Purpose To evaluate the effect of the size of zona pellucida opening by laser assisted hatching for frozen cleaved embryo that were thawed after both fresh and frozen cleaved embryo transfer cycles failed and were cultured to blastocyst after thawing in patients with multiple implantation failures. Materials ands methods Of 101 consecutive procedures (October 2003 to June 2006), 30 patients declined to perform assisted hatching and were selected as control group, 40 patients had 40 mu m opening of the zona (October 2003 to January 2005), 31 patients had 50% of the zona opening (February 2005 to June 2006). Results The pregnancy, implantation and delivery rates were significantly higher in 50% opening group (74%, 52% and 65%) compared to control group (17%, 10% and 13%; P < 0.01) and 40 mu m opening group (43%, 27% and 38%; P < 0.04). Conclusions The size of the zona pellucida opening may affect the outcome of frozen cleaved embryo transfer.