carson glossary+index v04 0407 · 2016-04-20 · for grimelius argyrophil stain, 264 for luxol fast...

372 Index

AABC method. See avidin-biotin enzyme complex method.absolute alcohol

and celloidin embedding, 40for Luxol fast blue staining, 213rapid processing with, 38

absolute alcohol hematoxylinfor Russell modifi cation of the Movat pentachrome stain, 172

absolute alcohol-chloroform solutionfor Holzer staining method, 209

acetate buff erfor peroxidase-antiperoxidase (PAP) immunohistochemical

technique, 298, 300for phosphorylase stain, 326composition, 125

acetate formalin, eosinophil fi xation, 11formulation, 12

acetic acid, 9comparative characteristics, 18composition, 125for Gomori 1-step trichrome stain, 166for Grimelius argyrophil stain, 264for Luxol fast blue-cresyl echt violet staining, 214for Masson trichrome staining, 163pH control with, in eosin counterstaining, 114preparation for alcian blue technique, 145preparation for alcian blue-PAS-hematoxylin technique, 148preparation for Müller-Mowry colloidal iron technique, 150preparation for thiofl avin T technique, 155rate of penetration, 7stability of, 100for thiofl avin S demonstration of Alzheimer disease, 206for Turnbull blue stain preparation, 258

acetic acid waterfor Giemsa staining with Diff -Quik solution, 233

acetic alcoholfor nucleic acid fi xation, 8

acetic water, composition, 127acetic zinc formalin

in immunohistochemical studies, 280acetone

for clearing, 36comparative characteristics, 18for dehydration, 34as fi xative, 22–23in immunohistochemical studies, 279and protein hardening, 3

achromatic objectivesof microscope, 54

acidfor decalcifi cation, 47

acid alcoholfor Fite acid-fast staining technique, 228for Kinyoun acid-fast stain, 224for microwave auramine-rhodamine fl uorescence technique, 230for microwave variation of Ziehl-Neelsen method, 227stability of, 100for Ziehl-Neelsen method, 226

acid dyes. See anionic dyes., 108acid fast bacilli, troubleshooting mounting, 131acid fuchsin

stability of, 100for van Gieson staining, 167

acid fuchsin-picric acid, stability of, 100acid phosphatase reaction, marking of infl ammatory muscle cells with,

320–322, 322acid-fast bacilli, fl uorescent dyeing, 56acid-fast bacteria

demonstration with Fite staining technique, 228–230, 229demonstration with Kinyoun acid-fast stain, 224–226, 225demonstration with microwave variation of Ziehl-Neelsen method,

227–228, 228demonstration with Ziehl-Neelsen method, 226–228detection with microwave auramine-rhodamine fl uorescence

technique, 230–231, 231acid-fast stains

for demonstration of bacteria, 222, 224-231, 225acidic dichromate solution

for PTAH staining without mercuric solutions, 181acidic potassium permanganate solution

for PTAH staining without mercuric solutions, 181acidifi ed water

for Churulian-Schenk method, 265acidophilia, 106acidulated water

for Warthin-Starry technique, 244acquired immunodefi ciency syndrome (AIDS). See AIDS, 82Actinomyces, demonstration with Brown-Hopps method, 233additive, fi xation, 2adenocarcinoma

Cam 5.2 staining of colon, 291subtypes, 290

adenosine triphosphatase (ATPase)demonstration of muscle types with, 319muscle atrophy demonstration, 320reaction in muscle, 320diff erentiation of muscle types and diseases with, 318–320, 319, 320pH histochemical reactions of human muscle types, 309

Index

Histotechnology 3rd Edition 373

adhesion problemswith smears, 358

adhesivesfor slide mounting, 69

adipose cellsconnective tissue types and structures, 160 processing with water-soluble waxes, 40

AECfor peroxidase-antiperoxidase (PAP) immunohistochemical

technique, 298, 300aerosols, infectious hazards from, 82–83agar

embedding with, 41for slide mounting, 69

agar method, loose cellular material prepared with, 360–361, 361agranular endoplasmic reticulum, ultrastructure of, 105AIDS (acquired immunodefi ciency syndrome), infectious hazards, 82air bubbles

in liver section, 185trapped under section, 69, 71

air dryingof small intestine, 73

albuminexcess, on slide, 71for slide mounting, 69

albumin method, loose cellular material prepared with, 360, 361alcian blue staining, 145-149

demonstration of C neoformans and B dermatitidis with, 244epithelial and connective tissue mucin demonstration, 147–148for Russell modifi cation of the Movat pentachrome stain, 172hyaluronidase digestion and, 147, 148mold demonstration with, 223mucopolysaccharide demonstration with, 145–146on intestinal specimen, 69pH 1.0, 146, 147pH 2.5, 145, 146stability of, 100sulfated mucosubstance demonstration with, 146–147with PAS and hematoxylin, 148, 149

alcian yellow solutionfor alcian yellow-toluidine blue staining method, 234

alcian yellow–toluidine blue staining method, 234–235, 235demonstration of H pylori, 235

alcohol, absolute, 40rapid processing with, 38

alcoholas fi xative, 22–23in immunohistochemical studies, 279for Luxol fast blue-cresyl echt violet staining, 214

alcohol fi xation, 66with incomplete formalin fi xation, 281

alcohol, acid, stability of, 100alcohol, saline, preparation for alkaline Congo red staining, 153alcoholic formalin, 13

overnight processing with, 38alcoholic hematoxylin

for Verhoeff elastic stain, 168for Weil staining method, 211

alcoholic safran solutionfor Russell modifi cation of the Movat pentachrome stain, 173

alcoholic salinefor prefi xation, 354

alcoholic uranyl nitratefor Dieterle staining method, 247

alcoholic zinc chloride formalin, formulation and applications, 22alcohols

for dehydration, 33–34denatured (or reagent), 33for fi xation, 3overnight processing with, 38rapid processing with, 38See also specifi c types.

aldehyde fuchsin elastic stainfor pathologies of elastic tissue, 170, 171

aldehyde fuchsin solutionfor aldehyde fuchsin elastic stain, 170, 171for Gridley staining, 238

aldehyde fuchsin, stability of, 100aldehydes, for fi xation, 3aliphatic hydrocarbons

attribute comparisons, 37for clearing, 36

Alipia felis, demonstration with microwave variation of Warthin-Starry technique, 246, 247

alizarin red, stability of, 100alizarin red calcium stain

calcium demonstrated in kidney and artery with, 271calcium demonstration with, 270, 271

alkaline alcohol solutionfor Russell modifi cation of the Movat pentachrome stain, 172

alkaline Congo red, staining of amyloid, 152–154alkaline phosphatase reaction

for immunohistochemical studies, 283in muscle fi ber regeneration, 322–323, 323

alkaline phosphatase red chromogenbreakdown of, 283on melanoma of the eye, 283

alkaline salt solution, preparation for alkaline Congo red staining, 153alkanes

for clearing, 36allergic reactions, diagnosis with toluidine blue, 188α-naphthyl acetate esterase stain

for muscle biopsies, 314–316, 316for skeletal muscle, 316

α-naphthyl acetate in acetonefor α-naphthyl acetate esterase stain, 315

alum, defi ned, 110alveolar sacs, fi xation illustrations, 6Alzheimer disease

demonstration with Bielschowsky-PAS staining, 202–204, 204demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 204–205, 205demonstration with thiofl avin S staining, 205–207, 207diagnosis with Bielschowsky-PAS staining, 200–205, 204senile plaques demonstrated, 204, 207

American Society of Clinical Oncologyguidelines for HER2 testing, 6, 67

fi xation time, 6microwave processing, 67

amino acids, pH of, 107amino group

and cytoplasmic staining, 107and fi xatives, 3and stain charge, 10

374 Index

aminoalkylsilanecoating on slides, 70for slide mounting, 69

ammoniacal silver, explosive hazard, 86ammoniacal silver solution

for Bielschowsky-PAS staining, 201for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178for Sevier-Munger modifi cation of Bielschowsky method, 204

ammonium bromide formalin, formulation, 12ammonium hydroxide

for microwave variation of Bielschowsky-PAS staining, 203for testing of decalcifi cation, 48

ammonium oxalatefor testing of decalcifi cation, 48

ammonium sulfi de solutionfor ATPase stain, 319

amyloidin crystal-violet-stained kidney section, 155demonstration with Bielschowsky-PAS staining, 202demonstration with thiofl avin S staining, 207demonstration with thiofl avin T, 155–156, 155fl uorescent dyeing, 56in kidney specimen, 55in kidney stained with thiofl avin T technique, 156staining, 152–156, 153, 155staining with Congo red, 153staining with crystal violet, 154–155, 155

anaplasia, defi ned, 289, 374anaplastic tumor workup, 290angle, bevel, 59angle, clearance, 59

of microtomes, 58and ribbon formation problems, 60

anhydrous substances, percentage solutions of, 96aniline blue

for Bodian staining method, 198for counterstaining with Bodian method, 198for Masson trichrome staining, 163overstaining of cortex with, 199stability of, 100

aniline dyes, 108anionic dyes, 108

diff erentiation with, 109anionic heteroglycans, types, occurrence, and reactivity, 136anions. See ions.antibodies

defi ned and described, 278, 366direct immunohistochemical staining with, 284evaluation and validation, 285–289for peroxidase-antiperoxidase (PAP) immunohistochemical

technique, 298quality control for, 290–291, 293storage considerations, 285–286, 287universal (multilink), 289validation form, 288and zinc formalin fi xation, 16–17

antibody control log, 293antigenicity restoration

in immunohistochemical studies, 280

antigensdefi ned and described, 278, 366for indirect immunohistochemical testing, 284preservation with zinc sulfate fi xation, 16zinc formalin fi xation advantages, 21

anti-roll plateof cryostat, 65

aortaincorrect orientation of, 45Verhoeff -van Gieson staining of, 169

apathy mounting medium, 154, 156with crystal violet staining, 154

apochromatic objectivesof microscope, 54

appendixhematoxylin and eosin staining of, 115specimen orientation, 43, 44

applicator stickfor embedding, 339for ribbon handling, 70specimen preparation with, 360

aqua regiafor Bodian staining method, 197

aqueous fi xatives, 9–17aqueous formalin, formulation, 12aqueous mounting media, 129aqueous phosphomolybdic acid

for Holzer staining method, 209aqueous silver nitrate

for Bielschowsky-PAS staining, 201for Holmes silver nitrate staining method, 199for Luxol fast blue-Holmes silver nitrate staining, 216

aqueous solutionsfor fi xation, 3

Araldite, embedding with, 41argentaffi n cells, 5, 262argentaffi n granules

in colon and gastrointestinal tract stained with Schmorl technique, 260

demonstrated with Schmorl technique, 259–260demonstration with Churukian-Schenk method, 266demonstration with Fontana-Masson stain, 260–263, 262, 263demonstration with Grimelius stain, 264demonstration with microwave variation of Churukian-Schenk

method, 267demonstration with microwave variation of Fontana-Masson stain,

261–263, 263argyrophil granules, 263-267, 263–264, 266

demonstration with Churukian-Schenk method, 265–267demonstration with microwave variation of Churukian-Schenk

method, 267in pancreas stained with microwave Churukian-Schenk method, 267tumor demonstration with Grimelius stain, 264

artery, calcifi cation demonstrated with Dahl procedure, 271artery, Verhoeff -van Gieson staining of, 169asbestos birefringence, 254aspirations, fi ne needle

collection and handling of, 353fi ne needle, smear preparation, 357


astrocytes 194, 210-211demonstration with Cajal stain, 210–211, 210

ATPase. See adenosine triphosphatase (ATPase).atrophy, of muscle, demonstration with ATPase, 320auramine O-rhodamine B solution

stability of, 100for microwave auramine-rhodamine fl uorescence technique, 230

auramine-rhodamine, fl uorescent dyeing, 56auramine-rhodamine fl uorescence technique,

microwave variation, for mycobacteria detection, 230–231, 231autofl uorescence, 56autoimmune disorders, diagnosis with toluidine blue, 188autolysis

fi xation troubleshooting, 24–26, 25Mayer mucicarmine staining aft er, 144of tissue specimens, 2, 5

autonomic nervous system, 194autopsies

and fi xation, 5auxochrome, role in staining, 108aviation gasoline

as clearing agent, 37avidin-biotin complex-immunoperoxidase procedure, 299–301avidin-biotin enzyme complex method

of immunohistochemical staining, 284, 285axons 194, 367

demonstration in Luxol fast blue-Holmes silver nitrate staining, 217demonstration of degeneration with Luxol fast blue, 212–213demonstration with Bielschowsky-PAS staining, 202, 204demonstration with Holmes silver nitrate method, 200

BB-5 solution, B-5 vs formalin fi xation, 19

and decalcifi cation testing, 48for fi xation, 16formulation and applications, 17–19in immunohistochemical studies, 280

bacillidemonstration with Brown-Hopps method, 232demonstration with Giemsa staining, 234

bacilli, acid fast, fl uorescent dyeing, 56bacteria, 222, 367

acid-fast, 222, 224–231demonstration with Fite staining technique, 228–230demonstration with Kinyoun acid-fast stain, 224–226, 225demonstration with microwave variation of Ziehl-Neelsen method,

227–228, 229demonstration with Ziehl-Neelsen method, 226–228, 228detection with microwave auramine-rhodamine fl uorescence

technique, 230–231, 231defi ned and classifi ed, 222demonstration through staining, 222demonstration with Dieterle staining method, 248, 249demonstration with Giemsa staining, 233–234, 234demonstration with Kinyoun acid-fast stain, 224–226, 225demonstration with microwave variation of Warthin-Starry

technique, 245–247, 247demonstration with Warthin-Starry technique, 245fi xative eff ect on, 2gram-negative, demonstration with Brown-Hopps method, 231–233,

232gram-positive

demonstration with Brown-Hopps method, 231–233, 232, 233

bag constructionfor cell suspensions, 347

balances, 74balsam-xylene mixture

for cresyl echt violet staining, 195barbital acetate

for ATPase stain, 318basement membranes, 161

demonstration with Luxol fast blue-PAS-hematoxylin staining, 218–219

demonstration with periodic acid-methenamine silver procedure, 183

PAS reaction testing for, 137–141perivascular feet of astrocytes on, 210staining of, 182–185, 183, 184 Schiff reaction to stain, 201separation from by autolysis, 5

basic (cationic) dyeing, 107, 108, 109basic fuchsin

for aldehyde fuchsin elastic stain, 171for Brown-Hopps modifi cation of gram staining, 232stability of, 100

basophiliaof plasma cell cytoplasm, 106

batch controlsand quality control, 296

baths, fl otation, 69–71benign, defi ned, 289benzene

fl ash point, 87for clearing, 35

benzoyl peroxide, explosive hazard, 86Best carmine, glycogen demonstration, 141Biebrich scarlet-acid fuchsin solution

for Masson trichrome staining, 163Bielschowsky-PAS staining, 200–205

demonstration of plaques in cortex section, 204of nerve fi bers and neurofi brillary plaques and tangles, 200–205Sevier-Munger modifi cation, 204–205

bile stain, 268–269bilirubin, 255

demonstration with bile stain, 268–269, 269Fouchet action on, 269

biliverdin, 255from bilirubin oxidation by Fouchet reagent, 269

biohazards. See also biological hazards.symbol, 90

biological hazards, 82–83biological waste disposal, 83birefringence

aft er alkaline Congo red staining, 152, 153bis-chloromethyl ether

for decalcifi cation, 47black acid hematin, 11

See also formalin pigment.

376 Index

bladesartifacts caused by, 59disposable, 58high- and low-profi le, 65microtome, 58

Blastomyces dermatitidisdemonstration with Gridley staining, 239demonstration with Mayer mucicarmine technique and alcian blue

staining, 244block holder

adjustment of, 60overextension of, 63and washboarding of section, 61, 62

blocking (embedding), 359–361for electron microscopy, 340

blocking reactions, 287blood cell, preparation for electron miscroscopy, 346–347blood smears, methyl alcohol fi xation for, 34blood vessels, demonstration with Luxol fast blue-PAS-hematoxylin

staining, 219bloodborne pathogen standard, infectious hazards, 82bloody specimens, smear preparation, 357–358Bodian method

for staining nerve tissues, 197–198white and gray matter demonstration, 198

body fl uidscollection and handling of, 352osmolality of, 7–8

body substance isolation, infectious hazards, 82bone

decalcifi ed, 48plastic embedding, 41specimen orientation, 43undecalcifi ed, 48–49

bone dust, troubleshooting, 49bone marrow

Giemsa staining of, 128iron demonstration with Prussian blue, 258plastic embedding, 41section thickness for, 58

boraxfor Grocott staining, 240for microwave variation of methenamine-silver nitrate staining, 242for periodic acid-methenamine silver microwave procedure, 182

borax solutionfor Holmes silver nitrate staining method, 199for Luxol fast blue-Holmes silver nitrate staining, 216

boric acid solutionfor Holmes silver nitrate staining method, 199for Luxol fast blue-Holmes silver nitrate staining, 216

Bouin solutionand dye binding, 109and fi xative selection, 7formulation and applications, 19for Gomori 1-step trichrome stain, 165in immunohistochemical studies, 279– 280for Masson trichrome staining, 163protein fi xation, 4specimen removal from, 7

brain specimensacetone fi xation for, 22autolysis in, 2cryostat processing temperature, 64holes caused by knife, 60–61ice crystal artifact in, 51infectious hazards from, 83precipitate in, 121SMI-31 staining pattern, 287specimen orientation, 43

breast, adenocarcinoma in, 290breast cancer, guidelines for HER2 testing, 6breast discharges

collection and handling of, 352smear adhesion problems, 358

Brij-35in fl otation bath processing, 70

bronchial specimens, collection and handling of, 352bronchiole, fi xation illustrations, 6Brown-Hopps modifi cation

Actinomyces demonstration with, 233of gram staining, 231–233S aureus, E coli, and liver disease demonstration, 232

bubbles, artifact in liver section, 185bubbling, nuclear

as drying artifact, 73from excessive heat, 71fi xation troubleshooting, 24–26, 26from formalin fi xation, 6

buff er solutions, 289defi ned, 98for epithelial and connective tissue mucin demonstration, 147pH monitoring, 295

buff ered PAF, for electron microscopy, 336buff ered substrate solution, for HRP enzyme labeled polymer

procedure, 302butanol (butyl alcohol)

for clearing, 36for dehydration, 34

CCajal stain, astrocyte demonstration with, 210–211, 210calcium

demonstrated with von Kossa stain, 270demonstration in artery with Dahl procedure, 271demonstration with alizarin red, 270–271, 271demonstration with Gomori methenamine-silver technique, 268demonstration with von Kossa technique, 269–270knife line caused by, 59

calcium chloridefor ATPase stain, 319

calcium-formalin solution, 12for Sudan black B staining, 186

Cam 5.2adenocarcinoma staining on colon, 291anaplastic tumor workup, 290normal colon staining, 291

cancer, demonstration on prostate with P504S antibody, 302CAP. See College of American Pathologists.


carbohydratesfi xative comparative characteristics, 18fi xative eff ect on, 2, 9and formaldehyde fi xation, 10and glyoxal fi xation, 14types, characteristics, and staining methods, 136–152

carbol fuchsin solutionfor microwave variation of Ziehl-Neelsen method, 227stability of, 100for Ziehl-Neelsen method, 226

carbon bisulfi deas clearing agent, 37

carbon pigment, 254carbon tetrachloride

as clearing agent, 37Carbowax

for processing, 40carboxyl group

and cytoplasmic staining, 107staining and, 3

carcinogens, 86carcinoid tumors

demonstration with Fontana-Masson stain, 260–263demonstration with Grimelius stain, 264demonstration with microwave variation of Fontana-Masson stain,

261–263carcinoma

anaplastic tumor workup, 290small cell neoplasm panel, 290

carmine solution, preparation for glycogen demonstration, 141Carnoy solution, 358

formulation and applications, 23in immunohistochemical studies, 279for nucleic acid fi xation, 8

carryover, of tissue, troubleshooting embedding, 46cationic dyeing, 107, 108, 109cations. See ions.CD20 B-cell, normal lymph node staining, 291CD3 T-cell, normal lymph node staining, 291CD45RB LCA, anaplastic tumor workup, 290CD56, small cell neoplasm panel, 290cedarwood, oil of

and celloidin embedding, 40for clearing, 36

celestine blue, composition, 113cell blocks, 359–361cell membranes, fat demonstration with oil red O, 184cell suspensions

bag construction for, 347preparation for electron miscroscopy, 347

celloidinembedding with, 40for overdehydrated specimens, 70

cellsfi xative eff ect on, 2reactions to fi xatives, 8ultrastructure of, 104–106, 104

cells, goblet. See goblet cells.Celsius, temperature, 97–98central nervous system, 194

celloidin embedding, 40chatter on section, 63demonstration with Luxol fast blue-PAS-hematoxylin staining, 219holes in section, 61knife lines in, 59processing with water-soluble waxes, 40

centriole, 104, 105, 106cerebellum

with Bodian stain and aniline blue counterstain, 198SMI-31 staining pattern, 287

cerebrospinal fl uids, collection and handling of, 352ceroid, 255cervix

demonstration of myelogenous leukemic cells in, 317stained with alcian blue-PAS technique, 149staining with Gill hematoxylin, 112

charge, electrostaticand slide mounting, 69

charge. See ions.chatter

on immunohistochemical section, 281on section, 61, 63

chelating agentsfor decalcifi cation, 47–48

chemicalsdisposal, 88 hazards, 85–91spills, 87 storage, 87, 88 testing of decalcifi cation, 48

chemical fi xation vs dehydration, 279chlorofl uorocarbons, for frozen sectioning, 50chloroform

and celloidin embedding, 40for clearing, 36

chloromas, demonstration with naphthol AS-D chloroacetate esterase technique, 316–317

chromaffi n granules, demonstration with microwave variation of Fontana-Masson stain, 263

chromatic aberration, of microscope, 54chromatin

as component of nucleus, 104demonstration with Pap staining, 363fi xation troubleshooting, 24–26formalin eff ect on, 8methyl green-pyronin Y staining, 126patterns with Giemsa staining, 128staining of, 105ultrastructure of, 104

chrome pigment artifact, 254chromic acid

for chromic acid-Schiff stain, 237for Gridley staining, 238for Grocott staining, 240for lipid fi xation, 8for microwave variation of methenamine-silver nitrate staining, 242stability of, 100

chromic acid-Schiff stain, demonstration of fungi with, 236–238chromidal substance, 194chromium potassium sulfate, coating on slides, 70chromium potassium sulfate alum, for slide mounting, 69

378 Index

chromogen, for immunohistochemical studies, 283chromophore, defi ned, 108Churulian-Schenk method, demonstration of argyrophil granules in

neurosecretory tumors, 265–267cilia, demonstration with Pap staining, 363circulating water bath

for temperature control of specimen, 72cirrhosis, diff erential diagnosis with Gordon and Sweets staining,

177–179citric acid

for Churulian-Schenk method, 265for microwave variation of Churukian-Schenk method, 266for Warthin-Starry technique, 244

CK20, adenocarcinoma panel, 290CK7, adenocarcinoma panel, 290clamping

of specimen, 61, 62Clark fi xative, 358Clarke fl uid, formulation and applications, 23clearance angle, 59, 342

of microtomes, 58and ribbon formation problems, 60

clearing, 35–37Clinical and Laboratory Standards Institute

and CAP accreditation, 292clots, specimen preparation with, 360clove, oil of, for clearing, 36coagulant fi xatives

eff ect on protein, 3, 4rate of penetration, 7

cobaltous chloridefor ATPase stain, 319

Coccidioides immitis, demonstration with Gridley staining, 239collagen, 160, 308

autofl uorescence in, 56demonstration with Gomori 1-step trichrome stain, 166demonstration with Gomori stain, 176demonstration with Mallory PTAH technique, 181demonstration with Masson trichrome staining, 163–165demonstration with Russell modifi cation of Movat pentachrome

stain, 172–174, 173diff erentiation with Gomori 1-step trichrome stain, 165–166poor diff erentiation of, 121swelling of, and pH, 9van Gieson staining, 167in Verhoeff -van Gieson staining of aorta and artery, 169

collection, of specimens, 352–353College of American Pathologists

guidelines for HER2 testing, 6laboratory accreditation, 292–296

colon, adenocarcinoma staining with Cam 5.2, 291argentaffi n demonstration with Schmorl technique, 260colloidal iron staining, 151demonstration with Russell modifi cation of Movat pentachrome

stain, 173lymphoma demonstrated with cytoplasmic IgA, 299normal Cam 5.2 staining, 291PAS staining of, 138stained with alcian blue at pH 1, 147staining with Congo red, 153

colorectal, adenocarcinoma, 290

columnar cells, of small intestine, Mayer mucicarmine staining of, 144compression, of section, 62, 63concentrated antibodies, storage considerations, 286concentration, of fi xative, and penetration rate, 7condenser, of microscope, 54Congo red

on kidney specimen, 55conidia, demonstration with Gridley staining, 239connective tissue, Gomori trichrome procedure with, 328–329connective tissue, mucin demonstration in, 147–148connective tissue, staining techniques, 188–189connective tissue, types, structures, and staining methods, 159–189control slides, storage of, 292controls

in immunohistochemical studies, 285positive and negative, on microarray cores, 291validation form, 288

copperdemonstration with microwave rhodanine method, 272–273, 273demonstration with rhodanine method, 271–273, 272

cork, for freezing muscle specimens, 313–314, 313corn-fl aking artifact, 130–131, 131corpora amylacea, demonstration with Luxol fast blue-PAS-hematoxylin

staining, 218–219corrosive substances, 86corrosivity, description, 88cortex

with Alzheimer disease, Bielschowsky-PAS-stained, 202overstained with aniline blue, 199

counterstainingwith Feulgen reaction, 124for Masson trichrome staining, 164

covalent bonding, 107coverslippers, automated, 68coverslipping, safety precautions, 84coverslips

compression artifact from, in liver section, 185mounting with, 129warping problems, 131

cresyl echt violet stainingcombined with Luxol fast blue, 214–215, 215of neurons, 196of Nissl substance, 195–197, 197

Creutzfeldt-Jakob disease, infectious hazards from, 83crocein scarlet-acid fuchsin solution

for Russell modifi cation of the Movat pentachrome stain, 173cross contamination

of cytology specimens, 364crosshatch method

of smear preparation, 354, 355cross-linking

by formaldehyde fi xation, 10with glutaraldehyde fi xation, 13with glyoxal fi xation, 14zinc formalin fi xation advantages, 21

crush methodof smear preparation, 354, 355, 356

cryogenic sprays, infectious hazards from, 82–83cryostat, 64–65

for frozen sectioning, 50maintenance of, 65


Cryptococcus neoformansdemonstration with Mayer mucicarmine technique, 143, 144demonstration with Mayer mucicarmine technique and alcian blue

staining, 244demonstration with microwave Müller-Mowry colloidal iron

technique, 151demonstration with Müller-Mowry colloidal iron technique, 151demonstration with Russell modifi cation of Movat pentachrome

stain, 174crypts of Lieberkühn, 5crystal size

and embedding, 44crystal violet

for Brown-Hopps modifi cation of gram staining, 231kidney stained with, 155for rapid amyloid screening, 154–155stability of, 100

cubic centimeter, defi ned, 97cumulative trauma disorders, 84CyclinD1 mouse monoclonal antibody staining vs CyclinD1 rabbit

monoclonal antibody staining, 279cysts, specimen orientation, 43cytocentrifuge preparation of smears, 354, 357, 358cytokeratin AE1/AE3, anaplastic tumor workup, 290cytology

gynecological, collection and handling of, 352liquid-based, 358–359nongynecological, collection and handling of, 352staining, 361

cytoplasm, demonstration with Diff -Quik Giemsa staining, 234demonstration with Gomori 1-step trichrome stain, 166demonstration with Masson trichrome staining, 163–165demonstration with Müller-Mowry colloidal iron technique, 151demonstration with Pap staining, 363methyl green-pyronin Y staining, 188–189, 189in nerve cell, 194staining, 107–108troubleshooting hematoxylin and eosin staining, 120ultrastructural components, 105–106van Gieson staining, 167

cytoplasmic granules, 256cytoplasmic IgA

in colon lymphoma using PAP technique, 299cytoplasmic neurofi brils, demonstration with Bielschowsky-PAS

staining, 204cytopreparatory techniques, 351–365, 359cytospin preparations, microwave variation of methenamine-silver

nitrate staining for, 242–244

DDAB (3,3'-diaminobenzidine chromogen)

demonstration of cancer on prostate, 302for HRP enzyme labeled polymer procedure, 302trapped beneath specimen tissue, 281

DAB reaction, 289Dahl alizarin red procedure, demonstration of calcium in artery with, 271dealcoholization. See clearing.decalcifi cation, 46–50

in bone specimen, 48electrolytic, 47safety precautions, 47testing, 48troubleshooting, 49–50

decayof tissue specimens, 2

dehydration, 33–35. See also drying.accidental, troubleshooting, 42vs chemical fi xation, 279for electron microscopy, 337incomplete, 122troubleshooting, 41–42

dehydrogenases, 312Delafi eld hematoxylin, composition, 111demineralization, 46–51denaturation

of protein, by fi xatives, 2denatured alcohol, 33dendrites, 194denervation

of muscle tissue, demonstration with α-naphthyl acetate esterase stain, 316

denervationof muscle tissue, 316

deoxyribonucleic acid. See DNA.deparaffi nation

and hematoxylin and eosin staining, 118dermis,

connective tissue types and structures, 160demonstration with Gomori aldehyde fuchsin procedure, 171

desiccation. See dehydration; drying.desmin, small cell neoplasm panel, 290developer solution

for Bielschowsky-PAS staining, 201for Dieterle staining method, 248for microwave variation of Bielschowsky-PAS staining, 203for Warthin-Starry technique, 245

diagnosis, 352diamond knives

for electron microscopy, 341–342diaphorases, 312diaphragm, iris, of microscope, 54diastase digestion

with PAS reaction, 139–141diastase of malt

preparation for PAS reaction with diastase digestion, 140stability of, 100

Dieterle staining methoddemonstration of L pneumophila with, 247–249demonstration of spirochetes with, 247–248

diff erentiating solutionfor Holzer staining method, 209for Weil staining method, 211

diff erentiationdefi ned, 289staining methods for, 109troubleshooting hematoxylin and eosin staining, 120–121, 121

Diff -Quik Giemsa stain, demonstration of H pylori, 233–234, 234digestion, proteolytic enzyme, 282dilution

of acids, 9percentage calculations, 94–97

dioxanefor clearing, 36for dehydration, 34

380 Index

disposalof biological waste, 83of chemicals, 88xylene, limonene, and aliphatic carbon comparisons, 37

DNAdemonstration with Feulgen reaction, 123methyl green-pyronin Y staining, 126,and picric acid fi xation, 15and potassium dichromate fi xation, 16reactions to fi xatives, 8and zinc formalin fi xation, 16–17

documentation, and quality control, 289–290Drierite, 33dry ice, for frozen sectioning, 50, 65dryers, 71drying, as fi xation method, 3. See also dehydration.duration, of fi xation, for electron microscopy, 335dye content, gravimetric factor and, 95dyes

fi xative eff ect on, 2for staining, 108–116

dystrophic neurites, demonstration with Bielschowsky-PAS staining, 202

EEA counterstain, 362

for Pap staining, 363ectocervix, glycogen demonstration in alcian blue-PAS technique, 149egg white, fi xative eff ect on, 3, 4Ehrlich hematoxylin, composition, 111EIER. See enzyme-induced epitope retrieval.elastic fi bers

connective tissue types and structures, 160demonstration with Gridley staining, 239

elastic tissuedemonstration with aldehyde fuchsin elastic stain, 171demonstration with alkaline Congo red staining, 153demonstration with Gomori aldehyde fuchsin procedure, 171demonstration with Russell modifi cation of Movat pentachrome

stain, 172–174, 173electrocautery, and heat artifact, 122electrolytic decalcifi cation, 47electron microscopy, 333–350

fi xative comparative characteristics, 18and glutaraldehyde fi xation, 13–14

electrostatically charged, slides, 69Elmer’s glue, for slide mounting, 69embedding, 37–41, 42–46

applicator stick for, 339centers, 74–75Epon, 339–340media for electron microscopy, 337–340safety precautions, 84Spurr, 337–339troubleshooting, 44–46

emboli, fatty, oil red O detection of, 184endocervix, mucin demonstration in alcian blue-PAS technique, 149endocrine cells

Bouin solution for, 19fi xation with Hollande solution, 20

endogenous deposits, 255–256

endogenous pigments, classes and formation, 254endometrial scrapings, cryostat processing temperature, 64endometrium, fi xation troubleshooting, 25endoplasmic reticulum, 106endospores, demonstration with Gridley staining, 239enzyme digestion, proteolytic, 282enzyme histochemistry, 307–332enzyme immune complex immunohistochemical staining, 284enzyme immunohistochemistry, 283enzyme-induced epitope retrieval, 282–283enzymes

attack of specimens, 2classifi cation of, 310–312fi xative comparative characteristics, 18fi xative eff ect on, 2preservation of, 310properties of, 310–312

eosinand amino acid staining, 10counterstain on intestine, 114formulas for, 113and potassium dichromate fi xation, 16

eosin Yfor Pap staining, 363stability of, 100

eosinophilfi xation with Hollande solution, 20formalin fi xation, 11

eosinophilia, 106eosin-phloxine counterstain

composition, 114overstaining, 121

ependymal cells, 195epidermis, demonstration with Gomori aldehyde fuchsin procedure, 171epithelial cells

mucin demonstration in, 147–148ultrastructure of, 104

epitheliumcorn-fl aking artifact on, 131defi ned and illustrated, 5fi xation troubleshooting, 25of lung, 6tumor demonstration with Grimelius stain, 264

epitope, 278retrieval, 281–283

Epon embedding, 41, 339epoxy resin block, 41

for electron microscopy, 340ergonomics, 84eriochrome black

for microwave auramine-rhodamine fl uorescence technique, 230erythrocytes

Carnoy solution lysing of, 23demonstration with microwave variation of Ziehl-Neelsen method,

228demonstration with microwave variation of Warthin-Starry

technique, 247and endogenous pigments, 255poor diff erentiation of, 121Zenker solution lysing of, 20


Escherichia coli, demonstration with Brown-Hopps method, 232esophageal specimens, collection and handling of, 352essential oils, for clearing, 36esterase solution, 311

for naphthol AS-D chloroacetate esterase technique, 316ethanol. See ethyl alcohol.ether

and celloidin embedding, 40ether, bis-chloromethyl

for decalcifi cation, 47ethyl alcohol

in Best carmine demonstration of glycogen, 142comparative characteristics, 18for dehydration, 33and dye binding, 109eff ect on protein, 3, 8fl ash point, 87rate of penetration, 7

euchromatin, ultrastructure of, 104evaluation, of antibodies, 285–286evaporation, xylene, limonene, and aliphatic carbon comparisons, 37examinations

and quality control, 294exogenous pigments, 254explosion hazard, 16, 86eyepiece, of microscope, 54

FFahrenheit, temperature, 97fallopian tube

fi xation troubleshooting, 26good vs poor diff erential staining, 122incorrect orientation of, 45specimen orientation, 43, 44troubleshooting hematoxylin and eosin staining, 119, 120

fast greenfor chromic acid-Schiff stain, 237for Hotchkiss-McManus PAS reaction, 236

fast-twitch muscle, 308fats

demonstration with oil red O, 185demonstration with osmium tetroxide paraffi n procedure, 187demonstration with Sudan black B, 186frozen sectioning, 49–50

fats. See also lipids.fatty tissue, incompletely processed, 66ferric ammonium sulfate

for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178for Weil staining method, 211

ferric chloridefor bile stain, 269preparation for Müller-Mowry colloidal iron technique, 150for Russell modifi cation of the Movat pentachrome stain, 172for Schmorl technique, 259stability of, 100for Verhoeff elastic stain, 168

ferric ions, absorption by carboxylated and sulfated mucosubstances, 149

ferric ironin bone marrow section stained with Prussian blue, 258demonstration with Prussian blue stain, 256–258

ferrous iron, demonstration with Turnbull blue stain, 258–259in spleen stained with Turnbull blue, 259

Feulgen reaction, 123–124, 124

fi brindemonstration with Mallory PTAH technique, 179–181demonstration with Russell modifi cation of Movat pentachrome

stain, 172–174, 173fi brinoid, demonstration with Russell modifi cation of Movat

pentachrome stain, 173

fi broblasts, connective tissue types and structures, 160

fi lters, in fl uorescence microscope, 56

fi ne needle aspirationscollection and handling of, 353smear preparation, 357

fi re, classifi cation, 87

fi re, hazards, 86

fi re class, xylene, limonene, and aliphatic carbon comparisons, 37

fi re extinguishers, 86

FITC (fl uorescein isothiocyanate), 56direct immunohistochemical staining with, 284for immunohistochemical studies, 283

Fite acid-fast staining technique, demonstration of mycobacteria with, 228–230

fi xation, 1–29defi nition, 2for diagnostic cytology, 353–354with electron microscopy, 334–337microwave, 2troubleshooting, 24–26chemical vs dehydration, 279for immunohistochemical studies, 279–280and quality control, 294

fi xatives, eff ect on protein, 8additive, 2, 3, 4for alcian blue technique, 145–147for alcian blue-PAS-hematoxylin technique, 148for alcian yellow-toluidine blue staining method, 234for aldehyde fuchsin elastic stain, 170for alizarin red calcium stain, 270for alkaline red staining, 152for α-naphthyl acetate esterase stain, 315for Best carmine demonstration of glycogen, 141for Bielschowsky-PAS staining, 201for bile stain, 268for Bodian staining method, 197for Brown-Hopps modifi cation of gram staining, 231cell reactions to, 8

382 Index

fi xatives. See also specifi c types.aqueous, 9–18choosing, 7for chromic acid-Schiff stain, 237for Churulian-Schenk method, 265coagulant, 3, 4

eff ect on protein, 8rate of penetration, 7

comparative characteristics, 18for cresyl echt violet staining, 195, 196for crystal violet staining, 154defi nition, 2for Dieterle staining method, 247for electron microscopy, 334–337factors aff ecting, 4–8for Fite acid-fast staining technique, 228for Fontana-Masson stain preparation, 261for Giemsa staining with Diff -Quik solution, 233for Gomori 1-step trichrome stain, 165for Gomori methenamine-silver technique for urates, 267for Gomori staining of reticulin, 174for Gordon and Sweets staining technique, 178for Gridley staining, 238for Grimelius argyrophil stain, 263for Grocott staining, 239for Holmes silver nitrate staining method, 199for Holzer staining method, 208for Hotchkiss-McManus PAS reaction, 235for HRP enzyme labeled polymer procedure, 301in immunohistochemical studies, 279–280for Kinyoun acid-fast stain, 224for Luxol fast blue staining, 213for Luxol fast blue-cresyl echt violet staining, 214for Luxol fast blue-Holmes silver nitrate staining, 215for Luxol fast blue-PAS-hematoxylin staining, 218for Mallory PTAH technique, 180, 207for Masson trichrome staining, 162, 164for Mayer mucicarmine technique, 142for microwave auramine-rhodamine fl uorescence technique, 230for microwave variation of Bielschowsky-PAS staining, 202for microwave variation of Churukian-Schenk method, 266for microwave variation of Fontana-Masson stain, 262for microwave variation of methenamine-silver nitrate staining, 242for microwave variation of rhodanine method, 273for microwave variation of Warthin-Starry technique, 245for microwave variation of Ziehl-Neelsen method, 227for mineral deposit demonstration, 256for Müller-Mowry colloidal iron technique, 150for naphthol AS-D chloroacetate esterase technique, 316nonadditive, 2, 8noncoagulant, 3, 4for oil red O lipid demonstration, 184for osmium tetroxide paraffi n procedure, 187for Pap staining, 362for PAS reaction testing, 137for periodic acid-methenamine silver microwave procedure, 182for peroxidase-antiperoxidase (PAP) immunohistochemical

technique, 297, 299for phosphorylase stain, 326for Prussian blue stain preparation, 257for rhodanine copper demonstration, 272for Russell modifi cation of the Movat pentachrome stain, 172for Schmorl technique, 259for Steiner and Steiner procedure, 249for Sudan black B staining, 186

for thiofl avin S demonstration of Alzheimer disease, 205for thiofl avin T technique, 155for toluidine blue wet fi lm technique, 364for Turnbull blue stain preparation, 258urate crystal preservation with absolute alcohol, 256for Verhoeff elastic stain, 168for von Kossa calcium stain, 269for Warthin-Starry technique, 244for Weil staining method, 211for Ziehl-Neelsen method, 226

fl ammable materialclassifi cation, 87symbol, 90

fl ash points, 87fl otation, of section

and water-soluble waxes, 40fl otation baths, 69–71fl uids

preparation of, 355transitional

and plastic embedding, 41fl uorescein isothiocyanate (FITC), 56

direct immunohistochemical staining with, 284for immunohistochemical studies, 283

fl uorescence, 56fl uorocarbon sprays, and parched earth artifact, 69fl uorochrome, 56

for immunohistochemical studies, 283fl ying, of sections, troubleshooting, 64focus irregularities, troubleshooting mounting, 130folds, in section, from fl otation baths, 69Fontana-Masson stain, demonstration of cytoplasmic granules with,

260–263, 262forced air dryer, 72

overdehydration from, 73forceps artifact, compression artifact from, in liver section, 185forceps metastasis, 43formaldehyde, 9–11. See also formalin.

comparative characteristics, 18for decalcifi cation, 47and dye binding, 109in immunohistochemical studies, 279morphologic preservation with, 17for nucleic acid fi xation, 8with phosphate buff er, for electron microscopy, 335rate of penetration, 7warning sign, 85

formaldehyde standard, 85–86formaldehyde-glutaraldehyde, for electron microscopy, 336formalin. See also formaldehyde.

acetate, 11, 12alcoholic, 13, 38

and protein hardening, 4ammonium bromide, formulation, 12aqueous, formulation, 12calcium, formulation, 12fi xation of gastrointestinal tract, kidney, and spleen, 10and fi xative selection, 7for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178in immunohistochemical studies, 280


incomplete fi xation on lymph node, 281and lipid demonstration in liver section, 185Millonig, 11, 12neutral buff ered, 11

and bubbling artifact, 6fi xation with, 11formulation, 12in immunological studies, 280and lymphoma fi xation, 19and protein hardening, 4and tissue storage, 7

nuclear bubbling from, 8overnight processing with, 38personal protective equipment with, 13pigment, 11, 12safety precautions, 13saline, formulation, 12for Sevier-Munger modifi cation of Bielschowsky method, 204stability of, 100storage of, 13unsuitability for immunohistochemistry, 279zinc, and decalcifi cation testing, 48zinc, for immunohistochemical studies, 16–17

formalin pigment, 10, 254demonstrated with Schmorl technique, 259on Gomori-stained muscle, 329

formaline-saline solutionfor succinic dehydrogenase (SDH) reaction, 326

formic acidfor decalcifi cation, 47for Dieterle staining method, 248

formulasfor percentage solutions, 94–95

Fouchet reagentaction on bilirubin, 269for bile stain, 269stability of, 100

freeze thaw artifactfrom frozen sectioning, 65

freezers, for reagent storage, 72freezing

of antibody stocks, 286of muscle biopsy specimens, 312–314, 313, 314

freezing artifact, troubleshooting, 50–51, 50, 51frozen sectioning, 47, 49–50, 50

for immunohistochemical studies, 279troubleshooting, 50–51, 50

frozen sections, staining, 117–118fuchsin, acid, stability of, 100fuchsin, basic

for aldehyde fuchsin elastic stain, 171stability of, 100

fuchsin, carbol, stability of, 100fungi

defi ned and classifi ed, 222–223demonstration with chromic acid-Schiff stain, 236–238demonstration with Gridley staining, 238demonstration with Grocott staining, 239–242, 241demonstration with Hotchkiss-McManus PAS reaction, 235–236, 236demonstration with Luxol fast blue-PAS-hematoxylin staining,

218–219

demonstration with microwave variation of methenamine-silver nitrate staining, 242–244, 243, 244

pathologic, stains for demonstrating, 223

Ggall bladder, specimen orientation, 43Gallego solution

for Brown-Hopps modifi cation of gram staining, 232gasoline, aviation

as clearing agent, 37gastric section

with microwave variation of Warthin-Starry technique, 247with Steiner and Steiner microwave method, 250

gastric specimens, collection and handling of, 352gastrointestinal tract, argentaffi n demonstration with Schmorl

technique, 260Bouin solution for, 19fi xation troubleshooting, 24–26, 25fi xation with Hollande solution, 20fi xed in formalin, 10incorrect orientation of, 45muscle of, 162specimen orientation, 43, 44Zenker solution fi xation of, 21

gelatinembedding with, 41in fl otation bath processing, 70for microwave variation of Warthin-Starry technique, 246for slide mounting, 69for Warthin-Starry technique, 245

Gendre solution, formulation and applications, 19GFAP, anaplastic tumor workup, 290Giemsa stains

application and procedure, 127–128on bone marrow, 128composition, 127for demonstration of bacteria, 222for demonstration of bacteria, rickettsia, and T gondii, 233–234stability of, 100

Gill hematoxylincervix stained with, 112composition, 112for Pap staining, 361

glass knivesfor electron microscopy, 341

glia, 194–195cresyl echt violet staining, 196demonstration with Holzer method, 208–209, 209demonstration with Mallory PTAH technique, 207–208, 208

glial fi bers, demonstration with Mallory PTAH technique, 179–181glioma, anaplastic tumor workup, 290gliosis

demonstration with Holzer method, 208–209demonstration with Mallory PTAH technique, 207–208

glomerular basement membrane, detection with periodic acid-methenamine silver microwave procedure, 182

glomerulithrough electron microscope, 57in kidney blocked for electron microscopy, 340in kidney stained with thiofl avine T technique, 156staining with periodic acid-methenamine procedure, 184

384 Index

glues, for slide mounting, 69glutaraldehyde, 13–14

comparative characteristics, 18phosphate-buff ered, formulation, 14safety precautions, 14specimen removal from, 7

glycinefor microwave variation of Churukian-Schenk method, 266

glycine-acetic acid solutionfor microwave variation of Warthin-Starry technique, 246

glycogenα-0 glucose unit, 136demonstration in cervix with alcian blue-PAS technique, 149demonstration with Best carmine, 141, 142demonstration with PAS reaction and diastase digestion, 141PAS reaction with diastase digestion, 139–141and picric acid fi xation, 15storage sites and staining methods, 136–152trapping with formaldehyde, 10

glycol methacrylateembedding with, 41for undecalcifi ed bone, 48–49

glycolipids, types, occurrence, and reactivity, 136–137glycoproteins,

demonstration with Müller-Mowry colloidal iron technique, 149–152PAS reaction, 138stained with alcian blue at pH 1, 147types, occurrence, and reactivity, 136

glyoxal, 14, 18GMA. See glycol methacrylategoblet cells, 5

argentaffi n demonstration with Schmorl technique, 260demonstration with Russell modifi cation of Movat pentachrome

stain, 173demonstration with Schmorl technique, 260Mayer mucicarmine staining of, 144methyl green-pyronin Y staining, 126staining with Gill hematoxylin, 112

gold chloridefor Bielschowsky-PAS staining, 201for Bodian staining method, 197for Fontana-Masson stain preparation, 261for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178for Grocott staining, 240for Holmes silver nitrate staining method, 200for Luxol fast blue-Holmes silver nitrate staining, 216for microwave variation of Fontana-Masson stain, 262for microwave variation of methenamine-silver nitrate staining, 243for periodic acid-methenamine silver microwave procedure, 182for periodic acid-methenamine silver procedure without

microwave, 183stability of, 100

Golgi complex, 104, 106Gomori 1-step trichrome stain

for collagen and smooth muscle diff erentiation, 165–166Gomori aldehyde fuchsin procedure, skin stained with, 171Gomori methenamine-silver technique

urate crystal demonstration with, 256for urate demonstration, 267–268

Gomori stainfor reticular fi bers, 174–179reticulin demonstration in liver section, 176

Gomori trichromestability of, 100for detection of morphology changes in muscle and connective

tissue, 328–329for detection of morphology changes in muscle and connective

tissue, 329Gomori variation, oxidizer, sensitizer, impregnating solution, and

reducing solution, 175Gordon and Sweets variation

for diff erential diagnosis of liver diseases, 177–179oxidizer, sensitizer, impregnating solution, and reducing solution,

175reticulin demonstration in liver section, 179

gouty tophi, 255–256gram idodine

for Brown-Hopps modifi cation of gram staining, 231for Mallory PTAH technique, 180for phosphorylase stain, 327stability of, 100

gram molecular weight, 96gram stains

Actinomyces demonstration with, 233Brown-Hopps modifi cation, 231–233for demonstration of bacteria, 222mold demonstration with, 223S aureus, E coli, and liver disease demonstration, 232

gram-equivalent weight, 96gram-negative and gram-positive bacteria, demonstration with Brown-

Hopps method, 231–233, 232, 233granules, argentaffi n. See argentaffi n granules.granules, argyrophil, 263–264

demonstration with Churukian-Schenk method, 265–267demonstration with microwave variation of Churukian-Schenk

method, 267in pancreas stained with microwave Churukian-Schenk method, 267tumor demonstration with Grimelius stain, 264

granules, cytoplasmic, 256granules, neurosecretory

demonstration with Fontana-Masson stain, 260–263demonstration with microwave variation of Fontana-Masson stain,

261–263granulocytes

demonstration with Mayer hematoxylin, 317–318demonstration with naphthol AS-D chloroacetate esterase technique,

316, 317gravimetric factor, 95gray matter

demonstration with Bodian staining method, 198demonstration with Luxol fast blue, 213demonstration with Weil method, 212indistinct diff erentiation with Luxol fast blue, 214

green birefringenceaft er alkaline Congo red staining, 152, 153

Gridley stainingdemonstration of C immitis and B dermatitidis, 239demonstration of fungi with, 238mold demonstration with, 223

Grimelius argyrophil stain, 263–264tumor demonstration in intestine, 264

Grocott stainingdemonstration of fungi with, 239–242demonstration of P jirovecii, 243, 244H capsulatum demonstration with, 241mold demonstration with, 223

ground substance, demonstration with Russell modifi cation of Movat pentachrome stain, 173


gum masticfor Dieterle staining method, 248stability of, 100for Steiner and Steiner procedure, 249

gum tragacanth, for freezing muscle specimens, 313–314, 313

HH&E staining. See hematoxylin and eosin staining.handling, of solutions, 98Hansen’s disease

demonstration with Fite staining technique, 228–230demonstration with Kinyoun acid-fast stain, 225

hardening by fi xative, 2Harris hematoxylin

composition, 110demonstration of muscle fi ber regeneration, 322for Pap staining, 361

hazardsbiological or infectious, 82–83chemical, 84–90explosive, 86fi re, 86mechanical, 83–85symbols for, 90

hazy water, troubleshooting hematoxylin and eosin staining, 121heat

and penetration by fi xatives, 7for protein stabilization, 2artifact on specimen, 122

heat-induced epitope retrieval, 282HEIR artifact

on brain biopsy, 287Helicobacter pylori, demonstration with alcian yellow-toluidine blue

staining, 234–235, 235Helicobacter pylori, demonstration with Giemsa staining, 233–234, 234Helicobacter pylori, demonstration with microwave variation of Warthin-

Starry technique, 246, 247Helicobacter pylori, demonstration with Steiner and Steiner microwave

method, 249–250, 250Helly solution

formulation and applications, 20labeling, 89specimen removal from, 7

hemapoietic tissue, and B-5 solution, 19hematein vs hematoxylin, 110hematin, black acid, 11hematoidin, 255hematoxylin

combined with Luxol fast blue and PAS, 218–219counterstain for PAS reaction, 138vs hematein, 110for immunohistochemical studies, 283for Pap staining, 361stability of, 100staining, 109–113

hematoxylin, alcoholic, for Verhoeff elastic stain, 168hematoxylin and eosin stained tissue, processing for electron

microscopy, 347–348

hematoxylin and eosin staining, 114–123with automatic stainers, 67dehydration preparation, 71demonstration of phagocytosis in muscle, 309of diseased muscle, 309fi xation troubleshooting, 26of normal muscle, 308processing problems, 42section thickness for, 58troubleshooting, 118–123with urate crystals, 256

hematoxylin solution, for Movat pentachrome stain, 172hemochromatosis, ferrous iron in spleen stained with Turnbull blue, 259hemofuchsin, demonstration with Prussian blue stain, 257hemosiderin

in bone marrow fi xed with Zenker solution, 258demonstration with Prussian blue stain, 257in spleen stained with Turnbull blue, 259

hepatitis, infectious hazards from, 83hepatocellular fi brosis, with Gordon and Sweets staining, 177–179HER2, guidelines for testing, 6heterochromatin, 104, 105HIER. See heat-induced epitope retrieval.histiocytes, connective tissue types and structures, 160histochemical reactions, of human muscle types, 309Histoplasma capsulatum, demonstration with Grocott staining, 241HIV (human immunodefi ciency virus), infectious hazards from, 83HMB-45/MART-1, anaplastic tumor workup, 290holes in section, troubleshooting, 60, 61Hollande solution, formulation and applications, 20Holmes silver nitrate staining method

combined with Luxol fast blue, 215–217, 217nerve fi ber and neurofi bril demonstration, 199–200

Holzer staining methoddemonstration of gliosis with, 208–209, 209

horseradish peroxidase, for immunohistochemical studies, 283Hotchkiss-McManus PAS reaction, demonstration of fungi with,

235–236, 236HRP enzyme labeled polymer procedure, 301–302human epidermal growth factor receptor 2. See HER2.human immunodefi ciency virus. See HIV.hyaluronic acid, alcian blue demonstration of, 145hyaluronidase, epithelial and connective tissue mucin demonstration,

147–148, 147hydrates, percentage solutions of, 96hydrochloric acid

for acid phosphatase detection in muscle, 321for α-naphthyl acetate esterase stain, 315for chromic acid-Schiff stain, 237for decalcifi cation, 47demonstration of muscle fi ber regeneration, 322in Feulgen reaction, 123for Hotchkiss-McManus PAS reaction, 236for naphthol AS-D chloroacetate esterase technique, 316preparation for alcian blue technique, 146preparation for Müller-Mowry colloidal iron technique, 150preparation for PAS reaction testing, 137preparation for PAS reaction with diastase digestion, 140Prussian blue stain preparation, 257stability of, 100for Turnbull blue stain preparation, 258

386 Index

hydrochlorofl uorocarbon refrigerants, for frozen sectioning, 65hydrogen bonding, 107hydrogen peroxide

for blocking reactions, 287for HRP enzyme labeled polymer procedure, 302for thiofl avin S demonstration of Alzheimer disease, 206

hydrolases, 311hydronium ions, 74hydroquinone

for microwave variation of Warthin-Starry technique, 246for Steiner and Steiner procedure, 249for Warthin-Starry technique, 245

hydroxyl ions, 74hypertonicity

and fi xative selection, 7–8, 8

Iice crystal artifact, preventing, in muscle, 312–314ice crystals

in brain section, 51from frozen sectioning, 50, 65troubleshooting, 50–51, 50

ignitability, description, 88illumination, of microscope, 54imidazole, 289immunoblasts

methyl green-pyronin Y staining, 126, 188–189, 189immunofl uorescence, 56

in frozen sectioning, 50for immunohistochemical studies, 283obsolescence as method, 279PBS-sucrose preservation of, 23

immunohistochemical studiesPBS-sucrose transport solution for, 23staining techniques, 297–302zinc formalin fi xation advantages, 21zinc salt fi xatives for, 16–17

immunohistochemistry, 277–305quality control for, 292slides, dehydration of, 71

immunolabeling, with LR white, 339–340immunology, 278–279immunoperoxidase techniques, troubleshooting, 296–297impregnating solution

for Holmes silver nitrate staining method, 199for Luxol fast blue-Holmes silver nitrate staining, 216variations for reticulin staining, 175

infectious hazards, 82–83infi ltration, 37–41infl ammation

in muscle cells, acid phosphatase reaction in, 320–322, 322instrumentation, 53–80

quality control, 75–79interalveolar septa, fi xation illustrations, 6intermyofi brillar sarcoplasm, demonstration with Gomori trichrome

procedure, 329interstitial connective tissue, demonstration with Gomori trichrome

procedure, 329

intestinal metaplasia, detection with alcian blue-PAS-hematoxylin, 148intestine, 63

alcian blue stain on, 69colloidal iron staining with hematoxylin counterstaining, 152demonstrating cytoplasmic granules with Fontana-Masson staining,

262demonstration of argentaffi n in, with microwave Fontana-Masson

technique, 263eosin-counterstained, 114incorrect orientation of, 45incorrect polychromatic staining, 127Mayer mucicarmine staining of, 144overdehydration from forced air dryer, 73overstaining, with eosin-phloxine, 121specimen orientation, 43troubleshooting hematoxylin and eosin staining, 120tumor demonstration with Grimelius stain, 264

iodine-glycerine mounting medium, for phosphorylase stain, 327iodine-iodide solution, for Russell modifi cation of the Movat

pentachrome stain, 172ion-exchange resins, for decalcifi cation, 47ions, 74

and fi xation, 3and staining, 10, 11, 107

iris diaphragmof microscope, 54

iron hematoxylin staining, diff erentiation with, 109iron, ferric

in bone marrow section stained with Prussian blue, 258demonstration with Prussian blue stain, 256–258

iron, ferrousdemonstration with Turnbull blue stain, 258–259in spleen stained with Turnbull blue, 259

irritants, xylene, limonene, and aliphatic carbon comparisons, 37isoelectric point, 108isopentane, 313, 314

fl ash point, 87for freezing muscle specimens, 313–314in frozen sectioning, 50, 65

isopropyl alcohol (isopropanol)fl ash point, 87for dehydration, 33, 34

isotonicity, and fi xative selection, 7–8

Jjamming, during sectioning, troubleshooting, 62Jenner solution, composition, 127juxtanuclear region, ultrastructure of, 105

Kkeratin

demonstration with Gomori 1-step trichrome stain, 166demonstration with Masson trichrome staining, 163–165demonstration with Pap staining, 363HMW small cell neoplasm panel, 290

Kernechtrot solutionfor Churulian-Schenk method, 265for Gomori staining of reticulin, 176for Gordon and Sweets staining technique, 178for Grimelius argyrophil stain, 264for microwave variation of Churukian-Schenk method, 266for microwave variation of Fontana-Masson stain, 263preparation for alcian blue technique, 145Prussian blue stain preparation, 257for Turnbull blue stain preparation, 258for von Kossa calcium stain, 270


kidneybasement membrane in, 161biopsy examination with immunofl uorescence, 283blocked for electron microscopy, 340calcium demonstration with alizarin red stain, 271calcium demonstration with von Kossa stain, 270crystal violet staining, 155through electron microscope, 57fi xed in formalin, 10incomplete diff erentiation of eosin staining, 121through light microscope, 55PAS reaction with fresh vs old Schiff reagent, 139plastic embedding, 41through polarizing microscope, 55section thickness for, 58stained with thiofl avin T technique, 156staining with periodic acid-methenamine procedure, 184

killing, of tissue specimens, by fi xative, 2Kinyoun acid-fast stain

acid-fast bacteria demonstration in lung section, 225leprosy demonstration with, 225on lung section, 73mast cell demonstration, 189mycobacteria demonstration with, 224–226

Kinyoun carbol-fuchsin solution, for Kinyoun acid-fast stain, 224knife guard, 84knives

artifacts caused by, 59 diamond, for electron microscopy, 341–342for electron microscopy, 341–343glass for electron microscopy, 341

knives, glass, 58knives, Ralph, 58knives. See also microtomes.

Llabeling, 85

of hazardous materials, 89and quality control, 295of Zenker and Helly solutions, 89

laboratory standard, 84Laidlaw variation, oxidizer, sensitizer, impregnating solution, and

reducing solution, 175lake, defi ned, 110lamina propria

defi ned and illustrated, 5pseudomelanin in colon stained with Schmorl technique, 260

latex sensitivity, 82LCA, small cell neoplasm panel, 290Legionella pneumophila

demonstration with Dieterle staining method, 247–248, 249demonstration with microwave variation of Warthin-Starry

technique, 246demonstration with Steiner and Steiner microwave method, 249–250

lens, of microscope, 54leprosy


leukemias, demonstration with naphthol AS-D chloroacetate esterase technique, 316, 317

leukocyte precursors, Giemsa staining of, 128

leukocytes, connective tissue types and structures, 160light green solution

for aldehyde fuchsin elastic stain, 170for Gomori methenamine-silver technique for urates, 268for Grocott staining, 240for microwave variation of methenamine-silver nitrate staining, 243for Masson trichrome staining, 164for Pap staining, 363for periodic acid-methenamine silver microwave procedure, 183

limonene reagentsattribute comparisons, 37for clearing, 36

Lindquist rhodanine method, copper demonstration with, 272lipid stains, 108lipids. See also fats.

demonstration with oil red O, 185demonstration with osmium tetroxide paraffi n procedure, 187demonstration with Sudan black B, 186fi xative comparative characteristics, 18fi xative eff ect on, 2, 8and formaldehyde fi xation, 10osmium tetroxide fi xation, 15and potassium dichromate fi xation, 16processing with water-soluble waxes, 40staining techniques, 184–188ultrastructure of, 105

lipofuscin, 106, 255demonstration with Bielschowsky-PAS staining, 202, 204

lipoid capsuledemonstration with Fite staining technique, 228–230demonstration with Kinyoun acid-fast stain, 225

liposarcomas, fat demonstration with oil red O, 184liquid nitrogen

for freezing muscle specimens, 313–314, 313, 314for frozen sectioning, 50, 65for freezing antibodies, 72, 286

liquid-based cytology, 358–359liter, defi ned, 97lithium carbonate

in decalcifi cation processing, 48for Luxol fast blue staining, 213for Luxol fast blue-cresyl echt violet staining, 214for Luxol fast blue-Holmes silver nitrate staining, 216for Luxol fast blue-PAS-hematoxylin staining, 218for picric acid neutralization, 15stability of, 100

liver, autolysis in, 2Best carmine demonstration of glycogen in von Gierke disease, 142copper demonstration with microwave rhodanine method, 272–273cryostat processing temperature, 64demonstrating bilirubin from bile staining, 269holes caused by knife, 60Masson trichrome staining of, 162necrotic

Brown-Hopps gram stain on, 232diff erential diagnosis with Gordon and Sweets staining, 177–179

oil red O staining of, 185PAS reaction with and without diastase digestion, 141reticulin demonstration with Gomori stain, 176reticulin demonstration with Gordon and Sweets staining, 179reticulin staining with Snook variation, 177staining with osmium tetroxide paraffi n procedure, 187Sudan black B staining, 186Wilson disease diagnosis with rhodanine method, 271–273, 272, 273

388 Index

liver diseasesdiff erential diagnosis with Gordon and Sweets staining, 177–179silver staining techniques for, 174–179

log, antibody control, 293LR white processing

for electron microscopy, 339–340lugol iodine solution

formulation and applications, 24for Mallory PTAH staining of glia, 207stability of, 100for Verhoeff elastic stain, 168

lungadenocarcinoma in, 290colloidal iron staining, 151demonstration with Hotchkiss-McManus PAS reaction, 236detection with microwave auramine-rhodamine fl uorescence

technique, 231fi xation illustrations, 6Kinyoun acid-fast technique on, 73

Luxol fast blueon central nervous system specimen, 59combined with cresyl echt violet, 214–215, 215combined with Holmes silver nitrate staining, 215–217, 217combined with PAS and hematoxylin, 218–219demonstration of myelin sheath, 212–213, 214stability of, 100staining on medulla, olivary nucleus, 213

Luxol fast blue-PAS-hematoxylin staining, demonstration with Luxol fast blue-PAS-hematoxylin staining, 219

lymph nodesalcohol fi xation in, 66cryostat processing temperature, 64Feulgen reaction on, 124fi xation troubleshooting, 25, 26hematoxylin and eosin staining of, 115holes caused by knife, 60incomplete formalin fi xation, 281melanin demonstration with Schmorl technique, 260methyl green-pyronin Y staining, 189with microwave variation of Warthin-Starry technique, 247normal CD20 B-cell staining, 291normal CD3 T-cell staining, 291plastic embedding, 41showing melanin with Fontana-Masson stain, 262specimen orientation, 44

lymphocytes, hematoxylin and eosin staining of, 105lymphoma

anaplastic tumor workup, 290B-5 vs formalin fi xation, 19on colon demonstrated with cytoplasmic IgA, 299small cell neoplasm panel, 290

lymphoreticular tissueand B-5 solution, 19

lyophilized antibodies, storage of, 287lysosomes, 106

acid phosphatase as marker for, 320demonstration with α-naphthyl acetate esterase stain, 316ultrastructure of, 105

Mmacrophages

connective tissue types and structures, 160demonstration with α-naphthyl acetate esterase stain, 316

maintenanceof balances, 74of cryostat, 65history chart, 78–79of microscope, 54of tissue processors, 66

malignant, defi ned, 289Mallory PTAH technique

demonstration with Mallory PTAH technique, 208gliosis demonstration with, 207–208for muscle staining, 179–181

malt diastase solutionpreparation for PAS reaction with diastase digestion, 140stability of, 100

marrowGiemsa staining of, 128plastic embedding, 41section thickness for, 58

Masson trichrome stainwith osmium tetroxide paraffi n procedure, 187in tumor diff erentiation and collagenous tissue identifi cation,

162–165mast cells, connective tissue types and structures, 160

demonstration with Mayer hematoxylin, 317–318demonstration with microwave variation of Ziehl-Neelsen method,

228demonstration with toluidine blue, 188, 188methylene blue counterstaining in Kinyoun acid-fast technique, 189in skin tumor, 189

mastocytomasdemonstration with toluidine blue, 188diagnosis with toluidine blue, 188

Material Safety Data Sheets (MSDSs), 85mathematics, laboratory, 93–102Mayer hematoxylin

composition of, 111demonstration of granulocytes and mast cells, 317–318demonstration of muscle fi ber regeneration, 322for immunohistochemical studies, 283for microwave variation of rhodanine method, 273preparation for thiofl avin T technique, 155for rhodanine copper demonstration, 272

Mayer mucicarminedemonstration of C neoformans and B dermatitidis with, 244epithelial mucin demonstration with, 142–144, 144mold demonstration with, 223on small intestine, 144for Schmorl technique, 259

May-Grunwald Giemsa stain, application and procedure, 127–128measuring technique, 94mechanical hazards, 83–85mechanical testing of decalcifi cation, 48media, aqueous mounting, 129medulla

demonstration with Luxol fast blue, 213demonstration with Weil method, 212


melanin, 255demonstrated with Schmorl technique, 259, 260demonstration with Fontana-Masson stain, 260–263, 262demonstration with microwave variation of Fontana-Masson stain,

261–263in lymph node section stained with Fontana-Masson technique, 262in lymph node section stained with Schmorl technique, 260

melanomaanaplastic tumor workup, 290in lymph node section stained with Fontana-Masson technique, 262in lymph node section stained with Schmorl technique, 260on eye, marked with alkaline phosphatase red chromogen, 283

melting pointand embedding technique, 42and ribbon formation problems, 60

mercuric chloride, 14–15, 18comparative characteristics, 18and dye binding, 109in immunohistochemical studies, 279, 280rate of penetration, 7

mercury pigment artifact, 254mesenchymal cells, connective tissue types and structures, 160mesothelioma, birefringence, 254metal mordants, in nuclear staining, 107metanil yellow

for Gridley staining, 238Mayer mucicarmine staining of, 144preparation for Mayer mucicarmine technique, 143for Schmorl technique, 259stability of, 100

metastasis, defi ned, 289methacarn solution

in immunohistochemical studies, 279formulation and applications, 23

methanol. See methyl alcohol.methenamine silver nitrate

for periodic acid-methenamine silver microwave procedure, 182for periodic acid-methenamine silver procedure without microwave,

183stability of, 100

methenamine solutionfor Gomori methenamine-silver technique for urates, 267for Grocott staining, 240for microwave variation of methenamine-silver nitrate staining, 242

methenamine-silver nitrate staining, demonstration of fungi with, 239–244

methyl alcoholin Best carmine demonstration of glycogen, 142comparative characteristics, 18for dehydration, 34eff ect on protein, 8fl ash point, 87rate of penetration, 7

methyl green solutionfor acid phosphatase detection in muscle, 321composition, 125

methyl green-pyronin Y staining, 124–126, 125, 126of mast cell tumor of skin, 189plasma cell staining, in lymph node, 189plasma staining with, 188–189solution composition, 125

methylene blueacid-fast bacteria demonstration in lung section, 225counterstain for mast cell demonstration, 189for demonstration of bacteria, 222for Fite acid-fast staining technique, 229for Kinyoun acid-fast stain, 224for microwave variation of Ziehl-Neelsen method, 227stability of, 100for Ziehl-Neelsen method, 226

metric system, 97–98metric units, US equivalents, 97Michel transport solution

formulation and applications, 23for specimen storage, 8

microglia, 195microliter, defi ned, 97micrometer pipettes, 75micrometry, with microscope, 54microorganisms, 221–249microscopes, 54–57

darkfi eld, 55electron, 56–57fl uorescence, 56light, 54, 55phase-contrast, 55polarizing, 55, 55

microtomes, 57–64. See also knives.chatter from, 281clinical freezing, 57–58diamond for electron microscopy, 341–342for electron microscopy, 341–343glass for electron microscopy, 341rotary, 57safety precautions, 58, 84sliding, 57

microtomy, 314for electron microscopy, 341–343for immunohistochemical studies, 280–281troubleshooting, 58–64

microwave auramine-rhodamine fl uorescence technique, demonstration of M tuberculosis, 230–231, 231

microwave oven processing, 2, 39of rectal tumor specimen, 40

microwave processors, 66–67microwave rhodanine method, copper demonstration with, 273microwave staining oven, 67–68microwave variation

of Bielschowsky-PAS staining, 202–204of Churulian-Schenk method, 266–267

pancreatic islet stained with, 267of Fontana-Masson technique, 261–263

argentaffi n in intestine demonstrated with, 263of Masson trichrome staining, 164–165of methenamine-silver nitrate staining, demonstration of P jirovecii,

243, 244of methenamine-silver nitrate staining, 242–244of Müller-Mowry colloidal iron technique, 151–152of periodic acid-methenamine silver microwave procedure, 182–187of rhodanine copper demonstration, 272–273of Warthin-Starry technique, 245–247

bacteria demonstration with, 247of Ziehl-Neelsen method, 227–228

390 Index

microwavesfor protein stabilization, 2

milky water, troubleshooting hematoxylin and eosin staining, 121Miller technique, 171milliliter, defi ned, 97Millipore-fi ltered deionized water

for fl otation bath processing, 70Millonig fi xative, modifi ed

for electron microscopy, 335Millonig formalin, eosinophil fi xation, 11Millonig formalin, formulation, 12minerals, 255–256mitochondria, abnormality demonstration with Gomori trichrome

procedure, 329, 329mitochondria, abnormality demonstration with NADH diaphorase

reaction, 323–325, 324mitochondria, 105

and potassium dichromate fi xation, 16succinic dehydrogenase (SDH) reaction with, 325–326, 326ultrastructure of, 104, 105

modifi ed Millonig fi xativefor electron microscopy, 335

molar solutions, 96molds, 222–223

fi xative eff ect on, 2monoclonal antibodies, defi ned and described, 278–279monocytes, connective tissue types and structures, 160mordant dyes

for diff erentiation, 109mordanting

and fi xative selection, 7for Masson trichrome staining, 164of muscle section in Mallory PTAH staining, 181

mordantsfi xatives as, 2metal

composition, 110–113in nuclear staining, 107

motor endplates, demonstration with α-naphthyl acetate esterase stain, 316, 316

mountingstained sections, 128–131troubleshooting, 130–131, 130

mounting medium retraction, 131, 131Movat pentachrome stain, Russell modifi cation of, 172–174MSDSs (Material Safety Data Sheets), 85Müller colloidal iron solution, preparation for Müller-Mowry colloidal

iron technique, 150Müller-Mowry colloidal iron, carboxylated and sulfated

mucopolysaccharide demonstration with, 149–152mucicarmine

stability of, 100for Mayer mucicarmine technique, 143

mucins, demonstration with alcian yellow-toluidine blue staining, 235, 235demonstration in cervix with alcian blue-PAS technique, 149demonstration with Gomori 1-step trichrome stain, 166demonstration with Gridley staining, 239demonstration with Grocott staining, 241demonstration with Masson trichrome staining, 163–165demonstration with Mayer mucicarmine technique, 143demonstration with Russell modifi cation of Movat pentachrome

stain, 172–174, 173demonstration with Schmorl technique, 260staining with Gill hematoxylin, 112types, occurrence, and reactivity, 136

mucoid specimens, smear preparation, 355mucopolysaccharides, alcian blue demonstration of, 145–146

demonstration with Müller-Mowry colloidal iron technique, 149–152stained with alcian blue at pH 1, 147types, occurrence, and reactivity, 136

mucoproteins, types, occurrence, and reactivity, 136mucosa, defi ned and illustrated, 5, 5mucosubstances

absorption of ferric ions, 149diff erentiating neutral and acidic, 148PAS reaction testing for, 137–141types, occurrence, and reactivity, 136sulfated alcian blue demonstration of, 146–147

mucus, cellular material trapped in, 360multilink biotinylated secondary antisera, 289muscle

acid phosphatase reaction in, 320, 322demonstration with Masson trichrome staining, 163–165demonstration with Russell modifi cation of Movat pentachrome

stain, 172–174, 173fast-twitch, 308of gastrointestinal tract, 162Gomori trichrome procedure with, 328–329, 329histology, 308holes in section, 61Mallory PTAH staining in, 181pathology, 308periodic acid-Schiff technique on, 328phagocytosis demonstration with hematoxylin and eosin staining,

309phosphorylase stain, 326–328, 327poor diff erentiation of, 121sectioning of, 314skeletal, 308

α-naphthyl acetate esterase stain, 316atrophy demonstration with ATPase, 320demonstration of normal pattern with ATPase, 319demonstration with α-naphthyl acetate esterase stain, 316frozen sectioning of, 65hematoxylin and eosin staining of, 308neuropathic disease in, 309vs cardiac, 161

slow-twitch, 308smooth

demonstration with Russell modifi cation of Movat pentachrome stain, 173

diff erentiation with Gomori 1-step trichrome stain, 165–166in Verhoeff -van Gieson staining of aorta and artery, 169

stained with oil red O, 328staining techniques, 179–181types and characteristics, 161van Gieson staining, 167

muscle biopsy specimensα-naphthyl acetate esterase stain, 314–316freezing, 312–314, 313, 314

muscle fi bers, demonstration with Gomori 1-step trichrome stain, 166muscle regeneration, alkaline phosphatase reaction in, 322–323, 323muscle type diff erentiation

with ATPase, 318–320muscularis mucosa, defi ned and illustrated, 5musculoskeletal disorders, 84mushy tissue, troubleshooting, 44–45, 45, 58


mycelia, demonstration with Gridley staining, 239mycobacteria

demonstration with Fite staining technique, 228–230demonstration with Kinyoun acid-fast stain, 224–226demonstration with microwave variation of Ziehl-Neelsen method,

227–228demonstration with Ziehl-Neelsen method, 226–228detection with microwave auramine-rhodamine fl uorescence

technique, 230–231, 231Mycobacterium leprae


Mycobacterium tuberculosis, detection with microwave auramine-rhodamine fl uorescence technique, 230–231

myelin, 195demonstration with Luxol fast blue, 213demonstration with Luxol fast blue-PAS-hematoxylin staining, 219demonstration with Mallory PTAH technique, 179–181, 208fat demonstration with oil red O, 184indistinct diff erentiation with Luxol fast blue, 214

myelin sheath, 194demonstration with Luxol fast blue, 212–213demonstration with Luxol fast blue-PAS-hematoxylin staining,

218–219demonstration with Weil method, 211, 212simultaneous demonstration with nerve fi bers, 215–217, 217simultaneous demonstration with Nissl substance, 214–215, 215

myelinated nerve twigs, demonstration with Gomori trichrome procedure, 329

myelogenous leukemic cells, in cervix, demonstrated with naphthol AS-D chloroacetate esterase technique, 317

myofi brils, demonstration with Gomori trichrome procedure, 329myopathic disease, ATPase stain to distinguish from neuropathy,

318–320

NNADH, histochemical reactions of human muscle types, 309naphthol AS-D chloroacetate esterase technique, demonstration of

myelogenous leukemic cells with, 317naphthol AS-D chloroacetate esterase technique, 316, 317Nasher and Shankin variation, oxidizer, sensitizer, impregnating

solution, and reducing solution, 175National Fire Protection Association (NFPA), hazard rating system, 89NBT solution

for NADH diaphorase reaction, 324for succinic dehydrogenase (SDH) reaction, 325

necrotic liver disease, diff erential diagnosis with Gordon and Sweets staining, 177–179

nemaline rodsdemonstration with Gomori trichrome procedure, 329demonstration with Mallory PTAH technique, 179–181

neoplasm panel, small cell, 290nephrocalcinosis

demonstrated with alizarin red stain, 271demonstrated with von Kossa stain, 270

nerve endingsBodian staining method, 197–198demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 205

nerve fi bersBodian staining method, 197–198demonstration with Bielschowsky-PAS staining, 200–205demonstration with Bodian staining method, 198demonstration with Holmes silver nitrate method, 200demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 204Holmes silver nitrate staining method, 199–200simultaneous demonstration with myelin sheath, 215–217, 217

nerve tissue, 194–220staining techniques, 195–219

nervous system, 194–195neurites, dystrophic, demonstration with Bielschowsky-PAS staining, 202neurites, peripheral

demonstration with Bielschowsky-PAS staining, 202demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 205neuroblastoma, small cell neoplasm panel, 290neurofi brillary plaques and tangles

demonstration with Bielschowsky-PAS staining, 200–205, 202, 204demonstration with Luxol fast blue-PAS-hematoxylin staining,

218–219demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 204–205, 205demonstration with thiofl avin S staining, 205–207, 207fl uorescent dyeing, 56

neurofi brils, 194Bodian staining method, 197–198demonstration with Bielschowsky-PAS staining, 204demonstration with Holmes silver nitrate method, 200demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 205Holmes silver nitrate staining method, 199–200

neurofi lament, small cell neoplasm panel, 290neuroglia, 194–195neurologic eff ects, xylene, limonene, and aliphatic carbon

comparisons, 37neuromelanin, demonstration with Bielschowsky-PAS staining, 204neuronal nuclei, hematoxylin and eosin staining of, 105neurons, 194

cresyl echt violet staining, 196demonstration with Holmes silver nitrate method, 200demonstration with Mallory PTAH technique, 208staining of, 105

neuropathic diseaseATPase stain to distinguish from myopathy, 318–320in muscle, 316

neurosecretory granulesdemonstration with Fontana-Masson stain, 260–263demonstration with microwave variation of Fontana-Masson stain,

261–263neurosecretory tumors, demonstration with Churukian-Schenk method,

265–267neutral buff ered formalin. See formalin, neutral buff ered.neutralization, of picric acid, 15NFPA (National Fire Protection Association), hazard rating system, 89nickel method, of smear preparation, 354nicotinamide adenine dinucletide phosphate tetrazolium reductase

(NADH diaphorase) reaction, abnormality demonstration in muscle with, 323–325, 324

nipple discharges, collection and handling of, 352

392 Index

Nissl substance, 194demonstration with cresyl echt violet staining, 195–197, 197simultaneous demonstration with myelin sheath, 214–215, 215

nitric acidfor decalcifi cation, 47for microwave variation of Bielschowsky-PAS staining, 203

nitrocellulose, embedding with, 40nitrogen, liquid, 313, 314

for freezing muscle specimens, 313–314for frozen sectioning, 50, 65

Nocardia, Fite technique variation for, 229Nocardia asteroides, demonstration with microwave variation of

Warthin-Starry technique, 246node of Ranvier, 194nonadditive fi xation, 2nonaqueous fi xatives, 22–23noncoagulant fi xatives, 3, 4nonionic homoglycans, types, occurrence, and reactivity, 136normal solutions, 96NSE, small cell neoplasm panel, 290nuclear basophilia

from acid decalcifi cation, 47in decalcifi ed bone, 48

nuclear bubblingas drying artifact, 73from excessive heat, 71fi xation troubleshooting, 24–26, 26from formalin fi xation, 6

nuclear dyes, 109–113nuclear envelope, ultrastructure of, 105nuclear fast red

for Churulian-Schenk method, 265for Gomori staining of reticulin, 176for Gordon and Sweets staining technique, 178for Grimelius argyrophil stain, 264for microwave variation of Churukian-Schenk method, 266for microwave variation of Fontana-Masson stain, 263preparation for alcian blue technique, 145Prussian blue stain preparation, 257stability of, 100for Turnbull blue stain preparation, 258for von Kossa calcium stain, 270

nuclear membraneas component of nucleus, 104

nuclear pores, 104, 105nuclear staining, 103–134, 107nuclei, 104–106, 104, 105

alcian blue demonstration of, 145, 147cresyl echt violet staining, 196demonstration with alcian yellow-toluidine blue staining, 235demonstration with alkaline Congo red staining, 153demonstration with Best carmine technique, 142demonstration with Bodian staining method, 198demonstration with Brown-Hopps method, 232demonstration with chromic acid-Schiff stain, 238demonstration with Churukian-Schenk method, 266demonstration with cresyl echt violet staining, 196demonstration with Diff -Quik Giemsa staining, 234demonstration with Fontana-Masson stain, 261demonstration with Gomori 1-step trichrome stain, 166demonstration with Gomori trichrome procedure, 329

demonstration with Grimelius stain, 264demonstration with Masson trichrome staining, 163–165demonstration with Mayer hematoxylin, 317–318demonstration with Mayer mucicarmine technique, 143demonstration with microwave rhodanine method, 273demonstration with microwave variation of Fontana-Masson stain, 263demonstration with microwave variation of Warthin-Starry

technique, 247demonstration with Müller-Mowry colloidal iron technique, 151demonstration with Prussian blue stain, 257demonstration with rhodanine method, 272demonstration with Russell modifi cation of Movat pentachrome

stain, 173demonstration with Sudan black B, 186Luxol fast blue-cresyl echt violet staining, 215methyl green-pyronin Y staining, 126, 126, 189of muscles, 308, 308in nerve cell, 194

demonstration with Luxol fast blue-PAS-hematoxylin staining, 219Mallory PTAH staining, 208

reactions to fi xatives, 8troubleshooting hematoxylin and eosin staining, 119–120, 119, 120van Gieson staining, 167

nucleic acidsfi xative comparative characteristics, 18reactions to fi xatives, 8staining of, 123–126

nucleoli, 104demonstration with Pap staining, 363methyl green-pyronin Y staining, 126, 189in nerve cell, 194staining of, 105ultrastructure of, 104

nucleoproteinsacetic acid and, 9and picric acid fi xation, 15

nucleus. See nuclei.

Oobjective lens, of microscope, 54Occupational Exposure to Hazardous Chemicals in the Laboratories, 84ocular lens, of microscope, 54odor, xylene, limonene, and aliphatic carbon comparisons, 37OG-6 counterstain, 362oil red O, stability of, 100oil red O staining

of lipids, 184–186, 185on muscle, 328

oligodendroglia, 194olivary nuclei, demonstration with Luxol fast blue, 213orange G, for Pap staining, 362orcein fuchsin stain, 171organ capsules, connective tissue types and structures, 160organelles, demonstration in liver with Gomori stain, 176orientation, of specimen, 42–43, 43

in frozen sectioning, 51troubleshooting, 45

origanum, oil offor clearing, 36

Orth solution, formulation and applications, 21


OSHA Hazard Communication Standard, 84osmium tetroxide, 289

with cacodylate buff er, for electron microscopy, 336comparative characteristics, 18and dye binding, 109for electron microscopy, 334in frozen sectioning, 50for lipid fi xation, 8morphologic preservation with, 17with phosphate buff er, for electron microscopy, 336rate of penetration, 7safety precautions, 15

osmium tetroxide paraffi n procedure, lipid demonstration with, 187osmolality, and fi xative selection, 7–8, 8ovarian, adenocarcinoma, 290ovens, 71overdecalcifi cation, troubleshooting, 49overdehydration

holes in section from, 61troubleshooting, 41–42

overfi xation, and washboarding of section, 61overstaining, with eosin-phloxine counterstain, 121oxalic acid

for Bodian staining method, 197for Gordon and Sweets staining technique, 178for Holmes silver nitrate staining method, 200for Luxol fast blue-Holmes silver nitrate staining, 216for Mallory PTAH staining of glia, 208for Mallory PTAH technique, 180for PTAH staining without mercuric solutions, 181stability of, 100for thiofl avin S demonstration of Alzheimer disease, 206

oxidases, 312oxidation

defi ned, 310of hematoxylin, 110

oxidizer variations for reticulin staining, 175oxidizers, for diff erentiation, 109oxireductases, 312

PP504S, demonstration of cancer on prostate, 302PAF. See Zamboni solution.pancreas

adenocarcinoma, 290autolysis in, 2stained with microwave Churukian-Schenk method, 267

pancreatic duct, ultrastructure of, 104Paneth cells, defi ned and illustrated, 5PAP (peroxidase-antiperoxidase) immunohistochemical technique,

lymphoma demonstrated on colon with cytoplasmic IgA, 299PAP (peroxidase-antiperoxidase) immunohistochemical technique, 284,

297–299Pap smears, 352, 359Pap (Papanicolaou) stain, 361–362

composition and technique, 362–363on cytoplasm and neutrophils, 362

paraffi nfor embedding, 37–40, 42melting point for immunohistochemical studies, 280processing with water-soluble waxes, 40rapid processing with, 38artifacts, 71

paraffi n-processed tissue, fi xatives for, 280paraformaldehyde

with cacodylate buff er, for electron microscopy, 335comparative characteristics, 18with phosphate buff er, for electron microscopy, 335

pararosaniline solutionfor acid phosphatase detection in muscle, 321for α-naphthyl acetate esterase stain, 315

parched earth artifact, 69parfocality

of microscope, 54Parlodion, embedding with, 40PAS staining (PAS reaction). See also alcian blue-PAS-hematoxylin

technique; Bielschowsky-PAS staining; Hotchkiss-McManus PAS reaction

of carbohydrates, 137–141on central nervous system specimen, 59in colon section, 138combined with Luxol fast blue and hematoxylin, 218–219on kidney section, 139mold demonstration with, 223on muscle, 328

pathology, in muscles, 308PBS buff er

for peroxidase-antiperoxidase (PAP) immunohistochemical technique, 297, 300

PBS solutions, formulation and applications, 23penetration

by fi xative, 2fi xative comparative characteristics, 18by formaldehyde fi xation, 10of glutaraldehyde fi xation, 13osmium tetroxide fi xation, 15rate of, by fi xatives, 7with zinc formalin fi xation, 17

percentage, calculation, for solutions, 94–97perikaryon, demonstration with Bielschowsky-PAS staining, 202periodic acid

for alcian yellow-toluidine blue staining method, 234for Bielschowsky-PAS staining, 201for Hotchkiss-McManus PAS reaction, 235for Luxol fast blue-PAS-hematoxylin staining, 218for osmium tetroxide paraffi n procedure, 187for periodic acid-methenamine silver microwave procedure, 182preparation for alcian blue-PAS-hematoxylin technique, 149preparation for PAS reaction testing, 137preparation for PAS reaction with diastase digestion, 140stability of, 100

periodic acid-methenamine silver microwave procedurefor basement member delineation, 182–186

periodic acid-methenamine silver procedure, kidney section stained with, 184

periodic acid-Schiff stain. See PAS reaction.peripheral nervous system, 194peripheral neurites

demonstration with Bielschowsky-PAS staining, 202demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 205peroxidase eff ects, on red blood cell specimen, 287peroxidase-antiperoxidase (PAP) immunohistochemical technique, 284,

297–299lymphoma demonstrated on colon with cytoplasmic IgA, 299

394 Index

peroxidases, 312personal protective equipment

with formalin, 13for infectious hazards, 82

pH, 74of amino acids, 107and dye binding, 109and fi xation for electron microscopy, 335of fi xatives, 7meters, 72–73monitoring of buff ers, 295and quality control, 294and tissue swelling with acetic acid, 9

phagocytosisin muscle, 309

phosphatase, 311phosphate buff er

for α-naphthyl acetate esterase stain, 315for NADH diaphorase reaction, 324preparation for PAS reaction with diastase digestion, 140for succinic dehydrogenase (SDH) reaction, 325

phosphate-buff ered glutaraldehyde, formulation, 14phosphate-buff ered saline. See PBS.phosphomolybdic acid-alcohol solution, for Holzer staining method,

209phosphomolybdic-phosphotungstic acid solution, for Masson trichrome

staining, 163phosphorylase, histochemical reactions of human muscle types, 309phosphorylase, 312phosphorylase stain

for muscle, 326–328, 327phosphotungstic acid

for Russell modifi cation of the Movat pentachrome stain, 173stability of, 100

phosphotungstic acid hematoxylin. See PTAH.phosphotungstic acid-hematoxylin technique. See PTAH technique.physiological saline, for succinic dehydrogenase (SDH) reaction, 326picric acid, 15–16

comparative characteristics, 18and dye binding, 109explosion hazard, 16, 86neutralization, 15rate of penetration, 7safe storage, 88safety precautions, 16saturated aqueous, stability of, 100for van Gieson staining, 167

picric acid-acetonefor Brown-Hopps modifi cation of gram staining, 232

picric acid-acetone, stability of, 100pigment artifact, 254

fi xation troubleshooting, 26with formalin, 12with mercuric chloride, 15with potassium dichromate fi xation, 16removal of, 23–24with Zenker and Helly solutions, 20with zinc formalin fi xation, 21

pigments, 254–255pipettes, micrometer, 75

placenta, Brown-Hopps gram stain on, 232plaques, neurofi brillary. See neurofi brillary plaques and tangles; senile

plaques.plasma cell cytoplasm, methyl green-pyronin Y staining, 126plasma cells

basophilia of, 106connective tissue types and structures, 160methyl green-pyronin Y staining, 188–189, 189

plasma stains, 113–114plasmalemma, 105plastics, embedding with, 41Pneumocystis jirovecii, demonstration with microwave variation of

methenamine-silver nitrate staining, 242–244, 243, 244poison, symbol, 90polarization

with mercuric chloride, 14and Zenker solution, 15

polarized lighton kidney specimen, 55

polychrome methylene blue, 127polychromic staining, 126–128polyclonal antisera, defi ned and described, 278polyethylene glycol, for processing, 40poly-l-lysine, coating on slides, 70polymeric detection in immunohistochemical staining, 285polyp, PAS staining of, 138polysaccharides

as marker for fungi, 235, 236, 239PAS reaction testing for, 137–141, 138types, occurrence, and reactivity, 136

portal vein, Masson trichrome staining of, 162postfi xation, and fi xative selection, 7potassium bromide solution

for Holzer staining method, 209potassium dichromate

comparative characteristics, 18and dye binding, 109safety precautions, 16

potassium ferrocyanidepreparation for Müller-Mowry colloidal iron technique, 150Prussian blue stain preparation, 257for Schmorl technique, 259stability of, 100for Turnbull blue stain preparation, 258

potassium hydroxidefor Gomori staining of reticulin, 175

potassium metabisulfi tefor Gomori staining of reticulin, 175preparation for PAS reaction testing, 138preparation for PAS reaction with diastase digestion, 140stability of, 100for thiofl avin S demonstration of Alzheimer disease, 206

potassium permanganatefor enhancing silver staining of nuclei, 177for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178for Mallory PTAH staining of glia, 208for Mallory PTAH technique, 180stability of, 100for thiofl avin S demonstration of Alzheimer disease, 206


potassium phosphatefor epithelial and connective tissue mucin demonstration, 147

power setting, on microwave staining oven, 68precipitate

troubleshooting, 41troubleshooting hematoxylin and eosin staining, 121

prediluted antibodies, storage considerations, 285–286prefi xatives, 354preparation, of solutions, 98preservatives vs fi xatives, 17primary aldehyde fi xation, for electron microscopy, 334primary buff ered aldehyde fi xation, for electron microscopy, 334primary buff ered PAF fi xation, for electron microscopy, 334primary fl uorescence, 56primary site, defi ned, 289proargol, for Bodian staining method, 197procedures, standards for, 292–296processing, 31–52

for electron microscopy, 337–340for immunohistochemical studies, 280troubleshooting, 41–42

processors, microwave, 66–67processors, tissue, 65–67progressive staining, with hematoxylin and eosin staining, 114–115propylene glycol, for Sudan black B staining, 186propylene oxide, and plastic embedding, 41prostate, cancer demonstrated with P504S antibody, 302protein solution, for blocking reactions, 287proteins

coagulation with mercuric chloride fi xation, 14fi xative eff ect on, 2fi xative comparative characteristics, 18and formaldehyde fi xation, 10and glyoxal fi xation, 14pH of, 107reactions to fi xatives, 8

protocolsand quality control, 294

protozoans, defi ned, 224Prussian blue stain

on bone marrow with iron, 258demonstration of ferric iron with, 256–258

pseudomelaninin colon and gastrointestinal tract stained with Schmorl technique,

260PTAH solution

for Mallory PTAH staining of glia, 207Mallory variation, 179–181, 180without mercuric solutions, 181

pull-apart methodof smear preparation, 354, 355

pyridinefor Holmes silver nitrate staining method, 199for Luxol fast blue-Holmes silver nitrate staining, 216

Qquality control, 289–296

for antibody, 290–291antibody log, 293chart, equipment, 77chart, reagents, 39for immunohistochemistry, 292of instrumentation, 75–76of paraffi n processing, 39for tissue block, 292

Rrabbit monoclonal antibodies, 279rabbit PAP, for peroxidase-antiperoxidase (PAP) immunohistochemical

technique, 298radiation hazard, symbol, 90radiographic, testing of decalcifi cation, 48Ralph knives, 58RBCs. See red blood cells.reactions, blocking of, 287reactivity, description, 88reagent alcohol, 33reagents, limonene, for clearing, 36rectum, fi xed in formalin and microwave processed, 40red blood cells

Bouin solution lysing of, 19demonstration with Pap staining, 363fi xation with Hollande solution, 20lysing by acetic acid, 9peroxidase eff ects on specimen, 287

reduction, defi ned, 310reducing rinse, preparation for alcian blue-PAS-hematoxylin technique,

149reducing solution

for Bodian staining method, 197for Churulian-Schenk method, 265for Grimelius argyrophil stain, 264for Holmes silver nitrate staining method, 200, for Luxol fast blue-Holmes silver nitrate staining, 216for microwave variation of Churukian-Schenk method, 266for Steiner and Steiner procedure, 249variations for reticulin staining, 175

reducing substances in tissue, demonstrated with Schmorl technique, 259–260

refractive index, fi xative eff ect on, 2refrigerators, for reagent storage, 72regressive staining

for diff erentiation, 109with hematoxylin, 112with hematoxylin and eosin staining, 115–116

regulations, safety, 82renal cell, adenocarcinoma in, 290repetitive stress injuries, 84reproductive toxins, 86RER. See rough endoplasmic reticulum.resolving power, of microscope, 54resorcin fuchsin stain, 171reticular fi bers

connective tissue types and structures, 160demonstration with Gordon and Sweets staining, 177–179, 179silver staining techniques for, 174–179staining variations, 175

396 Index

reticulin, demonstration in liver with Gomori stain, 176demonstration in liver with Snook variation, 177demonstration with Gomori stain, 176demonstration with Gordon and Sweets staining, 179

rhabdomyosarcomademonstration with Mallory PTAH technique, 179–181, 181small cell neoplasm panel, 290

Rhinosporidium seeberi, demonstration with Mayer mucicarmine technique and alcian blue staining, 244

rhodamine, for immunohistochemical studies, 283rhodanine method, copper demonstration with, 271–273, 272rhodanine solution, for microwave variation of rhodanine method, 273ribbon formation

on fl otation baths, 69optimal, 64with paraffi n, 37and paraffi n melting point, 42specimen orientation, 44troubleshooting, 59, 60

ribonucleic acid. See RNA.ribosomes, 105, 105Rickettsia, demonstration with Giemsa staining, 233right to know, 85, 89ripening, of hematoxylin, 110RNA, methyl green-pyronin Y staining, 126RNA

and picric acid fi xation, 15reactions to fi xatives, 8and zinc formalin fi xation, 16–17

Romanowski-type stains, 127rosebud, 396rough endoplasmic reticulum, 194

basophilia of, 106methyl green-pyronin Y staining, 126, 188–189ultrastructure of, 104, 105

Russell modifi cation of the Movat pentachrome stain, for demonstration of mucin and other elastic fi bers, 172–174, 173

SSaccomano fl uid, for prefi xation, 354Saccomano method of smear preparation, 356safety, 81–92

with alcohols and universal solvents, 34with decalcifi cation, 47with formalin, 13with glutaraldehyde fi xation, 14with mercuric chloride, 14with microtome blades, 58with osmium tetroxide, 15with picric acid, 16with potassium dichromate, 16with zinc salt fi xatives, 17

saline alcohol, preparation for alkaline Congo red staining, 153saline solution

for NADH diaphorase reaction, 323for specimen storage, 8

saline, physiological, for succinic dehydrogenase (SDH) reaction, 326salt linkage, 107salts, and dye binding, 109sandalwood, oil of, for clearing, 36

sarcoma, anaplastic tumor workup, 290sarcoplasmic reticulum, abnormality demonstration with NADH

diaphorase reaction, 323–325scales, 74Schiff reagent

for Bielschowsky-PAS staining, 201for chromic acid-Schiff stain, 237in Feulgen reaction, 123and glutaraldehyde fi xation, 13for Gridley staining, 238for Hotchkiss-McManus PAS reaction, 235for Luxol fast blue-PAS-hematoxylin staining, 218preparation for PAS reaction testing, 137preparation for PAS reaction with diastase digestion, 140

Schmorl techniquedemonstration of reducing substances in tissue, 259–260melanin demonstration with, 260

Schwann cell, 194Scott solution, composition, 112scratches, troubleshooting microtomy, 58SDH, histochemical reactions of human muscle types, 309secretory granules, 106, sectioning, 314

for electron microscopy, 340–341troubleshooting, 342–343uneven, 60

senile plaquesdemonstration with Bielschowsky-PAS staining, 200–205, 202, 204demonstration with Luxol fast blue-PAS-hematoxylin staining,

218–219demonstration with Sevier-Munger modifi cation of Bielschowsky

method, 204–205demonstration with thiofl avin S staining, 205–207, 207

sensitizer variations for reticulin staining, 175sensitizers, xylene, limonene, and aliphatic carbon comparisons, 37SER. See smooth endoplasmic reticulum.serosal surfaces, defi ned, 5sharps containers, 83shelf life, of solutions, 99, 100short term exposure limit (STEL), 85shrinkage

with formaldehyde fi xation, 10sialomucins

alcian blue demonstration of, 145demonstration with Müller-Mowry colloidal iron technique, 151stained with alcian blue at pH 1, 147

silver nitratefor Churulian-Schenk method, 265for Dieterle staining method, 248for Fontana-Masson stain preparation, 261for Gomori methenamine-silver technique for urates, 267for Gomori staining of reticulin, 175for Gordon and Sweets staining technique, 178for Grimelius argyrophil stain, 264for Grocott staining, 240for microwave variation of Bielschowsky-PAS staining, 203for microwave variation of Churukian-Schenk method, 266for microwave variation of Fontana-Masson stain, 262for microwave variation of methenamine-silver nitrate staining, 242for microwave variation of Warthin-Starry technique, 246for Sevier-Munger modifi cation of Bielschowsky method, 204for Steiner and Steiner procedure, 249for von Kossa calcium stain, 270for Warthin-Starry technique, 244


silver staining techniques, for reticular fi bers, 174, 175sinusoids, demonstration with Gordon and Sweets staining, 179, 176size, of specimen, and fi xation, 4skeletal muscle, 308

α-naphthyl acetate esterase stain, 316atrophy demonstration with ATPase, 320demonstration of normal pattern with ATPase, 319demonstration with α-naphthyl acetate esterase stain, 316frozen sectioning of, 65hematoxylin and eosin staining of, 308neuropathic disease in, 309staining with Mallory PTAH technique, 181

skinbiopsy examination with immunofl uorescence, 283demonstrating cytoplasmic granules with Fontana-Masson staining,

262mast cell demonstration, 188, 189specimen orientation, 43stained with Gomori aldehyde fuchsin procedure, 171troubleshooting hematoxylin and eosin staining, 119Verhoeff -van Gieson staining of, 169waviness in, 73

slidescoatings for, 70electrostatically charged, 69control, storage of, 292

slow-twitch muscle, 308small cell neoplasm panel, 290smear preparation, methods, 354–355SMI-31 antibody, staining pattern on brain biopsies, 287smooth endoplasmic reticulum, ultrastructure of, 105smooth muscle, in Verhoeff -van Gieson staining of aorta and artery, 169Snook variation

oxidizer, sensitizer, impregnating solution, and reducing solution, 175

reticulin demonstration in liver section, 177sodium acetate

composition, 125for Grimelius argyrophil stain, 264

sodium acetate-formalin solutionfor microwave variation of rhodanine method, 273

sodium barbitalfor ATPase stain, 318for naphthol AS-D chloroacetate esterase technique, 316

sodium bisulfi tefor Grocott staining, 240for microwave variation of methenamine-silver nitrate staining, 243stability of, 100

sodium boratefor Gomori methenamine-silver technique for urates, 268for microwave variation of rhodanine method, 273for periodic acid-methenamine silver microwave procedure, 182for rhodanine copper demonstration, 272stability of, 100

sodium carbonatefor Sevier-Munger modifi cation of Bielschowsky method, 204

sodium hydroxidefor α-naphthyl acetate esterase stain, 315for Gordon and Sweets staining technique, 178for thiofl avin S demonstration of Alzheimer disease, 206

sodium metabisulfi tefor alcian yellow-toluidine blue staining method, 235for Hotchkiss-McManus PAS reaction, 236preparation for alcian blue-PAS-hematoxylin technique, 149

sodium nitritefor acid phosphatase detection in muscle, 321for α-naphthyl acetate esterase stain, 315

sodium phosphatefor epithelial and connective tissue mucin demonstration, 147

sodium thiosulfatefor Bielschowsky-PAS staining, 201for Bodian staining method, 197for Gomori methenamine-silver technique for urates, 268for Gomori staining of reticulin, 176for Gordon and Sweets staining technique, 178for Grocott staining, 240for Holmes silver nitrate staining method, 200for Luxol fast blue-Holmes silver nitrate staining, 216for Mallory PTAH technique, 180for microwave variation of Bielschowsky-PAS staining, 203for microwave variation of Fontana-Masson stain, 262for microwave variation of methenamine-silver nitrate staining, 243for periodic acid-methenamine silver microwave procedure, 182for Russell modifi cation of the Movat pentachrome stain, 172for Sevier-Munger modifi cation of Bielschowsky method, 205stability of, 100for Verhoeff elastic stain, 168for von Kossa calcium stain, 270

soft mushy tissue, troubleshooting, 44–45, 45, 58solution preparation, 93–102solutions, molar, 96solutions, stability of, 99, 100solvent power, xylene, limonene, and aliphatic carbon comparisons, 37solvent recycler, 75solvents, transitional, for electron microscopy, 337somatic nervous system, 194sparsely cellular specimens, smear preparation, 356specifi cation sheet, antibody, 286specimen, collection, 352–353specimen orientation, in frozen sectioning, 51spherule, demonstration with Gridley staining, 239spills, chemical, 87spinal cord, demonstration with Weil method, 212spirochetes

demonstration with Dieterle staining method, 247–248demonstration with microwave variation of Warthin-Starry

technique, 245–247demonstration with Steiner and Steiner microwave method, 249–250demonstration with Warthin-Starry technique, 244–247, 245

spleencryostat processing temperature, 64ferrous iron in, demonstrated with Turnbull blue stain, 259fi xed in formalin, 10M leprae demonstration in, with Fite staining technique, 229

splits, troubleshooting microtomy, 58sponge artifact, troubleshooting, 42Spurr embedding, 41, 337–339squamous cell

contamination in fl otation bath, 70demonstration with Pap staining, 363

398 Index

stability of solutions, 99, 100stabilization of specimens, 2stage, of microscope, 54staining

automated, 67–68, 116–117of basement membranes, 182–184of carbohydrates, 137–152of connective tissue cells, 188–189of connective tissues, 162–189cytoplasmic, 107–108fi xative eff ect on, 2of frozen sections, 117–118and ions, 10of lipids, 184–188of nerve tissue, 195–219nuclear, 107of nucleic acids, 123–126of plasmas, 113–114principles and methods, 107, 129

staining techniquesfor electron microscopy, 343immunohistochemical, 284–285for immunohistochemical studies, 297–302for molds, 223for thin sections, 345–346, 345

staining, cytology, 361staining. See also specifi c methods and stains.stains, ionic bonding of, 107standards, for immunohistochemical procedures, 292–296Staphylococcus aureus, demonstration with Brown-Hopps method, 232static electricity, troubleshooting sectioning problems, 64Steiner and Steiner procedure

demonstration of H pylori with, 250demonstration of spirochetes, H pylori, and L pneumophila with,

249–250STEL (short term exposure limit), 85sticking, of sections, troubleshooting, 64storage

of acetic acid, 9of antibodies, 287of B-5 solution, 19of chemicals, 87, 88of control slides, 292of formalin, 13with glyoxal fi xation, 14and quality control, 294of solutions, 98solutions and fi xatives for, 8of specimens, 7

striations, in muscle, demonstrated with Mallory PTAH technique, 179–181, 181

submucosahematoxylin and eosin staining of, 115tumor demonstration with Grimelius stain, 264

substage, of microscope, 54succinic dehydrogenase (SDH) reaction, for narrowing diagnosis of

NADH reaction, 325–326, 326sucrose, embedding with, 41Sudan black B, lipid demonstration with, 186Sudan black B, liver staining with, 186sulfated acid mucopolysaccharides, stained with alcian blue at pH 1, 147

sulfurous acidfor chromic acid-Schiff stain, 237in Feulgen reaction, 123for Hotchkiss-McManus PAS reaction, 236

SurePath System, 359, 359, 360swine antirabbit linking serum, for peroxidase-antiperoxidase (PAP)

immunohistochemical technique, 298symbols, for hazardous materials, 90synapse, 194

Ttalc, in frozen sectioning, 50tangles, neurofi brillary. See neurofi brillary plaques and tangles.tattoo ink, specimen marking, 43tattoo pigments, 254tears, in section, 64temperature

for control slide storage, 292and dye binding, 109and fi xation, 4

for electron microscopy, 335metric, 97–98and quality control, 77and ribbon formation problems, 60

tendons, connective tissue types and structures, 160tertiary butanol

for clearing, 36for dehydration, 34

testing, decalcifi cation, 48tetrahydrofuran

for clearing, 36for fi rming soft mushy tissue, 46for dehydration, 34

thickness, of sectiontroubleshooting, 62and fi xation, 4

thin sections, staining techniques, 345–346, 345Th inPrep System, 359, 359, 360thiofl avin S, fl uorescent dyeing, 56thiofl avin S staining, demonstration of neurofi brillary plaques and

tangles, 205–207, 207thiofl avin T, fl uorescent dyeing, 56

for amyloid demonstration, 155–1563,3'-diaminobenzidine chromogen. See DAB (3,3'-diaminobenzidine

chromogen).threshold limit value, xylene, limonene, and aliphatic carbon

comparisons, 37thyroid, adenocarcinoma, 290tigroid substance, 194time, and fi xation, 5, 6

for electron microscopy, 335tissue, fi xative eff ect on, 2tissue block, quality control for, 292tissue fi xation, for immunohistochemical studies, 279–280tissue handling, 279–283tissue microarray cores, with positive and negative controls, 291tissue processors, 65–67

maintenance of, 66tissue waste, handling, 83


tissue, soft mushy, troubleshooting, 44–45, 45, 58TLV. See threshold limit value.toluene

for clearing, 35fl ash point, 87for testing water content, 33

toluidine blue stainingfor alcian yellow-toluidine blue staining method, 235for electron microscopy, 344of kidney under electron microscope, 345for mast cell demonstration, 188mast cell demonstration, 188mold demonstration with, 223

toluidine blue wet fi lm, for determining cellularity, 363–364toluidine blue-basic fuchsin procedure

for electron microscopy, 343–344of kidney under electron microscope, 344

tonicity, and fi xation for electron microscopy, 335toxicity, description, 88Toxoplasma gondii, demonstration with Giemsa staining, 233training and quality control, 294transferases, 312transitional fl uids, and plastic embedding, 41transport solutions, 23trapping of chromogen beneath specimen, 281Tris buff er, demonstration of muscle fi ber regeneration, 322tris-buff ered saline solution, for HRP enzyme labeled polymer

procedure, 301Triton X-100, in fl otation bath processing, 70Trizma base, demonstration of muscle fi ber regeneration, 322troubleshooting, autolysis from fi xation, 25

chromatin autolysis, 24–26decalcifi cation, 49–50embedding, 44–46fi xation, 24–26frozen sectioning, 50–51hematoxylin and eosin staining, 118–123immunoperoxidase techniques, 296–297microtomy, 58–64mounting, 130–131, 130nuclear bubbling, 24–26, 26pigment artifact, 26processing, 41–42sectioning, 342–343

TTF-1, adenocarcinoma panel, 290tuberculosis exposure, 82tubular structures, specimen orientation, 44tumors

B-5 vs formalin fi xation, 19defi ned, 289diff erential diagnosis with Gordon and Sweets staining, 177–179carcinoid, demonstration with Fontana-Masson stain, 260–263carcinoid, demonstration with Grimelius stain, 264carcinoid, demonstration with microwave variation of Fontana-

Masson stain, 261–263fi xation illustrations, 6fi xed in formalin and microwave processed, 40neurosecretory, demonstration with Churukian-Schenk method,

265–267silver staining techniques for, 174–179

tunica media, in Verhoeff -van Gieson staining of aorta and artery, 169Turnbull blue stain, demonstration of ferrous iron with, 258–259, 259Tzank smears, collection and handling of, 352

Uultrastructure, of cells, 104–106umbilical cord, alcian blue staining with and without hyaluronidase

digestion, 148underdecalcifi cation, troubleshooting, 49, 49undulations, in section, 61, 62universal precautions, with infectious hazards, 82universal secondary antibodies, 289universal solvents

for dehydration, 34for clearing, 36

uranyl nitratestability of, 100for Steiner and Steiner procedure, 249

urates, 255–256urates

demonstration with Gomori methenamine-silver technique, 267–268hematoxylin and eosin staining of, 256

urine specimenscollection and handling of, 353smear adhesion problems, 358

uterusfi xation troubleshooting, 25washboarding in, 61, 62

Vvacuum

for decalcifi cation, 47for paraffi n embedding, 38

validationof antibodies, 286form for antibodies and controls, 288

Van der Waals forces, 107van Gieson picric acid-acid fuchsin stain, counterstaining with, 166–167van Gieson solution

for bile stain, 269 solution, composition, 167for Verhoeff elastic stain, 168

veinMasson trichrome staining of, 162van Gieson staining, 167

Verhoeff elastic stainfor pathologies of elastic tissue, 168–169stability of, 100

veronal acetate buff er, for acid phosphatase detection in muscle, 321villi, of small intestine, defi ned and illustrated, 5Vimentin

anaplastic tumor workup, 290small cell neoplasm panel, 290

viral lesion scrapings, collection and handling of, 352viruses, defi ned, 223volume, of specimen

and fi xation, 4and fi xation, 5

volumetric glassware, 94von Gierke disease

and Best carmine demonstration of glycogen, 142von Kossa calcium stain, calcium demonstration with, 269–270, 270

400 Index

Wwarning sign, for formaldehyde, 85Warthin-Starry technique, demonstration of spirochetes with, 244–247, 245washboarding, of section, 61, 62water, Millipore-fi ltered deionized, for fl otation bath processing, 70water bath, circulating, for temperature control of specimen, 72water beads, on mounted section, 69water content, testing for, 33wattage setting, on microwave staining oven, 68waviness, in section, 73waxes, water-soluble, for processing, 40Weigert hematoxylin, composition, 113Weigert iron hematoxylin

for Gomori 1-step trichrome stain, 165for Masson trichrome staining, 163preparation for Mayer mucicarmine technique, 143for van Gieson staining, 167

Weil staining methodmyelin sheath demonstration with, 211on spinal cord and medulla, 212

white matter, demonstration with Bodian staining method, 198demonstration with Luxol fast blue, 213demonstration with Weil method, 212indistinct diff erentiation with Luxol fast blue, 214

Wilder variation, oxidizer, sensitizer, impregnating solution, and reducing solution, 175

Wilms tumor, small cell neoplasm panel, 290Wilson disease

copper demonstration with microwave rhodanine method, 272–273, 273

copper demonstration with rhodanine method, 271–273, 272Wright staining

diff erentiation with, 109drying preparation for, 3

wrinkling, of sectionfrom fl otation bath processing, 69on lung section, 73troubleshooting, 62

WT-1, small cell neoplasm panel, 290

Xxylene

attribute comparisons, 37for clearing, 35for dehydrating soft mushy tissue, 45fl ash point, 87for microscope maintenance, 54overnight processing with, 38rapid processing with, 38for testing water content, 33

xylene-peanut oilfor Fite acid-fast staining technique, 228

Yyeasts, 223

ZZamboni solution

for electron microscopy, 334, 336formulation and applications, 21

Zenker solutionon bone marrow section stained with Prussian blue, 258and decalcifi cation testing, 48and dye binding, 109as fi xative in Mallory PTAH-stained muscle, 181and fi xative selection, 7formulation and applications, 20for gastrointestinal tract, 21in immunohistochemical studies, 279labeling, 89and nuclear staining, 15and polarized light, 15specimen removal from, 7stability of, 100

Ziehl-Neelsen carbol-fuchsin solutionfor Fite acid-fast staining technique, 229

Ziehl-Neelsen method, mycobacteria demonstration with, 226–228zinc chloride

for fi xation, 16zinc formalin

and decalcifi cation testing, 48zinc formalin, eff ect on protein, 4zinc formalin, formulation and applications, 21–22zinc formalin

for immunohistochemical studies, 16–17zinc formalin

in immunohistochemical studies, 280zinc salts, comparative characteristics, 18zinc salts

for fi xation, 16–17zinc salts, safety precautions, 17zinc sulfate

for fi xation, 16–17

carson glossary+index v04 0407 · 2016-04-20 · for grimelius argyrophil stain, 264 for luxol fast...

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