-
ing 0.1 % cupric sulfate, with 100 mg calcium carbonate at PERIODIC ACID-SCHIFF STAIN (PAS)
the bottom of the staining jar. This solution should be
freshly made each day. 0.5% Aqueous solution periodic acid 10 min
5. Develop in a solution of 1% hydroquinone and 5% sodium Rinse in distilled water.
sulfite. Use two baths, the first for 10 sec and the second Schiff reagene: 30 min
for 3-5 min. Discard solutions when cloudy. Dissolve 1 g basic fuchsin in 200 ml boiling distilled water.
6. Fix for 2-5 min in 5% sodium thiosulfate. Use large (1000 ml) Pyrex Ehrlenmeyer flask. Solution may
7. Tone for 5 min in 0.2% sodium tetrachloroaurate (gold boil over when dye is added, so flask should be placed in
chloride) containing one drop of glacial acetic acid per 100 a sink. Shake vigorously to dissolve dye and cool in run-
136 Staining Methods

ning water to 50°C. Add 20 ml normal HCl. Cool to 25°C Distilled water* 95 ml
and add 1 g of potassium metabisulfite (Na 2SzOs). Allow Rinse rapidly in 2 changes distilled water.
solution to stand in dark 18-24 hr. Add 2 g activated neu- Stain in Nuclear Fast Red solution. 2-5 min
tral charcoal and shake thoroughly. Filter. Solution Nuclear Fast Red (Kernechtrot) (Roboz Surgical 0.1 g
should be water-clear. Store in brown bottle in Instrument Co., Washington, D.C.)
refrigerator. 5% aluminum sulfate (5 g/lOO ml water) 100 ml
Rinse in 1% aqueous potassium metabisulfite. Dissolve with heat, cool, and filter.
Wash in running water 5-10 min (red color is enhanced). Can be used for 3-4 wk. Staining intense in freshly made
Counterstain in Harris's hematoxylin. 2-3 min stain. Reaction is reduced when solution separates into
Rinse in tap water. dark and light red areas.
Lithium carbonate saturated aqueous solution 2-3 sec Rinse in 2 changes of distilled water.
Rinse in distilled water. Dehydrate through 95% ethanol, then absolute ethanol (2
Dehydrate in 95% ethanol. changes).
Rinse in absolute ethanol. 2 changes Clear in xylene and mount.
Clear in xylene and mount. Results: Myelinated nerve fibers: Greenish-blue
Nuclei of neurons and neuroglial cells: Bright red

LUXOL FAST BLUE4 -NUCLEAR FAST RED

ALCIAN BLUE-TARTRAZINE STAIN 6
Fix in 10% formalin or Williams's fixative.
Fix tissues 24 hr in 10% formal saline.
Deparaffinize to 95% ethanol.
Deparaffinize and bring sections to water.
Stain in Luxol Fast Blue at 57 -60°C overnight.
Oxidize 5 min in freshly prepared equal volume mixture of
Luxol Fast Blue MSBN (E. I. DuPont de Nemours 0.1 g
0.3% aqueous potassium permanganate and 0.3% aqueous
Co., Wilmington, Delaware)
sulfuric acid. Bleach in 3% aqueous sodium metabisulfite.
95% Ethanol 100 ml
Wash in running tap water 5 min.
10% Aqueous acetic acid 0.5 ml
Rinse in 2 changes of 95% ethanol to remove excess stain.
Wash in distilled water. 2-3 min
Dip in lithium carbonate. 3-4 sec 'This step in differentiation is important: White matter of nerve fibers is
LiC0 3 1% aqueous solution 5 ml greenish blue. Gray matter (nerve cell bodies) is colorless.
 Staining Methods 137

Stain in celestine blue. 3-5 min Mayer's hematoxylin

Celestine blue (Roboz Surgical Instrument 0.25 g Hematoxylin 1g
Co., Washington, D.C) Distilled water 1000 ml
Ferric ammonium sulfate (iron alum) 50 ml Ammonium or potassium alum 50 g
(J. T. Baker Co., Phillipsburg, New Jersey), Sodium iodate 0.2 g
5 g/lOO ml water Citric acid 1g
Glycerine 7 ml Chloral hydrate 50 g
Rinse in water. Dissolve the alum in water, warming gently. Dissolve hema-
Harris's or Mayer's hematoxylin 2-3 min toxylin in this solution. Add citric acid, sodium iodate, and
Blue in running water. 5-10 min chloral hydrate. Shake well to dissolve all components. Final
Stain in Alcian Blue solution. 30 min color of solution is reddish-violet. Keeps well for months.
Alcian Blue 8 GS or 8 GX (Roboz Surgical 1g
Instrument Co., Washington, D.C)
Distilled water 100 ml MASSON CONNECTIVE TISSUE STAIN'
Rinse in distilled water.
Wash in running water. 5 min Fixative: Any general fixative; Bouin's and Zenker-formol
Stain in tartrazine solution. 30 sec recommended.
Tartrazine (Roboz Surgical Instrument Co., 2g Following deparaffinization: stain in celestine blue-hematox-
Washington, D.C)
ylin (see under Alcian Blue-Tartrazine stain).
1 % Aqueous acetic acid 100 ml
Wash in running water to improve bluing.
Rinse in distilled water.
Stain in Biebrich scarlet-acid fuchsin. 5 min
Dehydrate rapidly through
Biebrich scarlet, 1% aqueous solution 90 ml
95% Ethanol
Acid fuchsin, 1% aqueous solution 10 ml
Absolute ethanol 2 changes
Glacial acetic acid 1 ml
Clear in xylene and mount.
Rinse in distilled water. 2 changes
Results: Nuclei: Blue-black Phosphomolybdic-phosphotungstic acid 5 min
Epithelial lining: Green Phosphomolybdic acid 5g
Mucous neck cells: Pale green or bluish Phosphotungstic acid 5g
Zymogenic granules of chief cells: Dark green Distilled water 200 ml
Parietal cells: Yellow (Transfer directly to Fast Green solution.)
Erythrocytes: Bright yellow Fast Green FCF, 0.5% aqueous solution 1-2 min
138 Staining Methods

1 % Acetic acid rinse Rinse in acid alcohol.

Distilled water rinse Rinse in distilled water. 2 changes
Dehydrate through 95% ethanol, absolute 2 changes Transfer to phosphotungstic acid. 2 hr
ethanol. Phosphotungstic acid 5g
Clear in xylene and mount. Distilled water 100 ml
Rinse in distilled water.
Results: Nuclei: Deep mauve-blue
Stain in Aniline Blue solution. 2 hr
Cytoplasmic elements: Varying shades of red
Aniline Blue, water solution 0.5 g
Muscle: Red
Orange G 2g
Collagen: Green
Oxalic acid 2g
Distilled water 100 ml
5% Aqueous phosphotungstic acid 1 ml
MALLORY-HEIDENHAIN'S AZAN STAIN 6
Rinse in distilled water.
Treat with 5% aqueous solution 3-5 min
Fixative: Zenker-formol, Bouin's; mordant formalin-fixed tis-
phosphotungstic acid.
sues overnight in 3% potassium dichromate to improve
Rinse in distilled water.
staining.
Rinse in aqueous 1% acetic acid.
Treat with aniline alcohol. 45 min Dehydrate through 95% ethanol. 2 changes
Aniline 5-10 drops Dehydrate through absolute ethanol. 2 changes
95% Ethanol 100 ml Clear in xylene and mount.
Treat with acid alcohol. 1-2 min
Results: Nuclei: Brilliant red
Glacial acetic acid 1 ml
Collagen and reticulum: Blue
95% Ethanol 100 ml
Muscle: Red and yellow
Rinse in distilled water.
Basophilic cytoplasm:
Stain in Azocarmine at 55-57°C. 1 hr
Blue
Azocarmine G 0.5 g
Acidophilic cytoplasm:
Distilled water 100 ml Anterior pituitary cells
Orange-red
Glacial acetic acid 1 ml
Chromophobes:
Rinse in distilled water. 2 changes
Colorless or light gray
Differentiate in aniline-alcohol. *
By overstaining with Aniline Blue solution (18-24 hr) striking
"Nuclei, brilliant red; cytoplasm, very light red. Check with microscope. staining effects may be achieved.
 Stammg Methods 139

SNOOK'S METHOD FOR RETICULUM! Rmse m 2 changes dIstilled water

Dehydrate through 95% ethanol 2 changes
FIxatIVe WIllIams's fIxative, or 10% neutral buffered formalm, Dehydrate through absolute ethanol 2 changes
recommended Clear m xylene 2 changes, 5 mm each
Mount In Permount or sUItable mounting medIUm
OXIdIze In 025% aqueous solution potassIUm 5mm
permanganate Results Reticular fIbers Gray to black
RInse m dIstilled water NucleI Red
OxalIc aCId 5% aqueous solution (until brown 3-5 mm
sections are clear)
Wash m runnIng tap water 2-3mm
Mordant In 1% aqueous uranIum nItrate 5-10 sec BODIAN METHOD FOR NERVE FIBERS 6
solutIOn
Wash m runnmg water 3-5 mIn FIX overnIght In WIllIams's fIxative or 10% neutral buffered
AmmonIacal slIver solution 1 mm formalIn
To 20 ml of 5% aqueous sIlver nItrate solution m a 125-ml Formaldehyde 35-40% 40 ml
aCId-cleaned Ehrlenmeyer flask add 20 drops of GlaCIal acetic aCId 10 ml
aqueous 10% sodIUm hydroxIde solution Heavy brown 80% Ethanol 100 ml
precIpItate forms DIssolve by addmg ammOnIum Plcnc aCId 2g
hydroxIde, drop by drop, shakmg gently, leavmg only Place tissues In and store In
a few granules at the bottom of the flask Add dIstilled 70% ethanol, 2 changes, to remove most of 10-20 mm
water to make 60 ml and use at once plcnc aCId
Rmse In 2 changes dIstilled water Stam paraffin sectIOns In
Develop m 1% aqueous formalIn solution 1 mm Strong slIver protem (Roboz SurgIcal Instrument 1g
Rmse m 2 changes dIstilled water Co , Washmgton, 0 C )
Tone m 1% aqueous gold chlonde solution until 1-2 mm DIstilled water 100 ml
section IS graYIsh black Place slIdes In staInIng dIsh contaInIng water, dlstnbute 5-6
Rmse In 2 changes dIstilled water g clean copper (wIre, shot, or sheet) between slIdes and
SodIUm thIOsulfate, 5% aqueous solutIOn 5 mm around bottom of dIsh Spnnkle SlIver protem on top of
Rmse m 2 changes dIstilled water water evenly and place stamlng dIsh In oven at 37°C for
Counterstam m Nuclear Fast Red (see Luxol 2-3 mm 12-24 hr (Do not dIssolve by stIrrmg ) Cover wIth glass top
Fast Blue-Nuclear Fast Red Stam) DIstilled water (2 changes), rmse 8-10 times
140 Staining Methods

Place in reducing solution. 5-10 min taglomerular granules and mitochondna in the kidney stain
Hydroquinone 1g bright red.
Sodium sulfite 5g Stock solutions:
Distilled water 100 ml 1. 0.13 g methylene blue in 100 ml deionized water
Rinse in 3 changes of water. 1 min total 2. 0.13 basic fuchsin in 100 ml deionized water
Tone in 1-2 min 3. 0.2 M phosphate buffer, pH 7.2-7.8
Gold chloride 1g Staining solution: Mix
Distilled water 100 ml Methylene blue solution 12 ml
Rinse in 3 changes of water. 1 min total Basic fuchsin solution 12 ml
Develop in 2% aqueous oxalic acid. 2-4 min 0.2 M phosphate buffer 21 ml
Examine slide with microscope. Fibers become dark brown, Ethanol (95% or absolute) 15 ml
then gray to black. This filtered solution is useful for 4 or 5 days.
Wash in distilled water, 3 changes. 1 min each
Fix in 5% aqueous sodium thiosulfate. 5 min Staining procedure: The sections are immersed in the staining
Wash in running water. 5-10 min solution for 10-15 sec.* The slides are rinsed briefly in
Rinse in distilled water. deionized water and allowed to dry. The dried slides are
Dehydrate through 95% ethanol, 2 changes absolute ethanol. mounted under a cover slip.
Clear in xylene (2 changes,S min each) and mount. Phosphate buffer:
A. Sodium phosphate, monobasic 13.8 g
Results: Nerve fibers: Dark brown to black
(NaH 2 P0 4 • H 2 0)
Distilled water 500 ml
B. Sodium phosphate, dibasic (Na 2 HP0 4 ) 26.8 g
METHACRYLATE SECTIONS Distilled water
For pH 7.2, add
Lee's Methylene Blue-Basic Fuchsin Stain 7 : Monobasic sodium phosphate 10 ml
Dibasic sodium phosphate 40 ml
This general histological stain is used to differentiate cell
'It has been our expenence that thIs IS too short a hme for vanous tissues
types in the pituitary and endocrine cells in the intestinal and
By stammg a test shde for about 1 mm the nuclear components should take
stomach epithelium. It displays C cells of the thyroid and the blue stam By allowmg the shde to stay m the stam 1-2 mm more, the
clearly differentiates between smooth muscle and collagen. baSIC fuchsm should stam the connective tIssue and erythrocytes bnlhant
Elastic fibers and mitochondria are clearly revealed, and jux- red
 Staining Methods 141

PROCEDURE FOR PREPARING METHACRYLATE Cutting: Sections routinely cut at 1.5 p,m using glass knife with
SECTIONS JB-4 microtome (Sorvall)
Staining: See Lee's Methylene Blue-Basic Fuchsin stain.
Reagents from JB-4 Embedding Kit (Polysciences, Inc., Paul
Valley Industrial Park, Warrington, Pennsylvania 18976)

Fixation: Neutral buffered formalin at room temperature 4-24 ALDEHYDE FUCHSIN TRICHROME STAIN 8
hr depending upon size
Distilled water 900 ml Fixative: Zenker-formol; Bouin's
Formaldehyde, 35-40% 100 ml
Na 2 HP0 4 6.5 g Deparaffinize through xylenes.
NaH 2 P0 4 • H 2 0 4g Hydrate through absolute and 95% alcohols.
80% Ethanol plus iodine 5 min
One lobe of a mouse lung (1.25 em X 0.63 em thick) can be 70% Ethanol 5 min
fixed in 4 hr. Water Rinse
If tissues cannot be processed immediately after fixation, Oxidize in 0.3% potassium permanganate. 2-3 min
store in 70% ethanol. Freshly prepare by mixing equal parts of 0.6% KMN0 4 and
Other common fixatives may be used, except glutar- 0.6% H 2S0 4 .
aldehyde. Rinse in distilled water.
Decolorize in 4% aqueous sodium bisulfite 1-3 min
Dehydration: 70% Ethanol for 30 min
(NaSO}).
80% Ethanol for 30 min
Wash in running water. 5 min
95% Ethanol for 30 min
Stain in aldehyde-fuchsin solution. 1-3 min
Absolute ethanol, 2 changes, 30 min each
Add 3 ml paraldehyde and 1 ml concentrated HCl to 0.3%
Catalyzed Solution A 2 hr, under vacuum, at solution of basic fuchsin in 60% ethanol.
room temperature Staining solution should mature for 3 days before use. Dis-
card after 30 days.
Embedding: Catalyzed Solution A 25 parts Rinse in 95% ethanol. 3 changes
Solution B 1 part Wash in running water until background is pale. 1 hr
Mix well, pour into mold, and orient tissue. Stain in celestine blue-hematoxylin (see Alcian Blue-Tartra-
Should harden within 1 hr, ready to section. zine Stain).
142 Staining Methods

Rinse in water. Rinse in water.

"Blue" in lithium carbonate. 5-10 sec Bleach in 5% oxalic acid. 5 min
Rinse in water. Wash in running water. 5 min
Stain in Ponceau-fuchsin mixture. 2 hr Mordant in 4% iron alum. 1 hr
1% Ponceau-xylidine 4 parts Wash in running water. 10 min
1% Acid fuchsin 1 part Stain in PTAH (4 hr usually is sufficient). 3-24 hr
1% Fuchsin S 1 part Distilled water 1000 ml
Dilute 1:10 in 1% acetic acid. Hematoxylin 1g
Rinse in 1% acetic acid. Phosphotungstic acid 20 g
Rinse in distilled water. Add potassium permanganate 0.18 g
Stain in 1% solution of light green in 1% acetic 5-15 min to aid immediate ripening
acid. Transfer sections directly to 95% ethanol for brief rinse.
Rinse in distilled water. Absolute ethanol, rapid rinse
Differentiate in 0.5% phosphomolybdic acid. 15 min Clear in xylene.
Place in 1% acetic acid. 20 min Mount in Permount or suitable mounting medium.
Dehydrate rapidly through absolute ethanol. 3 changes
Results: Nuclei, neuroglial fibers, contractile elements of
Clear in xylene and mount.
striated muscle, fibrin: Blue
Results: B cell granules: Purple Collagen, elastin, cartilage, bone ground substance:
A cells: Red Yellowish to brownish red
Elastic tissue and mucus: Purple
Collagen: Green
WRIGHT'S STAIN
PHOSPHOTUNGSTIC ACID-HEMATOXYLIN (PTAH)
Place smears on staining rack and stain 2 min with stock
Wright's stain.
Fixative: Zenker or any fixative with mercuric chloride Wright's stain, dry 0.1 g
recommended.
Absolute methanol 60 ml
Following deparaffinization, down to water 5 min The dry stain may be mixed with the alcohol by grinding,
through graded alcohols, tissues are oxidized in using a mortar and pestle, or, to avoid well-stained fin-
freshly prepared 0.5% potassium permanganate. gers, the alcohol and dry stain may be mixed in a suitable
 Stammg Methods 143

flask and placed on an automatic shaker for 4-6 days A Dehydrate and embed m Paraplast or SImIlar embeddmg
satisfactory solutIOn IS obtamed medIum
Flood the slIde wIth dIstilled water, thus 4 mm Followmg deparaffimzmg, sectIOns are stamed m 2-3mm
dilutmg the stammg solution Harns's hematoxylIn
Buffered dilutmg solution may be necessary If the dIstilled RInse In tap water
water IS too aCId "Blue" m saturated aqueous LIC0 3 1 mIn
Phosphate buffer DlsodIUm phosphate (Na 2HP0 4 ) 71g RInse In water
25% Methanol m dIstilled water 250 ml Stam m 0 5% aqueous aCId fuchsm 3-5 mm
CItnc aCId CItnc aCId 525 g DIfferentiate m tap water, aCIdophIl granules
25% Methanol m dIstilled water 250 ml should be bnght red
Test stamed smear under mIcroscope If too red add 3-4 drops Place m 1% aqueous phosphotungstic aCId 3-5 mm
phosphate buffer per ml of dIlutmg flUId Pass slIdes dIrectly mto Mallory's Amlme Blue 1-2 hr
Lymphocytes should have reddIsh-purple nucleus and lIght Orange G stam
blue cytoplasm Eosmophiis should exhIbIt bnck red cyto- AmlIne Blue, water soluble 05g
plasmIC granules and dark, reddIsh-purple nucleus Orange G 20g
Phosphotungstic aCId, 1 % aqueous solutIOn 100 ml
Apply cover glass wIth mountmg medIUm for permanent
DIfferentiate m 95% ethanol
preparation
Blue granules are Intense but fuchSIn-staIned
granules are clearly VISIble
Rmse m two changes absolute ethanol
MODIFIED MARTIN-MALLORY STAIN FOR MOUSE Clear m two changes xylene
PITUITARY GLAND Mount WIth Permount or sUItable mountmg medIUm

FIx m Zenker-formol (95% Zenker flUId, 5% formaldehyde) Results NucleI Dark blue
overmght ACIdophIl granules Bnght red
Wash tissue m runmng tap water 8-10 hr BasophIl granules Blue
For pItUItary gland In sztu decalcIfy m 10% 2-3 days Chromophobes Gray cytoplasm
formIc aCId Erythrocytes Orange to red
Wash In runnmg tap water to remove aCId 24 hr Bone Intensely blue
144 Staining Methods

NEW METHYLENE BLUE STAIN FOR RETICULOCYTES GIEMSA STAIN

AND HEINZ BODIES
Smears: Fix in absolute methanol. 15 min
Place 2 drops oxalated or heparinized whole blood on a clean Air dry.
glass slide. Stain in working solution. 15-30 min
Add 3-4 drops of New Methylene Blue to the blood, mix well, Giemsa stock* 3 ml
and allow to stand 10-15 min. Absolute methanol 3 ml
Distilled water 100 ml Distilled water 100 ml
Potassium oxalate 1.6 g Adjust staining solution to pH 6.8-7.0.
Dye 0.5 g Buffers: 25% Methanol 250 ml
Rinse with distilled water and apply cover glass for viewing Disodium phosphate (Na 2 HP0 4 ) 7.09 g
under the microscope or
25% Methanol 250 ml
Air dry.
Citric acid 5.25 g
Apply cover glass with Permount or any suitable mounting
medium for permanent preparation. Rinse in tap water
Results: Reticulocytes have distinct, blue reticular network. Results: Lymphocytes: Nucleus-dark purple; cytoplasm-sky
blue
Eosinophils: Nucleus-reddish purple; cytoplasmic
granules-brick red

TOLUIDINE BLUE STAIN

On a warming plate at 60°C flood epon section with staining

solution. 4-5 min
1% aqueous solution toluidine blue 1 part
2.5% aqueous solution sodium carbonate (Na 2C0 3 ) 20 parts
Rinse in tap water.
Methacrylate sections should be stained vertically 4-5 min in
a Coplin jar to prevent residue from accumulating on *Glemsa Blood Stalfl, Azure B type, Harleco, GIbbstown, New Jersey Glemsa
slide. wIth Azure A MC/B Manufactunng ChemIsts, Norwood, OhIO
 References

Lee G Luna (ed), Manual of Histological Stamlllg Methods of the Armed 7 Polysclences, Inc, JB4 Embeddmg Kit, May 1976, Warrenton, Pennsyl-
Forces Instllute of Pathology, 3rd ed, McGraw-HIll, 1968 vama 18976, Data Sheet 123
2 Alan Pestronk and Damel B Drachman, A new stain for quantitative 8 Sydney S Lazarus and Bruno W Volk, The Pancreas III Human and E-r:per
measurement of sprouting at neuromuscular Junchons, Muscle and Nerve Imcntal Dzabetes Grune and Stratton, New York, 1962
Jan.-Feb.: 70-74 1978
3 AGE Pearse, Hlstocherll1stry, Theorellcal and Applied, J & A ChurchIll,
London, 1960
4 Edward Gurr, Stallllllg, Practical and Theoretical, Wllhams and WIlkms,
RECOMMENDED READING
Baltimore, 1962
WIlham Bloom and Don W Fawcett, A Tntbook of Histology, 9th ed, W B
5 T A AsoJo, AIClan blue tartrazme A simple differential stain for the
Saunders, Philadelphia, 1968
fundiC gland cells of mammahan stomach, Stam Technology 50:55-57,
M B L Cralgmyle, Color Atlas of Histology, Year Book Medical Pubhshers,
1975
Chicago, 1975
6 Gretchen L Humason, Ammal Tissue Teclmlques, 3rd ed ,W H Freeman,
Earl L Green (ed), BIOlogy of the Laboratory MOll'C, 2nd ed, McGraw-HIll,
San FranClsco, 1972
New York, 1966 Chapter 13, Anatomy, Kathanne P Hummel et al

145
 Index

Lightface numbers are text page numbers; boldface numbers are color figure numbers. Figure A will be found on p. 70.

Acetylcholinesterase, 18, 134 Bronchiole, 25, 131 Chromaffin reaction, 11

Acidophils, 9, 33,35 Brown degeneration, 11, 39 Chromophobes, 9, 33, 35
Acinar cells, 19, 99,100 Brown fat, 30, 122 Ciliary body, 6, 23
Adipose tissue, 139 Brunner's glands, 18,90,91 Clara celis, 25, 131
Adrenal gland, 9, 10, 36-39 Brush border, 21, 107 Clitoral gland, 47
Alpha cells, 19,100 Bulbourethral glands, 16, A, 71 Coagulating gland, A, 74-76
Alveolar duct, 25, 127-129 Cochlear duct, 6
Alveolar sac, 25, 127-130 C cells, 10 Collagen fibers, 112-114
Ameloblast layer, 149 Capillaries, 33 Conjunctiva, 21, 23-28
Amnion, 50 Cardiac muscle, 32, 153, 155 Cornea, A, 20, 21, 27
Ampullary gland, 16, A Cartilage Corpora cavernosum, 22, 64
Anal region, 18, 98 elastic, 29, 31 Corpus luteum, 13, 44
Anterior chamber, 24 fibrocartilage, 29, 141,143 Cowper's glands, 16
Anterior horn, 14 hyaline, 29, 40, 124, 126, 141 Crypts of Lieberkuhn, 18, 93, 94
Axon, 3, 5, 8, 12 Cecum, 96
Cells of Leydig, 15, 66 Decidua basalis, 14, 50-52
Basket celis, 17 Central arteriole, 27, 136 Dendrites, 3, 7, 8
Basophils, 9, 33 Cerebellum, 1,2,10,10-12,19 Dental pulp, 148-149
Beta celis, 19, 100 Cerebral cortex, 1,3, 7 -9, 10, 11, 19 Dentine, 147 -149
Bile canaliculi, 19,103 Cerebrospinal canal, 15 Deparaffinizing sections, 133
Bile duct, 19 Chief cells, 10, 18,43,88,89 Dermis, 23, 31, 61
Bladder, 16,21,22,109-111 Chorionic gonadotrophin, 14 Descemet's membrane, 6, 21
Bone,29,142 Choroid, 6, 20, 22-24 Diestrus, 14,53
Bowman's capsule, 21, 106 Choroid plexus, 2, 4, 10, 11 Distal convoluted tubule, 106, 107

147
148 Index

Dorsal prostate, 16, A Esophagus, 1, 18,81,82,123 Gallbladder, 19,104

Duodenum, 18, 90, 91 Estrogen, 14 Ganglia, 3, 3, 6, 13, 16
Estrus, 14, 57 Germinal centers, 27, 135, 139
Ear, 5, 29-31 Eye, 6,20 Germinal epithelium, 13, 44, 45
Elastic cartilage, 29, 31 Giant cells, 14
Elastic fiber, 130, 155, 156 Fat cells, 44, 47, 60-62, 64 Glands
Embryo, 50 Femur, 144 adrenal, 9, 10,36-39
Enamel, 147, 149 Fibers ampullary, 16, A
Endometrium, 13,48,50,52 collagen, 112-114 Brunner's, 18, 90, 91
Endothelial cells, 21, 33, 108, 115 elastic, 130, 155, 156 bulbourethral (Cowper's), 16, A, 71
Enterochromaffin system, 19 myelinated (nerve) 3, 3, 9, 14, 15 coagulating, 16, A, 74-76
Eosinophils, 34, 165-168 non myelinated (nerve) 3,14 Harderian, 7, 20,23,28
Ependymal cell layer, 4, 4 reticular, 27, 129 lacrimal, 7, 20, 28
Epidermis, 23, 61, 117, 119 Fibroblast, 113-115 mammary, 14,61,62
Epididymis, 15, A, 69 Fibrocartilage, 29, 141, 143 mucous, 77, 120, 124-126
Epiglottis, 29, 123 Filiform papillae, 17, 79 parathyroid, 9, 10, 40-43
Epiphyseal plate, 29, 144 Fimbria, 49 parotid, 17,83,84
Epithelium Fixatives pineal, 9, 10, 19
ciliated columnar, 5, 49,124, Bouin's, 131 pituitary, 1, 9, 32-35
126 Carnoy's, 131 preputial, 16, A, 63, 64
pseudostratified ciliated columnar, 5, formalin-mercuric chloride, 132 prostate, 16, 72, 73
25, 125 formalin, neutral buffered, 131 salivary, 17, 83, 84
simple cuboidal, 4, 5, 9, 14, 15, 17, 18, formol-alcohol-acetic (FAA), 132 sebaceous, 5, 18, 23, 31, 47, 60, 98,
19, 21, 42, 106 formol-saline, 132 117,119
simple columnar, 4, 15, 16, 18, 19,48, Vandergriff's, 132 sublingual, 17, 18, 83, 138
70, 72, 76, 90, 91, 95, 96, 104 William's, 131 submandibular, 17, 18,83-86,138
simple squamous, 5, 15, 21, Zenker-formol (Helly's), 131 thyroid, 9, 40-42
stratified squamous, 6, 13, 15, 16, 17, Foliate papillae, 17 vesicular (seminal vesicle), 16, A, 74-76
18, 21, 22, 23, 25, 25 Follicular cells, 9, 10, 44-46 Glomeruli, 21, 36-39, 106
transitional, 21, 22, 109-111 Fungiform papillae, 17 Glucagon, 19,38
 Index 149

Goblet cells, 18, 93-95 Lacrimal gland, 7, 20, 28 Muscle

Great alveolar cell, 25, 132 Lactating breast, 60 cardiac (striated), 32, 33, 153, 155
Growing follicle, 13, 44, 45, 46 Lactiferous duct, 60, 62 smooth, 19,31,68,72,87,90,91,96,
Gustatory cell, 78 Lactogen, 14 97,104,111
Lens, 6, 20, 23, 24, 28 striated (skeletal), 17, 18, 31, 40, 42,
Hair, 23 Liver, 19,101-103 61,62,71,77,79,81-82,98, 118,
Hair cells, 6, 29, 30 Lung, 25, 127-132 145, 150, 151, 153
Hair follicles, 31, 60, 61, 117, 118 Lymph nodes, 27, 83, 138, 139 Myelinated nerve fibers, 3, 3, 9, 14, 15
Harderian gland, 7, 20, 23, 28 Lymphocytes, 34, 164 Myoepithelial cells, 7, 17
Hassall's corpuscle, 27, 134 Myometrium, 13,48,52
Heart, 33 Macrophage, 113
Heart valves, 157 Mammary glands, 14, 61, 62 Nasal passages, 25
Hepatic artery, 19 Mast cell, 112, 113, 115 Neuroepithelial cells, 17
Hepatic cells, 19,101-103 Megakaryocytes, 27, 136, 137, 140, 158, Neurofibrils, 3, 5, 8
Howship's lacunae, 29, 142 159 Neuroglia, 2, 7, 12
Hyaline cartilage, 29, 40, 124, 126, 141 Meibomian gland, 25, 27 Neuroglial cells, 3,4, 9, 10, 14
Hypophysis, 9, 34 Melanocytes, 7 Neurohypophysis, 9, 34
Meninges, 4, 15 Neuron, 3
Ileum, 18, 93 Meniscus, 145 Neutrophils, 34, 158,159,162
Insulin, 19 Metestrus I, 14, 58 Nipple, 14, 15, 60
Intercalated disks, 32, 153 Metestrus II, 14, 59 Nissl substance, 3, 9
Interstitial cells of Leydig, 15, 65-67 Methacrylate sections, 21, 85, 95, 115, Nonglandular stomach, 18,87
Iris, 6, 20, 23, 24, 26-28 131,132 Nonlactating mammary gland, 61
Islets of Langerhans, 9, 19,99,100 Metrial gland, 50 Nonmyelinated nerve fibers, 14
Molecular layer, 10-12 Normoblast, 158, 159
Jejunum, 18, 94 Monocytes, 34, 158, 161
Motor cell,S, 7 Odontoblasts, 148, 149
Kidney, 10,21,36,38, A, 105-108 Motor end plate, 17, 18 Olfactory cells, 25, 125
Knee joint, 145 Mucous glands, 77, 120, 124-126 Optic nerve, 22
Kupffer cells, 19, 101, 103 Mucous neck cells, 88 Organ of Corti, 5,29,30
150 Index

Os penis, 22, 63, 64 Posterior lobe, 9 Skeletal muscle, 31, 16

Osteoblasts, 29, 141, 142 Preputial gland, 16, A, 63, 64 Skin, I, 60-64, 98
Osteoclasts, 29, 142 Primary spermatocytes, 15,66,67 Small intestine, 18, 92, 93
Osteocytes, 29, 142 Primordial follicle, 44, 45 Smooth muscle, 19,31,68,72,73,87,90,
Ovary, 9, 13,44-46 Proestrus, 14, 54, 55 91, 94, 96, 97, 104, 109, 111, 152
Oviduct, 49 Proestrus / estrus, 55, 56 Spermatids, 15,66
Ovum, 45, 46 Progesterone, 14 Spermatogonia, 15, 66, 67
Oxyphil cell, 10, 41, 43 Proximal convoluted tubule, 21, 106, Spermatozoa, 15,67-70
Oxytocin, 9 107 Spinal cord, 4, 13-15
Pulmonary vein, 33, 154-156 Spiral ligament, 6
Pancreas, 9, 19,90,99,100 Pulvinus, 118, 119 Spleen, 27, 135-137, 140
Paneth cells, 18, 93 Purkinje cell, 10 Stains
Parafollicular cells, 10 aldehyde-fuchsin, 32, 33, 35, 100,
Parathyroid glands, 9, 10, 40-43 Rectum, 73 113, 114, 120, 130, 141
Parietal cells, 18, 89 Red pulp, 27, 135-137,140 alcian blue-tartrazine, 88, 136
Parotid gland, 17,83,84 Reticular fibers (black), 27, 129 Bodian silver, 5, 7, 8, 12, 13, 16, 17, 139
Pars dis talis, 9 Reticulocyte, 160 Giemsa, 144, 158, 159
Pars intermedia, 9 Retina, 6,20,22-24 Heidenhain's Azan, 27, 30, 49, 60, 64,
Pars nervosa (neurohypophysis), 9 67,76,80,101, 107, 112, 138, 151,
Penis, 16, 21, 22, A, 63, 64 Salivary glands, 17, 83,84 153,155
Peyer's patches, 18, 92, 93 Sclera, 6, 22, 24 hematoxylin and eosin (H & E), I, 11,
Pharynx, 25, 123 Sebaceous glands,S, 18, 23, 31, 47,60, 19, 20, 22-24, 29, 31, 36, 37, 39, 41-
Pia mater, 4, 10,15 98,117,119 47, 50-55, 57 -59, 61-63, 65, 66,
Pineal gland, 9, 10, 19 Sebum, 23 68,70-75,77,78,81-83,87,89,92,
Pituitary gland, 9, I, 32-35 Secondary spermatocytes, 15, 66, 67 93, 96, 98, 99, 102-106, 109, 111,
Placenta, 9, 14, 50-52 Seminal vesicles, 16, A 116-118,121-123,125,127,133-
Plasma cell, 158, 159 Seminiferous tubules, 15, 65 139,141-149,152,157
Platelets, 34, 163 Serosa, 13, 19, 48, 87, 92, 97, 104 Lee's methylene blue-basic fuchsin,
Portal triad, 102 Serous glands, 25, 77, 124, 126 21, 85, 86, 94, 95, 108, 131, 140
Portal vein, 19 Sertoli cell, 15, 66, 67 Luxol Fast Blue-Nuclear Fast Red, 3,
Posterior chamber, 6, 24 Sinusoids, 19, 101, 103 14,15, 136
 Index 151

Stains (cont.) Submandibular gland, 17, 18, 83-86, Vagina, 13,47,53-59

Martin-Mallory, 33, 143 138 Vallate papillae, 17
Masson, 2, 6, 28, 34, 79, 110, 119, 120, Sustentacular cell, 17,78, 125 Valve leaflet, 157
124, 126, 137, 150 Vas deferens, 16, A, 72, 73
motor end plates, 17, 134 Taste buds, 17, 77,78,80 Vasopressin, 9
new methylene blue, 144, 160 Tectorial membrane, 6, 29, 30 Veins, 33
periodic acid-Schiff (PAS), 25, 26, 38, Testis, 9, 15, A, 65-67 Ventral prostate, 16, A, 72, 73
40,48,56,84,90,91,97, 135 Thick skin, 23, 121 Vesicular gland (seminal vesicle), 16, A,
phosphotungstic acid hematoxylin Thymus,27,133,134 74-76
(PTAH), 142, 154, 156 Thyrocalcitonin, 10 Vestibular membrane (Reissner's), 29, 30
Snook's silver, 129, 139, 140 Thyroid gland, 9, 40-42 Vibrissae, 23, 118, 119
toluidine blue, 128, 132, 144 Tongue, 17,1, 17,77-80 Villi, 18, 90-92, 94, 95
Wright's, 161-168, 142 Tooth, 1, 147-149 Vitreous humor, 6, 20
Stereocilia, 15, 70 Trabeculae, 27,133, 135-137,140,144,145
Stomach, 18, 87-91 Trachea,25,123,126 White fat, 30, 122
Stratified squamous epithelium, 6, 13, Tunica albuginea, 15 White pulp, 27, 135
15, 16, 17, 18,21,22, 25, 31, 40, 47, Type I epithelial cell, 25, 132
53-59,77,78,80-82,87,98, Type II (great alveolar) cell, 25, 132 X zone, 10, 36, 37
146
Stratum corneum, 23, 121 Ultimo branchial bodies, 10 Zona fasciculata, 10,36-39
Stratum germinativum, 23, 79, 121 Ureter, 21, A Zona glomerulosa, 10, 36, 37, 39
Stratum granulosum, 23, 79, 121 Urethra, 16, 21, 22, 47, 63, 64, 73 Zona pellucida, 13, 46
Striated muscle, 17, 18, 31, 40, 42, 61, 62, Urinary bladder, A, 73 Zona reticularis, 10
71,77,79,81,82,98,118,119,145, Urothelium, 21, 22 Zone of hypertrophy, 29, 144
150 Uterine glands, 48 Zone of maturation, 29
Sublingual gland, 17, 18, 83,84,138 Uterus, 13, 48 Zone of proliferation, 29, 144