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Histology Spec Stain
Histology ASCP Special Stains
| Question | Answer |
|---|---|
| Substances attracting basic dyes | Basophillic |
| Substances attracting acid dyes | Acidophillic |
| A group that confers the property of color | Chromophore |
| A benzene derivative containing chromophoric groups | Chromogen |
| Ionizing group required to enable the dye to link firmly to tissue | Auxochrome |
| A basic dye in which the charge on the dye ion is postive | Cationic dyes |
| An acidic dye with a negative charge | Anionic dyes |
| Progressive staining | Once desired intensity of color is achieved, the reaction is stopped |
| Substances or metals that act as a link between dye and tissue | Mordant |
| Regressive staining | The tissue is overstained and then differentiated, or decolorized, until only the desired element is left stained. |
| Basic, or cationic dyes, are differentiated by: | Weak acid solutions |
| Acidic, or anionic dyes, are differentiated by: | Weak alkaline solutions |
| Reagents: Hematoxylin, Ethanol, Ammonium Alumnium Sulfate, Distilled Water, Mercuric Oxide | Harris Hematoxylin |
| Mordant: Aluminum Ripener: Sodium Iodate | Harris Hematoxylin, Mayer Hematoxylin, Ehrlich Hematoxylin, Gill Hematoxylin |
| Reagents: Ammonium Aluminum Sulfate, Distilled Water, Hematoxylin, 95% Alcohol, Glycerol | Delafield Hematoxylin |
| Mordant: Aluminum Ripner: Light and air | Delafield Hematoxylin, Ehrlich Hematoxylin |
| Reagents: Hematoxylin, Distilled Water, Sodium Iodate, Ammonium or potassium Aluminum Sulfate, citric Acid, chloral Hydrate | Mayer Hematoxylin |
| Reagents: Hematoxylin, 95% Alcohol, Distilled Water, Glycerol, Ammonium or Potassium Aluminum Sulfate, Glacial Acetic Acid | Ehrlich Hematoxylin |
| Reagents: Distilled Water, Ehtylene Glycol, Hematoxylin, Sodium Iodate, Aluminum Sulfate, Glacial Acetic Acid | Gill Hematoxylin |
| Mucin, especially goblet cells, will be stained by which hematoxylin | Gill Hematoxylin |
| Reagents: Ferric Chloride, Distilled Water, Hydrochloric Acid, Hematoxylin, 95% Alcohol | Weigert Hematoxylin |
| Mordant and oxidizer: Ferric Chloride | Weigert Hematoxylin |
| Hematoxylin substitute | Celestine Blue |
| Bluing agent | Scott Solution |
| Fixatives that cause loss of basophilia | Bouin, Zenker or unbuffered formalin solutions. |
| Demonstration of DNA | Feulgen Reaction |
| Any fixative but Bouin | Feulgen Reaction |
| Reagents: Hydrochloric Acid, Schiff Reagent, Sulfurous Acid | Feulgen Reaction |
| DNA: Reddish Purple Cytoplasm (if counterstained): Light Green | Feulgen Reaction |
| Differentiation between DNA and RNA | Methyl Green-Pyronin Y |
| 10% NFB preferred. B5, Helly or Zenker satisfactory. | Methyl Green-Pyronin Y |
| Reagents: Acetic Acid, Sodium Acetate, Methyl Green Staining Solution, Methyl Green-Pyronin Y Staining Solution | Methyl Green-Pyronin Y |
| DNA: Green to blue-green RNA: Red to rose Goblet Cells: Mint green Background: Pale pink to colorless Immunoblast and plasma cell cytoplasm: Intense Red Nuclei Green to blue-green | Methyl Green-Pyronin Y |
| Process in which a dye forms other dyes spontaneously | Polychroming |
| Differentiation of cells present in hematopoietic tissue. | May-Grunwald Giemsa |
| Demonstration of microorganisms | May-Grunwald Giemsa |
| Zenker or B5 preferred. 10% NBF satisfactory. | May-Grunwald Giemsa |
| Control: Spleen | May-Grunwald Giemsa |
| Jenner Solution, Giemsa Solution, Acetic Water | May-Grunwald Giemsa |
| Nuceli: Blue Cytoplasm of leukocytes: Pink, gray or blue Bacteria: Blue | May-Grunwald Giemsa |
| Demonstration of polysaccharides, neutral mucosubstances and basement membranes | PAS |
| 10% NBF or Bouin | PAS, Alcian Blue pH 2.5, Alcian Blue pH 1.0 |
| Control: Kidney | PAS, Alcian Blue-PAS-Hematoxylin, Periodic Acid-Methenamine Silver |
| Reagents: Periodic Acid, Hydrochloric Acid, Potassium Metabisulfite, Schiffs | PAS |
| Glycogen, neutral mucosubstances, certain epithelial sulfomucins and sialomucins, colloid material of the thyroid and pars intermedia of the pituitary, basement membranes and fungal walls: Bright Rose | PAS |
| Demonstration of Glycogen | PAS D, Best Carmine |
| 10% NBF, formalin alcohol or absolute alcohol | PAS D |
| Control: Liver | PAS D, Best Carmine, Gomori Stain, Gordon and Sweets |
| Reagents: Periodic Acid, Hydrochloric Acid, Schiffs, Potassium Metabisulfite, Malt Diastase, Phosphate Buffer | PAS D |
| Glycogen: Bright Rose | PAS D |
| Absolute Alcohol preferred. Carnoy and Bouin satisfactory. | Best Carmine |
| Reagents: Carmine, Ammonium Hydroxide, Methanol | Best Carmine |
| Glycogen: Pink to red Nuclei: Blue | Best Carmine |
| Demonstration of epithelial mucin | Mayer Mucicarmine |
| Control: Unautolyzed colon, small intestine or appendix. | Mayer Mucicarmine, Alcian Blue pH 2.5, Alcian Blue pH 1.0, Muller-Mowry Colloidal Iron |
| Reagents: Mucicarmine, Weigert Hematoxylin, Metanil Yellow | Mayer Mucicarmine |
| Mucin: Deep rose to red Capsule of cryptococcus: Deep rose to red Nuclei: Black Other: Blue or yellow | Mayer Mucicarmine |
| Demonstration of acid mucopolysaccharides | Alcian Blue pH 2.5 |
| Reagents: Acetic Acid, Alcian Blue, Nuclear Fast Red | Alcian Blue pH 2.5 |
| Sulfated Mucosubstances: Dark Blue Hyaluronic Acid: Dark Blue Sialomucins: Dark Blue Background: Pink to red Nuclei: Red | Alcian Blue pH 2.5 |
| Demonstration of sulfated mucosubstances | Alcian Blue pH 1.0 |
| Reagents: Hydrochloric Acid, Alcian Blue, Nuclear Fast Red | Alcian Blue pH 1.0 |
| Sulfated mucosubstances: Pale blue Background: Pink to red Nuclei: Red | Alcian Blue pH 1.0 |
| Differentiation of epithelial and connective tissue mucins | Alcian Blue with Hyaluronidase |
| Control: Umbilical cord | Alcian Blue with Hyaluronidase |
| Reagents: Potassium Phosphate, Sodium Phosphate, Hyaluronidase Digestion Solution, Alcian Blue, Nuclear Fast Red | Alcian Blue with Hyaluronidase |
| Without digestion: acid mucopolysaccharides and sialomucins: Deep Blue With digestion: mucosubstances containing hyaluronic acid and chondroitin sulfates: Marked loss of staining | Alcian Blue with Hyaluronidase |
| Differentiation between neutral and acidic mucosubstances | Alcian Blue-PAS-Hematoxylin |
| 10% NBF or Zenker | Alcian Blue-PAS-Hematoxylin, Verhoeff |
| Reagents; Acetic Acid, Alcian Blue,Periodic Acid, Sodium Metabisulfite | Alcian Blue-PAS-Hematoxylin |
| Acid mucosubstances: Blue Neutral polysaccharides: Magenta Certain substances will be purple | Alcian Blue-PAS-Hematoxylin |
| Demonstration of carboxylated and sulfated mucopolysaccharides and glycoproteins | Muller-Mowry Colloidal Iron |
| Chemical reaction to demonstrate iron bound to tissue | Prussian Blue |
| 10% NBF, Carnoy Solution or alcoholic formalin | Muller-Mowry Colloidal Iron |
| Avoid chromate fixatives | Muller-Mowry Colloidal Iron, Gomori Aldehyde Fuchsin |
| Reagents: Ferric chloride, Muller Colloidal Iron, Potassium Ferrocyanide, Hydrochloric Acid, Acetic Acid, Nuclear Fast Red | Muller-Mowry Colloidal Iron |
| Acid mucopolysaccharides and sialomucins: Deep blue Nuclei: Pink-red Cytoplasm: Pink | Muller-Mowry Colloidal Iron |
| Demonstration of Cryptococcus: | Mayer Mucicarmine, Muller-Mowry Colloidal Iron, Russell Modification of the Movat Pentachrome Stain |
| Demonstration of Amyloid | Congo Red, Crystal Violet, Thioflavine T |
| Alcohol or Carnoy is preferred. 10% NBF, Bouin or Zenker satisfactory. | Congo Red |
| Control: Sections containing amyloid | Congo Red, Crystal Violet, Thioflavine T |
| Reagents; 80% Alcohol saturated with Sodium Chloride, Alkaline salt solution, Congo Red | Congo Red |
| Amyloid: Deep pink to red Elastic tissue: Pale Pink Nuclei: Blue | Congo Red |
| Birefringent bright apple green | Congo Red |
| 10% NBF or alcohol. | Crystal Violet, Prussian Blue Stain, Turnbull Blue, Von Kossa |
| Amyloid: Purplish Violet Other: Blue | Crystal Violet |
| Reagents: Thioflavine T, Acetic Acid, Mayer Hematoxylin | Thioflavin T |
| Amyloid: flouresces to yellow-green | Thioflavine T |
| Differentiate between collagen and smooth muscle | Masson Trichrome, Gomori 1-Step Trichrome |
| Bouin is preferred. 10% NBF satisfactory. | Masson Trichrome |
| Internal Control | Masson Trichrome, Gomori 1-Step Trichrome, Van Gieson Picric Acid-Acid Fuchsin, Oil Red O, Sudan Black B, Osmium Tetroxide |
| Reagents: Bouin, Weigert Iron Hematoxylin, Biebrich Scarlet-Acid Fuchsin, Phsophomolybdic/Phosphotungstic Acid, Aniline Blue, Acetic Acid | Masson Trichrome |
| Nuclei: Black Cytoplasm, keratin, muscle fibers; Red Collagen and Mucin: Blue | Masson Trichrome |
| Any well fixed tissue. | Gomori 1-Step Trichrome, Van Gieson Picric Acid-Acid Fuchsin |
| Bouin used as a mordant to intensify color reactions. | Gomori 1-Step Trichrome |
| Reagents: Bouin, Weigert Iron Hematoxylin, Gomori Trichrome, Acetic Acid | Gomori 1-Step Trichrome |
| Nuclei: Black Cytoplasm, keratin, muscle fibers: Red Collagen and mucin: Green or blue | Gomori 1-Step Trichrome |
| Considered primary connective tissue stain, rarely used, except as counterstain. | Van Gieson |
| Demonstration of connective tissue | Van Gieson Picric Acid-Acid Fuchsin |
| Weigert Iron Hematoxylin, Acid Fuchsin, Picric Acid, van Gieson | Van Gieson Picric Acid-Acid Fuchsin |
| Nuclei: Black Collagen: Brilliant Red Muscle and Cytoplasm: Yellow | Van Gieson Picric Acid-Acid Fuchsin |
| Demonstration of elastic fibers | Verhoeff, Gomori Aldehyde Fuchsin, Russell Modification of the Movat Pentachrome Stain |
| Reagents: Lugol Iodine, Ferric Chloride, Alcoholic Hematoxylin, Verhoeff, van Gieson, Sodium Thiosulfate | Verhoeff |
| Elastic = Blue-black to black Nuclei = Blue to black Collagen = Red Other = Yellow | Verhoeff |
| Control: Aorta or muscular artery | Verhoeff, Gomori Aldehyde Fuchsin |
| Elastic Fibers = Deep blue to purple Other = Green | Gomori Aldehyde Fuchsin |
| Reagents: Aldehyde Fuchsin, Light Green | Gomori Aldehyde Fuchsin |
| Demonstration of mucin | Russell Modification of the Movat Pentachrome Stain |
| Demonstration of fibrin | Russell Modification of the Movat Pentachrome Stain, Mallory PTAH |
| Demonstration of muscle | Russell Modification of the Movat Pentachrome Stain |
| Demonstration of collagen | Russell Modification of the Movat Pentachrome Stain |
| Control: Lung, skin or colon | Russell Modification of the Movat Pentachrome Stain |
| Reagents: Alcian Blue, Alkaline Alcohol, Iodine-Iodide, Absolute Alcoholic Hematoxylin, Ferric Chloride, Sodium Thiosulfate, Crocein Scarlet-Acid Fuchsin, Phosphotungstic Acid, Alcoholic Safran | Russell Modification of the Movat Pentachrome Stain |
| Nuclei = black Elastic fibers = black Ground substance and mucin = Blue Collagen = Yellow Fibrinoid, fibrin = Intense Red Muscle = Red | Russell Modification of the Movat Pentachrome Stain |
| Cryptococcus = Bright Blue | Russell Modification of the Movat Pentachrome Stain |
| Metallic impregnation step | Sensitization |
| Reduction step in silver impregnation | Developing |
| When bound metallic silver is treated with gold chloride and the color of the impregnated component is changed from brown to black | Toning |
| Most frequently used oxidizers | Phosphomolybdic acid, potassium permanganate and periodic acid |
| Demonstration of reticular fibers | Gomori Stain, Gordon and Sweets |
| Potassium permanganate is oxidizer | Gomori Stain, Gordon and Sweets |
| Ferric ammonium sulfate is sensitizer | Gomori Stain, Gordon and Sweets |
| Reducer in all reticular fiber silver methods | Formaldehyde |
| Reagents: Silver Nitrate, Potassium Hydroxide, Ammoniacal Silver, Potassium Permanganate, Potassium Metabisulfite, Ferric Ammonium Sulfate, Formalin, Gold Chloride, Sodium Thiosulfate, Nuclear Fast Red | Gomori Stain |
| Reticulin = Black Collagen = Taupe Other = Counterstain method dependent | Gomori Stain |
| Demonstration of muscle cross-striations | Mallory PTAH |
| Demonstration of glial fibers | Mallory PTAH, Holzer Method |
| Demonstration of myelin | Mallory PTAH, Weil Method |
| Zenker is preferred. 10% NBF is satisfactory. | Mallory PTAH |
| Control: skeletal or cardiac muscle | Mallory PTAH |
| Control: Section containing fibrin | Mallory PTAH |
| Reagents: PTAH (Hematoxylin, Phosphotungstic acid), Gram Iodine, Sodium Thiosfulfate, Potassium Permanganate, Oxalic Acid | Mallory PTAH |
| Cross striations, fibrin = Blue Nuclei = Blue Collagen = Red-brown | Mallory PTAH |
| Demonstration of basement membranes | PAS, Periodic Acid-Methenamine Silver, Luxol Fast Blue-PAS-Hematoxylin |
| Avoid mercuric fixatives | Periodic Acid-Methenamine Silver |
| Reagents: Methenamine Silver, Borax, Periodic Acid, Sodium Thiosulfate, Gold Chloride, Light Green | Periodic Acid-Methenamine Silver |
| Basement membranes = Black Background = Green | Periodic Acid-Methenamine Silver |
| Demonstration of Lipids | Oil Red O, Sudan Black B, Osmium Tetroxide |
| 10% NBF or calcium-formalin | Oil Red O, Sudan Black B |
| Use frozen sections | Oil Red O, Sudan Black B, Cajal Stain |
| Reagents: Oil Red O (Isopropanol and water) | Oil Red O |
| Fat = Red Other - Dependent on method | Oil Red O |
| Avoid alcoholic fixatives | Oil Red O, Sudan Black B, Fontana Masson |
| Reagents: Calcium Formalin, Sudan Black B (propylene glycol) | Sudan Black B |
| Fat = Blue-black Nuclei = Red | Sudan Black B |
| Reagents: Osmium Tetroxide, Periodic Acid | Osmium Tetroxide |
| Fat = Black Other Dependent on method | Osmium Tetroxide |
| Demonstration of mast cells | Methyl Green Pyronin Y, Toluidine Blue, Ziehl-Neelsen Method |
| When tissue stains a different color from the dye solution and the rest of the tissue | Metachromatic |
| Color shift | Metachromasia |
| Control: Section containing mast cells | Methyl Green Pyronin Y, Toluidine Blue, Ziehl-Neelsen Method |
| Reagents: Toluidine Blue, water | Toluidine Blue |
| Mast cells = Deep rose-violet Background = Blue | Toluidine Blue |
| Mast cells = orange-red | Methyl Green Pyronin Y |
| Demonstration of Nissl substance | Cresyl Echt Violet |
| Loss of Nissl substance | chromatolysis |
| Control: Spinal cord | Cresyl Echt Violet, Weil Method, Luxol Fast Blue |
| Reagents: Cresyl Echt Violet, Balsam-Xyelene | Cresyl Echt Violet |
| Nissl Substance = Blue to purple Nuclei = Blue to purple Background = colorless | Cresyl Echt Violet |
| Demonstration of nerve fibers, nerve endings and neurofibrils | Bodian Method, Holmes Silver Nitrate Method, Bielschowsky-PAS, Churukian Bielschowsky, Sevier-Munger, Thioflavin S |
| When an axon is severely or irreversibly injured, all of the axon distal to the injury disappears along with its myelin sheath | Wallerian degeneration |
| Control: peripheral nerve | Bodian Method |
| Reagents: Protargol, Reducing Solution (Hydroquinone, Formaldehyde), Gold Chloride, Oxalic Acid, Sodium Thiosulfate, Aqua Regia (Hydrochloric acid and Nitric Acid), Aniline Blue | Bodian Method |
| Nerve Fibers = Black Nuclei = Black Background = Light gray or blue | Bodian Method |
| Control: Cerebral cortex | Bodian Method, Holmes Silver Nitrate Method, Mallory PTAH, Holzer Method, Cajal Stain |
| Reagents: Impregnating Solution (Aqueous Silver Nitrate, Boric Acid, Borax, Pyridine), Reducing Solution (Hydroquinone, Sodium Sulfite Crystals), Gold Chloride, Oxalic Acid, Sodium Thiosulfate | Holmes Silver Nitrate Method |
| Axons and nerve fibers = Black Neurofibrils = Black | Holmes Silver Nitrate Method |
| Demonstration of senile plaques | Bielschowsky-PAS, Churukian Bielschowsky, Sevier-Munger, Thioflavin S, Luxol Fast Blue-PAS-Hematoxylin |
| Control: CNS | Bielschowsky-PAS, Churukian Bielschowsky, Sevier-Munger, Thioflavin S |
| Reagents: Aqueous Silver Nitrate, Ammonialcal Silver, Developer (Formaldehyde, Nitric Acid, Citric Acid), Gold Chloride, Sodium Thiosulfate, Periodic Acid, Schiffs | Bielschowsky-PAS |
| Neurofibrillary tangles = Dark Black Peripheral neurites of neuritic plaques = Dark Black Axons = Black Amyloid (plaque cores and vascular) = Magenta Lipofuchsin = Magenta | Bielschowsky-PAS |
| Reagents: Silver Nitrate, Nitric Acid, Developer (Formaldehyde, Citric Acid, Nitric Acid), Ammonium Hydroxide, Sodium Thiosulfate | Churukian Bielschowsky |
| Axons = Brown to black Cytoplasmic neurofibrils = Brown to Black Neurofibrillary tangles and plaques = Dark brown or black Neuromelanin = Black Lipofuchsin = Brown or black Background = Yellow | Churukian Bielschowsky |
| Reagents: Silver Nitrate, Formalin, Sodium Carbonate, Ammoniacal Silver, Sodium Thiosulfate | Sevier-Munger |
| Nerve endings and neurofibrils = Black Neurofibrillary tangles and periphera neurites of neuritic palques = Black Background = Yellow | Sevier-Munger |
| Reagents: Potassium permanganate, Potassium Metabisulfite-Oxalic Acid, Sodium Hydroxide-Hydrogen Peroxide Solution, Acetic Acid, Thioflavin S in 50% Alcohol | Thioflavin S |
| Neurofibrillary tangles, senile plaque neurites, neuropil threads, senile plaque amyloid and cerebrovascular amyloid = Bright green Diffuse plaques and extracellular tangles = Paler yellow green PSP tangles and Pick bodies = Not well demonstrated | Thioflavin S |
| Birefringent stains | Congo Red, Thioflavin T, Thioflavin S, Auramine-Rhodamine, Alizarin Red S |
| Glial Fibers = Blue Nuclei = Blue Neurons = Salmon Myelin = Blue | Mallory PTAH |
| Reagents: Aqueous Phosphomolybdic Acid, 95% Alcohol, Chloroform, Potassium Bromide, Crystal Violet, Differentiating Solution (Aniline Oil, Chloroform) | Holzer Method |
| Glial Fibers = Blue Background = Very pale blue to colorless | Holzer Method |
| Demonstration of astrocytes | Cajal Stain |
| Reagents: Formalin Ammonium Bromide, Gold Sublimate, Sodium Thiosulfate | Cajal Stain |
| Astrocytes with pervascular feet = Black Background = Magenta to purple | Cajal Stain |
| Demonstration of myelin sheath | Weil Method, Luxol Fast Blue, Luxol Fast Blue-PAS-Hematoxylin |
| Control = Medulla | Weil Method, Luxol Fast Blue |
| Reagents: Ferric Ammonium Sulfate, Alcoholic Hematoxylin, Differentiating Solution (Sodium Borate, Potassium Ferricyanide) | Weil Method |
| Myelin Sheath = Blue to blue-black Background = Light tan | Weil Method |
| Reagents: Luxol Fast Blue, Lithium Carbonate, 70% Alcohol | Luxol Fast Blue |
| Myelin = Blue to blue-green Background = Colorless | Luxol Fast Blue |
| Myelin = Blue Nissl substance = Violet Nuclei = Violet | Luxol Fast Blue-Cresyl Echt Violet Stain |
| Myelin Sheaths = Blue to green Axons and nerve fibers = Black | Luxol Fast Blue-Holmes Silver Nitrate Method |
| Demonstration of fungi | Luxol Fast Blue-PAS-Hematoxylin, Hotchkiss-McManus PAS (PAS F), Chromic Acid-Schiff (CAS), Gridley, Grocott-Methenamine Silver Nitrate (GMS) |
| Demonstration of corpora amylacea | Luxol Fast Blue-PAS-Hematoxylin |
| Reagents: Luxol Fast Blue, Lithium Carbonate, Schiffs, Periodic Acid, Harris Hematoxylin | Luxol Fast Blue-PAS-Hematoxylin |
| Capillary basement membranes = Rose Fungi = Rose Corpora amylacea = Rose Senile plaques = Rose Myelin sheath = Blue to blue-green Nuclei = Purple | Luxol Fast Blue-PAS-Hematoxylin |
| Demonstration of acid-fast mycobacteria | Kinyoun, Ziehl-Neelsen Method, |
| Avoid Carnoy Solution | Kinyoun, Ziehl-Neelsen Method, Fite |
| Control: Tissue containing acid-fast organisms | Kinyoun, Ziehl-Neelsen Method, Auramine-Rhodamine |
| Reagents: Kinyoun Carbol-Fuchsin, Acid Alcohol, Methylene Blue | Kinyoun |
| Acid fast bacteria = Bright Red Background = Light Blue | Kinyoun, Ziehl-Neelsen Method |
| Use Millipore-filtered water in water bath | Kinyoun, Ziehl-Neelsen Method, Fite, Auramine-Rhodamine |
| Reagents: Ziehl-Neelsen Carbol-Fuchsin, Acid Alcohol, Methylene Blue | Ziehl-Neelsen Method |
| Mycobacterium avium intracellulare = Pink Erythrocytes = Pink Mast Cells = Blue Other = Pale Blue | Churukian Ziehl-Neelsen Method |
| Demonstration of Mycobacterium leprae | Fite |
| Control: Tissue containing leprosy organisms | Fite |
| Reagents = Xylene-Peanut Oil Solution, Acid Alcohol, Ziehl-Neelsen Carbol-Fuchsin Solution, Methylene Blue | Fite |
| M leprae = Bright Red Other acid fast organisms = Bright Red Background = Light Blue | Fite |
| Demonstration of Mycobacterium tuberculosis | Auramine-Rhodamine |
| Reagents: Auramine O-Rhodamine B solution, Acid Alcohol, Eriochrome Black T | Auramine-Rhodamine |
| Acid Fast Organisms = Reddish-yellow flourescence Background = Black | Auramine-Rhodamine |
| Demonstration of gram-positive and gram-negative bacteria | Gram |
| Control: Tissue containing both gram-positive and gram-negative bacteria | Gram |
| Reagents: Crystal Violet, Gram Iodine, Basic Fuchsin, Gallego Solution, Picric Acid-Acetone | Gram |
| Gram positive = Blue Gram negative = Red Nuclei = Light Red Background = Yellow | Gram |
| Demonstration of H. Pylori | Diff-Quik Giemsa, Alcian Yellow-Toluidine Blue Method, Warthin-Starry, Steiner and Steiner |
| Control: Sections containing H. Pylori | Diff-Quik Giemsa, Alcian Yellow-Toluidine Blue Method, Warthin-Starry, Steiner and Steiner |
| Reagents: Diff Quik, Acetic Acid Water | Diff-Quik Giemsa |
| H Pylori = Dark Blue Other bacteria = Blue Nuclei = Dark Blue Cytoplasm = Pink | Diff-Quik Giemsa |
| Reagents: Periodic Acid, Alcian Yellow, Sodium Metabisulfite, Toluidine Blue | Alcian Yellow-Toluidine Blue Method |
| H Pylori = Blue Mucin = Yellow Background = Pale Blue | Alcian Yellow-Toluidine Blue Method |
| 10% NBF, Bouin or Zenker. | Hotchkiss-McManus PAS (PAS F) |
| Control: Section containing fungi | Hotchkiss-McManus PAS (PAS F), Chromic Acid-Schiff (CAS), Gridley, Grocott-Methenamine Silver Nitrate (GMS) |
| Reagents: Periodic Acid, Schiffs, Hydrochloric Acid, Sodium Metabisulfite, Sulfurous Acid Rinse, Fast Green | Hotchkiss-McManus PAS (PAS F) |
| Fungi = Rose Background = Green | Hotchkiss-McManus PAS (PAS F) |
| Reagents: Chromic Acid, Hydrochloric Acid, Schiffs, Sulfurous Acid Rinse, Fast Green | Chromic Acid-Schiff (CAS) |
| Fungi = Deep Rose to purple Nuceli = Blue Background = Green | Chromic Acid-Schiff (CAS) |
| Reagents: Chromic Acid, Schiffs, Aldehyde Fucchsin, Metanil Yellow | Gridley |
| Mycelia = Deep purple Conidia = Deep rose to purple Elastic Fibers and mucin = Deep Purple Background = Yellow | Gridley |
| Control: Pneumocystic jirovecii | Grocott-Methenamine Silver Nitrate (GMS) |
| Reagents: Chromic Acid, Silver Nitrate, Methenamine, Borax, sodium Bisulfite, Gold Chloride, Sodium Thiosfulfate, Light Green | Grocott-Methenamine Silver Nitrate (GMS) |
| Fungi = Black Mucin = Taupe to dark gray Background = Green | Grocott-Methenamine Silver Nitrate (GMS) |
| Demonstration of spirochetes | Warthin-Starry, Dieterle Method, Steiner and Steiner |
| Control: Tissue containing spirochetes | Warthin-Starry, Dieterle, Steiner and Steiner |
| Reagents: Citric Acid, Acidulated Water, Silver Nitrate 1%(Developer), Silver Nitrate 2% (Impregnation), Developer (Gelatin, Hydroquinone) | Warthin-Starry |
| Spirochetes = Black Other bacteria = Black Background = Pale yellow to light brown | Warthin-Starry |
| Demonstration of legionella organisms | Dieterle Method, Steiner and Steiner |
| Reagents: Alcoholic Uranyl Nitrate, Alcoholic Gum Mastic, Silver Nitrate, Developer (Hydroquinone, Sodium Sulfite, Acetone, Formaldehyde, Pyridine), Formic Acid | Dieterle Method |
| Spirochetes, bacteria = Brown to black Background = Pale yellow or tan | Dieterle Method |
| Control: Tissue containing legionella organisms | Dieterle Method, Steiner and Steiner |
| Reagents: Uranyl Nitrate, Silver Nitrate, Reducing Solution (Gum Mastic, Hydroquinone, Absolute Alcohol) | Steiner and Steiner |
| Spirochetes = Dark brown to black H Pylori - Dark Brown to black Legionella = Dark Brown to Black Other nonfilamentous bacteria = Dark Brown to Black Background = Light Yellow | Steiner and Steiner |
| Produced in tissue during processing and most commonly result from fixation. | Artifact pigment |
| List exogenous pigments | Carbon, asbestos fibers, tattoo pigments, metals |
| List endogenous pigments that are hematogenous | Hemoglobin, hemosiderin, biliverden and Hematoidin. |
| List endogenous pigments that are nonhematogenous and lipidic | Lipofuscin and ceroid |
| List endogenous pigments that are nonhematogenous and nonlipidic | Melanin |
| Has the ability to be impregnated by silver, but cannot reduce silver to a visible metallic form. | Argyrophil |
| Has the ability to both be impregnated with silver and to reduce silver. | Argentaffin |
| Demonstration of ferric iron | Prussian Blue Stain |
| Control: Tissue containing ferric iron | Prussian Blue Stain |
| Reagents: Potassium Ferrocyanide, Hydrochloric Acid Solution, Nuclear Fast Red | Prussian Blue Stain |
| Nuclei and hemofuchsin = Bright Red Hemosiderin = Blue Background = Pink | Prussian Blue Stain |
| Demonstration of ferrous iron | Turnbull Blue |
| Control: Tissue containing ferrous iron | Turnbull Blue |
| Reagents: Hydrochloric Acid, Potassium Ferricynaide, Acetic Acid, Nuclear Fast Red | Turnbull Blue |
| Ferrous Iron = Blue Background = Pink-red | Turnbull Blue |
| Demonstration of melanin | Schmorl Technique, Fontana-Masson |
| Control: Tissue containing melanin or argentaffin granules | Schmorl Technique |
| Reagents: Ferric Chloride, Potassium Ferricyanide, Mayer Mucincarmine, Metanil Yellow | Schmorl Technique |
| Reducing substances = Blue-green Goblet cells, mucin = Rose Background = Yellow-green | Schmorl Technique |
| Demonstration of formalin pigment | Schmorl Technique |
| Demonstration of argentaffin granules | Schmorl Technique, Fontana-Masson |
| Control: Skin, small intestine or appendix | Fontana-Masson |
| Reagents: Silver Nitrate, Fontana Silver, Gold Chloride, Sodium Thiosulfate, Nuclear Fast Red | Fontana-Masson |
| Melanin = Black Argentaffin granules = Black Nuclei = Pink | Fontana-Masson |
| Demonstration of argyrophil granules | Grimelius, Churukian-Schenk |
| Control: Argyrophil-positive carcinoid tumor preferred. Small intestine satisfactory. | Grimelius, Churukian-Schenk |
| Reagents: Silver Nitrate, Acetic Acid, Sodium Acetate, Acetic Acid-Sodium Acetate Buffer, Reducing Solution (Hydroquinone, Sodium Sulfite), Nuclear Fast Red | Grimelius |
| Argentaffin Granules = Dark brown to black Argyrophil Granules = Dark brown to black Nuclei = Red Background = Pale yellow-brown | Grimelius |
| Reagents: Citric Acid, Acidified Water, Silver Nitrate, Reducing Solution (Sodium Sulfite, Hydroquinone), Nuclear Fast Red | Churukian-Schenk |
| Argyrophil granules = Black Argentaffin granules = Black Nuclei = Red Background = Yellow-Brown | Churukian-Schenk |
| Demonstration of Urates | Gomori Methenamine-Silver |
| Absolute alcohol required. | Gomori Methenamine-Silver |
| Control: Tissue containing urates | Gomori Methenamine-Silver |
| Reagents: Silver Nitrate, Methenamine Solution, Sodium Borate, Sodium Thiosulfate, Light Green | Gomori Methenamine-Silver |
| Urates = Black Background = Blue-green | Gomori Methenamine-Silver |
| Demonstration of bilirubin | Hall |
| Control: Tissue containing bile | Hall |
| Reagents: Ferric Chloride, Fouchet Reagent, van Gieson | Hall |
| Bile or bilirubin = Emerald green to olive drab Background = Yellow | Hall |
| Demonstration of calcium | Von Kossa, Alizarin Red S |
| Control: Tissue containing calcium | Von Kossa, Alizarin Red S |
| Reagents: Silver Nitrate, Sodium Thiosulfate, Nuclear Fast Red | Von Kossa |
| Calcium salts = Black Background = Red | Von Kossa |
| Alcoholic Formalin or 10% NBF | Alizarin Red S |
| Reagents: Alizarin Red S | Alizarin Red S |
| Calcium deposits = Orange-red | Alizarin Red S |
| Demonstration of copper | Rhodanine |
| Control: Tissue containing copper | Rhodanine |
| Reagents: Rhodanine Solution, Mayer Hematoxylin, Sodium Borate | Rhodanine |
| Copper = Bright red to red yellow Nuclei = Light Blue | Rhodanine |
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