Jun 17, 2024

Public workspaceHematoxylin and Eosin (H&E) Staining

Nature Communications
DOI
https://dx.doi.org/10.17504/protocols.io.ewov19deklr2/v1
  • Núria Peñuelas1,
  • Ariadna Laguna1,
  • Miquel Vila1
  • 1VHIR-CIBERNED-ASAP
  • Vilalab Public
  • Nuria
Miquel Vila
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DOI: https://dx.doi.org/10.17504/protocols.io.ewov19deklr2/v1
External link: https://doi.org/10.1038/s41467-024-53168-7
Protocol Citation: Núria Peñuelas, Ariadna Laguna, Miquel Vila 2024. Hematoxylin and Eosin (H&E) Staining. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov19deklr2/v1
Manuscript citation:
Laguna, A., Peñuelas, N., Gonzalez-Sepulveda, M. et al. Modelling human neuronal catecholaminergic pigmentation in rodents recapitulates age-related neurodegenerative deficits. Nat Commun 15, 8819 (2024). https://doi.org/10.1038/s41467-024-53168-7
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: June 17, 2024
Last Modified: June 17, 2024
Protocol Integer ID: 101945
Keywords: staining hematoxylin, histological section, eosin
Abstract
Hematoxylin and Eosin (H&E) Staining for histological sections
Troubleshooting
DEPARAFFINIZE
Xylene - 5 minutes

Xylene - 6 quick immersions
HYDRATE
100º alcohol - 6 quick immersions
100º alcohol - 6 quick immersions
96º alcohol - 6 quick immersions
70º alcohol - 6 quick immersions
50º alcohol - 6 quick immersions
Distilled water (H₂O) - 2 minutes
STAINING
Harris Hematoxylin - according to preparation
Running tap water (H₂O) - 5 minutes
DIFFERENTIATE HEMATOXYLIN
Acid alcohol - 2 minutes. Acid alcohol is made by adding 0.3 ml of HCl to 250 ml of 70º alcohol.
Running tap water (H₂O) - 5 minutes
BLUEING
NaHCO₃ - 1 minute. Add one tablespoon of NaHCO₃ to 250 ml of distilled water (H₂O).
Running tap water (H₂O) - 5 minutes
COUNTERSTAINING
96º alcohol - 3 minutes.
Eosin - according to preparation
DEHYDRATE
96º alcohol - 6 quick immersions
96º alcohol - 6 quick immersions
100º alcohol - 6 quick immersions
100º alcohol - 6 quick immersions
CLEAR
Xylene - until the preparations are clear. Leave in xylene until mounting.