[Histonet] RE: H&E help

[Smith, Allen]

Some tap waters blue hematoxylin very well, some not so well.  I avoid the issue of differences in tap waters by blueing in 1.2% aqueous lithium carbonate (almost a saturated solution).
   - Allen A. Smith, Ph.D.
     Professor of Anatomy
     Barry University School of Podiatric Medicine
     Miami Shores, FL

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Charles, Roger
Sent: Friday, January 09, 2009 9:14 AM
To: Histonet (histonet <@t> lists.utsouthwestern.edu)
Subject: [Histonet] H&E help

Hello All,
TGIF!!!!
I'm trying to help out one of my veterinarian pathologist in getting some information on how to increase the bluing of the H&E staining.  Presently we are using pre made Mayers Hematoxylin from Sigma with an automated schedule as follows:

 1.  Drying 45min
 2.  citrisolve 3 min
 3.  citrisolve 30 sec
 4.  citrisolve 3min
 5.  100% 45 sec
 6.  100% 45 sec
 7.  100% 3 min
 8.  95% 30 sec
 9.  95% 1 min 30 sec
 10. Tap water 1 min
 11. Hematoxylin 8 min
 12. tap water 4 min
 13. eosin 1 min 30 sec
 14. 95% 1 min
 15. 95% 1 min
 16. 95% 1 min
 17. 100%1 min
 18. 100%1 min
 19. 100%1 min
 20. Citrisolve 1 min
 21. Citrisolve 1 min
 22. Citrisolve 1 min
 23. Citrisolve up to 120 min
The pathologist is stating the slides are too eosinic and would like greater bluing.  My question is "are there limitations to the Mayers Hematoxylin on how blue it will actually get and is there something else we can change in our schedule to increase the bluing or decrease the eosin to get the desired affect?
Thanks to all


Roger Charles
Microbiologist
Pennsylvania Veterinary Laboratory
2305 N Cameron St
Harrisburg, PA 17110
717-787-8808

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