[Histonet] Re: Staining on Alcohol Fixed Smears

[Beth Cox]

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