[Histonet] Adult Mouse Skin Cryosections

[Nige]

Hi,

I'm having trouble sectioning adult mouse skin prepared for cryosections.
The dorsal skin has been taken freshly and put into plastic dispomoulds with
Tissue Tek O.C.T, orientated and then slowly frozen by placing on a metal
block which is nearly submerged in liquid nitrogen.  The sample freezes
nicely with no cracks, but when it comes to sectioning on a cryostat the
tissue tends to scrunch up and come away from the tissue tek on the fatty
side.  Shaving the skin had no effect on the outcome. I've tried lots of
different temperatures/section thickness but no luck.

I'm assuming the problem has something to do with the fatty layer that is
present with the skin, but it is very difficult (if not impossible) to
separate this from the skin.  Can anyone suggest how to get nice adult skin
cryosections, any other processing needed to be done before mounting in
O.C.T?

Many thanks,


nige