Testicular biopsy has been traditionally used as a diagnostic tool in the management of infertility. With the advent of assisted reproductive technology, testicular biopsy is now used therapeutically to retrieve sperm for intracytoplasmic sperm injection (ICSI). The feasibility of testicular sperm extraction for ICSI is reviewed in this article. Although less invasive techniques such as testicular fine needle aspiration and percutaneous needle biopsy are efficacious, particularly in cases of obstructive azoospermia, studies in the current literature support that open testicular biopsy is a more reliable method to obtain testicular specimen. Different measures to reduce the complication rate of open testicular biopsy, including the application of microsurgical techniques, are addressed here. Other areas of recent development related to testicular biopsy that are reviewed include processing of testicular tissue, cryopreservation of retrieved spermatozoa, in vitro maturation of spermatozoa, and microinjection of immature spermatogenic precursor cells.