Objective: To assess the sperm yield and patient acceptability of Trucut needle testicular biopsy followed by seminiferous tubule milking.Design: Prospective case analysis.Setting: The Regional Fertility Center, Royal Maternity Hospital, Belfast, Northern Ireland, United Kingdom.Patient(s): Forty-one males with obstructive azoospermia (normal testicular volume and FSH and LH levels).Intervention(s): Trucut needle testicular biopsies under local anesthetic with milking of the seminiferous tubules.Main Outcome Measure(s): Quantitation of sperm retrieved per biopsy core and patient follow-up by questionnaire.Result(s): A mean of 105,634 sperm (range, 5,000-427,800) were retrieved, and the mean biopsy weight was 9.17 mg. Twenty-six subjects found the biopsy painless and 15 were pain-free after biopsy.Conclusion(s): The Trucut needle can be used in combination with seminiferous tubule milking to obtain large numbers of sperm in men with obstructive azoospermia.