Intra-cytoplasmic sperm injection (ICSI) has revolutionized the treatment of male infertility by requiring only a single sperm to allow men whose infertility was previously considered to be uncorrectable to father a biological offspring. As a result, surgical sperm retrieval for assisted reproduction has developed to support this therapy. Microsurgical techniques have been applied to either identify areas of active spermatogenesis within the testis or to aspirate sperm-containing fluid or tissue. The combination of these techniques with in vitro fertilization (IVF)/ICSI has been shown to be a powerful approach to the treatment of azoospermic men. The availability of sperm cryopreservation offers an additional advantage, negating the need for synchronization of sperm retrieval and ovulation. Thus, the advanced methods for sperm retrieval discussed in this review also provide therapeutic options, as compared the traditional diagnostic testicular biopsy.