[Histonet] Re: Staining on Alcohol Fixed Smears

Tom McNemar TMcNemar <@t> lmhealth.org
Tue Dec 17 04:48:28 CST 2013


Thanks for the clarification.  I appreciate the info!

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar <@t> lmhealth.org
www.LMHealth.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Beth Cox
Sent: Monday, December 16, 2013 2:25 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Staining on Alcohol Fixed Smears

I agree that with no formalin exposure, there is no crosslinking that
needs to be broken, but a "then, do nothing" approach to antigen
retrieval oversimplifies the issue.   Alcohol fixation makes changes in
the protiens/antibodies also, although different changes than formalin
does, and these changes need to be accounted for.  In my research on
alcohol fixed smears, I found that most antibodies perform best with
some antigen retrieval, often gentler than formalin tissues require.

Beth Cox, HTL/SCT(ASCP)QIHC
Vagabond Histotech

------------------------------

Message: 4
Date: Mon, 16 Dec 2013 11:13:59 -0500
From: Tom McNemar<TMcNemar <@t> lmhealth.org>
Subject: RE: [Histonet] Re: Staining on Alcohol Fixed Smears
To: 'Beth Cox'<bethcoxx <@t> gmail.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>,"levi.fried <@t> gmail.com"
        <levi.fried <@t> gmail.com>
Message-ID:
        <E9A90E28259D2F4E84308C5E8EA8F7B4016951E94252 <@t> lmhs-exchange>
Content-Type: text/plain; charset="us-ascii"


Since there is no exposure to formalin, we do not do antigen retrieval on alcohol fixed specimens.  We use the Ventana Benchmark XT and copy the protocol to a new number and use the "Wet" option rather than paraffin.  No retrieval needed.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar <@t> lmhealth.org
www.LMHealth.org

-----Original Message-----
From:histonet-bounces <@t> lists.utsouthwestern.edu  [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Beth Cox
Sent: Saturday, December 14, 2013 6:30 PM
To:histonet <@t> lists.utsouthwestern.edu;levi.fried <@t> gmail.com
Subject: [Histonet] Re: Staining on Alcohol Fixed Smears

Levi,

I've worked up most of the more common antibodies on alcohol fixed
smears.  There are two things to remember here:  1) your specimens are
alcohol fixed, meaning antigen retrieval needs are different, and 2)
they are smears, meaning the cells weren't 'cut open' during microtomy,
which impeeds some nuclear staining.  Those are two different issues to
consider.

Some rules of thumb:
1)  most cell membrane antigens will stain nicely with minimal changes
to the protocols you use for FFPE specimens
2)  most cytokeratin antigens stain nicely with little change to your
FFPE protocols (if you need to change, start by cutting your antigen
retrieval time in half)
3)  most nuclear antigens require significant changes from FFPE
protocols, and some are essentially impossible to stain using standard
antibodies.

When I return to work on Monday, I will forward you my protocol
'adjustments' for the antibodies you requested.

Beth Cox, HTL/SCT(ASCP)QIHC
Vagabond Histotech




On 12/14/2013 12:00 PM,histonet-request <@t> lists.utsouthwestern.edu  wrote:

> Message: 4
> Date: Sat, 14 Dec 2013 18:25:08 +0200
> From: Levi Fried<levi.fried <@t> gmail.com>
> Subject: [Histonet] Staining on Alcohol Fixed Smears.
> To:histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>        <CA+puqxTXL85pEwvR=sqPQA36tMJPm=sx2Kv3GYEbRbu0He3C2g <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Hi Everyone.
>
> I am looking to stain alcohol fixed smears with a group of antibodies.
>
> The antibodies of particular interest are p63, p53 and ki67.
> Along with these antibodies I am looking for positive nuclear and
> cytoplastic staining antibodies.
>
> If anyone has any experience in staining alcohol fixed slides your
> information is very appreciated.
>
> All the best.
>
> -- Sincerely, - Levi Fried

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you.



More information about the Histonet mailing list