Mar 20, 2023
Hematoxyllin-Eosin ( HE ) Staining
- Freda Halim1,2
- 1Doctoral Programme, Faculty of Medicine, Universitas Padjajaran;
- 2Surgery Department, Faculty of Medicine, Pelita Harapan University
Protocol Citation: Freda Halim 2023. Hematoxyllin-Eosin ( HE ) Staining. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l27zo3g1y/v1
Manuscript citation:
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
We use this protocol and it's working
Created: March 18, 2023
Last Modified: March 20, 2023
Protocol Integer ID: 79019
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Abstract
This is a protocol to stain tissue with Hematoxyllin-Eosin ( HE ) Staining. We use human breast cancer tissue for running this protocol.
Deparaffinization
Deparaffinization
15m
15m
Incubate slides in Xylol
5m
Incubate slides in Xylol
5m
Rehydration
Rehydration
25m
25m
Incubate slides in 100% Alcohol
5m
Incubate slides in 96% Alcohol
5m
Incubate slides in 80% Alcohol
5m
Incubate Slides in 70% Alcohol
5m
Rinse slides with Aquadest for 5 minutes
5m
Nuclear Staining
Nuclear Staining
10m
10m
Incubate Slides in Hematoxyllin Reagent ( Hematoxyllin JT Baker )
5m
Rinse slides with running tap water
5m
Defferentiation
Defferentiation
10m
10m
Incubate slides in bluing reagent @ 1 minutes, 5 times
5m
Rinse slides with running tap water
5m
Cytoplasm staining
Cytoplasm staining
1m
1m
Incubate slides in Eosin reagent 1 minute
1m
Dehydration
Dehydration
20m
20m
Incubate Slides in 70% Alcohol
5m
Incubate Slides in 80% Alcohol
5m
Incubate Slides in 96% Alcohol
5m
Incubate Slides in 100% Alcohol
5m
Clearing
Clearing
15m
15m
Incubate Slides in Xylol
5m
Incubate slides in Xylol
5m
Incubate Slides in Xylol
5m
Mounting and Finishing
Mounting and Finishing
Wipe excess fluid,cover the slides with cover glass
Protocol references
Fischer AH, Jacobson KA, Rose J, Zeller R. Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc. 2008 May 1;2008:pdb.prot4986. doi: 10.1101/pdb.prot4986. PMID: 21356829.