[Histonet] Rolling sections

[Garrison, Becky]

Are you freezing muscle?  If so, for our muscle biopsies we use OCT
to adhere the base of the tissue to a piece of cork (which is frozen in
the isopentane); the tissue is not surrounded by OCT.    
The tissue section is guided onto the blade holder with a brush and then
picked up on the slide.   

All our other tissues are frozen surrounded by OCT.

Becky Garrison
Shands Jacksonville
904-244-6237

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Dearolf,
Jennifer
Sent: Tuesday, April 28, 2009 6:52 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Rolling sections

Greetings, Histonetters!

First, I wanted to thank all of you that responded to my e-mail a few
years back about freezing small pieces of muscle tissue.  We have found
a method that works for us, and if anyone is interested, I would be
happy to share.  It still involves the wonderfully explosive isopentane,
but it allows us to freeze fetal guinea pig muscle without artifact.

I am writing today to ask a question about cutting frozen sections with
a cryostat.  We are having problems with the sections rolling once they
come off the knife and before we can get them on a slide.  We have a
Microm 505E cryostat, and we cut our OCT mounted specimens at around -25
degrees C.  We use Accuedge high profile blades, cut sections between 8
and 12 microns thick, and use a brush to pull the sections off.  But,
when we remove the brush, the sections roll up.  Sometimes, they just
arc up and other times they completely roll into a jellyroll.  

I have tried putting 70% EtOH in a beaker in the cryostat.  This method
was suggested to us by a vendor, but it doesn't seem to work
consistently.  We can also flatten the sections with a brush, but unless
we are really quick, the sections roll up before we can get them on the
slide.  It makes it difficult to get serial sections.  

Any advice would be appreciated.  Thanks again for all your help so far.

Sincerely,
Jenn

Jennifer Dearolf, Ph.D.
Associate Professor
Biology Department
Hendrix College 
1600 Washington Ave.
Conway, AR 72032
(501) 450-4530 (office)
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