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Issue 8, 2002
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Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy

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Abstract

Low power laser irradiation is regarded to have a significant role in triggering cellular proliferation and in treating diseases of diverse etiologies. The present work contributes to the understanding of the mechanisms of action by studying low power laser effects in human fibroblasts. Confocal laser scanning microscopy is used for irradiation and observation of the same area of interest allowing the imaging of laser effects at the single cell level and in real time. Coverslip cultures were placed in a small incubation chamber for in vivo microscopic observation. Laser stimulation of the cells was performed using the 647 nm line of the confocal laser through the objective lens of the microscope. Mitochondrial membrane potential (ΔΨm), intracellular pH, calcium alterations and generation of reactive oxygen species (ROS) were monitored using specific fluorescent vital probes. The induced effects were quantified using digital image processing techniques. After laser irradiation, a gradual alkalinization of the cytosolic pH and an increase in mitochondrial membrane potential were observed. Recurrent spikes of intracellular calcium concentration were also triggered by laser. Reactive oxygen species were generated as a result of biostimulation. No such effects were monitored in microscopic fields other than the irradiated ones.

Graphical abstract: Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy

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Publication details

The article was received on 08 Nov 2001, accepted on 17 Jun 2002 and first published on 03 Jul 2002


Article type: Paper
DOI: 10.1039/B110213N
Citation: Photochem. Photobiol. Sci., 2002,1, 547-552
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    Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy

    E. Alexandratou, D. Yova, P. Handris, D. Kletsas and S. Loukas, Photochem. Photobiol. Sci., 2002, 1, 547
    DOI: 10.1039/B110213N

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