Histology H/E Colour staining protocol

1 Deparaffinize slides by incubating:

9 mins in Xylene
3x 3 mins in 100% EtOH
1x 3 mins in 95% EtOH
1x 3 mins in 80% EtOH

2 Hydrate with distilled water by shaking for 30 sec by hand

3 Stain in Hematoxylin for 3 mins

“Fountain” for 5 mins in tap water (Continuous flow of water in and out of
chamber without touching slides directly)

Dip slides in 0.3% acid ethanol (280ml 100% EtOH, 120ml H20, 1.2ml conc.
HCl)

2x 1 min rinse in tap water

1x 2 mins deionized water

4 Stain in Eosin for (3) mins

Fountain for 5 mins in tap water

5 Wash in:

2x 5 mins in 95% EtOH

2x 5 mins in 100% EtOH
1x 10 mins Xylene

6 Mount slides with a drop of glue (Make sure no air bubbles) and leave to dry
overnight in incubator 37o