Harris' Hematoxylin Protocol

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.

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