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Questions and Answers
What is the purpose of the Ziehl-Neelsen method?
What is the purpose of the Ziehl-Neelsen method?
Which color indicates the presence of acid-fast bacilli, including Mycobacterium avium intracellulare, using this method?
Which color indicates the presence of acid-fast bacilli, including Mycobacterium avium intracellulare, using this method?
What type of fixative is NOT suitable for the detection of acid-fast mycobacteria?
What type of fixative is NOT suitable for the detection of acid-fast mycobacteria?
Why are alcoholic solutions of acid preferred over aqueous solutions for decolorization?
Why are alcoholic solutions of acid preferred over aqueous solutions for decolorization?
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What is the final mounting medium used after the clearing steps in the procedure?
What is the final mounting medium used after the clearing steps in the procedure?
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What is the color code for the background when using the Ziehl-Neelsen staining method?
What is the color code for the background when using the Ziehl-Neelsen staining method?
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During the FITE acid-fast stain procedure, what temperature is applied for the hot solution phase?
During the FITE acid-fast stain procedure, what temperature is applied for the hot solution phase?
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What is the primary purpose of using 0.1% eriochrome black T during the staining process?
What is the primary purpose of using 0.1% eriochrome black T during the staining process?
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Why is it necessary to rinse sections in multiple changes of distilled water after staining?
Why is it necessary to rinse sections in multiple changes of distilled water after staining?
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What color do mast cells appear after staining with the mentioned method?
What color do mast cells appear after staining with the mentioned method?
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What color indicates the presence of Gram positive bacteria in the staining procedure?
What color indicates the presence of Gram positive bacteria in the staining procedure?
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What is a characteristic result of the acid fast staining procedure?
What is a characteristic result of the acid fast staining procedure?
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Which solution is used to decolorize the slides in the staining process?
Which solution is used to decolorize the slides in the staining process?
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What is the preferred fixative used in the described staining procedure?
What is the preferred fixative used in the described staining procedure?
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What is the primary purpose of this staining procedure?
What is the primary purpose of this staining procedure?
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How should the sections appear after counterstaining with methylene blue?
How should the sections appear after counterstaining with methylene blue?
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What is the purpose of deparaffinizing sections with a xylene-peanut oil mixture?
What is the purpose of deparaffinizing sections with a xylene-peanut oil mixture?
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What color represents the background tissue in this staining protocol?
What color represents the background tissue in this staining protocol?
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How long should the sections be stained with crystal violet?
How long should the sections be stained with crystal violet?
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What should be the appearance of sections after differentiation with 1% acid alcohol?
What should be the appearance of sections after differentiation with 1% acid alcohol?
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After staining with gram iodine, what is the next step?
After staining with gram iodine, what is the next step?
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What color does the background appear when examining acid fast organisms under UV light?
What color does the background appear when examining acid fast organisms under UV light?
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What should be used to mount the sections after staining?
What should be used to mount the sections after staining?
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What happens to acid-fast bacteria during the staining process?
What happens to acid-fast bacteria during the staining process?
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What is the primary purpose of using stain sections with schiff reagent?
What is the primary purpose of using stain sections with schiff reagent?
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What are the primary components of the background stain in this procedure?
What are the primary components of the background stain in this procedure?
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How long should sections be stained in aldehyde fuchsin?
How long should sections be stained in aldehyde fuchsin?
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Which dye is used to stain mucin yellow?
Which dye is used to stain mucin yellow?
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What is the purpose of rinsing the slides in distilled water after staining?
What is the purpose of rinsing the slides in distilled water after staining?
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After how many changes of alcohol should the slides be dehydrated?
After how many changes of alcohol should the slides be dehydrated?
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What color indicates the presence of conidia after staining?
What color indicates the presence of conidia after staining?
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Which fixative is used in this staining procedure?
Which fixative is used in this staining procedure?
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What is the purpose of rinsing slides in 1% sodium bisulfite?
What is the purpose of rinsing slides in 1% sodium bisulfite?
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After preheating, how long should the slides be placed in the methenamine silver solution?
After preheating, how long should the slides be placed in the methenamine silver solution?
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What color indicates that spirochetes are present after the staining process?
What color indicates that spirochetes are present after the staining process?
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What should be the condition of the chromic acid solution for reuse?
What should be the condition of the chromic acid solution for reuse?
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What step follows the 2% sodium thiosulfate treatment?
What step follows the 2% sodium thiosulfate treatment?
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What color indicates the background after the staining process?
What color indicates the background after the staining process?
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Which process should be done before mounting sections with synthetic resin?
Which process should be done before mounting sections with synthetic resin?
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Which solution is recommended for checking adequate silver impregnation?
Which solution is recommended for checking adequate silver impregnation?
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What is the purpose of the described method?
What is the purpose of the described method?
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Which chemical is noted for oxidizing polysaccharides to aldehydes?
Which chemical is noted for oxidizing polysaccharides to aldehydes?
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What role does sodium borate play in the procedure?
What role does sodium borate play in the procedure?
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What is used as a toning solution in the described method?
What is used as a toning solution in the described method?
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At what temperature should the silver nitrate impregnating solution be maintained?
At what temperature should the silver nitrate impregnating solution be maintained?
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What is the purpose of using sodium thiosulfate in the procedure?
What is the purpose of using sodium thiosulfate in the procedure?
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What is the function of hydroquinone in this staining method?
What is the function of hydroquinone in this staining method?
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What type of fixative is used for the procedure?
What type of fixative is used for the procedure?
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Study Notes
Special Stains for Microorganisms
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Kinyoun Acid-Fast Stain: Detects acid-fast mycobacteria in tissue sections.
- Purpose: To identify lipoid-coated acid-fast organisms that resist decolorization.
- Principle: Acid-fast organisms retain carbol-fuchsin, unlike other organisms.
- Fixative: 10% neutral buffered formalin, preferred.
- Procedure: Deparaffinize, hydrate, stain with carbol-fuchsin, decolorize with acid alcohol, counterstain with methylene blue, dehydrate, and mount.
- Result: Acid-fast bacteria appear bright red, background is light blue.
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Microwave Ziehl-Neelsen Method for Acid-Fast Bacteria: Alternative acid-fast stain using microwave energy.
- Procedure: Deparaffinize, hydrate, stain with carbol-fuchsin in a microwave, decolorize with acid alcohol, counterstain with methylene blue, dehydrate, and mount.
- Result: Acid-fast bacteria are bright red, background is blue.
Fite Acid Fast Stain for Leprosy Organisms
- Purpose: Detect Mycobacterium leprae in tissue sections.
- Principle: Leprosy organisms retain carbol-fuchsin.
- Fixative: 10% Neutral buffered formalin, preferred.
- Procedure: Deparaffinize, hydrate, stain in carbol-fuchsin, decolorize, counterstain with eriochrome black T, dehydrate and mount.
- Result: Leprosy organisms appear bright red, background is light blue
Microwave Auramine-Rhodamine Fluorescence Technique
- Purpose: Detect Mycobacterium tuberculosis or other acid-fast organisms.
- Principle: Uses fluorescence to enhance visibility.
- Procedure: Deparaffinize, hydrate, stain with auramine O-rhodamine B solution in microwave, dehydrate and mount using cover slip
- Result: Acid-fast organisms fluoresce reddish yellow, background is black.
Brown-Hopps Modification of Gram Stain
- Purpose: Differentiate between gram-positive and gram-negative bacteria in tissue sections.
- Procedure: Deparaffinize, hydrate, stain with crystal violet, gram iodine and basic fuchsin and differentiate in acetone/gallego solution, dehydrate and mount.
- Result: Gram-positive bacteria appear blue, gram-negative bacteria appear red, background is yellow.
Gridley Fungus Stain
- Purpose: Detect fungi in tissue.
- Procedure: Deparaffinize, hydrate, oxidize with chromic acid, stain with schiff reagent, stain with aldehyde fuchsin, counterstain with meta-nil yellow, dehydrate and mount.
- Result: Fungi appear deep purple, conidia as deep rose to purple, background as yellow.
Alcian Yellow - Toluidine Blue Method for H. pylori
- Purpose: Detect H. pylori in tissue sections.
- Principle: Alcian yellow stains mucin, Toluidine blue stains H. pylori and nuclei.
- Procedure: Deparaffinize, hydrate, stain with alcian yellow, stain with toluidine blue, dehydrate and mount
- Result: H. pylori appears blue, mucin appears yellow, background is blue.
Grocott Methenamine Silver Nitrate Fungus Stain
- Purpose: Identify fungi.
- Procedure: Deparaffinize and hydrate sections, oxidize in chromic acid, treat with methenamine silver solution, tone with gold chloride, remove unreduced silver, counterstain, dehydrate, and mount.
- Result: Fungi appear black, mucin as taupe to dark gray, background appears green.
Warthin-Starry Technique for Spirochetes
- Purpose: Detect spirochetes in tissue secretions.
- Principle: Spirochetes bind silver, treated with hydroquinone to visibly reduce silver.
- Procedure: Deparaffinize, hydrate, oxidize in chromic acid, treat with silver nitrate solution, reduce with developer, wash, dehydrate, and mount.
- Result: Spirochetes appear black, other bacteria also appear black, background is a pale yellow to light brown.
Dieterle Method for Spirochetes and Legionella Organisms
- Purpose: Detect spirochetes and Legionella organisms.
- Procedure: Deparaffinize, hydrate, treat with silver solution, develop with hydroquinone, wash, dehydrate and mount.
- Result: Spirochetes and Legionella appear black, background appears pale yellow to light brown.
Microwave Steiner & Steiner Procedure for Spirochetes, Helicobacter, and Legionella Organisms
- Purpose: Detect spirochetes, Helicobacter and Legionella using a microwave to enhance the silver impregnation.
- Procedure: Deparaffinize, hydrate, oxidize in chromic acid, place in silver nitrate solution in a microwave, reduce with developer, wash, dehydrate and mount.
- Result: Positive reaction shows organisms in dark brown/black, background is pale yellow to tan.
Perl's Prussian Blue Method for Hemosiderin
- Purpose: Identify hemosiderin (ferric iron)
- Procedure: Deparaffinize, hydrate, treat with acid ferrocyanide solution, wash, remove excess silver, counterstain, dehydrate, and mount.
- Result: Hemosiderin and ferric salts appear deep blue, other pigments retain natural color, tissue and nuclei stain in accordance with counterstain.
Leuco Patent Blue V Stain for Hemoglobin
- Purpose: Identify hemoglobin in peroxidase (RBCs and neutrophils).
- Procedure: Take sections to distilled water, stain with patent blue, counterstain with neutral red, dehydrate, and mount.
- Result: Hemoglobin peroxidase appears dark blue, nuclei appear red.
Gomori's Prussian Blue Stain for Iron
- Purpose: Detect iron pigments.
- Procedure: Immerse in hydrochloric acid and potassium ferrocyanide, wash, counterstain with nuclear fast red, rinse, dehydrate and mount.
- Result: Iron appears bright blue, nuclei appear red, cytoplasm pink/rose.
Turnbull's Blue for Ferrous Iron
- Purpose: Detect ferrous iron.
- Procedure: Deparaffinize, hydrate, treat with potassium ferricyanide solution, rinse, dehydrate, and mount.
- Results: Ferrous iron appears blue.
Modified Fouchet's Technique for Liver Bile Pigments
- Purpose: Identify bile pigments in liver.
- Procedure: Deparffinize, hydrate, treat with Fouchet's solution, wash, counterstain with van Gieson's solution, dehydrate, and mount.
- Results: Bile pigments appear emerald to blue, muscle appears green-yellow, collagen appears red.
Gmelin Technique for Bile and Hematoidin
- Purpose: Identify bile and hematoidin pigments.
- Procedure: Deparaffinize, hydrate, apply concentrated nitric acid to section to generate color changes, remove solution, observe color changes.
- Result: Bile pigments progressively change color from yellow-green-blue-purple, to red.
Fontana Masson Method for Melanin
- Purpose: Identify melanin.
- Procedure: Deparaffinize, hydrate, treat with silver solution, treat with thiosulfate, counterstain, dehydrate and mount.
- Result: Melanin appears black, argentaffin granules appear black, nuclei are red.
Modified Von Kossa Method for Calcium
- Purpose: Identify calcium.
- Procedure: Deparaffinize, hydrate, treat with silver nitrate solution, wash, dehydrate, counterstain, dehydrate, and mount.
- Result: Mineralized bone appears black, osteoid tissue appears red, nuclei appear blue.
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Description
Test your knowledge on Ziehl-Neelsen and FITE staining methods. This quiz covers critical concepts including the purpose of these techniques, color indicators, and proper procedures for detecting acid-fast bacilli. Perfect for students of microbiology wanting to solidify their understanding of staining methods.
