Hematoxylin and Eosin Staining
Staining of Sections for Histology
Place slides in slide holder and have all of solutions pre-made and in dipping trays prior to starting staining procedure. First two xylene washes are very important for complete de-paraffinization of the sections.
- Xylene: 3-5 minutes, 2X
- 100% Ethanol: 3-5 minutes, 2X
- 95% Ethanol: 3-5 minutes
- 85% Ethanol: 3-5 minutes
- 70% Ethanol: 3-5 minutes
- Eosin*: 15-30 seconds
- 50% Ethanol: 3-5 minutes
- 30% Ethanol: 3-5 minutes
- H2O: 3-5 minutes
- Hematoxylin: 10 seconds to 2 minutes, depending on tissue
- H2O: 3-5 minutes
- 30% Ethanol: 3-5 minutes
- 50% Ethanol: 3-5 minutes
- 70% Ethanol: 3-5 minutes
- 85% Ethanol: 3-5 minutes
- 95% Ethanol: 3-5 minutes
- 100% Ethanol: 3-5 minutes, 2X
- Xylene: 3-5 minutes, 2X
*Eosin
- 2.5 ml Acetic Acid
- 0.5 g Eosin
- 497.5 ml 70% Ethanol
Fixing Slides
- Blot away excess xylene, add 2-3 drops Permount and place coverslip on slide. Push out air bubbles
This should dry in ~10 min, but wait until the next day to place on microscope. - You can also clean away extra Permount using xylene