[Histonet] H&E help

[Weems, Joyce]

Also, warm water works best for bluing, if you have that option..

Joyce Weems 
Pathology Manager 
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE 
Atlanta, GA 30342 
678-843-7376 - Phone 
678-843-7831 - Fax 

 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lee &
Peggy Wenk
Sent: Saturday, January 10, 2009 11:00 AM
To: 'Charles, Roger'; 'Histonet'
Subject: RE: [Histonet] H&E help

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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