Background. To confirm the impact of near-infrared thoracoscopy with intravenous injection of indocyanine green (ICG) during video-assisted thoracic surgery (VATS)-based segmentectomy. Materials and Methods. We retrospectively screened the perioperative data in total 21 patients who underwent segmentectomy by VATS. The segmental arteries and bronchi were identified with the help of preoperative 3-dimensional computed tomography images. Among them, clinical effectiveness and postoperative complications were analyzed. VATS segmentectomy was performed using a 3-port approach with systemic intravenous injection of ICG. Results. A total of 21 patients underwent VATS-based segmentectomy with ICG injection. The mean operation time was 126.19 ± 15.32 minutes, and the mean bleeding volume was 158.10 ± 39.95 mL. In addition, the average drainage volume 1 day after surgery was 153.81 ± 32.19 mL, and mean duration of drainage was 1.62 ± 0.59 days. Complications occurred in 6 of the 21 patients. Two patients had pneumonia, 3 had arrhythmia, and 1 had prolonged air leak. There were no complications resulting from ICG angiography. Conclusions. Near-infrared thoracoscopy with intravenous injection of ICG is a safe, fast, simple, and highly accurate method that can be used to identify the intersegmental plane and facilitate the quality of VATS-based segmentectomy.