[Histonet] Re: Formalin Fixation issues

[Elizabeth Chlipala]

There is a real nice article on fixation and handling of breast lumpectomy samples in the Journal of Histotechnology by Dr. Stephen Ruby.  It goes over a nice technique we used to use on those samples, worked great.  It did use alcoholic formalin which I suspect can't be used currently if you have not validated for that fixative for ER/PR etc.  But the overall technique can be used with 10% NBF.  The thickness of samples is key as Bob pointed out.  We had a new pathologist on gross and the next day we had about 20 or 30 recuts on their breast samples.  That number of reacts was uncommon.  I needed to figure out what was the issue and we found out it was the size of tissue that was placed into the cassettes.  I explained that they needed to place smaller and thinner pieces of tissue in the cassette.  Once I did that we did not have anymore problems with their samples.  The article information is below.

Paper Towel "Sandwich" - Alcoholic Formalin Fixation for Breast Biopsies
Author: Ruby, Stephen G.

Source: Journal of Histotechnology, Number 1, March 1999 , pp. 49-51(3)

Publisher: Maney Publishing


Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bob Richmond
Sent: Monday, July 09, 2012 11:51 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Formalin Fixation issues

At this point I'm not sure I understand what Jill Cox's problem is.
Forgive me if I keep blaming the pathologist - since I am one! - but
in my experience most of these breast problems originate at the gross
desk.

Is your pathologist cutting the fatty breast tissue thin enough? This
is always a challenge, even after you've done it as long as I have. If
your pathologist is cramming the cassettes full of fat, then that's
where the problem lies.

Do you prepare your own neutral buffered formalin, or buy it? I
prepared a lot of it years ago, and supervised others who did, and I
learned the hard way that mistakes are easy to make when you brew your
own. If you buy your NBF ready made, as most people do nowadays, then
read the label carefully. Have you changed brands recently?

Bob Richmond
Samurai Pathologist
Knoxville TN
***************************************************
On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox <jcox90 <@t> yahoo.com> wrote:
> I think you misunderstood what I meant. I know it's not supposed to be fixed
> before grossing, it's also not getting fixed after processing. But thanks
> for your two cents..
>
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Monday, July 9, 2012 10:25 AM
> Subject: [Histonet] Re: Formalin Fixation issues
>
> Jill Cox asks: >>Is anyone having problems with breast fixation prior
> to processing in formalin? For a couple of months now our breast
> specimens aren't fixing very well before gross. Our pathologist thinks
> they have changed something in the formalin itself. We utilize 15/1
> ratio and in some cases let fix over the weekend. This has only been
> happening over the last couple of months and can't seem to figure this
> out. Any advice or similar problems, would love to hear from you.<<
>
> Breast specimens shouldn't be expected to fix before they're grossed,
> or at least before they've been cut into thin slices. Delaying
> fixation compromises immunostaining, to say nothing of H & E. They
> should be grossed as promptly as possible after they're received, and
> should never sit over the weekend without dissection.
>
> I seriously doubt that anything in the formalin has changed. It's your
> technique that needs to change. Unfortunately, this is a difficult
> task to delegate.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN

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