PAPANICOLAOU STAIN

1. Preserve the smear by covering with a liberal amount of hair spray or cyto-spray while the specimen is still wet, DO NOT AIR DRY.

2. Fix in methacarn.......................................15 min.

3. Rinse in 100% ETOH.................................1 min.

4. Hydrate in 95% ETOH................................1 min.

5. Rinse in DH2O...........................................2 min.

6. Stain in Harris hematoxylin (non-acidified).....3 min.

7. Rinse in H2O.............................................until clear

8. Differentiate in 0.25% HCl............................3 dips

9. Rinse in DH2O............................................1 min

10. Blue in Scott's tap water.............................1 min.

11. Rinse in DH2O...........................................1 min.

12. Dehydrate in 70% ETOH.............................1 min.

13. Dehydrate in 95% ETOH.............................1 min.

14. Counterstain in OG6...................................1 min. & 30 Sec.

15. Differentiate in 95% ETOH...........................3 dips

16. Differentiate in 95% ETOH...........................4 dips

17. Counterstain in EA50..................................2 min.

18. Differentiate in 95% ETOH...........................3 dips

19. Differentiate in 95% ETOH...........................5 dips

20. Dehydrate in 100% ETOH...........................10 dips

21. Dehydrate in 100% ETOH............................1 min.

22. Clear in xylene............................................1 min.

23. Clear in xylene............................................1 min.

24. Mount with permount and coverslip.