Low-power laser effects at the single-cell level: a confocal microscopy study

Eleni Alexandratou; Dido M. Yova; Vassilis Atlamazoglou; Panagiotis Handris; Dimitris Kletsas; Spyros Loukas

doi:10.1117/12.405919

3 November 2000 Low-power laser effects at the single-cell level: a confocal microscopy study

Eleni Alexandratou, Dido M. Yova, Vassilis Atlamazoglou, Panagiotis Handris, Dimitris Kletsas, Spyros Loukas

Proceedings Volume 4159, Effects of Low-Power Light on Biological Systems V; (2000) https://doi.org/10.1117/12.405919
Event: EOS/SPIE European Biomedical Optics Week, 2000, Amsterdam, Netherlands

Abstract

Confocal microscopy was used for irradiation and observation of the same area of interest, allowing the imaging of low power laser effects in subcellular components and functions, at the single cell level. Coverslips cultures of human fetal foreskin fibroblasts (HFFF2) were placed in a small incubation chamber for in vivo microscopic observation. Cells were stimulated by the 647 nm line of the Argon- Krypton laser of the confocal microscope (0.1 mW/cm²). Membrane permeability, mitochondrial membrane potential ((delta) Psi_m), intracellular pH_i, calcium alterations and nuclear chromatin accessibility were monitored, at different times after irradiation, using specific fluorescent vital probes. Images were stored to the computer and quantitative evaluation was performed using image- processing software. After irradiation, influx and efflux of the appropriate dyes monitored changes in cell membrane permeability. Laser irradiation caused alkalizatoin of the cytosolic pH_i and increase of the mitochondrial membrane potential ((delta) Psi_m). Temporary global Ca²⁺ responses were also observed. No such effects were noted in microscopic fields other than the irradiated ones. No toxic effects were observed, during time course of the experiment.

Citation Download Citation

Eleni Alexandratou, Dido M. Yova, Vassilis Atlamazoglou, Panagiotis Handris, Dimitris Kletsas, and Spyros Loukas "Low-power laser effects at the single-cell level: a confocal microscopy study", Proc. SPIE 4159, Effects of Low-Power Light on Biological Systems V, (3 November 2000); https://doi.org/10.1117/12.405919

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