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

[Sherwood, Margaret]

Actually, when I read about the problem, my thought was that if you don't circle
your tissue with an immunopen or wax pencil, then the reagents (esp. primary
antibody)might not cover the tissue evenly as you say.  So I tend to agree with
you.

Peggy 

-----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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