OBJECTIVE To develop a noninvasive procedure that employs image processing of power Doppler ultrasound (PDUS) images of several orthogonal cross-sections of the testis to construct a three-dimensional (3D) mapping of preferential testicular regions in which spermatozoa are most likely to be found in nonobstructive azoospermic testes. METHODS Clinical study. METHODS Ultrasound and andrology units in a large university-affiliated municipal hospital. METHODS Twenty-four nonobstructive azoospermic men. METHODS Before testicular sperm extraction was performed, PDUS images were acquired at seven cross-sections to reconstruct a 3D testicular vascularity index (TVI) matrix for spatial mapping of testicular regions in which spermatozoa are most likely to be found. The predictions based on TVI values of 107 regions were compared with the biopsy results. METHODS Prediction of presence or absence of spermatozoa by TVI values. RESULTS The prediction rate of the TVI matrix for the presence or absence of spermatozoa was 74.8%. The positive predicted value was 72%, negative predicted value was 75.6%, and specificity was 89.8%, but sensitivity was 47.3%. CONCLUSIONS Our technique may obviate the need for arbitrary multiple biopsies that inflict some degree of damage upon testicular tissue and may increase the success rate of identifying viable spermatozoa in testicular tissue.