IHC Documentation....& OCD

From:Amos Brooks

    There is a lot of variation with this in different labs. Currently we do
the initial titers of the antibodies (at least three) and often we try
various antigen retrieval methods. (DON'T TRUST THE DAMN SPEC SHEETS!!) We
save these test slides along with a list of the processes we took in getting
them (pretreatment, dilutions, incubation times, detection methods and
chromogen used). Unless the results are blazingly obvious as to what works
and doesn't work, we usually have a pathologist look at the dilutions and
initial the data sheet which we keep with the spec sheet.
    We retiter each new lot, which I often grumble loudly about (especially
when we get a large order) and save the same information as above with the
new antibodies spec sheet while filing the old one with retired antibodies.
This is actually a good thing though. Even though you will probably come up
with the same procedure as you had, it forces you to look at each antibody
individually and re-evaluate how it stains and weather it can be improved.
    For new antibodies we haven't used before (CAP says something about
verifying the results on 40 separate cases or some crazy nonsense like
that!), we stain one microarray with 40 controls in it and keep this on
    For proficiency ... if you can keep up with all that, you've proven
yourself ... congratulations! This is a self testing process. If you can do
all this, then you've proven your ability to read a spec sheet, titer an
antibody develop a procedure, evaluate results, and prove diligence in
record keeping. All this with out even touching patient tissue.
Who says OCD is a disorder,
Amos Brooks

----- Original Message -----
From: "Tom T. McNemar" 
To: "Histonet (E-mail)" 
Sent: Wednesday, October 03, 2001 8:04 AM
Subject: IHC Documentation....

> Howdy folks,
> I was wondering what kind of documentation is being kept with regard to
> things as the initial workup of an antibody, controls, reactivity, and
> specificity.  Also, what kind of proficiency is being done by those doing
> IHC?
> I'd appreciate any and all input.  Thanks.
> Tom Mc Nemar
> Licking Memorial Hospital
> Newark, Ohio

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