Objective To compare the differences between the up-lobectomy by single-microport assisted micro-uni-port thoracoscopy surgery and traditional uni-portal video assisted thoracic surgery, summarize and analyze the technical points of single-microport assisted micro-uni-port thoracoscopy surgery, and explore the surgical effect and value of promotion. Methods We retrospectively analyzed the clinical data of patients who underwent radical upper lobectomy at the Thoracic Surgery Department of Xi’an International Medical Center Hospital from March 2023 to June 2024. The patients were divided into two groups according the surgical procedure: a single-micromini-assisted group (patients underwent up-lobectomy by single-microport assisted micro-uni-port thoracoscopy) and a traditional uniportal group (patients underwent traditional uniportal thoracoscopic lobectomy). Clinical outcomes were compared between the two groups. Results We finally included 62 patients. There were 30 patients with 16 males and 14 females at an average of 57.4±10.8 years in the the single-micromini-assisted group and 32 patients with 20 males and 12 females at an average age of 57.6±8.7 years in the traditional uni-port thoracoscopy group. Both groups successfully completed minimally invasive surgery.The baseline data were consistent between the two groups. The operation time was shorter in the single microport assisted group [(146.03±30.79) min vs. (171.41±36.41) min, P=0.004] than that in the traditional uni-port thoracoscopy group with a statistical difference. There was no statistical difference between the two groups in terms of intraoperative blood loss, postoperative pain score, dissected lymph node number, postoperative drainage volume, postoperative tube time, postoperative hospital stay, hospitalization cost, or the incidence of postoperative complications (P>0.05). Conclusion Single-microport assisted micro-uni-port thoracoscopy surgery can maximize the advantages of three-portal and uni-portal VATS, and effectively avoid the disadvantages of three-portal and uni-portal VATS, which can significantly shorten the operation time, without increasing the postoperative pain and complications, is a more minimally invasive, safer and more convenient surgical method.