On Tue, 13 Feb 1996, Michael Stanley wrote:
>
> We have a couple of users who want to use the confocal to look at calcium
> and/or pH changes on suspension cells (culture cells).  Does anyone know of
> specific tricks for trying to stabilize single/suspension cells in a
> perfusion chamber.
 
We find, like Ellen Lumpkin, that cells (in our case epithelial cells)
attache well to clean glass. We simply run a bunch of coverslips through an
autoclave (which seems to enhance the inherent surface charge or some
resident "attaching factor" in the glass) and then use them. We place a
drop of suspension on the glass, and then let it sit for 5-10 minutes
before starting the perfusion. We lose a significant fraction of the
cells when the perfusion starts, but the ones which are left are the
healthy ones. We perfuse at about 1 ml/minute through a chamber which is
about 100ul. Chip
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  Chip Montrose
   Johns Hopkins University     tel: 410-955-9681
    Ross 930                      FAX: (410) 955-9677
     720 Rutland Avenue            email: [log in to unmask]
      Baltimore, MD 21205
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