RE: cryostat

From:Barry R Rittman

Jocelyn
Advantages
Cryostat sections are much faster to prepare than paraffin sections.
Adjacent sections can be subjected to variety fixatives, IHC, histochemistry etc. Eliminates the secondary fixation that occurs in paraffin processing in the alcohols etc.
Gold standard for some types of histochemisty such as lectin binding with unfixed sections.
Lipids retained.
Sections can be picked up directly on glass microscope slides.
Tissue for cryostat sectioning can be fresh tissue or fixed tissue.

Disadvantages
Technically more difficult and you do need a cryostat
Freezing must be carefully controlled to prevent ice crystal formation.
Only relatively small pieces of tissue can be frozen
Sometimes difficult to maintain tissue orientation
Costly to store both the blocks and the sections.
Due to the much smaller amount of shrinkage (2-5%), stained sections have a more open appearance and some pathologists may not be able to directly compare this with the  appearance of stained paraffin sections.
Barry

--Original Message-----
From: Jocelyn Torcolini [mailto:jmt175@psu.edu] 
Sent: Wednesday, July 02, 2003 12:33 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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