[Histonet] cyrosectioning

[Gayle Callis]

Steven,

What species?  Our preference is to put one slice of liver per block, or 
one whole spleen.  If you get a lot of curling with sections right at 
tissue location,  then you may want do one piece per block or use a whole 
or half lobe.

As long as the tissues have the same cryosectioning characteristics at a 
desired cryostat temperature, then you could put more than one piece per 
block   It has been a disaster use for to put liver with intestine.  We do 
put murltiple tiny lymph nodes (as many at 12) or peyers patches in one 
block or spinal cord cut into lengths for multiple pieces destined for 
cross sectioning but we would never put liver in with any of these tissues 
those - just liver with liver.   Cryosectioning liver has been best at -17C 
while nodes or patches works at colder, usually -20C.

One can "bread slice? liver lobes, and embed on cut edges.  We have done 
the more flat orientation of whole or partial lobe - both work, obviously 
the whole/half lobe gives a bigger section and more information.    If you 
need to see the outer capsule of liver, on edge works well but you don't 
have a generous section.  Try it both ways.

As for critical orientation of slices, I would embed them in some kind 
sequence so you know where the slices are from any given lobe, say and 
outer slice, middle slice, then another outer slice - that type of 
thing.  Marking ink helps with who belongs to who!!    We keep track of 
where we collect spinal cord, and know which end it being cut first, 
usually proximal end of any given slice - we even do cervical, thoracic and 
lumbar portions.

Bet I have confused people with this one!!!   Good luck!!



At 12:10 PM 11/14/2005, you wrote:
>
>We do allot of cryostat sectioning of both fresh and  fixed sucrose 
>cryoprotected tissue.  I was wondering what other techs have found to 
>result in better sections, one larger piece of tissue or several smaller 
>pieces within one section.  When sectioning several pieces of smaller 
>tissue, such as liver is orientation critical?
>
>Thanks everyone.
>
>Steve

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)