[Histonet] Rolling sections

[Watson, Linda]

May I suggest trying the following. I too have experienced cryosectioned slices rolling up.

1) try cutting sections at -20 instead of -25 (it may be too cold)
2) try cutting sections at 6 microns instead of 8-12 microns
3) allow some frost to build up on the metal plate surface and gently with two brushes hold either side down until it gently sticks then quickly pick the specimen up with your slide. This can be very tedious and slow going but at least I was able to get the sections I needed. 
4) the "breath" suggestion also worked, although not consistently, for me.
5) take your gloved "thumb" and quickly place over your sample then immediately section. This works too, but again not consistently-is worth a try.

Good luck,
Linda

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez
>Sent: Wednesday, April 29, 2009 9:04 AM
>To: Dearolf, Jennifer; histonet <@t> lists.utsouthwestern.edu
>Subject: RE: [Histonet] Rolling sections
>
>Jennifer,
>It sounds like the sections are a little thick.  Have you tried just
>barely blowing on the tissue with your breath to warm the OCT?
>
>-----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 3:52 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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