Determination of the rate constants of the fluorescent pH probe C.SNAFL-1 in the excited state

Jan Jaap ter Horst; Marcel Ameloot; Jacques Gallay; Michel Vincent; Frans C. De Schryver; Pol Steels

doi:10.1117/12.198738

4 January 1995 Determination of the rate constants of the fluorescent pH probe C.SNAFL-1 in the excited state

Jan Jaap ter Horst, Marcel Ameloot, Jacques Gallay, Michel Vincent, Frans C. De Schryver, Pol Steels

Proceedings Volume 2324, Optical Biopsy and Fluorescence Spectroscopy and Imaging; (1995) https://doi.org/10.1117/12.198738
Event: International Symposium on Biomedical Optics Europe '94, 1994, Lille, France

Abstract

A proper interpretation of the signals from fluorescent indicators in imaging technology implies a knowledge of the processes in the excited state. This work focuses on the pH probe C.SNAFL-1. The kinetics of the excited-state processes are investigated by global compartmental analyses of fluorescence decay surfaces obtained by time-correlated single photon counting. Within the pH range 5-12 only two species have to be considered in the ground and the excited state. The two fluorescent decay times do not depend on pH. Therefore, the process of protonation in the excited state is very slow as compared to the deactivation rate of the excited states. A proper identifiability study has been performed to determine the rate constant of the deprotonation process. The rate of deprotonation is also small and its upper value is estimated to be 0.05 ns^-1. It can be concluded that there is negligible interference of the excited-state reaction on the determination of intracellular pH by C.SNAFL-1.

Citation Download Citation

Jan Jaap ter Horst, Marcel Ameloot, Jacques Gallay, Michel Vincent, Frans C. De Schryver, and Pol Steels "Determination of the rate constants of the fluorescent pH probe C.SNAFL-1 in the excited state", Proc. SPIE 2324, Optical Biopsy and Fluorescence Spectroscopy and Imaging, (4 January 1995); https://doi.org/10.1117/12.198738

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