Preparing sections from bone - (Nov/19/2009 )
I haven't done any histology before and now I have to make thin sections from frozen bone without decalcification.
Does anyone know if there is some special blade for the cryotome to cut thin sections from bone? All the protocols that
I have found so far rely on decalcification.
We have cut bone at 6-8um in the cryostat with just a regular disposable low-profile blade, but we through a lot of blades though.
It's most important, in my experience, to make sure that the cryostat is operating at the right temperature, Too warm, and the tissue pulls out of the OCT, too cold the tissue shreds. I typically cut at -22Cabinet temp/-20object Temp, with at window of only about 3`C before the aforementioned problems arise. And make sure when turning the cutting wheel, that you keep it steady; this is not a problem if you have a motorized cryostat. Good luck!
I have heard of a process whereby bones are decalcified by heating in EDTA, rather than the usual decalcification products. Not much good for frozen tissue though.