Gridley's Method for Fungi

EMS26120

Components:

Chromic Acid 4%

Coleman's Feulgen

Aldehyde Fuchsin

Metanil Yellow 0.25%

Fixation:

10% Buffered Neutral Formalin

Sections:

Paraffin at 6µm sections

Staining:

Use a control slide

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

Stain Results:

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.