Follicular fluid (FF) is a dynamic medium rich in steroids, polypeptide hormones, and growth factors. Preovulatory FF can stimulate spermatozoal acrosome reaction. Moreover short preincubation of washed sperm with FF improves sperm performance in the hamster egg penetration assay. In the current study, FF was used to capacitate sperm and as a transfer medium in 131 gamete intrafallopian transfer (GIFT) procedures. Ham's F-10 medium (GIBCO, Grand Island, NY) with 50% maternal serum was used in another 29 GIFT procedures. In the Ham's F-10 group, 29 GIFT procedures were performed in 25 patients with a pregnancy rate of 21% per laparoscopy and 24% per patient. In the FF group, 131 GIFT procedures were performed in 100 patients with a pregnancy rate of 50% per laparoscopy and 66% per patient, with a 59% ongoing pregnancy rate per patient. The most significant improvement was in the male factor group with a pregnancy rate of 0% in the Ham's F-10 group and 44% per procedure in the FF group. We conclude that the use of FF for sperm capacitation and as a gamete transfer medium significantly improves GIFT pregnancy rates and outcome.