[Histonet] staining of negative controls with IHC

Tarango, Mark mtarango <@t> nvcancer.org
Tue Nov 13 10:24:00 CST 2007

You probably don't even need to get up to boiling for ki-67.  Try 30
minutes at 95 C in your pressure cooker.  Even on a "negative" control
you're bound to see something stain positive for ki-67, since it's a
proliferation marker... although it shouldn't be cytoplasmic.  I'm just
guessing there's got to be a cell or two in the tissue that's dividing.

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of MaryAnn
Sent: Tuesday, November 13, 2007 5:18 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] staining of negative controls with IHC

We are currently retrieving animal tissue in a pressure cooker with a
program of 125 degrees for 30 secs.  This protocol was taken from
biocare but I'm sure this is for human tissue. Most of the tissue is
retrieved with Biocare's Reveal (pH 6.0).  We are receiving some
background staining of our negative controls whereas if we did not
retrieve them, there is none.  We were staining for KI-67 which is a
nuclear stain. I am using a tonsil for the control. The negative control
showed some sporatic cytoplasmic staining on the Universal negative
control from biocare as well as deionized water. Since then we have
taken the temperature down to 115 degrees and increased the time to 25
minutes.  It seems to have decreased the cytoplasmic staining but there
is still some lingering.  Anyone out there have any ideas??? I'm sure
there is a better protocol for retrieving animal tissues. It has to be a
time and temperature thing!
MaryAnn Dixon
Biological Scientist
Anatomic Pathology
University of Florida
School of Veterinary Medicine
352-392-2235 ext 4517
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu

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