ca++ flux calibration

KUKURUGA@medmail.med.umich.edu
Tue, 24 Sep 1996 20:02:20 -0500 (EST)

We're attempting quantification fo Ca++ using indo1, and also using
MultiTime program. There are questions . . .
Some of the terms for the calibration equation are somewhat confusing:\
I think we have an approach for determining Rmax and Rmin, but
the R term . . . according to the manual, R is the fluor. intensity
ratio for the "experimental" Ca++ -- but, is that not a continuum as a
function of time? If this is true, R varies (unless R is in fact something
else)
Also . . . the correction term "Ssub-f2/Ssub-b2" . . . is the "ratio of
the blue or green fluorescence intensity of the calcium-free and bound
dye, respectively." How is this different from R?

I'm taking this directly from the MultiTime V3.0 manual . . . can't make
sense of it. Plus, I've read several papers describing the process --
all use a variation of the same calibration function. What am I missing?

Any help (explanations, or references which describe the logic behind
these terms, whatever . . . anyone have a web page with this technique
described?) will be greatly appreciated.

MAK.\

Mark A. KuKuruga
University of Michigan Flow Cytometry
kukuruga@medmail.med.umich.edu

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