Pigment & mineral Word Scramble
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| Question | Answer |
| Artifact pigment | Formed as a result of chemical action - fixation, processing, usually on tissue, not w/i cell, e.g. formalin, mercury, chrome pigments |
| Exogenous pigment | From outside the body, carbon pigment, asbestos fibers, tattoo pigments, metals - normal (iron) or pathologic (copper) |
| Carbon pigment | Seen in lung & lymph node, resists bleaching & extration, insol. in conc'd sulfuric acid - use to differentiate |
| Asbestos fibers | Birefringent, magnesium silicate, in body fibers coated w/ iron-containing protein, known as asbestos bodies, lose birefringence demo w/ Prussian blue |
| Endogenous hematogenous pigments | From blood, e.g. hemoglobin, hemosiderin, biliverdin |
| Hemoglobin | Conjugated protein, anionic, stains w/ acid dyes (eosin), hemoglobin breaks into globin protein & heme, heme splits into iron & biliverdin |
| Hemosiderin | Yellow to brown pigment, stored iron (Fe2+), usu. in marrow, pathologic elsewhere (liver), differentiate from other yellow-brown pigments w/ Prussian blue rxn |
| Hemochromatosis | Disease caused by excessive absorption of dietary iron, excessive hemosiderin deposits in liver, pancreas, damaging to tissue |
| Biliverdin | Greenish bile pigment, prod'd from destruction of rbc, liver red. to bilirubin - remove from circulation, abnormal accumulation - jaundice, pigment in bile canaliculi, cytoplasm of Kupffer cells & hepatocytes |
| Hematoidin | Similar to bilirubin, ox. to bilirubin, formed as result of hemorrhage & red'd O2 tension |
| Endogenous nonhematogenous pigments | Lipid - e.g. lipofucsin, ceroid, nonlipidic - e.g. melanin |
| Melanin pigment | Formed by ox. tyrosine, histochem. demo melanocytes, bleach w/ H2O2, potassium permanganate & oxalic acid, dissolved by strong alkali, insol. weak acid, base, organic solvents, argentaffin, positive Schmorl |
| Lipofuscin pigment | "Wear & tear" pigment, yellow-brown, in more permanent cells (heart, liver, neurons) |
| Ceroid pigment | Brownish-yellow, hepatocytes & macrophages of rats w/ cirrhosis, rarely seen in humans |
| What stains for lipofuscin & ceroid pigments? | Oil red O, Sudan black B, & PAS, differentiate - ceroid is acid-fast, lipofsucin is not |
| Engodenous deposits | Urate crytals (gouty tophi), chronic gout may deposit in soft tissue too, sol. in water, alcoholic fix., birefringent w/ polarizing, demo w/ argentaffin rxn |
| Minerals | Special stains can demo: Ca2+, Fe2+/Fe3+, Cu2+, PO43+, CO32+, silver, lead, copper, gold, microincineration can be used to ID inorganic components |
| Cytoplasmic granules | Adrenal chromaffin granules, pancreatic endocrine cells, GI enterochromaffin cells, "C" cells of thyroid, some pituitary cells, are APUD cells demo w/ argyrophil or argentaffin rxns, fixative v. important |
| APUD cells | Have high uptake of amine-precursors & ability to decarboxylate |
| Best fixative for chromaffin granules? | Primary chromate fixatives, e.g. Orth |
| Argentaffin granules of GI are destroyed by? | Alcoholic fixatives |
| Paneth cell granules are destroyed by? | Acetic acid |
| Prussian blue stain purpose | Detect ferric (Fe3+) iron, usu. small amount in marrow & spleen, large deposits - hemochromatosis, hemosiderosis |
| Prussian blue stain facts | Fixative: alcohol or 10% NBF, QC: section w/ Fe3+, don't want excess iron, chemically clean glassware, iron-free reagents |
| Prussian blue stain solutions | potassium ferrocyanide 2% (bind Fe3+) HCl 2% (acidify) nuclear-fast red |
| Prussian blue stain results | nuclei & hemofuchsin - bright red hemosiderin (iron) - blue background - pink |
| Nuclear-fast red solution | aluminum sulfate nuclear-fast red thymol (preservative) dH2O |
| Fixation for iron demo: | Some acidic fixatives & decal. solns. dissolve iron, can demo iron after Zenker w/ acetic acid, but mercury - bad, best to use another fixative & decal briefly in acetic or formic acid |
| Turnball blue stain purpose | Detect ferrous (Fe2+) iron, ferrous iron v. toxic, not normally stored, easily absorbed by intestine but converted to ferric, used w/ Schmorl technique |
| Turnball blue stain facts | Fixative: alcohol or 10% NBF QC: section w/ Fe2+ |
| Turnball blue stain solutions | potassium ferricyanide staining soln. (bind Fe2+) acetic acid 1% (post-stain rinse) nuclear-fast red |
| Turnball blue stain results | ferrous iron - blue background - pink-red |
| Potassium ferricyanide staining solution | potassium ferricyanide HCl 0.06N |
| Schmorl technique purpose | Indicate reducing substances - will red. ferric ions, stains melanin, argentaffin granules, formalin pigment, mostly replaced by IHC |
| Schmorl technique facts | Fixative: 10% NBF QC: section w/ melanin or argentaffin granules |
| Schmorl technique solutions | ferric chloride-potassium ferricyanide working soln. pH 2.4 Mayer mucicarmine soln. or metanil yellow |
| Schmorl technique results | reducing/argentaffin substances - blue-green goblet cells, mucin - rose background - yellow-green (less better) |
| Mayer mucicarmine solution | carmine alum lake aluminum hydroxide anhy. aluminum chloride 50% ethanol 25% ethanol |
| Fontana-Masson & microwave Fontana-Masson stains purpose | Demo argentaffin substances, e.g. melanin, argentaffin granules of carcinoid tumors, some neurosecretory granules, demos other red'ing substances (formalin pigment) |
| Fontana-Masson & microwave Fontana-Masson stains facts | Fixative: 10% NBF, avoid alcohol QC: skin for melanin, small intestine or appendix for argentaffin granules, Melanoma, tumor w/ argentaffin granules, chemically clean glassware, nonmetallic instruments |
| Fontana-Masson stain solutions | Fontana silver nitrate w/ NH4OH gold chloride sodium thiosulfate nuclear-fast red |
| Fontana-Masson stain results | melanin, argentaffin granules - black nuclei - pink overstaining gives gray background & loss of contrast |
| Microwave Fontana-Masson stain solutions | Fontana silver nitrate w/ NH4OH gold chloride sodium thiosulfate nuclear-fast red |
| Microwave Fontana-Masson stain results | argentaffin cell granules, chromaffin granules, melanin, other argentaffin substances - black nuclei - pink |
| Grimelius stain purpose | Demo argyrophil granules in neurosecretory tumors, also stains argentaffin substances |
| Grimelius stain facts | QC: argyrophil-positive carcinoid tumor preferred, can use small intestine, chemically clean glassware, nonmetallic forceps |
| Grimelius stain solutions | working silver solution reducing solution nuclear-fast red |
| Grimelius stain results | argentaffin & argyrophil granules - dark brown to black nuclei - red background - pale yellow-brown |
| Grimelius working silver solution | acetic acid-acetate buffer pH 5.6 silver nitrate 1% dH2O |
| Grimelius, Churukian-Schenk, microwave Churukian-Schenk reducing solution | hydroquinone sodium sulfite dH2O |
| How to differentiate between argentaffin & argyophil substances? | Stain w/ both Grimelius (argyophil) & Fontana-Masson (argentaffin) & compare |
| Churukian-Schenk & microwave Churukian-Schenk methods purpose | Demo argyrophil granules in neurosecretory tumors |
| Churukian-Schenk & microwave Churukian-Schenk methods facts | QC: argyrophil-positive carcinoid tumor preferred, can use small intestine, chemically clean glassware, nonmetallic forceps |
| Churukian-Schenk method solutions | acidified water pH 4.0 to 4.2 silver nitrate 0.5% reducing solution nuclear-fast red |
| Churukian-Schenk method results | argyophil granules, argentaffin substances - black nuclei - red background - yellow-brown |
| Microwave Churukian-Schenk method solutions | citric acid-glycine soln. silver nitrate 0.5% reducing solution nuclear-fast red |
| Microwave Churukian-Schenk method results | argyrophil & argentaffin cells - black nuclei - orange to red background - light yellow-orange |
| Gomori methenamine-silver method purpose | Demo urates, may demo large calcium deposits |
| Gomori methenamine-silver method facts | Fixative: abs. alcohol QC: section w/ urates |
| Gomori methenamine-silver method solutions | working methenamine-silver nitrate sodium thiosulfate light green w/ acetic acid |
| Gomori methenamine-silver method results | urates - black background - green |
| Bile stain purpose | Detect bilirubin in tissue, aka Hall stain, aka Fouchet rxn |
| Bile stain solutions | Fouchet reagent van Gieson |
| Bile stain results | bile, bilirubin - emerald green to olive drab background - yellow |
| Fouchet reagent solution | trichloracetic acid ferric chloride dH2O |
| van Gieson solution | acid fuchsin picric acid |
| von Kossa stain purpose | ID calcium in tissue, detects anions bound to calcium, not the calcium itself |
| von Kossa stain facts | Fixative: alcohol preferred, 10% NBF ok |
| von Kossa stain solutions | silver nitrate sunlight sodium thiosulfate nuclear-fast red |
| von Kossa stain results | calcium salts - black background - red |
| Alizarin Red S stain purpose | ID calcium in tissue, reaction product is birefringent |
| Alizarin Red S stain facts | Fixative: alcoholic formalin or 10% NBF |
| Alizarin Red S staining solution | Alizarin red S (chelate w/ calcium) dH2O NH4OH to pH 4.1 to 4.3 - critical |
| Alizarin Red S stain results | calcium deposits - orange-red |
| Rhodanine & microwave Rhodanine methods purpose | Detect copper in tissue, esp. liver - Wilson disease, aka Lindquist rhodanine |
| Rhodanine & microwave Rhondanine methods facts | Fixative: 10% NBF Sections: paraffin 6-8 µm QC: section w/ copper (fetal liver) |
| Rhodanine method solutions | working rhodanine soln. Mayer hematoxylin w/ dH2O borax (blue Mayer) |
| Rhodanine method results | copper - bright red to red yellow nuclei - light blue If copper conc. low, stain may fade after coverslipping |
| Microwave Rhodanine method solutions | working Rhodanine w/ sodium acetate-formalin Mayer hematoxylin w/ dH2O borax |
| Microwave Rhodanine method results | copper - bright red or rust-red intracytoplasmic granules nuclei - blue |
| Working Rhodanine solution | 5-(p-dimethylamino-benyzlidine) rhodanine abs. ethanol dH2O filter |
| Pigment | A substance that has enough color to be visible w/o any further staining |
| What are some diseases associated w/ melanin? | Melanomas, Addison's disease |
| Prussian blue vs. Turnball blue | Prussian - ferrocyanide for ferric ions Turnball - ferricyanide for ferrous ions |
| Fontana Masson w/ melanin bleach | To compare, deparaffinize & hydrate 2 slides, leave 1 in dH2O, 1 in potassium permanganate, wash w/ tap & dH2O, in oxalic acid, wash w/ tap & dH2O, stain both w/ Fontana Masson protocol |
| Argentaffin stains | Gomori methenamine-silver, Grocott methenamine-silver, Fontana Masson, von Kossa |
| Argyrophil stains | Reticulin, Holmes, Bielschowsky, Warthin-Starry, Dieterle, Steiner & Steiner, Grimelius, Churukian-Schenk |
| Formalin pigment | Is a reducing substance, may give positive rxn to Schmorl & Fontana-Masson |
| Chloroacetate Esterase (Leder) stain purpose | Detection of neutrophils |
| Stained starch granules show: | A characteristic Maltese cross configuration when polarized, may be introduced from talcum powder of gloves |
| Ferrous ions | Fe2+, toxic, not normally stored, Turnbull blue |
| Ferric ions | Fe3+, normally present in tissue, usu. bound to protein (hemosiderin), in marrow, spleen, decaying, hemorrhagic material, excess - hemachromatosis, Prussian blue |
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CCF
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