The frequency of in vitro fertilization of mouse eggs using sperm obtained weekly (for 1-9 weeks) from epididymis after testicular X-irradiation (200 rad) was observed. The sperm concentrations for insemination were 100, 200 and 300/mm3. The number of fertilized eggs seemed to remain constant almost at control level (90-98%) until the 4th week after X-irradiation. Thereafter, the number declined to reach a minimum level (about 30-50%) in the 6th week; they then recovered completely in the 8th week. This tendency was found at all sperm concentrations used. This result indicates that the most sensitive stage for the fertilization capacity of sperm to X-rays was the early spermatocytes or the late spermatogonia. The high radiosensitivity of the fertilization capacity of sperm irradiated during the early-spermatocyte to late-spermatogonial stage, corresponded well with the rate of induction of sperm abnormalities and spermatogenetic cell killing. The in vitro fertilization rate was not restored by an increase in sperm concentration at insemination; it is, therefore, likely that the in vitro fertilization frequency depends on the proportion of sperm with abnormal morphology and abnormal fertilization ability, and sperm number seems to have hardly any influence on the fertilization rate, as far as the in vitro fertilization experiment is concerned.