[Histonet] HELP- Cryosectioning FAT!

[Balasubbramanian, Dakshnapriya]

Hi all,

I've been having problems with cryosectioning fatty tissue for a long time now. The section leaves a hole in the middle. I know this is probably a much discussed topic, but I've tried a lot of strategies with no luck. The tissue is of mouse origin and has micro-lesions buried inside fat. So the fat needs to be cut in order to get to the lesion. I started with a temperature of -17C and tried upto -25C and also at -50C; didn't work at any of these temperatures. I tried rubbing the block, blade and anti-roll plate with dry ice-again,no luck. 

I rapid-freeze the tissue with OCT in LN2 and then store the block at -80C. Could I try sectioning the block directly from -80C? (never done that). 

Any other suggestions on how to tackle this?

Any advice is much appreciated!!

Thanks,
Dakshna