[Histonet] Re: ER/PR

[Reynolds,Donna M]

I don't work with these antibodies but when we were doing bone marrow bx pressure cooker, high pH retrieval we lost a lot of tissue. This was what we had previously work out for our antibody. We replaced the pressure cooker with a 70ºC water bath, kept the high pH retrieval and cooked for 2 hours. The stain and the morphology were beautiful. We lost none of our tissue. We also tried 4 hours with the same conditions but decided the 2 hours best for our tissue and antibody.  
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Message: 9
Date: Tue, 19 Jul 2011 11:20:54 -0400
From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
Subject: [Histonet] RE: ER/PR
To: Joanne Clark <jclark <@t> pcnm.com>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<C27AA2A01CEF31469813089E226F582E055F27B6F2 <@t> URMCMS7.urmc-sh.rochester.edu>
	
Content-Type: text/plain; charset="us-ascii"

We heat ours to 99C for 20 minutes with a cool down.  Using the Dako PT link.  (water bath)
BUT....before you change your pretreatment 
Make sure that your tissue is fixed properly, processed properly. 
After it is cut, make sure that the slides dry before you baked them in the oven.  
Any water left under those sections will lift the tissue off for sure. 
Hope that helps. 

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Joanne Clark [jclark <@t> pcnm.com]
Sent: Tuesday, July 19, 2011 11:11 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] ER/PR

Hi All,

I would like some input on ER/PR by IHC.  We have a lot of problems with tissue detachment from the slides.  We use a DAKO autostainer and do the heat retrieval in the Pascal pressure cooker.  We use high pH retrieval buffer and heat to 120 degrees and hold for 1 minute, but when you look microscopically at the slides the heat has really played a number on the tissue and it looks chewed up.  The slides have been in the slide dryer for at least an hour before running, sometimes longer.  Any ideas how I can reduce the effects of the heat on the tissue without compromising the staining?

Thanks for your help.
Joanne Clark, HT
Histology Supervisor
PCNM
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End of Histonet Digest, Vol 92, Issue 24
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