Hematoxylin and Eosin Stain - FOR PARAFFIN-EMBEDDED TISSUE SECTIONS
Michael X. Henderson
Published: 2023-12-02 DOI: 10.17504/protocols.io.kxygx3q9zg8j/v1
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Abstract
This protocol details Hematoxylin and Eosin Stain for paraffin-embedded tissue sections.
Attachments
911-2361.pdf
Steps
Hematoxylin and Eosin Stain
1.
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 |
2.
Filter undiluted Harris Hematoxylin and Eosin using Whatman #4 filter paper.
3.
Immerse sections in filtered Harris Hematoxylin for 0h 5m 0s.
4.
Rinse in 2 changes of ddH2O.
5.
Differentiate in 0.1% acid alcohol solution:
| A | B |
|---|---|
| Compound | Time |
| EtOH fixed tissue | 2 sec |
| NBF fixed tissue | 4 sec |
6.
Rinse in tap H2O for 10- 0h 15m 0s or until sections turn bright blue.
7.
Immerse in Eosin for 0h 1m 0s.
8.
Briefly rinse in tap H2O.
9.
Dehydrate in:
| A | B |
|---|---|
| Compound | Time |
| 95% EtOH | 2 x 30 sec |
| 100% EtOH | 2 x 1 min |
| Xylene | 2 x 5 min |
10.
Coverslip with Cytoseal.
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