Background: Ureteral injury is one of the most common anatomical complications during total laparoscopic hysterectomy (TLH). And most ureteral injuries resulted from electrosurgery and were recognized postoperatively. To avoid thermal damage to the ureter during TLH, we evaluated the feasibility and safety of vessel ligation using the Hem-o-lok clip. Methods: A retrospective analysis of 480 patients who underwent TLH. Hem-o-lok clips were used to ligate both uterine vessels. If the patient underwent unilateral or bilateral salpingo-oophorectomy, the gonadal vessels were also ligated by using Hem-o-lok clips. Clinical outcomes including perioperative and postoperative complications were evaluated. Results: Perioperative complications were reported in six patients, with four cases of stump dehiscence and two cases of bladder injury. All bladder injuries were found during the operation and repaired. None of the patients experienced ureter injury or vessel rebleeding. Conclusions: Vessel ligation by Hem-o-lok clips in TLH is a safe method for ligating vessels and avoiding unexpected ureteral injury. Prospective studies are needed to confirm the real clinical benefit of this surgical approach.