Centre Optical Instrumentation Laboratory

Haematoxylin & Eosin Staining

Frozen Sections

1. Rehydrate slides in tap water (1 min)
2. Full strength Ehrlichs haematoxylin (40 secs)
3. Gently rinse in tap water
4. Dip 3 times in acid alcohol
5. Gently rinse in tap water (5 mins)
6. Aqueous eosin (30ml eosin, 300ml water) for 15 mins
7. Rinse gently in tap water (5 mins)
8. Dehydrate
2 x 70% alcohol
2 x absolute alcohol
2 x xylene or histoclear
9. Mount in DPX

Paraffin Sections

1. Rehydrate
2 x xylene or histoclear
2 x absolute alcohol
2 x 70% alcohol
2. Ehrlichs haematoxylin (5 mins)
3. Rinse in tap water
4. Differentiate in acid alcohol (15 sec)
5. Blue in tap water (5 mins)
6. Aqueous eosin (30ml eosin, 300ml water) for 15 mins
7. Rinse in tap water
8. Dehydrate
2 x 70% alcohol
2 x absolute alcohol
2 x xylene or histoclear
9. Mount in DPX