Description
The mucopolysaccharide components of the fungal cell wall are oxidized to release aldehyde groups. The aldehyde groups then react with the silver nitrate, reducing it to a metallic silver, rendering them visible. VitroView™ Grocott-Gomori’s Methenamine Silver (GMS) Stain Kit is designed to identify fungi on FFPE section or frozen sections.
Kit Contents
| VB-2003-1 | Chromic Acid Solution | 100 ml |
| VB-2003-2 | Sodium Bisulfite Solution | 100 ml |
| VB-2003-3 | 0.2% Silver Solution | 50 ml |
| VB-2003- 4 | Methenamine/Borax Solution | 50 ml |
| VB-2003-5 | Gold Chloride Solution | 100 ml |
| VB-2003-6 | Sodium Thiosulfate Solution | 100 ml |
| VB-2003-7 | Light Green solution | 100 ml |
Storage
Store at 2-8°C: Silver solution, methenamine/borax solution and gold chloride solution.
Store at room temperature: The other reagents
Protocol
- Deparaffinize in xylene I for 6 minutes and II for 6 minutes.
- Rehydrate
- Ethanol 100% (2 minutes)
- Ethanol 100% (2 minutes)
- Ethanol 95% (2 minutes)
- Ethanol 95% (2 minutes)
- Ethanol 70% (2 minutes)
- Rinse in DI water (2 minutes).
- Place in 2% chromic acid, microwave with Hi power for 45 seconds and allows standing for 5 minutes. Or 60°C water bath for 1 hour.
- Wash in tap water, rinse in distilled water.
- 1% Sodium bisulfite, 1 minute, room temperature.
- Wash in tap water, rinse in distilled 3 changes.
- Fresh prepare methenamine silver working solution:
- 2% silver solution 25.0 ml
- Methenamine/Borax Solution 25.0 ml
- Mix just before use, discard
- Place in methenamine silver working solution, microwave with Hi power for 70 seconds, tissue should be the color of a brown paper bag. Or 60°C water bath for 1 hour. Agitate the slides in the hot solution.
- Rinse in distilled water, 2 changes.
- Place in 0.5% Gold chloride for 1 minute or until gray.
- Wash in distilled water.
- Place in 5% % Sodium Thiosulfate for 3 minutes.
- Wash in tap water, rinse in distilled.
- Place in light green solution for 1 minute.
- Rinse in distilled water.
- Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minute per change).
- Clear with 3 changes of xylene (5 minute per change).
- Mount coverslip onto glass slide with Permount or some other suitable organic mounting medium.
Result
- Fungi: black
- Background: green
Control Tissue
Fungus infected tissue section (provided)
Note
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Precautions
Keep hot solutions in fume hood. Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.
User Manual and Material Safety Data Sheet (MSDS) (PDF)
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