Gridley's Stain
for Fungi
Materials
- Schiff’s reagent
- Aldehyde fuchsin
- Chromic acid
Material Amount Chromium trioxide 5 g Distilled water 500 mL - Bleach
Material Amount Sodium metabisulfite 5 g Distilled water 500 mL - Metanil yellow
Material Amount Metanil yellow 1 g Distilled water 400 mL Acetic acid, glacial 2 drops
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.
Protocol
- Bring sections to water via xylene and ethanol.
- Place in chromic acid for 1 hour.
- Wash well with tap water.
- Treat with the metabisulfite bleach for 1 minute.
- Wash well with tap water.
- Rinse with distilled water.
- Place in Schiff’s reagent for 20 minutes.
- Wash well with tap water.
- Rinse with 70% ethanol.
- Place in aldehyde fuchsin 30 minutes.
- Rinse off excess with 95% ethanol.
- Wash well with tap water.
- Counterstain with metanil yellow for 1 minute.
- Rinse well with distilled water.
- Dehydrate, clear and mount in a resinous medium.
Expected Results
- Fungi – purple
- Background – yellow
Notes
- At step 2, a 10% solution of chromic acid applied for 10 minutes will give similar results.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Bancroft, J.D. and Stevens A. (1982)
Theory and practice of histological techniques Ed. 2
Churchill Livingstone, Edinburgh & London, UK. - Humason, G. L., (1967)
Animal tissue techniques, Ed. 2
W. H. Freeman and Company, San Francisco, Ca, USA
