[Histonet] Rolling sections

[Dearolf, Jennifer]

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)