ABSTRACT Objectives To describe the retrieval spermatozoa technique for cryopreservation after death, including the proximal part of vas deferens. Material and Methods A 28-years old man, with previous history of infertility, who had died 12 hours before, was submitted to spermatozoa retrieval for cryopreservation, with surgical bilateral resection in bloc of the proximal part of vas deferens, testicle and epididymis. At the laboratory, by milking the epididymis and vas deferens, the extracted fluid was collected; also, three samples of each testicle parenchyma were also harvested. Results The fluid from the vas deferens showed spermatozoa, mostly with in situ motility. Testicular fragments also presented spermatozoa, mostly with small tail movements or immobile. Conclusion The inclusion of the proximal part of vas deferens during spermatozoa retrieval after death must be performed, since it contains high concentration of spermatozoa, and even in the presence of previous infertility, as was with this patient, it is possible to retrieve spermatozoa.