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Histology Spec Stain

Histology ASCP Special Stains

QuestionAnswer
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
Created by: impossiblesum