Abstract: The survival situation and rate of sperms of pearl oyster Pinctada martensii was detected by being stained with eosin Y stain and typan blue respectively, the suitable ways were selected by comparing the effect of different sampling method, staining duration and dye concentration. The results showed that (1) there were significant differences of staining characters between dead and live spermatozoas: the live ones were transparent, spherical and became bigger, while the dead ones were stained easily by either dye; (2) the optimum staining duration of both dyes was 15min, and the optimum dye concentration was 0.04% eosin Y and 0.2 % trypan blue respectively; (3) the theoretical and real mortalities were correlated with coefficient R=0.99 (P0.01) and the rates detected by both eosin Y stain and trypan blue showed the same result; (4) the test of dye toxicity to live sperms showed that the mortality did not increase significantly with P0.05 with as staining duration prolonging to 45min. These results meant that dye solution was less toxic to sperms, and it was more credible to detect the survival rate of sperms within staining duration of 45min. All these results suggested that staining with either eosin Y or trypan blue was suitable for evaluation of sperm survival rate of Pinctada martensii and the results were sensitive and accurate.