[Histonet] H&E help

[Lee & Peggy Wenk]

It might not be a hematoxylin problem. It might be too much eosin.

Try one of the following:
- cut the eosin time down to 1 minute, and if it's still too pink/not blue
enough, cut the eosin time down to 30 seconds.
- and/or, change the first 95% alcohol after the eosin to 70% alcohol. That
will help to put out more eosin than 95%. 

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----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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