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Ray Hestor writes:
>Dear Group,
>An investigator wants to look (using flow) at Ca++ responses that occur
>within seconds (as in 1 or 2 seconds) after the addition of a particular
>agent to dissociated endothelial/epithelial cells. Can anyone tell us
>the fastest transit time from sample uptake to laser intercept
>with either a Coulter Elite or FACSVantage? I also seem to remember a
>commercially available device that could be attached to (any?) flow
>cytometer and that increased the speed of sample injection. Does anyone
>know the particulars about this device - manufacturer, cost, and how well
>it works? Can it can be attached to both the Coulter Elite and the
>FACS Vantage?
There are two articles: Cytometry 10:796(1989) & Cytometry 12:464(1991)
which describe some methods for reducing the transit time and at the same
time providing mixing and "real-time" kinetics. PLEASE note the quotes,
we don't need to have a discussion of real-time by flow. This was done at the
expense of my acquarium pump initially and considerably less than one
thousand dollars eventually. (see texts for details). You might still be
able to get your hands on one of these chambers through the instrumentation
shop at Yale University (Nigel Cox, 203-785-4677). Oh, yeah, I forgot to
mention above that the chambers also provide temperature control so tight that
when measuring the temperature inside the vial used to hold the sample with
a sensitive recording device you could see the cycling of the thermal unit.
The success of these chambers is directly proportional to the tinkering ability
of the user but it does show that one can do some reasonable kinetics with
what you have laying around. Especially if it is going to be a one-shot deal
or an infrequent investigation. I would be happy to address questions if you
can reach me at 203-812-5303.
Keith
**********************************************************************
*
Keith A. Kelley kelley@wh.bayer.com *
Institute for Research Technologies * "No tree is too big
Bayer Corporation * for a short dog to
400 Morgan Lane Ph 203-931-5303 * lift his leg on"
West Haven CT 06516 FAX 203-937-5467 *
* Texas Bix Bender
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web