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

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

 


 

 

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