CRESYL VIOLET -FROZEN SECTIONS (UCLA)

 

1. 95% EtOH 3-5 min. 13. D-H2O 1-3 min.
2. 95% EtOH 3-5 min. 14. D-H2O 1-3 min
3. 100% EtOH 3-5 min. 15. 70% EtOH 1-3 min.
4. 100% EtOH 3-5 min. 16. 95% EtOH 1-3 min.
5. 100% EtOH 3-5 min. 17. 95% EtOH 1-3 min.
6. Chloroform: 100% EtOH, 1:1 20 min. 18. 95% EtOH + Glacial Acetic Acid (add drop- wise to desired strength) 1-3 min. if necessary
7. 95% EtOH 3 min 19. 100% EtOH 3-5 min.
8. 95% EtOH 3 min. 20. 100% EtOH 3-5 min.
9. 70% EtOH 3-5 min. 21. 100% EtOH 3-5 min.
10. 50% EtOH (opt.) 3-5 min. 22. Xylene 3-5 min.
11. D-H2O 2 min. 23. Xylene 3-5 min.
12. Stain 1-10 min. 24. Coverslip with Permount  
       

Cresyl violet acetate:

Cresyl violet acetate 1.0 g Adjust pH to 3.5 with acetic acid of Na acetate;
filter before each use;
store in dark
D-H2O 1000.0 ml
Glacial Acetic Acid 3.0 ml
Sodium Acetate 0.2 g

Results: Nissl substance (cytoplasm of soma & proximal dend- rites) = reddish blue/purple; nucleus = clear; nucleolus = dark blue; axon hillock = clear.

Cresyl echt violet:

Cresyl echt violet 5.0 g Filter before each use;
store in dark
D-H2O 1000.0 ml
10% Glacial Acetic Acid 3.5 ml (50 drops)

Results: same as Cresyl violet acetate but not as red