To report our initial experience with laparoendoscopic single-site (LESS) surgery in 100 patients in urology. Between October 2007 and December 2008, we performed LESS urologic procedures in 100 patients for various indications. These included nephrectomy (N = 34; simple 14, radical 3, donor 17), nephroureterectomy (N = 2), partial nephrectomy (N = 6), pyeloplasty (N = 17), transvesical simple prostatectomy (N = 32), and others (N = 9). Data were prospectively collected in a database approved by the Institutional Review Board. All procedures were performed using a novel single-port device (r-Port) and a varying combination of standard and specialized bent/articulating laparoscopic instruments. Robotic assistance was used to perform LESS pyeloplasty (N = 2) and simple prostatectomy (N = 1). In addition to standard perioperative data, we obtained data on postdischarge analgesia requirements, time to complete convalescence, and time to return to work. In the study period, LESS procedures accounted for 15% of all laparoscopic cases by the authors for similar indications. Conversion to standard multiport laparoscopy was necessary in 3 cases, addition of a single 5-mm port was necessary in 3 cases, and conversion to open surgery was necessary in 4 cases. On death occurred following simple prostatectomy in a Jehovah's Witness due to patient refusal to accept transfusion following hemorrhage. Intra- and postoperative complications occurred in 5 and 9 cases, respectively. Mean operative time was 145, 230, 236, and 113 minutes and hospital stay was 2, 2.9, 2, and 3 days for simple nephrectomy, donor nephrectomy, pyeloplasty, and simple prostatectomy, respectively. The LESS surgery is technically feasible for a variety of ablative and reconstructive applications in urology. With proper patient selection, conversion and complications rates are low. Improvement in instrumentation and technology is likely to expand the role of LESS in minimally invasive urology.