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Electron Microscopy Sciences

Technical Data Sheets

Gridley's Method for Fungi

EMS Catalog Number: 26120-Series

Fixation:

10% Buffered Neutral Formalin (#15740).

Sections:

Paraffin at 6 µm sections

Staining:

Use a control slide

  1. Deparaffinize and hydrate to distilled water.
  2. Oxidize in Chromic Acid, 4% (#26120-01) for one hour. Wash in running water for five minutes.
  3. Coleman’s Feulgen Reagent (#26120-02) for 15 minutes. Wash in running tap water for 15 minutes. Rinse in several changes or 70% Alcohol.
  4. Aldehyde Fuchsin (#26120-03) for 30 minutes. Rinse off excess stain with 95% alcohol Rinse in distilled water.
  5.  Counterstain lightly with Metanil Yellow (#26120-04). Rinse in distilled water.
  6. Dehydrate in 95% alcohol, absolute alcohol, and clear in two changes of Xylene (#23400)
  7. Mount.

Stain Results:

Mycelia, Conidia Deep purple
Elastic Tissue & Mucin Background Yellow

Filaments of Nocardia & Actinomyces are not stained by this method.

References:

Gridley, M.F. – Amer. J. Clin. Path., 23:303 (1953)

AFIP – Manual of Histologic Staining Techniques: 3rd, ED. ED. L. Luna; New York: McGraw-Hill Publications, c 1968, p. 229.

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Gridley's Method for Fungi