[Histonet] RE: Eliminating the edge effect in IHC/IF

[Della Speranza, Vinnie]

I think we'll agree that there are different scenarios so that the solution is not a one size fits all. For example, darkened staining around the periphery of needle core biopsies is not uncommon with even tinctorial stains, often thought to be the result of drying of the tissue during the collection of the sample. Years ago I came across an article maintaining that the dark staining on the periphery of needle cores was in fact due to the "trauma" of the needle cutting into the sampled organ. I've since forgotten the author's name and wish I could get my hands on that reference.

So I agree with Chris that there doesn't appear to be one simple answer to prevent this artifact and while fixation may contribute in some circumstances it's unlikely to be the remedy for all.

Vinnie Della Speranza
Manager for Anatomic Pathology Services
165 Ashley Avenue Suite 309
Charleston, SC 29425
tel. 843-792-6353
fax. 843-792-8974
 
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos
Sent: Tuesday, January 12, 2010 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF

Hi all,We also have observed this phenomenon many times. But sorry Colleen and Rene, I don't believe that an fixation issue is the explanation why the edges are sometimes stronger than the rest. To my opinion this is a bit too easy. One of my explanations is that the immuno reagents tend to stick to the edges of the tissue section, especially when no Tween20 is involved. As a result the outer edges might become a little dry during incubation times and cause darker mostly non-specific staining. Always try to cover a section completely, including a rim around the section. However, to be honest, I am sure my explanation is certainly not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone:  +31 20 5665631
 From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>

Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren� J.

--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:


From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM


Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?

Your kind help is greatly appreciated,


Thanks in advance,

Best,
Karen

Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
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