Objective: Evaluate the IVF ( in vitro fertilization) rate of the post-thawed oocytes and the post-thawed punched oocytes of C57BL/6J fertilized with fresh or frozen sperm of 9 different strains. Methods: (1) The changes of nuclear, microfilament and zona pellucida glycoprotein-2 of C57BL/6J oocytes before and after cryopreservation were studied By immunofluorescence technique. (2)The IVF rate of C57BL/6J post-thawed oocytes or laser-punched oocytes fertilized with fresh or frozen sperms of 9 strains were compared.Results：(1) there were no obvious changes of nuclear before or after cryopreservation, microfilament were changed slightly when handled with cryopretectant or after cryopreservation. Zona pellucida glycoprotein-2 were hurted after cryopreservation or handled with cryopreservant ; (2) the IVF rate of fresh sperm of 9 strains with post-thawed oocytes were 17.6～42.9%(averaged 29.6%), which were significantly lower than the IVF rate of fresh sperm with laser-punched oocytes, 29.1～72.3% (average 49.7%); the IVF rate of post-thawed sperms of 9 strains with post-thawed oocytes were 5.4～23% (averaged 15.5%), also significantly lower than the IVF rate of post-thawed sperms fertilized with laser-punched oocytes, 16.7～48.6% (average 28.8%). Conclusions: (1) Cryopreservation would have a remarkable effect on microfilament and zona pullucida glycoprotein-2, but have not obvious effect on nuclear; (2) The IVF rate of oocytes would decreased remarkably after cryopreservation, but if the laser-punched system were used, the IVF rate would increased apparently. It showed that the laser-punched system would be useful for oocytes cryopreservation.