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Date: | Wed, 14 Feb 1996 07:30:47 -0500 |
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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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