[Histonet] Cutting frozen decalcified mouse bones

[Adam .]

Hi all,

I had a very difficult time cutting my very precious triple transgenic
frozen decalcified mouse bones this morning. As I cut into them, the
sections scrunched up into a mess of OCT pretty much as soon as the blades
hit the block. These were 4% PFA perfused and post-fixed for 24 hours,
decalcified with 14% EDTA using weight loss/weight gain as an endpoint,
cryoprotected overnight in 30% sucrose, embedded in OCT using liquid
nitrogen cooled isopentane vertically (i.e. I was cutting from one growth
plate to the other), and cut at 7 uM. Changing to a new blade didn't help.
Changing the cutting temperature from -10C to -15C didn't help. Changing the
cutting speed didn't help. I can't change the cutting angle without a
special tool, and this isn't my device so I don't want to mess with it.

The only way I could get reasonable sections was to use a tape transfer
system, which seemed like overkill to me, but I need these sections ASAP.
Ideas? I have a whole bunch more bones I need to cut over the next few
weeks...

Thanks,
Adam