RE: cryostat

From:

Margaret Blount

Dear Jocelyn,

Cryosectioning is required for some purposes, e.g. enzyme histochemistry, lipids, some antibodies etc. Technique recipes should indicate where frozen sections are required and whether they should be fixed en block, as sections and what type of fixation is appropriate. Sections will adhere to the slides as you hold them close to the section, careful not to touch the knife as the section will stick to the knife giving an artefact that I like to describe as "lacey"! Slides should be at room temperature and sections should be flat before picking them up. Whether you place the slides at room temperature for a while after picking up sections or keep them frozen will depend upon the requirements of subsequent staining (e.g. for in situ hybridisation, you will need to keep the slides frozen). For some techniques you will definitely require either polylysine coated slides or Plus slides. You will need to practise to get the best sections and nothing is better than being trained by an experienced cryotomist.

I hope this helps.

Regards

Margaret Blount

Dept of Clinical Biochemistry

University of Cambridge

-----Original Message-----
From: Jocelyn Torcolini [mailto:jmt175@psu.edu]
Sent: Wednesday, July 02, 2003 6:33 PM
To: histonet@pathology.swmed.edu
Subject: cryostat

Hi Histonetters,

I have a few questions about cryostat sectioning.

In what situations is a cryostat better than regular paraffin processing, and why?

How do you get cryostat sections to stick to a slide for staining?

Do you need to fix tissue for cryostat in paraformaldhyde/sucrose solution? Can you section fresh/frozen tissue?

Thanks in advance,

Jocelyn Torcolini

Electron Microscopy Facility

1 Frear South Building

University Park , Pa 16802

Phone (814) 865-0212

www.lsc.psu.edu

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