Histotechnology Technical methods

Stain for Fungi

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1. Bring sections to water.

2. Oxidise in 5% chromic acid for 1 hour.

3. Wash in running tap water.

4. Rinse well in distilled water.

5. Place in incubating solution at 60°c in the dark for approx 1 hour - needs checking microscopically. (Use coplin jar in a water bath and preheat solution before use).

6. Wash well in distilled water.

7. Tone in 0.1% gold chloride 2 mins.

8. Place sections in 3% sodium thiosulphate 2 mins.

9. Wash well in tap water.

10. Counterstain using sat alcoholic picric acid 1/2 hour.

11. Rinse in absolute alcohol.

12. Clear in xylene . Mount sections in DPX


--Fungal hyphae and yeast bodies black

--Background yellow


5% Borax in distilled water.

Silver Solution

--5% Silver nitrate in distilled water 5ml

--3% methenamine (hexamine) in distilled water 100ml

Add the methenamine solution to the silver, a white precipitate forms, but clears on shaking. Both Solutions keep well at 4°C

Incubating solution ( make up fresh)

--Borax solution 5ml --Distilled water 25ml --Silver solution 25ml

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.

© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk