Introduction: The totally thoracoscopic approach for mitral valve (MV) disease is a minimally invasive method. We investigated the procedure's feasibility, safety and effectiveness when it was performed by an experienced operator. Methods: We retrospectively analysed 96 consecutive patients with MV disease treated between March 2016 and November 2019 by minimally invasive procedures. The procedures were performed on a femoral artery-vein bypass through two ports, including a main operation port and a thoracoscopic port. The clinical data of patients were collected, including preoperative cardiac function, operative data, postoperative complications, and follow-up. Results: A total of 96 patients (57 male patients; average age, 49.7 +/- 14.5 years; left ventricular ejection fraction, 65.6 +/- 7.7%) were enrolled in this study. No intraoperative conversion incision or death occurred. The cardiopulmonary bypass and aortic cross-clamp times were 163.8 +/- 50.6 minutes and 119.7 +/- 38.9 minutes, respectively. Postoperative chest tube drainage in the first 24 hours was 232.8 +/- 108.1 ml. The ventilation time and length of intensive care unit stay were 13.2 +/- 6.2 hours and 2.9 +/- 2.2 days, respectively. One patient died of disseminated intravascular coagulation and prosthesis thrombosis 3 days after the operation, fearing anticoagulant-related hemorrhage. The overall success rate of valve repair during 1-year follow-up was 97.9%. Conclusion: The totally thoracoscopic procedure on mitral valves by an experienced surgeon is technically feasible, safe, effective and worthy of widespread adoption in clinical practice.