[Histonet] H&E help

[Bryan Llewellyn]

Mayer's hemalum is a nuclear selective progressive formula so the staining 
is largely confined to nuclei.  If the pathologists want more blue tinge in 
other parts of the tissue, then you should change the hemalum, or simply 
increase the hematoxylin content in the solution.  Cole's does stain darker 
than Mayer's, but has 1.5 grams hematoxylin per litre, fully oxidised.  Try 
that.  You might want to decrease the time in eosin as well.  Try one 
minute.

----- Original Message ----- 
From: "Charles, Roger" <rcharles <@t> state.pa.us>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, January 09, 2009 6:13 AM
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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