RE: fixation??

From:"Sebree Linda A."

I know this would be expensive Ann, but you could switch to Dako's
polyclonal CD117 that doesn't need HIER (I'm assuming you are using
something else).  We use it at 1:100 on Ventana instruments with a 32"
incubation and AB block.  We switched from a monoclonal a while back because
a particular clinical trial some of our patients were being randomized to
required Dako's antibody.  

Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792-2472
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: ANN MARUSKA [mailto:AMARUSK1@FAIRVIEW.ORG]
Sent: Friday, October 26, 2001 10:30 AM
To: histonet@pathology.swmed.edu
Subject: fixation??


Hi Histonetters!

I really need your help on this one.....

I got a lung wedge resection in for IHC stains.....everything stayed on but
my CD117 which uses a heat retrieval (Tris-Urea Buffer pH9.5 in
steamer)......it kept falling off no matter if I increased the drying time
(up to 2hrs 60C and then room temp over nite)....used new retrieval
buffer...nothing helped.  Even tried different blocks thinking maybe it was
the tissue itself. The pathologist assured me it was properly fixed and sent
for processing (in our lab).  I was never able to get anything to stay on
and the pathologist sent out his report without the CD117.

OK....I got past that, and everything else in the lab went fine with tissue
staying on when I did other CD117s...when low and behold it happened again!
When I called the pathologist  (same one) to explain my predictment about
not being able to do CD117.......he told me it was the same patient, but who
came in for an axillary resection.  I have tried other blocks on this
patient and everything that requires a heat retrieval, no matter what my
retrieval buffer, FALLS OFF!!!  What is going on??........it's just this
patient that is posing these problems.

FYI....staining is done on NexES or Benchmark....falls off on both
instruments.

All comments welcome!

TIA

Ann



Thanks.

Ann Maruska
Fairview-University Medical Center
Mpls. MN  55454
amarusk1@fairview.org
612-273-9119





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