[Histonet] Cryostat help

Setlak, Lisa LSetlak <@t> childrensmemorial.org
Tue Jan 11 11:00:56 CST 2011

We have a similar frozen section protocol but we fix the slide in 95% alcohol for ~1minute and then stain with Gill's Hematoxylin for 1 minute, rinse, blue and then alcohol, foloowed by Eosin for ~ 15secs.- dehydrate and mount.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Tuesday, January 11, 2011 10:29 AM
To: histonet <@t> lists.utsouthwestern.edu; KimberlyBlundon
Subject: Re: [Histonet] Cryostat help

That depends on what staining you need to use.
If an IHC, just fix the sections in acetone, air dry and proceed without HIER
If H&E fix quickly with formalin 10%, wash → hematoxylin for 30 secs without differentiation → eosin 15 sec→ dehydrate and mount.
Other HC procedures all will require fixation and using the specific protocol.
Always remember that it is a frozen section and therefore more delicate and less adhered than a paraffin section.
René J.

--- On Tue, 1/11/11, Blundon, Kimberly <Kimberly.Blundon <@t> drdc-rddc.gc.ca> wrote:

From: Blundon, Kimberly <Kimberly.Blundon <@t> drdc-rddc.gc.ca>
Subject: [Histonet] Cryostat help
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, January 11, 2011, 10:28 AM

Hello Histonetters,

I am new to the histology world and I was hoping to get some feedback
about staining after cutting sections in the cryostat. 

Does anyone have a protocol they use for staining after sectioning
tissue in a cryostat?

I have never done it before so I thought I would come to the experts.

Thanks for your help!


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