H&E staining on paraffin sections
Mel Feany
Published: 2022-10-04 DOI: 10.17504/protocols.io.4r3l24on4g1y/v1
Abstract
This protocol describes H&E staining on fly heads paraffin sections for neuronal counting experiments.
Attachments
Steps
For neuronal counting experiments
1.
Fix fly heads in 4%.
2.
Embed in paraffin.
3.
Cut 4 micron sections.
4.
Dry at 42°C .
5.
Deparaffinize and bring through ethanols to water (xylene x 2, 100% x 4, tap H2O x 2).
6.
Stain with Harris’ hematoxylin for 0h 10m 0s .
7.
Rinse 3x tap H2O.
8.
Differentiate in acid alcohol (1% in 70%) – one quick dip.
9.
Rinse 3x tap H2O.
10.
Blue in tap H2O for 0h 5m 0s or to desired color.
11.
Rinse 3x tap H2O.
12.
Stain with eosin (10% in 100%) – one quick dip or to desired color.
13.
Rinse 2x 100%.
14.
Dehydrate through ethanol series and xylenes.
15.
Mount with Permount.
