[Histonet] edge effect on IHC

Tue Sep 24 10:43:25 CDT 2013

That artifact cannot be fixation related because if there is some part of the sample that is well fixed is the outer limit, where the NBF enters.
It does not matter what stainer you use, they all can deliver incorrect amounts of reagents.
René J. 

________________________________
From: Caroline Miller <cmiller <@t> gladstone.ucsf.edu>
To: "Manfre, Philip" <philip_manfre <@t> merck.com> 
Cc: Rene J Buesa <rjbuesa <@t> yahoo.com>; Mary Benoit <mbplab <@t> yahoo.com>; "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Tuesday, September 24, 2013 11:13 AM
Subject: Re: [Histonet] edge effect on IHC

However, I use the Shandon sequenza, and I still see edge effect. I always thought it was a fixation related. Maybe even that the edge staining is 'more real' than the rest!!

Histology, it's all voodoo!!

Caroline 

Caroline Miller
Gladstone Institutes
www.gladstoneinstitutes.org

Tel: 415 7342566
Cell: 415 2187297

On Sep 24, 2013, at 7:51 AM, "Manfre, Philip" <philip_manfre <@t> merck.com> wrote:

> 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.
> 
> Phil.
> 
> 
> Philip Manfre, B.A., HT (ASCP)
> Associate Principal Scientist
> Merck Research Laboratories
> WP45-251
> PO Box 4
> West Point, PA 19486
> 
> 215-652-9750
> 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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