AIM:To determine the change of intracellular free Ca2+ in trabecular meshwork cells (TMC) under the influence of genistein(Gen) through laser scanning confocal microscopy(LSCM).METHODS: TMC were obtained through the method of tissue cultured. Loaded with Fluo-3/AM, the value of [Ca2+]i was obtained by dynamically scanning changes of intracellular fluorescent intensity after application of genistein with different density.RESULTS:The application of genistein (10-6,10-5,10-4mol/L, final concentration) led to an decrease in [Ca2+]i and especially 10-5mol/L genistein led to a significant decrease in [Ca2+]i: from 328.62±16.77 to 309.69±16.57 (n=8,P<0.05). CONCLUSION:Genistein with a certain density may decrease the resistance to the outflow of aqueous by relaxing TMC and lead to an decrease of IOP.