[Histonet] Re:cryosectioning liver/sucrose

[Gayle Callis]

Steve,

You did not say what temperature you were cutting at?  However, with 
sucrose cryoprotected tissue, we prefer -26C (colder)  or the sucrose 
begins to ooze out of the block like Karo syrup. That is a bit colder than 
what we would cut unfixed, fresh liver, at -17C for this very homogenous 
tissue.  So play with the temperatures a bit.

  We also blot excess sucrose from the tissue a bit before snap freezing 
just to have a good interface with the OCT.  With the cryoprotected tissue, 
cutting too arm may help cause the folding.  We do use brush technic, and 
the sections should come off perfectly flat - sharp blade (new edge) is a 
good idea too.

Good luck

  At 01:27 PM 7/22/2005, you wrote:
>Something a "tad" new for me here.  I've never sectioned 4%Para liver 
>infiltrated with 30% sucrose.  They sectioned well but the slide appear to 
>be loaded with what appears to be folds throughout.  Not what I routinely 
>see on regular unfixed FS.  Is this normal for fixed sucrose infiltrated 
>sections.  Are there any special techniques when dealing with this special 
>tissue.
>

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)