Gridley's Stain
for Fungi

Solutions

Schiff's reagent

Aldehyde fuchsin

Chromic acid
Chromium trioxide	5	g
Distilled water	500	mL

Bleach
Sodium metabisulphite	5	g
Distilled water	500	mL

Metanil yellow
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.

Method

1. Bring sections to water via xylene and ethanol.
2. Place in chromic acid for 1 hour.
3. Wash well with tap water.
4. Treat with the metabisulphite bleach for 1 minute.
5. Wash well with tap water.
6. Rinse with distilled water.
7. Place in Schiff's reagent for 20 minutes.
8. Wash well with tap water.
9. Rinse with 70% ethanol.
10. Place in aldehyde fuchsin 30 minutes.
11. Rinse off excess with 95% ethanol.
12. Wash well with tap water.
13. Counterstain with metanil yellow for 1 minute.
14. Rinse well with distilled water.
15. Dehydrate, clear and mount in a resinous medium.

Expected results

• Fungi  –  purple
• Background  –  yellow

Notes

1. At step 2, a 10% solution of chromic acid applied for 10 minutes will give similar results.

 

Reference
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