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Although both groups of dyes are use- 279 2 Calculating Absolute Ca Values After obtaining the Rmin, Rmax, sf, and sb, the intracellular concentration can be calculated using the following formula (4): Ca2 [(R-Rmin)(Rmax-R)] × (sfsb) × Kd in which R is the ratio value for which the concentration is desired, and Kd is the dis- sociation constant of the dye (250 nM for indo-1).
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