Oct 03, 2022
H&E staining on paraffin sections
- Mel Feany1
- 1Harvard Medical School, Brigham and Women's Hospital
Protocol Citation: Mel Feany 2022. H&E staining on paraffin sections. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l24on4g1y/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
The protocol is in use and works as intended.
Created: October 29, 2020
Last Modified: May 31, 2024
Protocol Integer ID: 44019
Keywords: paraffin, neuronal counting, h&e staining, ASAPCRN
Abstract
This protocol describes H&E staining on fly heads paraffin sections for neuronal counting experiments.
Attachments
Safety warnings
For hazard information and safety warnings, please refer to the SDS (Safety Data Sheet).
For neuronal counting experiments
For neuronal counting experiments
15m
15m
Fix fly heads Overnight in 4 % formalin .
Embed in paraffin.
Cut 4 micron sections.
Dry at 42 °C Overnight .
Deparaffinize and bring through ethanols to water (xylene x 2, 100 % ethanol x 4, tap H2O x 2).
Stain with Harris’ hematoxylin for 00:10:00 .
10m
Rinse 3x tap H2O.
Differentiate in acid alcohol (1 % hydrochloric acid in 70 % alcohol ) – one quick dip.
Rinse 3x tap H2O.
Blue in tap H2O for 00:05:00 or to desired color.
5m
Rinse 3x tap H2O.
Stain with eosin (10 % in 100 % ethanol ) – one quick dip or to desired color.
Rinse 2x 100 % ethanol .
Dehydrate through ethanol series and xylenes.
Mount with Permount.