[Cytometry] Indo-1 on an Aria?

Elizabeth R. Simons esimons at bu.edu
Fri Feb 20 21:37:59 EST 2009


Hi, Ray,
	We don't have an Aria but have done (and published) lots of rapid  
calcium flux measurements with indo-1 on a FACS 440 and a MoFlo. The  
easiest way is not to remove the sample tube but to use a vacutainer  
stopper, than add your stimulus via a syringe, piercing that stopper,  
to the flowing cells. If you don't have an available stirring set-up  
you can put a small magnetic "flea" (Fisher Scientific) in the tube  
and hold one of the small magnetic stirring units next to the tube.  
We use a home-built set-up encompassing one of those, a spiral of  
copper tubing (through which we circulate water from a 37o water  
bath) which holds the sample tube outfitted with the above-described  
vacutainer stopper. The first readings depend on the diameter and  
length of the tube to the tip - we can do 4-7 sec. depending on flow  
rate.
	I hope this helps.
	Elizabeth R. Simons

On Feb 19, 2009, at 10:58 AM, Ray Hester wrote:

> Hi,
>
> Has anyone demonstrated calcium flux on a FACS Aria or Aria II  
> (_not_ a
> FACSVantage) where at some point after equilibration of Indo1-loaded
> cells, the sample tube was removed from the sample uptake tube, an
> agonist (or some compund) added to the tube, the sample tube replaced
> and the experiment continued?
>
> If so, or if you know where this type of experiment on an Aria has  
> been
> published, can you point us to references (journal articles) detailing
> the experiment/s?
>
> Thanks.
>
> Ray Hester
> Research Cytometry Laboratory
> College of Medicine
> The University of South Alabama
> Mobile, AL 36688
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Elizabeth R. Simons, Ph.D.
Professor of Biochemistry
Boston University School of Medicine
80 E. Concord St.
Boston, MA 02118
(617) 638-4332 phone
(617) 638-5339 FAX
esimons at bu.edu
ersimons at earthlink.net



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