BackgroundIatrogenic ureteric injury remains a risk in laparoscopic pelvic procedures. Near‐infrared fluorescence (NIRF) imaging is a promising new technique for enhanced intraoperative visualization of anatomical structures that could improve the safety of laparoscopic surgery. A new dye, IRDye® 800‐BK, has been developed for intraoperative visualization of the ureters using NIRF. The present study was a first evaluation of the performance of IRDye® 800‐BK for ureteric imaging during NIRF laparoscopy.MethodsThis study consisted of three parts: real‐time in vivo NIRF imaging using IRDye® 800‐BK in pigs during laparoscopic surgery, ex vivo NIRF imaging of freshly explanted pig ureters and ex vivo NIRF imaging of explanted human ureters.ResultsIn all animals, both left and right ureters were visualized throughout the laparoscopic procedure for 120 min, with the best results at a dose of 0·15 mg dye per kg bodyweight. NIRF imaging was successful in all human and porcine ureters studied, with a range of dye concentrations.ConclusionNIRF imaging of the ureters using IRDye® 800‐BK was used successfully both in vivo in a porcine model, and ex vivo in porcine and human ureters.