Live cell microscopy, or live cell imaging, is a popular method in the field of cell biology. A great emphasis in live cell imaging is placed on reducing the photo-toxic effects of light ( bleaching).
At high light power densities, intense bleaching of fluorophores occurs, leading to the formation of free radicals and other reaction products that destroy molecules in the environment and alter the environment inside and outside the cells. The illumination power density at the sample plane, which is compatible with the lifetime of the cells being analysed, ranges from mW/cm2 for Widefield fluorescence microscopes to W/cm2 for SDCM (spinning disc confocal microscopy). Confocal microscopes (SDCMs) provide high scanning speeds due to the spinning disk with pinhole confocal set and thus low bleaching of fluorophores, due to illumination of the entire field of view at once and the use of cameras to detect the emission signal.