Purpose To evaluate the distances using ultrasound between the supericial peroneal nerve (SPN) and sural nerve along the peroneus longus tendon (PLT) autograft harvest path at diferent ankle or knee positions in order to minimize risk of iatrogenic nerve injury during PLT autograft harvest. Methods Twenty-four fresh-frozen human cadaveric lower extremities were used to harvest a full-thickness PLT autograft with a tendon stripper. Four specimens were utilized to validate correct identiication of nerves under ultrasound. Sonographically guided perineural injections were performed at the start point and end point of the PLT harvest path using coloured latex, followed by dissection with gross inspection. Using ultrasound, the distance from the peroneus brevis muscle to the sural nerve at diferent ankle positions (20° dorsilexion, neutral, and 20° plantarlexion) was measured, and the distance from the end of the tendon stripper to the SPN at diferent knee positions (full extension and 90° lexion) was also measured. Measurements were performed by two separate observers using ImageJ software. Results Cadaveric dissection showed the presence of latex around nerves in all four specimens. The average distance from the brevis muscle to the sural nerve increased signiicantly from dorsilexion to plantarlexion. The shortest distance from the tenodesis site to the sural nerve was 5.8 ± 1.7 mm. There was no signiicant diference from the end of the tendon stripper to the SPN between full extension or 90° lexion of the knee. Conclusion When harvesting the PLT, it is recommended to place the ankle at plantarlexion. The knee at full extension or 90° lexion had no efect. Joint positions at the time of graft harvest should be monitored to reduce risks of iatrogenic nerve injury.