Staining Procedure

1. Stain rehydrated sections in Hematoxylin solution for 20-40 minutes.
2. Wash in tap water for 1-5 minutes, until sections turn blue ("bluing").
3. Differentiate sections in 70% ethanol—containing 1% HCl—for 5 seconds. This removes excess dye, allowing nuclear details to emerge.
4. Wash 1-5 minutes in tap water until blue.
5. Stain in Eosin solution for 10 minutes.
6. Wash 1-5 minutes in tap water.
7. Dehydrate, clear and mount.

Note the use of tap water in the washing steps—tap water provides the alkanlinity necessary for the "bluing" process.