To explore the clinical application effect of preoperative localization technology combined with video-assisted thoracoscopic surgery for the resection of multiple nodules in both lungs and its influence on relevant surgical indexes and postoperative laboratory indexes. Methods: Eighty patients with multiple bilateral pulmonary nodules were randomly divided into a control group and an observation group, 40 cases in each one. In the control group, hook wire positioning was performed under the guidance of video-assisted thoracoscopic surgery before operation. In the observation group, medical glue was injected and positioned through the body surface under the assistance of laparoscopic surgery before operation. The puncture location, treatment effect and nodule pathological analysis of patients between the two groups were compared. Results: There were no significant differences in puncture depth, positioning time, and the distance between pulmonary nodules and locating point between the two groups (P>0.05);  There were no significant differences in the success rate of puncture and the disposable resection rate of pulmonary nodules between the two groups (P>0.05); Compared with the control group, the incidence of postoperative complications in the observation group was lower, making the difference statistically significant (P<0.05); There was little difference in the intraoperative and postoperative pathology between the two groups, maintaining a high goodness of fit. Conclusion: Preoperative localization by injection of medical glue through the body surface assisted with hook wire and laparoscopic surgery can provide a good adjuvant effect for the surgery, but laparoscopic adjuvant therapy can reduce postoperative complications and maintain a high safety