Abstract: Uses giemsa's dye and microscope observe bursa copulatrix sperm of silkworm directly, artificial insemination using the sperm, the studies shows: The seminal fluid is similar in the male moth seminal vesicle and in the female bursa copulatrix after mating 20 to 30 minute, and the seminal fluid is contains large number of eupyrene sperm bundles and not be activated. The result of microscope observation and artificial insemination using sperm treated with diffirent trypsin concentration shows that 0.2μg/ml trypsin in 50% semen solution, the fertilization rate is highest. The sperm treated with trypsin, plasmin, pepsin and succinic acid sodium respectively, only trypsin can dissociate the eupyrene sperm bundles and improve the motility of apyrene sperm synchronously , and olso only the trypsin-treated sperm have the ability of fertilization. The optimum concentration of sperm for artificial insemination is 2-fold diluted still be found in the research.