Design of ratiometric emission probe with visible light excitation for determination of Ca2+ in living cells.

An organic salt as a fluorescent probe based on intramolecular charge transfer for Ca(2+) determination is developed. Ca(2+) can be detected by ratiometric emission at 490 and 594 nm with an excitation wavelength of 405 nm. This probe is highly selective for Ca(2+) over other divalent metal cations and displays a large Stokes shift of 189 nm that can avoid interference of the excitation light beam and autofluorescence of biological samples. The dissociation constant for Ca(2+) is 2.25 ± 0.47 μM and pertinent to Ca(2+) detection in cellular resting and dynamic states. The probe demonstrates its application in monitoring Ca(2+) in living cells under confocal microscopic imaging.