Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes.

The influence of femtosecond near-infrared (NIR) microirradiation on cell vitality and cellular reproduction has been studied. Chinese hamster ovary cells exposed to a highly focused 150-fs scanning beam at 730, 760, and 800 nm (80 MHz, 80-mus pixel dwell time) of </=1 mW remained unaffected by the femtosecond microbeam. However, increased mean power led to impaired cell division. At >/=6 -mW mean power, cells were unable to form clones. They died or became giant cells. Complete cell destruction, including cell fragmentation, occurred at mean powers >10 mW. Cell death was accompanied by intense luminescence in the mitochondrial region. When we consider the diffraction-limited spot size in the submicrometer region, intensities and photon flux densities of 0.8-kW pulses (10-mW mean power) are of the order of terawatts per square centimeter (10(12)W/cm (2)) and 10(32) photons cm(-2) s(-1) , respectively. Extremely high fields may induce destructive intracellular plasma formation. The power limitations should be considered during NIR femtosecond microscopy of vital cells and in the design of compact NIR femtosecond solid-state lasers for two-photon microscopes.