This study investigated the feasibility of video-assisted thoracic surgery (VATS) performed under two-lung ventilation (TLV) and single-lumen endotracheal tube (SLET) intubation in patients with spontaneous pneumothorax. From January 2016 to December 2019, 344 patients who underwent VATS with spontaneous pneumothorax, whether primary or secondary, were enrolled. The surgery was performed through TLV using SLET intubation or one-lung ventilation (OLV) using double-lumen endotracheal tube (DLET) intubation. Patient data were collected retrospectively from medical records and compared with an emphasis on the time required for anesthesia and surgery. The average anesthesia time was 72.6±17.8 min for TLV and 89.9±24.3 min for OLV (P<0.001). The average operating time was 42.1±16.2 min for TLV and 54.7±23.8 min for OLV (P<0.001). The average time from the onset of anesthesia to incision was 23.6±7.0 min for TLV and 27.6±9.5 min for OLV (P<0.001). There was no case of conversion to OLV using DLET intubation during surgery with TLV using SLET intubation. Removal of the chest tube took 1.6±1.1 days for the TLV group and 2.3±3.6 days for the OLV group (P=0.017). Patients were discharged at 2.7±1.2 days after surgery for the TLV group and 3.2±2.3 days after surgery for the OLV group (P=0.009). TLV using SLET intubation could shorten the time required for anesthesia-related procedures and surgery. In addition, it can be a beneficial surgical and anesthetic option for pneumothorax.