Microsurgical spermatozoid extraction is a highly effective method for the acquisition of the spermatozoids in case of non-obstructive azoospermia of different etiology. Intraoperative microscopy allows the visualization of the extended seminal channels, which could contain spermatozoids, and their microdissection. The advantage of this technology is reduced invasiveness and high efficacy in the acquisition of the spermatozoids in case of secretory azoospermia. The aim of the study was the estimation of the microsurgical extraction efficacy in patients with non-obstructive azoospermia. Thirty patients were included with microsurgical testis biopsy with parallel stimulation of the hyperovulation and oocyte extraction in female counterparts in frame of the extracorporeal fertilization program. According to the results of the biopsy spermatozoids were successfully extracted in 12 patients (40%), which were feasible for the ICSI. The spermatozoids extracted were all used for the fertilization, in 8 patients cryoconservation of the remaining spermatozoids was carried out. Pathomorphologically testis tissue contained sclerotic changed seminal channels with signs of atrophy or hypotrophy of spermatogenic epithelia. Spermatogenesis was blocked at different levels. High efficacy of the microsurgical extraction in patients with non-obstructive azoospermia in absence of complications could be accounted for as the perspective method in the treatment of the male infertility.