[Histonet] Monitoring of IHC staining trends

Rene J Buesa rjbuesa <@t> yahoo.com
Tue Apr 30 15:14:41 CDT 2013


In your example you acknowledge that your pathologist is right. In consequence you go back to your protocol and, more importantly, to the lot that cause your pathologist's concern and analyze if it contains the same amount of antibody.
By the way, you should have tested the lot to avoid the problem in the first place.
But maybe the problem is not in the lot but in the tissue or the processing protocol and you have to go over this problem also.
I remember that when I switched from xylene to mineral oil, the IHC staining came stronger because I eliminated xylene that has an "extracting" effect on the epitopes causing a weaker IHC reaction. When it was eliminated the dilution used for tissue processed with xylene resulted in a stronger IHC staining.
My point is that you have to look to the whole process and try to find out "what is different" and probably causing the stronger reaction.
René J.

From: "Troutman, Kenneth A" <Ashley.Troutman <@t> Vanderbilt.Edu>
To: "Histonet <@t> lists.utsouthwestern.edu" <Histonet <@t> lists.utsouthwestern.edu> 
Sent: Tuesday, April 30, 2013 1:27 PM
Subject: [Histonet] Monitoring of IHC staining trends


Hello Histonet,

I have a question for the group at large.  How are labs monitoring drift in IHC staining over time?

Here's the scenario:  You do lot to lot testing and everything looks fine until one day your pathologists are telling you that the CAM5.2 is too dark.  Now, you've been looking at these slides every day for the last year and, sure enough, when you pull out a slide from last year's lot, it is significantly lighter.  So what do we do about it?  Do we revalidate the stain?  Does anyone have a mechanism to monitor this better?  What is the threshold for revalidation?

Feedback from techs as well as any pathologists would be greatly appreciated.

Regards,

Ashley Troutman BS, HT(ASCP) QIHC
Immunohistochemistry Supervisor
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4531
Nashville, TN  37232
ashley.troutman <@t> vanderbilt.edu<mailto:ashley.troutman <@t> vanderbilt.edu>
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