Hematoxylin and Eosin Stain - FOR PARAFFIN-EMBEDDED TISSUE SECTIONS

Deparaffinize slides and hydrate to ddH₂O:

A	B
Compound	Time
Xylenes	2 x 5 min
100% EtOH	2 x 1 min
95% EtOH	1 min each
80% EtOH	1 min each
70% EtOH	1 min each
ddH2O	1 min each

Filter undiluted Harris Hematoxylin and Eosin using Whatman #4 filter paper.

Immerse sections in filtered Harris Hematoxylin for 0h 5m 0s.

Rinse in 2 changes of ddH₂O.

Differentiate in 0.1% acid alcohol solution:

A	B
Compound	Time
EtOH fixed tissue	2 sec
NBF fixed tissue	4 sec

Rinse in tap H₂O for 10- 0h 15m 0s or until sections turn bright blue.

Immerse in Eosin for 0h 1m 0s.

Briefly rinse in tap H₂O.

Dehydrate in:

A	B
Compound	Time
95% EtOH	2 x 30 sec
100% EtOH	2 x 1 min
Xylene	2 x 5 min

Coverslip with Cytoseal.