Dr Piyush Tailor

Take test slide on which staining has to be performed along with positive control.

Deparaffinize section by keeping in oven for 5 minuteand clear it with2 changes of xylene each for 15 minutes.

Bring section to water.

Oxidize in 5% chromic acid for 1 hour.

Wash in distilled water for 1 minute.

Bleach with 2% sodium metabisulphite for 1 minute.

Wash in running tap water for 2 to 5 minute.

Wash in several changes of distilled water for 2minutes.

Place sections in preheated hexamine silver solutions in a coupling jar at 560C in a water bath.

Examine the sections after 10minute and then after 5min interval until the fungi are blackened and the background is clear.

Wash in several changes of distilled water for 2 minute.

Treat sections with gold chloride solution for 2minute.

Wash in several changes of distilled water for 3 minute

Treat section with hypo solution for 3minute

Wash in running water for 5 minute.

Counter stain with light green solution for 30 seconds

Wash in running tap water for 1 minute

Dehydrate in alcohol, clear in xylene, mount with DPX