CD34 for Brett, Gi Biopsies and burnt tissues for Julie and Rita

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From:"Jeff Silverman" <> (by way of Marvin Hanna)
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Hey Netters I'm back in digest form. Happy Millenium.

Rita, why use agar. Have each embedding tech stand at the grossing to learn
orientation of the biopsies. I use  Perfect Papers, used by hairdresers
doing permanent waves to wrap the tiny pieces, Also, I think if your final
alcohol has water, or our first paraffin has too much xylene and is hot
you will get a cooked appearance. I use a schedule on my old VIP shown
below that gives good results:
Formalin  5 hours 37 degrees C  change every day
70% alcohol   30 min RT
95%	         30 min RT
100% x 5 changes  30 min each Room Temp
Xylene            30 min  RT
Xylene            30 min	RT
Paraffin x4       30 MIN EACH  60 degrees C Rotate once weekly
We use plain Paraplast for both infiltration and embedding the blocks. The
DMSO that they add to Paraplast Plus, with its oyster taste,  makes me sick
as soon as I touch it.

Brett- Any immuno company sells the QBend-10 clone of CD34 and this reacts
well with formalin fixed human tissues staining most endothelia, numerous
primitive interstitial dendritic fibroblasts in . No antigen retreival is
necessary. The transmebrane molecule is highly conserved among species but
I don't know about cross reactivity.

Julie- with the above processing schedule, which works well on both
endoscopic biopsies and placentas and fatty breast tissue, GI stuff should
be no problem. The key is the microtomy. It is essential to use a slow,
smooth even stroke when cutting throught the block. I always find it
helpful to blow gently on the block with warm breath, as though I'm trying
to fog up a mirror. The humidity helps- it looks real dumb and it a wonder
I haven't cut off my nose getting so close. I never soak. Also, try cutting
them at  2
microns, no trouble with fresh Accuedge blades.
Anyone know of any temp work in Holland?
Jeff Silverman
Southampton Hospital
Southampton NY

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