[Histonet] IHC trouble shooting question

Katri Tuomala katri <@t> cogeco.ca
Tue Feb 20 18:27:20 CST 2007


We have noticed similar kind of artifact with some of the heat retrieved 
tissues. We seem to be able to control it by:
1. Rinsing the slides well in TBS/Tween buffer before putting them in the 
retrieval buffer.
2. We also add Tween in the home made retrieval buffers, commercial ones 
usually have a surfactant in them.

If we are careless with pre-rinsing the artifact returns.
The slides are very close to each other in the racks we use for retrieval.


Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "Lester Raff" <LRaff <@t> lab.uropartners.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, February 19, 2007 5:02 PM
Subject: [Histonet] IHC trouble shooting question

Hello to the Histonet,

We are having intermittent difficulties with IHC staining.  We are
staining well fixed prostate core biopsies for AMACR and p63 using the
PIN2 cocktail.  We do a pressure cooker HIER and stain on the Dako
Autostainer.  We use slides that have the control tissue on the top,
patient tissue on the lower 2/3rds.

We are running into a condition we call "train-tracking".  Looking at a
core longitudinally, the center will stain, but the edges will not. The
artifact is very linear, with sharp cut-off between staining and
non-staining.  We do not see this artifact with our CK stain, which we
do not use retrieval for.

Dako has been out to level the racks and check the probe tips, but the
problem keeps coming back.  Anyone else deal with something like this?


Lester J. Raff, MD
Medical Director
UroPartners, LLC Laboratory

2225 Enterprise Dr. Suite 2511

Westchester, IL 60154

ph:  708-486-0076
fax: 708-486-0080

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