Objective To investigate the effects of laser irradiation on intracellular ROS(reactive oxidant species), intracellular calcium concentration(［Ca2+］i，and cell membrane integrity in the process of live cell imaging with confocal laser scanning microscopy. Methods The effects of a given laser irradiation on ROS, intracellular calcium concentration (［Ca2+］i and cell viability were revealed respectively by stained ECV304 with H2DCFDA，Fluo4AM and calceinAM/PI, and visualized and analyzed using ultra view LCI(live cell image)confocal microscopy. Results The irradiation of 488nm laser induced fluorescent intensity of DCF to increase abruptly and attain the climax in about 80 seconds, afterwards the fluorescent intensity fell and returned to the baseline. In the 70 minutes of the irradiation, the fluorescent intensity of intracellular Fluo4 kept a slightly ascending tendency. The fluorescent intensity of calcein decreased 15minutes after the irradiation, and serval cells were PI positively stained.Conclusion 488nm laser irradiation