To report our experience with single-incision laparoscopic surgery (SILS) for cholecystectomy and to perform a retrospective comparison with conventional 4-port laparoscopic cholecystectomy. Data were prospectively collected for all patients undergoing SILS for cholecystectomy at St Mary's Hospital, Imperial College NHS Trust, London, England (n = 41 patients between June 13, 2008, and June 30, 2009) and compared with data for those who had undergone conventional 4-port laparoscopic cholecystectomy in the preceding year (n = 58 patients between June 26, 2007, and May 30, 2008). This included patient demographic data and intraoperative and postoperative outcomes. Four-port laparoscopic cholecystectomy and SILS for cholecystectomy. Operative time, conversion to open operation, and length of hospital stay. Operative time was longer with SILS for cholecystectomy compared with conventional laparoscopic cholecystectomy (P < .001). A correlation was seen between reducing SILS operative time and increasing experience (Spearman rank correlation coefficient, -0.29). Three patients in the SILS for cholecystectomy group required the addition of extra laparoscopic ports. No patients in the SILS for cholecystectomy group required conversion to open surgery compared with 4 patients in the standard laparoscopic cholecystectomy group. Patients stayed an average of 0.76 days following SILS for cholecystectomy and 1.53 days following conventional laparoscopic cholecystectomy. One patient in each group had a postoperative biliary leak. Single-incision laparoscopic surgery for cholecystectomy may be equal to conventional laparoscopic cholecystectomy in terms of safety and efficacy. Further randomized studies are required to investigate any significant advantages of this new and attractive technique.