[Histonet] Re: Cryostat vs decal sections of bone

[Johnson, Teri]

Hi Louise,

Remember me? It's been a while.

You asked if there were distinct advantages to using cryosections of bone over decalcified wax samples for IHC and ISH and the answer is yes, of course. Some antibodies and probes just won't work well in paraffin processed samples, either due to the decalcification process or the alcohol/solvent exposure. The ISH purists know that just paraffin processing can diminish signal greatly. Having said that, obtaining excellent sections of undecalcified, fresh frozen bone is tricky. Very tricky indeed, and what works one day doesn't work the next. You'll need to have at least one tungsten carbide knife for it, so make sure you also get the regular blade holder. I don't know if you still use steel knives on the cryostat, and if so you'll already have the correct blade holder, but we use disposables so we needed to order a separate one.

We've had pretty good luck getting beautiful sections of fixed and decalcified cryosections and the cryojane, and this would work well for samples that just flat don't like paraffin processing but the antibody staining does ok with formalin and formic acid treatment. Having the system opens up this possibility for your studies as well.

Best wishes always,

Teri Johnson, HT(ASCP)QIHC
Managing Director, Histology Facility
Stowers Institute for Medical Research
Kansas City, MO