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Gridley’s Stain for Fungi

Protocol

Gridley's Stain

for Fungi

15
steps
7
materials
print

Materials

  • Schiff’s reagent
  • Aldehyde fuchsin
  • Chromic acid
    MaterialAmount
    Chromium trioxide5g
    Distilled water500mL
  • Bleach
    MaterialAmount
    Sodium metabisulfite5g
    Distilled water500mL
  • Metanil yellow
    MaterialAmount
    Metanil yellow1g
    Distilled water400mL
    Acetic acid, glacial2drops

Tissue Sample

5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.

Protocol

  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 metabisulfite 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

  • 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

  1. Bancroft, J.D. and Stevens A. (1982)
    Theory and practice of histological techniques Ed. 2
    Churchill Livingstone, Edinburgh & London, UK.
  2. Humason, G. L., (1967)
    Animal tissue techniques, Ed. 2
    W. H. Freeman and Company, San Francisco, Ca, USA