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Everyone,

Thanks for the responses, they've been very helpful and given me points to
consider and discuss with our group. I'm new to biology, and I'm surprised
at how much of cell culture is still a 'black art'.

My understanding, which is limited at best, is that CO2 perfusion is
required to maintain pH, nothing more.  We will perform temperature control
by heating the objective and media.

As for simply perfusing with HEPES/equilibrated media, that's a new one to
me and I need to speak with others regarding it's feasibility. It sure
sounds good, and I admit it's a lot simpler.

Andy

At 08:52 AM 3/14/2005, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Slight misunderstanding here, I guess. The CO2-enriched air has a slightly
>higher density, therefore it will sink down in normal air (as long as the
>two haven't fully mixed). So superfusing the dish with CO2-enriched air
>will work in principle. Be aware though that even a slight draft/convection
>will cause problems. That's why it will not work (well) in a microscope
>incubator, for example. And it will work better with small dishes/chambers.
>
>Andy - can you perfuse your sample with equilibrated medium? In many cases,
>this is a lot simpler and far less expensive than any other solution (I
>admit this against my own commercial interest as a manufacturer of
>microscopy "live support" equipment). It also obviates the need to prevent
>evaporation.
>Apart from that, you're certainly welcome to contact me off-list to discuss
>the solutions we sell.
>
>Beat
>
>At 01:12 14-03-2005, you wrote:
>><snip rest>
>
>Andrew Resnick, Ph. D.
>Fellow
>Department of Physiology and Biophysics
>Case Western Reserve University
>216-368-6899 (V)
>216-368-4223 (F)