Hematoxylin & Eosin staining

Hematoxylin stains nuclei blue / purple. Eosin stains the cytoplasm pink. Several protocol variants exist for H & E staining. The most basic variant is described below.

Paraffin sections

For paraffin sections wash with Xylene for 5 minutes, followed by 100% ethanol for 2 minutes, 90% ethanol for 2 minutes, 70% ethanol for 2 minutes, and finally deionised water for 2 minutes. Go to step 1 below.

Cryo-sections

For cryo-sections, air dry slides then wash / hydrate slides in deionised water for 2 minutes. (May need to repeat the 2 minute water wash). Go to step 1 below.

Steps

1. Hematoxylin for 5 – 10 minutes

2. Wash with deionised water changing water periodically

3. Eosin for 0.5 – 2 minutes

4. Wash with 90% ethanol

5. Wash with 100% ethanol

6. Wash with Xylene

7. Mount