[Histonet] Problems with Frozen tissues

[Kim Donadio]

may I ask why you are not fixing them? Even the stain the 1st gentlman was doing(Beta Gal) recomended to have a formalin fixation apllied first. 
 
There are many ways to get to the same show as we all know, But in my  experiance air drying frozen sections will dry out and cells can autolyse as the process of autolysation has not been stopped, also vacouls in the tissue can develope from the process of dehydration without the use of fixation< sorry for spelling, on lap top on dark porch. 
 
Are you trying to minumize usage of alcohols or other fixatives? 
 
PLease explain to me if you would because I have cut just about all kinds of tissues for all kinds of procedures and never have I heard to keep a fresh section out for an hour before staining and without a fixative. I love ot learn new things )
 
Thanks
 
Kim


________________________________
From: Daniela Bodemer <daniela.bodemer <@t> mcri.edu.au>
To: Kim Donadio <one_angel_secret <@t> yahoo.com>; Igor Deyneko <igor.deyneko <@t> gmail.com>; Histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, November 1, 2011 6:16 PM
Subject: RE: [Histonet] Problems with Frozen tissues

Our tissues are frozen in OCT. I use Super Frost Plus slides from Thermo Scientific and store the sections in the fridge until I need them. After leaving them out on the bench for an hour I start my H&E with 1 min running water and so on. I rarely lose sections with these slides.

Daniela

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
Sent: Tuesday, 1 November 2011 3:02 AM
To: Igor Deyneko; Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Problems with Frozen tissues

Ive never just air dried my frozen sections. always put them in a fixative such as pen fix, a alcohol and or formalin mixture, something( depends on what your going to look for, test etc). That with using charged slides and never had too many problems with this. 
 
Kim


________________________________
From: Igor Deyneko <igor.deyneko <@t> gmail.com>
To: Histonet <@t> lists.utsouthwestern.edu
Sent: Monday, October 31, 2011 9:56 AM
Subject: [Histonet] Problems with Frozen tissues

I'm looking for some advice on frozen tissues. This is the first time I'm doing it. All the tissues: skin, lungs, spleen, liver, and pancreas cut well onto special Gold Plus slides from Fisher. Then, when I was ready to stain the slides, i air dried them fro an hour and wanted to do H&E and Beta-Gal, all the tissues fell off slides. Can anyone suggest any tips on preventing this mischief?
Thank you in advance.
Igor Deyneko
Infinity Pharmaceuticals
Cambridge, MA
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