Calculation of [Ca2+] with excitation at single wavelength

Reference: Grynkiewicz et al, 1985

Fura-2 Excitation Spectra



F: Fluorescence Intensity
Fmax: Maximum Fluorescence when all dyes are bound.
Fmin: Minimum Fluorescence when all dyes are free.

Sf: Proportionality Coefficient (Free)
Sb: Proportionality Coefficient (Ca2+-bound)

Cf: Free Dye Concentration
Cb: Ca2+-bound Dye Concentration

Kd: Dissociation Constant


At wavelength 380 nm,