Notre Dame Integrated Imaging Facility

H&E

Hematoxylin and Eosin

  1.              Xylene-  5 minutes
  2.             Xylene-  5 minutes
  3.             Xylene-  5 minutes
  4.            100% ETOH- 2 minutes
  5.            100% ETOH- 2 minutes
  6.            100% ETOH- 2 minutes
  7.            95% ETOH-  2 minutes
  8.            70% ETOH-  2 minutes
  9.            Distilled Water-  2 minutes  *
  10. Hematoxylin-  3-5 minutes
  11. Distilled Water-  Rinse x 3
  12. Clarifier-  30 seconds (20 dips)
  13. Distilled Water-  2 minutes
  14. Scott’s Tap Water-  30 sec (20 dips)
  15. Distilled Water-  1 minute
  16. 70% ETOH-  1 minute
  17. Eosin Y-  1-2 minutes
  18. 100% ETOH-  1 minute
  19. 100% ETOH-  1 minute   **
  20. 100% ETOH-  2 minutes
  21. 100% ETOH-  2 minutes
  22. Xylene-  2 minutes
  23. Xylene-  2 minutes
  24. Xylene-  2 minutes

 

  •  Stop at this point if you are deparrifinizing for immune staining.

**   Start here for dehydrating and cover slipping.