Grocott-Gomori’s Methenamine Silver Staining 50 FAQs and 30 MCQs
Grocott-Gomori’s Methenamine Silver Staining 50 FAQs
What is Grocott-Gomori’s Methenamine Silver (GMS) stain used for?
Primarily to detect fungal organisms and polysaccharide-rich microorganisms in tissue sections.
Who developed the GMS staining method?
György Gömöri, a Hungarian physician.
How does GMS compare to PAS (Periodic Acid-Schiff) and Gridley stains?
GMS has higher sensitivity for fungi due to its strong affinity for fungal cell wall polysaccharides.
What are the major components of GMS stain?
Chromic acid, methenamine-silver nitrate, and borax.
What types of samples can GMS stain be applied to?
Tissue sections, aspirates, and smears.
How does GMS stain fungi black?
Fungal cell wall polysaccharides are oxidized to aldehydes, which reduce silver nitrate to metallic silver (black precipitate).
What is the “argentaffin reaction”?
The reduction of silver solution to metallic silver by reactive groups (e.g., aldehydes) in fungal cell walls.
Why is chromic acid used in GMS staining?
It oxidizes polysaccharides to release aldehyde groups for silver reduction.
What counterstain is used in GMS, and what does it highlight?
Light green SF yellowish; stains the background pale green.
Can GMS stain bacteria or viruses?
No, it is specific to fungi and other carbohydrate-rich structures (e.g., mucin).
What is the role of sodium metabisulfite in GMS?
Removes excess chromic acid after oxidation.
Why is gold chloride used in the procedure?
Toning step enhances contrast by replacing unbound silver with gold.
How long is the methenamine silver working solution stable?
Must be prepared fresh before use (discard after staining).
What happens if the silver solution turns cloudy?
It indicates contamination; prepare a fresh solution.
Why is borax added to the silver solution?
Acts as a buffer to maintain alkaline conditions for silver reduction.
What are the two main GMS staining methods?
Conventional (room temperature/water bath) and microwave-assisted.
How long should sections oxidize in chromic acid (conventional method)?
1 hour at room temperature or 60°C.
What is the microwave modification for chromic acid oxidation?
Microwave at 150W for 2.5 minutes, then stand for 2 minutes.
How do you know when silver impregnation is complete?
Sections turn yellowish-brown; check microscopically for dark brown fungi.
Why is a water bath used for the silver solution?
Heat accelerates the reduction of silver by aldehydes.
Why are fungi not staining black?
Check freshness of chromic acid, sodium metabisulfite, or borax.
What causes tissue detachment during staining?
Over-oxidation with chromic acid or improper slide adhesion.
How to fix excessive background staining?
Adjust toning time with gold chloride or reduce silver incubation.
Why does the silver solution mirror the Coplin jar?
Contamination or outdated reagents; replace the solution.
What if Pneumocystis jiroveci stains slower than other fungi?
Extend silver incubation time (up to 20–30 minutes).
Why is chromic acid hazardous?
Corrosive, carcinogenic, and toxic to kidneys.
What PPE is required for GMS staining?
Gloves, goggles, lab coat, and fume hood use for hot solutions.
Is sodium thiosulfate toxic?
Yes; irritates skin, eyes, and respiratory tract.
How should silver nitrate waste be disposed of?
Follow hazardous waste protocols (reacts with organic matter).
Can light green SF yellowish cause cancer?
It is a possible carcinogen; avoid inhalation/contact.
What color are fungal hyphae in GMS?
Black cell walls with pink-red internal structures.
How does Histoplasma spp. appear with GMS?
Small, black yeast-like cells.
What does mucin look like in GMS-stained sections?
Dark grey.
Why is a control slide necessary?
Verifies staining efficacy and reagent functionality.
Can GMS detect Leishmania or Toxoplasma?
No; these organisms are GMS-negative.
Which fungi are commonly identified with GMS?
Pneumocystis jiroveci, Aspergillus, Candida, Histoplasma.
Can GMS differentiate fungal species?
Yes, based on morphology (hyphae, yeast forms).
What is the advantage of GMS over H&E for fungi?
Higher contrast and specificity for fungal elements.
What are the limitations of GMS staining?
Toxic reagents, lengthy procedure, and non-specific mucin staining.
Can GMS be used on frozen sections?
Typically no; paraffin-embedded sections are standard.
How does microwave staining save time?
Reduces oxidation and silver steps to minutes vs. hours.
What wattage is recommended for microwave steps?
150W for oxidation; 450W for silver solution heating.
Can any microwave be used?
Lab-grade microwaves are preferred for consistency.
Why agitate slides during microwave steps?
Ensures even heating and staining.
What if sections overdevelop in the microwave?
Reduce power or time; check microscopically every 30 seconds.
How long can stained slides be stored?
Indefinitely if properly mounted and sealed.
Can GMS stain glycogen?
Yes, but it’s nonspecific; PAS is preferred for glycogen.
What fixative is best for GMS?
10% neutral buffered formalin.
Why use acid-cleaned glassware?
Prevents contamination of silver solutions.
Where can I purchase GMS kits?
Companies like Newcomer Supply provide pre-made reagent kits.
Grocott-Gomori’s Methenamine Silver Staining 30 MCQs
- What is the primary purpose of GMS staining?
a) Detecting bacterial infections
b) Identifying fungal cell walls✔
c) Staining viral particles
d) Highlighting collagen fibers - Which component of the fungal cell wall reacts with GMS stain?
a) Lipids
b) Proteins
c) Polysaccharides✔
d) Nucleic acids - Which fungus is commonly detected using GMS in immunocompromised patients?
a) Escherichia coli
b) Pneumocystis jiroveci✔
c) Mycobacterium tuberculosis
d) Herpes simplex virus - What color do fungi appear after GMS staining?
a) Blue
b) Red
c) Black✔
d) Green - Which counterstain is used in GMS to highlight the background?
a) Hematoxylin
b) Light green SF yellowish✔
c) Eosin
d) Methylene blue
- Which chemical oxidizes polysaccharides in GMS staining?
a) Sodium thiosulfate
b) Chromic acid✔
c) Gold chloride
d) Methenamine - What is the role of sodium metabisulfite in GMS?
a) Oxidizing agent
b) Reducing agent✔
c) Counterstain
d) Toning agent - Which reagent tones the sections to improve contrast?
a) Silver nitrate
b) Gold chloride✔
c) Borax
d) Light green - How long should slides remain in chromic acid (conventional method)?
a) 5 minutes
b) 30 minutes
c) 1 hour✔
d) 2 hours - What indicates proper silver impregnation?
a) Slides turn blue
b) Slides turn yellowish-brown✔
c) Slides turn red
d) Slides become transparent
- Why might fungi fail to stain black?
a) Old chromic acid✔
b) Insufficient dehydration
c) Overuse of light green
d) Excessive gold toning - What hazard is associated with chromic acid?
a) Flammable
b) Radioactive
c) Corrosive and carcinogenic✔
d) Explosive - How should silver nitrate waste be handled?
a) Pour down the sink
b) Dispose as hazardous waste✔
c) Reuse for future stains
d) Incinerate immediately - What causes tissue to detach during staining?
a) Over-oxidation✔
b) Insufficient dehydration
c) Using tap water
d) Skipping the gold step - Which PPE is mandatory for GMS staining?
a) Gloves only
b) Lab coat and gloves
c) Goggles, gloves, lab coat, fume hood✔
d) No special precautions
- Which stain is more sensitive for fungi than GMS?
a) H&E
b) Gram stain
c) PAS (Periodic Acid-Schiff)
d) None—GMS is the most sensitive✔ - What does mucin look like in GMS-stained sections?
a) Black
b) Dark grey✔
c) Pink-red
d) Green - Which step is unique to microwave GMS staining?
a) Using a water bath
b) Heating chromic acid for 2.5 minutes✔
c) Overnight incubation
d) Freezing slides - What is the purpose of sodium thiosulfate?
a) Oxidize fungi
b) Remove unbound silver✔
c) Enhance gold toning
d) Counterstain nuclei - Which organism is GMS-negative?
a) Aspergillus
b) Candida
c) Toxoplasma gondii✔
d) Histoplasma
- How are hyphae visualized in GMS?
a) Black walls, pink interiors✔
b) Blue walls, clear interiors
c) Red walls, green interiors
d) Unstained - What indicates a successful GMS control slide?
a) Uniform green background
b) Clear unstained fungi
c) Black fungal elements✔
d) Blue nuclei - Why is borax added to the silver solution?
a) Lowers pH
b) Prevents precipitation
c) Accelerates oxidation
d) Acts as a buffer✔ - How long can methenamine silver stock solution be stored?
a) 1 week
b) 3 months (refrigerated)✔
c) 1 year
d) Indefinitely - Which fixative is optimal for GMS?
a) 95% ethanol
b) 10% formalin✔
c) Acetone
d) Bouin’s solution
- What wattage is used for microwave silver incubation?
a) 50W
b) 150W
c) 450W✔
d) 1000W - How long is chromic acid microwaved in the modified protocol?
a) 30 seconds
b) 2.5 minutes✔
c) 10 minutes
d) 1 hour - What is a drawback of microwave GMS?
a) Higher cost
b) Inconsistent heating✔
c) Longer procedure
d) Weaker staining - Which step is skipped in microwave GMS vs. conventional?
a) Chromic acid oxidation
b) Silver incubation
c) Gold toning
d) None—steps are identical✔ - Why use plastic Coplin jars in microwave GMS?
a) Cheaper than glass
b) Prevents metal interference✔
c) Faster heating
d) Better staining
Possible References Used
