Apoptosis in frozen sections

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From:"Patterson, Noelle" <PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL> (by way of histonet)
To:histonet <histonet@magicnet.net>
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I have been reading and tracking histonet discussions on the Tunnel assay
and apoptosis detection, but am unable to determine the alternatives/best
ways to detect apoptosis (as well as apoptosis in contrast to necrosis) in
frozen sections.

I suppose what I need help with is the different ways, protocols, company
kits, etc. out there to do the tunnel assay on frozen sections (paraffin if
need be)/detect apoptosis.

I will need to conclusively differentiate between necrosis and apoptosis in
kidney biopsy material generally embedded in OCT and snap frozen.  I gather
that the "standard" for assessing necrosis is the traditional H&E stain, but
this does not stain frozen sections as vibrantly as may be necessary.  Are
there other suggestions/possibilities?  I have tweaked the H&E for frozens
quite a bit...and am already pretty close to optimal, so I will need to
explore other ways.

Any ideas are welcome.  Vendors are certainly welcome in this discussion of
apoptosis vs necrosis detection methods/kits out there.  I am not, by
nature, a kit person though...so please explain the basis for the method.  I
understand apoptosis quite well, just never needed to detect it

Thanks, in advance, for your input.


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