[Objective] To introduce the surgical technique and primary clinical outcomes of navigation assisted Single-position lateral lumbar interbody fusion. [Methods] From October 2017 to May 2019, 27 patients underwent navigation assisted single-position lateral lumbar interbody and fusion for degenerative lumbar spondylolisthesis. The patients were placed in right lateral position by a left lumbar incision. Firstly completed the lateral approach discectomy and cage implantation. Secondly didn’t change the position, the navigation reference fixed on the left side of the illac crest. Obtained surgical segmental three-dimensions images by O-arm and transfer to navigation. The insertion point of the percutaneous pedicle screw was confirmed under navigation guidance. Opened circuit under real-time guidance of navigation and placed the guide wire, tapped along the guide wire. Percutaneous cannulated pedicle screw were inserted by navigation. [Results] All the 27 patients had operation performed smoothly without serious complications. All patients were followed up for 3-24 months. at the latest follow-up the ODI and VAS scores of low back pain and leg pain, as well as postoperative slippage rate, intervertebral space height and cross-sectional of the dural sac were significantly improved postoperatively compared with those preoperatively(P<0.05). The excellent and good rate of clinical results based on the modified MacNab criteria was 100% (27/27). 122 pedicle screws were placed in 27 patients with an average screw placement time of (9.22±3.84) min/screw. The accuracy of all pedicle screws was 93.4% according to the Gertzbein-Robbins classification standard. [Conclusion] The navigation assisted single-position lateral lumbar interbody and fusion is a safe and high-efiicient surgical method that avoids change position intraoperative.