H&E Staining on Paraffin Embedded Sections
1. Deparafinization in oven for 10 minutes
2. Rinse in xylene for 10 minutes
3. 100% Alcohol for 3 minutes
4. 95% alcohol for 3 minutes
5. 90% alcohol for 3 minutes
6. 70% alcohol for 3 minutes
7. Water for 5 minutes
8. Hematoxylin solution for 3-4 minutes
9. Rinse in tap water
10. Eosin solution for 30 seconds
11. Rinse in tap water
12. 70% Alcohol for 3 minutes
13. 90% alcohol for 3 minutes
14. 95% alcohol for 3 minutes
15. 100% alcohol for 3 minutes
16. Xylene for 3 minutes
17. Mount with coverglass
This method can also be used for frozen sections with following change.
Fix the frozen sections and wash with PBS 3 times 5 minutes each, then begin with step 8 above.