Intra Cellular Fluorescence for Monitoring The Physiological Changes and Biostimulation of Yeast Irradiated with Low Power 532nm CW Nd:YAG Laser

Abstract

The purpose of this study is to investigate the biostimulation effect of 532 nm CW laser on the metabolism of Saccharomyces cerevisiae yeast. Cells were irradiated by 532 nm Nd:YAG laser using 0.153 W/cm2 power density at 30, 45, 60,180 and 300 seconds exposure times in their respective orders. Intrafluorescence parameters were measured by detection the autofluorescence intensity, proliferation rate and Imaging the fluorescent mitochondria using confocal laser scanning microscope. The results showed that the 30 and 45 second exposure times seem to have stimulated changes in the cells that led to increase proliferation, viability and mitochondrial activity. Autofluorescence of cells increased after 45 and 60 seconds exposure time. After 300 seconds there seems to be very noticeable decrease in proliferation, viability and autofluorescence. Confocal microscopy images showed that here is a correlation between fluorescence intensity using mitochondrial probes and proliferation rates of cells.
Introduction
Saccharomyces cerevisiae is one of the most important fungi in the history of the world. This yeast is responsible for the production of alcoholic beverages and bread and a source of protein and was used in biotechnology and genetics as a host for the genes of other organisms (Madigan, et al. 2006). Low energy laser irradiation of which output power is in the range of mW modulates various biological effects and has been shown to have positive effect on living organisms both in vitro and in vivo. However, the true effect of low energy laser on cell proliferation is sill controversial, because of conflicting reports on the effects of visible laser light on the cells in culture (Antonio, et al., 2002).
There are many evidences that the most intracellular autofluorescence