[Histonet] Problems with cryosectioning snap-frozen mouse kidney

[Mira Krendel]

Hello Histonet members,

I was wondering if you could provide some input on a problem we have encountered recently.  We routinely section mouse kidneys for IF.  The kidneys are cut in half and snap-frozen in Tissue-Tek cryomold in OCT using isopentane/dry ice bath.  Lately we have been having problems with tissue not sectioning properly (i.e., the OCT will section fine but when the technician gets to the tissue itself, the blade starts "chattering", creating multiple folds).  We have tried a different cryostat and had a similar issue. Someone has mentioned to me that this problem may be caused by not using cryoprotection with sucrose prior to freezing, although we have not had issues with sectioning snap-frozen tissue before.

Any advice on how to troubleshoot this?

Thanks in advance for your time,
Mira

Mira Krendel
SUNY
Syracuse, NY