Rational use of fluorescent probes for the evaluation of intracellular ionic concentrations

Pierre M. Viallet; Jean Vigo; Bruno Morelle; Jean-Marie Salmon

doi:10.1117/12.198724

4 January 1995 Rational use of fluorescent probes for the evaluation of intracellular ionic concentrations

Pierre M. Viallet, Jean Vigo, Bruno Morelle, Jean-Marie Salmon

Proceedings Volume 2324, Optical Biopsy and Fluorescence Spectroscopy and Imaging; (1995) https://doi.org/10.1117/12.198724
Event: International Symposium on Biomedical Optics Europe '94, 1994, Lille, France

Abstract

The importance of the intracellular concentration and transmembrane flux of cations has led scientists to develop various techniques for the evaluation of intracellular concentrations of H⁺, Ca²⁺, Mg²⁺. Quantitative microfluorometry appears to be a powerful and noninvasive technique for it allows kinetic studies or mapping of ionic concentration in different cellular compartments as long as fluorescent markers specific of given cations are disposed of. Nevertheless, the previously published results sometimes look controversial. To minimize problems encountered by some scientists the following protocol is proposed: (1) careful and exhaustive determination of the physicochemical properties of the probe(s), (2) use of microspectrofluorometric techniques to resolve the complex intracellular fluorescence spectra into its components and calculate the molar fraction of each compound participating in the cellular fluorescence, (3) use of the same kind of algorithm to perform numerical image analysis.

Citation Download Citation

Pierre M. Viallet, Jean Vigo, Bruno Morelle, and Jean-Marie Salmon "Rational use of fluorescent probes for the evaluation of intracellular ionic concentrations", Proc. SPIE 2324, Optical Biopsy and Fluorescence Spectroscopy and Imaging, (4 January 1995); https://doi.org/10.1117/12.198724

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