H&E staining

H&E Stain---Kit

6.6.2013 Li

Components

Item Quantity
Mayer’s Hematoxylin Solution (#SH26-500D) 250 ml
Eosin Y Intensified (#23314631 or 23314630) 250 ml
1% Acid Alcohol 250ml (2.5ml HCl + 250ml 70% EtOH)
Acetic Acid Glacial 1 ml eosin Solution + 2ul Acetic Acid

Procedure

For paraffin sections

  1. Deparaffinize sections with xylene, 5 min ×2
  2. Re-hydrate with 100% alcohol, 2 min ×2
  3. Re-hydrate with 95% alcohol, 2 min ×2
  4. Rinse in running tap water for 2 min
  5. Stain in Mayer’s Hematoxylin Solution for 30 seconds
  6. Rinse in running tap water for 2 min
  7. Dip in 1% Acid Alcohol ×2
  8. Wash with running tap water for 2 min
  9. Counterstain in Eosin Solution (1ml Eosin Solution + 2ul Acetic Acid Glacial) for 2 -3 min
  10. Rinse with 3-4 washes of tap water
  11. Rinse quickly in 95% alcohol (20 dips ×2)
  12. Then dehydrate through 100% alcohol (10 dips ×2), Until dry
  13. Clear in xylene, 2 min ×2
  14. Add 2-3 small drops of Cytoseal, Slowly apply coverslip,avoiding air bubbles.