Transit time (fwd)

mcfarland david c (mcfarlnd@ux1.cso.uiuc.edu)
Fri, 7 Mar 1997 09:45:27 -0600 (CST)

We have encountered similar problems trying to assay ion flux, especially
if it is a transient change. We set up a kinetics experiment where the
sample is run to get a baseline. We add an agent to induce ion flux,
mix, and then get the sample running again as quickly as possible. A lot
of the time we would miss the major shift in fluorescence, which is
linked to the ion flux obviously, and just observe the fluorescence
returning to baseline. Upon further observation we discovered that mixing
is a major problem. 1) It takes too long to mix the sample in the sample
tube. 2)After mixing the sample with your ion flux inducer(or
inhibitor), when the sample starts running again, it is going to mix with
the untreated sample that is in the sample tubing. We are trying to
overcome this problem using some hardware and software from Alitea USA,
Medina, WA. 800-963-1101. It is computer driven peristaltic pumping
system that can deliver small volumes precisely and at
controlled
rates. However, it isn't cheap ($3-4,000?). We haven't invested a
whole lot of time in it so far (busy,busy). Gary Durack,the facility
director, would know more but he's out until the 17th. I'm sure that
Alitea would also be helpful.

David McFarland
University of Illinois Biotechnology Center
Flow Cytometry Facility

---------- Forwarded message ----------
Date: Thu, 6 Mar 1997 12:41:33 -0600 (CST)
From: Raymond B. Hester <rhester@jaguar1.usouthal.edu>
To: cyto-inbox
Subject: Transit time

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?

Thanks for any info.

Ray

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Raymond B. Hester, Ph.D.
Flow Cytometry Lab
CSAB 357, Biotechnical Services
The University of South Alabama
Mobile, AL 36688-0002 To err is human -
Email: rhester@jaguar1.usouthal.edu but it feels divine.
Voice: (334) 460-6029 Mae West
FAX: (334) 460-6073
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

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