EZQuant–ELN Knowledge Base

Hematoxylin and Eosin (H&E) staining	Table of Contents

Dehydrate and rehydrate sections:

• 2 x 5 min Xylene.
• 2 x 5 min ethanol 100%.
• 3 min ethanol 95%.
• 3 min ethanol 70%.
• 5 min ddH₂O.

Blot excess water from the slide holder before going into hematoxylin.

Hematoxylin staining:

• 3 min Mayers/Harris Hematoxylin (filtered).
• Wash briefly in ddH₂O.
• Wash 5 min in tap water (to allow stain to develop).
• Dip 8-12 times (fast) in acid ethanol (1ml concentrated HCl + 400ml ethanol 70%).
• Wash 2 x 1 min in tap water.
• Wash 2 min with ddH₂O.

Blot excess water from slide holder before going into eosin.

Eosin staining and dehydration:

• 30 sec Eosin.
• 1 x 2 min ethanol 70%.
• 1 x 2 min ethanol 95%.
• 1 x 2 min ethanol 100%.
• 2 x 5 min Xylene.

   Add 1-2 drops of mounting medium to the slide and cover with a coverslip.