ABSTRACT
TABLE 15.1 Solutions Commonly Used for Tissue Fixation
Stains Commonly Used in Histopathology
Stains Commonly Used in Histopathology
Stains Commonly Used in Histopathology
TABLE 15.3 Common Abbreviations and Codes Used in Histopathology
Frequently Used Grading Schemes for Histopathologya
TABLE 15.5 Goals for Protocol Tissue Availability during Histopathologya
Goals for Protocol Tissue Availability during Histopathologya
Adrenal gland, cortex Condensation of cells of zona glomerulosa (40 min*) Karyoklasis (clumping of chromatin) in epithelial cells of
zona fasciculata and zona reticularis (8 h)
Adrenal gland, medulla Nuclear condensation and pyknosis (2 h) Increased cytoplasmic vacuolation (2 h)
Ampullary gland Sloughed and disruption of epithelium (<20 min)
Blood Hemolysis and pigmentation of adjacent tissues (>16 h)
Blood vessel (major) Aorta-formation of clear and pale staining areas between
elastic laminae (<20 min) Aorta-darkening and condensation of smooth muscle
nuclei (16 h)
Bone (sternum) Bone and cartilage-no remarkable postmortem changes
at 16 h
Bone marrow Pyknosis of megakaryocytes (2 h)
Brain Increased clear space around oligodendroglia in white
matter (2 h) Formation of clear space around cortical astrocytes (4 h) Increased numbers of shrunken hyperchromatic neurons (8 h) Pyknosis of oligodendroglia in corpus callosum (8 h)
Goals for Protocol Tissue Availability during Histopathologya
Increased foamy and vacuolated cytoplasm of cortical
neurons (12 h)
Brown fat (perithymic) Granular cytoplasm (4 h) Blood-˜lled vessels (8 h) Condensed nuclei (8 h) Formation of intracellular soap, basophilic deposits (8 h)
Bulbourethral gland Increased granularity and lack of uniformity to the epithe-
lial cell cytoplasm (12 h)
Clitoral gland No remarkable postmortem changes at 16 h
Coagulating gland Pyknosis of villous epithelium (20 min) Pyknosis of basal epithelium (20 min) Disruption of epithelium (4 h)
Ductus (vas) deferens Formation of intercellular clear clefts between adjacent
epithelial cells (20 min)
Epididymis Formation of intercellular clear clefts between adjacent
epithelial cells (16 h)
Esophagus Pale cytoplasm of polyhedral epithelial cells (40 min) Formation of inter˜ber clear spaces in muscle (40 min) Formation of vesicular spaces within collagen of the sub-
mucosa (8 h) Pyknosis of ˜brocyte nuclei in submucosa (8 h) Absence of mitotic ˜gures in basal epithelium
layer (12 h)
Eye Shrinkage of nuclei and formation of clear spaces and
clefts in the substantia propria of cornea (20 min) Absence of mitotic ˜gures in anterior epithelium of the
cornea (40 min) Condensation of nuclei in the retina (8 h)
Fat No remarkable postmortem changes at 16 h
Fat, brown (see Brown Fat) Harderian gland
Pyknosis of glandular epithelial cells (4 h) Karyorrhexis of glandular epithelial cells (12 h)
Heart Condensation and darkening of endothelial nuclei (20 min) Formation of inter˜ber clear spaces in muscle (40 min)
Intestine, large-cecum Loss of super˜cial epithelial cells (<20 min) Saprophytic bacilli in the lamina propria (<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (1 h)
Basophilia of the lamina propria caused by the presence
of large numbers of bacilli (1 h) Loss of crypt epithelial cells (2 h) Loss of cells in lamina propria (8 h)
Intestine, large-colon Loss of super˜cial epithelial cells (<20 min) Saprophytic bacilli in the lamina propria (<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (40 min) Disruption and sloughing of crypt epithelium (2 h) Loss of crypt epithelial cells (4 h) Basophilia of the lamina propria caused by the presence
of large numbers of bacilli (4 h) Pyknosis of remaining epithelial cells (8 h) Loss of cells in lamina propria (12 h)
Intestine, small-duodenum Loss of villar epithelium (<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (40 min) Loss of cells in the villous lamina propria (4 h) Lateral separations in crypt epithelium (4 h) Loss of epithelium from crypts (12 h) Loss of cells in basal lamina propria (12 h)
Intestine, small-ileum Loss of villar epithelium (<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (40 min) Lateral separations in crypt epithelium (40 min) Loss of cells in the villous lamina propria (1 h) Loss of cells in basal lamina propria (2 h) Pyknosis of Peyer’s patch lymphocytes (4 h) Loss of epithelium from crypts (12 h) Total loss of mucosal epithelium (12 h)
Intestine, small-jejunum Loss of villar epithelium (<20 min) Loss of cells in the villous lamina propria (<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (40 min) Lateral separations in crypt epithelium (1 h) Loss of cells in basal lamina propria (12 h) Loss of epithelium from crypts (>16 h)
Islets, pancreatic Pyknosis of peripheral islet cells (4 h) Pyknosis of most of islet cells (16 h)
Kidney Pyknosis and vacuolated cytoplasm of distal convoluted
tubular epithelium (40 min) Pyknosis of majority of nuclei of ascending and descend-
ing tubules of loops of Henle at the corticomedullary junction (2 h)
Contraction of nuclei and increased granularity of cytoplasm of proximal convoluted tubular epithelium (4 h)
Majority of glomerular cell nuclei pyknotic (8 h) Formation of clear space around the loops of Henle deep
in the medulla (8 h) Pyknotic nuclei in the collecting tubule epithelium of the
cortex (12 h)
Lacrimal gland (exorbital) Nuclear condensation of acinar epithelium (12 h)
Liver Sinusoidal accumulation of blood (resembles congestion)
(<20 min) Presence of saprophytic bacilli (2 h)
Lung (immersion-˜xed) Parenchymal blood accumulation (resembles congestion)
(<20 min) Separation of bronchiolar epithelium from the lamina pro-
pria (<20 min) Presence of proteinaceous ©uid in alveolar spaces
(40 min) Pyknosis of a majority of alveolar cells (4 h) Pyknosis and sloughing of bronchiolar epithelium (16 h)
Lung (airway-perfused) Parenchymal blood accumulation (resembles congestion)
(40 min) Pyknosis of a majority of alveolar cells (40 min) Separation of bronchiolar epithelium from the lamina pro-
pria (1 h) Presence of proteinaceous ©uid in alveolar spaces (2 h)
Lymph node, mandibular Decreased density or rarefaction in cortical lymphocytes
(20 min) Cleavage lines resembling “dull-knife distortion” (1 h) Pyknosis of cortical lymphocytes (4 h)
Lymph node, mesenteric Decreased density or rarefaction in cortical lymphocytes
(20 min) Cleavage lines resembling “dull-knife distortion” (1 h) Pyknosis of cortical lymphocytes (2 h)
Mammary gland (female) Formation of clear spaces at base of acinar epithelial
cells (1 h) Pyknosis of acinar epithelium (2 h) Sloughing of epithelial cells into ducts (4 h)
Mammary gland (male) Increased foamy, vacuolated appearance of acinar cell
cytoplasm (4 h) Pyknosis of acinar epithelium (4 h)
Ovary Formation of clear spaces around follicles (40 min) Individualization of luteal cells (4 h)
Vacuolar spaces in acinar epithelium (40 min) Loss of central chromatin (hollow nuclei) in pancreatic
duct epithelium (1 h) Formation of clear interstitial spaces around acini
(4 h) Disappearance of eosinophilic, proteinaceous luminal
duct contents (4 h)
Parathyroid gland Formation of clear spaces at base of chief cells (1 h)
Peripheral nerve, sciatic nerve Slight nuclear contraction of Schwann cells (2 h)
Pituitary gland, pars distalis Increased pyknotic pituicytes (2 h) Pallor or disappearance of pituicyte cytoplasm (4 h)
Pituitary gland, pars intermedia Increased pyknotic pituicytes (4 h) Pallor or disappearance of pituicyte cytoplasm (4 h)
Preputial gland No remarkable postmortem changes at 16 h
Prostate, dorsal Pale or clear luminal contents (<20 min)
Prostate, ventral Pale or clear luminal contents (16 h)
Salivary gland, parotid Vacuole and clear space formation in the basal portion of
the ductular epithelium (40 min) Pyknosis of ductular epithelium (1 h) Pyknosis and karyoklasis (clumping of chromatin)
of acinar epithelium, which resembles very acute necrosis (12 h)
Salivary gland, sublingual Pyknosis and disruption of ductular epithelium (8 h)
Salivary gland, submaxillary Loss of tubuloalveolar (serous) cell nuclei (2 h) Tubuloalveolar (serous) cell dispersion that resembles
very acute necrosis (2 h) Disintegration and disruption of interlobular duct epithe-
lium that resembles very acute necrosis (4 h) Mucus-producing acini were unremarkable at 16 h
Seminal vesicles Pyknosis of epithelium (1 h) Detachment and change to cuboidal appearance of epithe-
lial cells with disruption of epithelial surface (2 h) Appearance of fractures and lucent or paler areas in lumen
contents (8 h)
Skeletal muscle Fiber disorganization and “fracturing” into small haphaz-
ard cleavage spaces (20 min)
Condensed and hyperchromatic endomysial nuclei (1 h)
Skin Chromatin clumping and karyorrhexis of sebaceous cells
(40 min) Pyknosis of dermal ˜brocytes (2 h) Epidermis and dermis were unremarkable at 16 h
Spleen Pyknosis of megakaryocytes (<20 min) Decreased density or rarefaction of lymphocytes in white
pulp (40 min) Cleavage lines resembling “dull-knife distortion” (40 min) Pyknosis of most white pulp lymphocytes (8 h) Generalized pyknosis of red pulp (12 h)
Stomach, forestomach Pyknosis of basal epithelial cells (4 h) Pyknosis of ˜brocytes in super˜cial lamina propria (4 h) Formation of clear spaces around ˜brocytes in super˜cial
lamina propria (4 h) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (8 h)
Stomach, glandular Loss of super˜cial epithelium from mucosal ridges
(<20 min) Separation of glandular epithelium from basement mem-
brane (40 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (1 h) Separation of crypt epithelium from basement mem-
brane (2 h) Lateral separations in glandular epithelium (4 h) Pyknosis of crypt epithelium (4 h)
Testes Individualization of interstitial cells (1 h) Pyknosis of interstitial cells (2 h) Formation of clear spaces around seminiferous tubules (8 h)
Thymus Large medullary extravasations of erythrocytes, which
resemble antemortem thymic hemorrhages and apparently result from the loss of blood from leaky thymic vessels (<20 min)
Cleavage lines resembling “dull-knife distortion” (4 h) Nuclear contraction of cortical lymphocytes (4 h) Decreased density or rarefaction in cortical lympho-
cytes (16 h)
Thyroid gland Pale, foamy, and/or bubbly colloid (<20 min) Pyknosis of parafollicular “C” cells (40 min) Epithelial cells free in colloid (1 h) Pyknosis of follicular epithelium (2 h) Disruption of follicular epithelium (4 h) Pyknosis of epithelium in largest follicles occurred later
than in small-to medium-sized follicles (8 h)
Formation of inter˜ber clear spaces in skeletal muscle
(40 min) Perinuclear clear spaces (halo) in polyhedral cell layer of
the mucosa (4 h) Contraction and darkening of muscle cell nuclei (4 h) Absence of mitotic ˜gures in basal epithelial cell layer (12 h)
Trachea Disruption or sloughing of epithelium into glandular
lumen (4 h) Pyknosis of super˜cial epithelium (8 h) Loss or sloughing of super˜cial epithelium (8 h) Loss of cilia (12 h) Pyknosis of glandular epithelium (12 h) Formation of intracellular soap, basophilic deposits in fat
cells, deep in the mucosa (16 h)
Urinary bladder Formation of vacuoles in transitional epithelium
(<20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (1 h) Disruption and sloughing of the surface transitional epi-
thelium (4 h)
Uterus Separation of glandular epithelium from basement mem-
brane (20 min) Formation of inter˜ber clear spaces and pyknosis of myo-
˜ber nuclei (4 h) Sloughing of super˜cial endometrial epithelium (8 h) Cervix had no remarkable postmortem changes at 16 h
Vagina No remarkable postmortem changes at 16 h
Adrenal gland, cortex Amyloidosis (amyloid deposition)—associated with sys-
temic amyloidosis (see Kidney) Atrophy, diffuse-low incidence (CD-1: males 5%,
females <1%) Accessory cortical nodules-common in adrenal capsule
or adjacent connective tissues (CD-1 mice, 2%–5%) Extramedullary hematopoiesis-usually occurs in asso-
ciation with leukemoid reactions and myeloid hyperplasia (see Bone marrow)
Pigmentation, ceroid (lipofuscin)—accumulates in residual X-zone cells
Hyperplasia, eosinophilic cell, focal (CD-1: 2000 males 10.8%, 2000 females 8.8%)
Hyperplasia, subcapsular (type A) spindle cells: common in mice of all ages; incidence up to 80% in aged CD-1 females (CD-1: 2000 males 16.6%, 2000 females 33.6%)
liferation of large polyhedral vacuolated cells; usually associated with spindle cell hyperplasia
X-zone (inner cortex)—normal basophilic cells in young mice of both sexes; involution complete by about 10 days of age in males, may be seen up to 30 weeks of age in females; characterized in females by marked vacuolation, especially in nonbred females
Tumors Cortical adenoma, all types-low incidence
(CD-1: 891 males 0.9%, 890 females 0.5%) (CD-1: 2000 males 1.3%, 2000 females <1%) (B6C3F1: 2240 males 2.4%, 2306 females 0.3%)
Type A cell adenoma Type B cell adenoma Cortical carcinoma (adenocarcinoma), all types: very
low incidence (B6C3F1: 3 reported in 2240 males 0.1%, 1 reported
in 2306 females <0.1%) (CD-1: none in 891 males, 3 in 890 females 0.3%) (CD-1 mice: 2000 males none, 2000 females <1%)
Adrenal gland, medulla Hyperplasia-low incidence (CD-1 mice, 1%–8%; males
> females) Hypertrophy Tumors
Ganglioneuroma-very rare; none reported in more than 10,000 CD-1 and B63F1 mice
Pheochromocytoma (benign)—low incidence (CD-1: 5 in 891 males 0.6%, 3 in 890 females 0.3%) (CD-l mice: 2000 males <1%, 2000 females <1%) (B6C3F1: 2240 males 1.2%, 2306 females 0.7%)
Pheochromocytoma, malignant-very low incidence (CD-1: none in 891 males, 1 in 890 females 0.1%) (B6C3F1: 2 reported in 2240 males 0.1%, none
reported in 2306 females 0.0%)
Blood vessels Amyloidosis-frequent vascular ˜nding especially in
lungs and liver (see Kidney). Angiectasis-common ˜nding especially in uterus, ovary,
liver, spleen, and lymph nodes. Arteritis (polyarteritis nodosa, periarteritis, perivasculi-
tis)—frequent ˜nding, most often in the kidney, heart, mesentery, uterus, testis, and urinary bladder (RF: 311 females 18.0%).
