Evaluation of human sperm morphology using strict criteria after Diff-Quik staining: correlation of morphology with fertilization in vitro.

New, very strict criteria were used after Diff-Quik staining for evaluating sperm morphology. The results of morphology scoring were correlated with the fertilization rate in vitro. Semen samples from 64 men participating in an in-vitro fertilization programme were used for this study. All men had to have a sperm concentration of greater than or equal to 20 million/ml and a progressive motility of greater than 30%. The morphology evaluation using strict criteria was performed on the same aliquot of semen as that used for in-vitro insemination. If strict criteria showed that normal morphology was less than or equal to 4%, the fertilization rate per oocyte was 23%. If normal morphology was greater than or equal to 11%, 77% fertilization occurred. For proportions of normal morphology between 4 and 11%, the fertilization rate per oocyte was 59% (P less than 0.000001). Among all these morphology groups, classical semen parameters, such as the mean volume, the mean concentration and the mean motility, did not differ significantly, except for the morphology evaluation using WHO criteria. The correlation with fertilization was better for morphology evaluation using strict criteria than for WHO. In conclusion, the method of evaluating sperm morphology based on very strict criteria allows a more accurate prediction of the chance of fertilization in vitro. Further studies should be done to establish the most appropriate cut-off points for severely impaired, intermediate and high fertilization rates.