Objective To study the clinical application of self-made specimen bags using intraoperative materials in single-port thoracoscopic lobectomy.Methods The clinical data of 40 patients who underwent single-port thoracoscopic lobectomy from October 2021 to July 2022 were retrospectively analyzed, and divided into two groups by different operation methods, the conventional method group and the new method group, 20 cases each group. The conventional method group used a medical sterile protective sheath, leaving about 20 cm on one side, knotting the bottom and turning it inward. The mouth of the bag was held with two oval forceps and inserted into the chest cavity to remove the specimen. The new method group followed the same procedure, with the addition of a drainage tube inserted into the bag mouth. Intraoperative specimens were taken out from selfmade specimen bags. The specimen loading time, one-time success rate, wound healing status, and short-term prognosis were compared between the two groups.Results No specimens fell off in the new method group, and all were successfully retrieved on the first attempt. In the conventional method group, specimens fell off twice (2/20), but the difference between the two groups was not significant. The average completion time of specimen loading in the new method group was (36.20 ± 6.08) s, which was significantly shorter in the conventional method group (57.95 ± 6.59) s, with a statistically significant difference (t = 10.85, P < 0.01). There were no ulcers in any specimen bags, and the surgical incisions of all patients healed well, with no incision infection, no tumor implantation, and no intrathoracic dissemination and metastasis within one year.Conclusion The selfmade thoracoscopic specimen bag is simple to make, low-cost, safe and convenient to use. It can save operation time and reduce medical costs.