Takifugu bimaculatus aquaculture industry has been largely developed in Fujian Province, however, its benign development was restricted by some of its reproductive characters like the unsynchronization of maturity of sperm and egg, and low fertility rate under nature mating conditions. Sperm cryopreservation method provides an effective solution for this restriction because it can preserve germplasm directly. In this study, the optimal parameters for sperm cryopreservation of T. bimaculatus were tested. The most suitable treatment for T. Bimaculatus sperm is to dilute the sperm by MPRS solution with 5 % DMSO at a ratio of 1:1, freeze at 4 ℃ for 30 min, fumigate at 10 cm above the surface of liquid nitrogen for 5 min and then at 5 cm above the surface of liquid nitrogen for 5 min, and finally preserve in liquid nitrogen. By this procedure, the motility of frozen-thawed sperm could reach (83±3) % and the hatching rate after artificial insemination was about 70 %, which can meet the requirement of lager scale breeding of T. bimaculatus. And our result could also be a good reference for relative researches in other fugu fishes.