Harris' Hematoxylin Protocol
- Stain rehydrated sections in Hematoxylin solution for 20-40 minutes.
- Wash in tap water for 1-5 minutes, until sections turn blue ("bluing").
- Differentiate sections in 70% ethanol—containing 1% HCl—for 5 seconds. This removes excess dye, allowing nuclear details to emerge.
- Wash 1-5 minutes in tap water until blue.
- Stain in Eosin solution for 10 minutes.
- Wash 1-5 minutes in tap water.
- Dehydrate, clear and mount.
Note the use of tap water in the washing steps—tap water provides the alkanlinity necessary for the "bluing" process.