Fluorescence imaging and dynamics of intracellular ionic concentrations in single living cells: application to pHi and Mgi variations

Pierre M. Viallet; Mohamed Yassine; Jean-Marie Salmon; Jean Vigo

doi:10.1117/12.239500

10 May 1996 Fluorescence imaging and dynamics of intracellular ionic concentrations in single living cells: application to pHi and Mgi variations

Pierre M. Viallet, Mohamed Yassine, Jean-Marie Salmon, Jean Vigo

Author Affiliations +

Proceedings Volume 2678, Optical Diagnostics of Living Cells and Biofluids; (1996) https://doi.org/10.1117/12.239500
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

The intracellular concentration of ions such as H+, Mg2+, Ca2+ is known to monitor the activity of many intracellular enzymes. Furthermore these ions are considered as intracellular messengers involved in signal transducing. Moreover recent technological progresses gave rise to the feeling that accurate data are instantly accessible on microvolumes. So the determination of ionic intracellular concentrations has been achieved using fluorescent specific probes and different equipments (Microspectrofluorometer, Flow Cytometer, Numerical Image Analyzer with or without Confocal system), without taking care of the physico-chemical properties of the probe. Unfortunately fluorescent probes are supposed to fill up conflicting requirements in terms of ionic affinity, specificity, fluorescence quantum yield of the free and ion-bound probe, absence of fading and diffusibility out of the cell. Because most of the probes are not so specific than it is claimed, unexpected interactions may obscure the interpretation of results and even make it difficult to get an intracellular calibration curve. Such a situation generally precludes the use of the popular simplest methods of data acquisition and treatment. The scope of this presentation is to point out some underestimated difficulties, to discuss different ways for bypassing some of them and to rationale the use of Videomicrofluorometry.

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Pierre M. Viallet, Mohamed Yassine, Jean-Marie Salmon, and Jean Vigo "Fluorescence imaging and dynamics of intracellular ionic concentrations in single living cells: application to pHi and Mgi variations", Proc. SPIE 2678, Optical Diagnostics of Living Cells and Biofluids, (10 May 1996); https://doi.org/10.1117/12.239500

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KEYWORDS

Luminescence

Ions

Calibration

Data processing

Image analysis

Magnesium

Calcium

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