Objective: In a general way, both X-ray fluoroscopy and ultrasound (US) are used for guiding access creation during percutaneous nephrolithotomy (PCNL). However, the use of X-ray inevitably incurs radiation exposure, not only for patients but also for doctors. Conventional US guidance, being radiation free, can serve the purpose, but it is difficult to visualize the needle tract during screening. It causes following problems: visceral injure, miss of target calix and severe hemorrhage. This study evaluated the feasibility of using US guidance assisted with laser to solve the above problems. Methods: From May 2015 to March 2016, we performed PCNL on 24 patients with US guidance assisted with laser (group 1). The procedure was done with US and laser. Using the overlap of laser beam, US plate and puncture needle, laser beam’s offset reflecting the way of needle migration, visualizing the needle pathway on ultrasound images, helped establishing puncture channel. And the clinical data of 24 patients performed with free-hand US guidance (group 2) by the same surgeon in the same period were compared with those of group 1. Results: The 22 patients in group 1, 91.7% (22/24) of cases had their punctures effectively done with a single attempt. The whole needle pathways were visualized. The other two needle pathways were immediately adjusted by deviation of laser beam. Respectively 66.7% (16/24) in group 2. The puncture time in group 1 was significantly shorter than that in group 2. Conclusion: US guidance assisted with laser can provide radiation-free guidance for renal access creation in PCNL. The technique is safe and effective.