[Histonet] RE: ER/PR

McMahon, Loralee A Loralee_Mcmahon <@t> URMC.Rochester.edu
Tue Jul 19 10:20:54 CDT 2011


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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