Objective: We aimed to explore the feasibility and safety of S²b+S³a combined subsegmentectomy for pulmonary nodules between S² and S³ in the right upper lung. Methods: We retrospectively analyzed the clinical data of segmentectomy in our center from 2015 to 2020. Under the preoperative three-dimensional computed tomography bronchography and angiography (3D-CTBA) planning and the real-time navigation of 3D-CTBA intraoperatively, the nodule between S² and S³ was identified using intersegmental vein V²c, and the lateral subsegment of S² (S²b) and the lateral subsegment of S³ (S³a) were combined as a new resection unit. We further analyzed the nodule characteristics and perioperative data of S²b+S³a resection. Results: A total of 46 cases of S²b+S³a resection were included. The size of nodules in the CT image was 11.57±3.65 mm. The pathological diameter was 9.48±3.06 mm. According to the consolidation to tumor ratio (CTR), the nodules were classified as: pure-GGO (Ground Glass Opacity, GGO) nodules (30 cases), partial-GGO nodules (11 cases), partial-solid nodules (3 cases), and pure-solid nodules (3 cases). There were 35 nodules located in the outside 1/3 of lung parenchyma and 11 nodules located in the middle 1/3 of lung parenchyma. The pathological diagnosis included: adenocarcinoma in situ (AIS, 8 cases), minimally invasive adenocarcinoma (MIA, 19 cases), invasive adenocarcinoma (IAC, 15 cases), benign (3 cases), and metastatic tumor (1 case). The number of lymph node sampling stations was 2.57±1.42 and resected lymph nodes was 4.50±3.06. The mean surgical margin was 18.21± 3.24 mm. The median postoperative hospital stay was 4 days, and the median chest tube drainage time was 2 days. Long-term air leakage (≥3 days) and postoperative hemoptysis occurred on 2 patients respectively. No perioperative death, postoperative recurrence, or postoperative death happened at the follow-up endpoint. Conclusion: S²b+S³a combined subsegmentectomy is a safe and feasible method to manage pulmonary nodules such as early non-small cell lung cancer (NSCLC) between S² and S³ under the navigation of 3D-CTBA.