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Simon,
We have tried
bubbling the media - however, if we ever have serum in it, we get
frothing. We can minimize the problem by using an aquarium
device.
For rapid changes of
pCO2 or pO2, I have bubble the gas through water (heated to 37) to saturate the
gas, and then blow this gas right over the imaging chamber.
We have been able to
step the pCO2 from 2% to 5% to 10% in < 2 seconds.
Two examples where
we have used this:
N. Jouvenet, P. D. Bieniasz,
and S. M. Simon. Imaging the biogenesis of individual HIV-1 virions in live
cells. Nature (London) 454 (7201):236-240,
2008.
S. M. Simon,
D. Roy, and M. Schindler. Intracellular pH and the control of multidrug
resistance. Proc.Natl.Acad.Sci.U.S.A. 91:1128-1132,
1994.
Contact me
off-list if you'd like further info.
Regards,
Sandy (the other Simon)
Sanford M. Simon
The Rockefeller University
1230 York Avenue
New York, NY 10065
212-327-8130
-----Original Message-----
From: Confocal Microscopy List [
[hidden email]] On Behalf Of Watkins, Simon C
Sent: Monday, 28 July 2008 11:13 PM
To: [hidden email]
Subject: bubbling media and imaging
=20
Hi folks, I in the middle of a bunch of experiments where I need to bubble
gas into media while imaging (Going from normoxic to hypoxic media). Obviously
the bubbles perturb the quality of the DIC image. I have tried all sorts of
homemade diffusers to minimize the effect but to no avail... So has anyone of
you developed a solution to this problem?
I could exchange the media, however regassing happens really quickly and the
effects we are measuring are subtle. Thus any changes in ionic concentrations or
temperature may lead to a similar effect.
Ideas anyone
simon
=20
Simon C. Watkins Ph.D, FRCPath
Professor and Vice Chair, Cell Biology and Physiology
Professor, Immunology
Director, Center for Biologic Imaging
BSTS 225, University of Pittsburgh
3500 Terrace St.
Pittsburgh PA 15261
Tel: 412-352-2277
Fax:412-648-2797
URL:
http://www.cbi.pitt.edu
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