[Histonet] edge effect on IHC

Manfre, Philip philip_manfre <@t> merck.com
Tue Sep 24 09:51:23 CDT 2013

I concur with Rene'.  That was my first thought. Edges drying up on an autostainer due to hydrophobia or insufficient amounts of solution. Stainers usually have the option to adjust the volume that is applied and sometimes even the location on the slide (zones).  Thorough deparaffinization is also essential to avoid the hydrophobic repelling of solutions and wash buffers.  Also be sure your reagents have some form of surfactant or detergent (e.g.Tween 20) in them to break the surface tension and allow the even spread of whatever is being applied.


Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
PO Box 4
West Point, PA 19486

215-993-0383 (fax)
philip_manfre <@t> merck.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Tuesday, September 24, 2013 10:36 AM
To: Mary Benoit; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] edge effect on IHC

Sometimes that "edge effect" is caused by drying when there is not enough reagents. If you are using an automated stainer, please check the amount of delivered reagents.
René J.

From: Mary Benoit <mbplab <@t> yahoo.com>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Monday, September 23, 2013 5:00 PM
Subject: [Histonet] edge effect on IHC

I have been aware of some "edge effect" on an occasional slide, but lately we noticed the effect is showing up in the area of the slide that is farthest from the labels. We use DAKO Link Autostainer and their pretreatment baths for everything. One case in particular we stained a large melanoma met to colon that was well fixed and processed that stained only the edge, however when we sent block to Neogenomics reference lab, their slide stained evenly over the entire tumor area. We both used a melanoma cocktail. Not sure what is going on. Any suggestions?
Thank you
Mary F Benoit, MT(ASCP)
The Pathology Laboratory
830 Bayou Pines Drive
Lake Charles, LA 70601
mbplab <@t> yahoo.com
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