Dear all,
The combination of Fura Red and Fluo-3 seems to work better than Calcium
Green/Fura Red, at least in a fluorometer. Fura Red is only weakly
fluorescent (maximum QY near 0.01 at zero Ca2+ (10 mM EGTA)). When we were
running the spectra for the M.Probes catalog, it was hard to get Calcium
Green and Fura Red on the same scale, so we switched to fluo-3 (maximum QY
near 0.1). The first reports of this combination in cells was in an
abstract by Xynaxis Cell Science - they found that when using Fluo-3
(loading at 1 uM) and Fura Red (loading at 5 uM) the resulting pseudo-ratio
(488/650 approx.) was useful for real time Ca2+ imaging.
Best regards,
Michael Kuhn, President
Helix Research Co.
www.helixresearch.com
----- Original Message -----
From: Flow Cytometry Facility <system@flocyt.int-med.uiowa.edu>
To: cyto-inbox
Sent: Wednesday, September 06, 2000 6:45 AM
Subject: Re: visible Ca dyes
>
> Greetings,
>
> We used a combination of calcium green and fura red for calcium flux
> studies with a FACScan on Jurkat and human PBL with good success. We used
> FlowJo to produce the ratio of the two fluorochromes. The changes between
> resting state and maximal flux were very similar to indo-1.
>
>
>
>
>
> At 04:01 PM 09/01/2000 -0400, you wrote:
> >
> > A number of years ago we used Fluro-3 red and Snarf-1 in order to use a
> >488 laser to measure Ca. Since then I know (or think I know) that I
have
> >seen other dyes used in this manner.
> > I would like to hear from others who have used other visible dyes for
> >measuring Ca.
> > Thanks in advance for any and all input.
> > Joan K
> >
> >
> > Flow Cytometry Lab Supervisor
> > VMRCVM
> > (540) 231-4115
> > FAX 540-231-7367
> > jkalnits@vt.edu
> >
> > "It is better to serve than to receive."
> > B. Borg
> >
> >
> ------------------------------------------------
> | Justin Fishbaugh |
> | University of Iowa |
> | Flow Cytometry Facility |
> | 48 EMRB |
> | Iowa City, IA 52242 |
> | |
> | justin-fishbaugh@uiowa.edu |
> | http://www.medicine.uiowa.edu/flowcytometry/ |
> ------------------------------------------------
>
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