OBJECTIVES: This phase I trial evaluated the RUS 3D virtual surgical navigation system's feasibility during robot-assisted partial nephrectomy. METHODS: In ten patients who underwent robot-assisted partial nephrectomy at Asan Medical Center, a 3D virtual anatomical model was reconstructed from preoperative computerized tomography scans. The 3D model was integrated with the surgeon's console through TilePro, providing manipulable virtual anatomy during surgery. Following standard robot-assisted partial nephrectomy procedures performed by a single experienced surgeon using the navigation system, perioperative parameters were compared with a control group of 30 patients matched for tumor and body morphometric characteristics. RESULTS: The navigation system was well integrated into the surgical workflow without technical issues, providing accurate anatomical visualization, particularly of the regional calyces and intrarenal vessel branches. The navigation group had shorter operative times (122 vs. 144.7 min) and less frequent collecting system entry (40% vs. 66.7%). For equivalent tumor size, resected specimen volume was significantly smaller in the navigation group (6.8 vs. 16.7 cm CONCLUSIONS: The integration of the 3D navigation system into robot-assisted partial nephrectomy was both feasible and safe, providing enhanced anatomical information while maintaining a consistent level of operative risk. The use of the navigation system resulted in reduced renal parenchymal volume resected, suggesting potential benefits in renal function preservation.