cell plug preps

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet)
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I learned some of the neatest tricks for handling cells in Barb Wright's
workshop, substituting for John Tarpley, and it works beautifully.

You can make the cells release or scrape off, whatever is done to get them
floating, put them in a 50 ml tube, centrifuge, spin down, pour off
supernate, do this a couple of time more with pure PBS, then add
OCT to top of plug, snap freeze the bottom, pop plug out, double embed it
in OCT.  Or better yet, take the cells and make a suspension, purchase
CollaPlugs (sorry, I'm at home and not near company info) which are collagen
plugs used by dentists for wound healing, the resorbable type, soak up the
cell suspension in this plug.  The plug can then be cut into 3 pieces,
and each piece fixed with a different fixative then paraffin process, or
or embed in OCT,  snap freeze for frozen sections.  You can
then stain the cells, which are maintained without being slammed against
a slide by a cytospin, or messed up, the cells looked wonderful!

Don't try to use a 15 ml conical, the tip is too narrow. The snap frozen
plug has a very high number of cells, so cut at 4 um for frozens, and
the paraffin embedded plug is treated just like a tissue.

Have fun, and boy did this ever work.

Gayle Callis

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