Hematoxylin and Eosin (H&E) Staining
Ariadna Laguna, Miquel Vila, Núria Peñuelas
Abstract
Hematoxylin and Eosin (H&E) Staining for histological sections
Steps
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.
