[Histonet] IHC validation

Liz Chlipala liz <@t> premierlab.com
Tue Feb 8 16:57:40 CST 2011


If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative). 


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Joe
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
    The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?

Joe (JTT)

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