BACKGROUND Situs inversus totalis (SIT) is a rare developmental anomaly. For many years, laparoscopic cholecystectomy has been a well-established and effective treatment for patients with cholecystolithiasis. However, when it comes to SIT patients with cholecystolithiasis, this surgery presents unique challenges. Due to its complete mirror inversion of viscera, it is uncomfortable to carry out standard laparoscopic cholecystectomy for some surgeons. Single-incision laparoscopic cholecystectomy (SILC) offers a minimally invasive alternative to traditional laparoscopic cholecystectomy, potentially enhancing aesthetic outcomes and minimizing postoperative discomfort. CASE REPORT A 67-year-old woman was admitted to the hospital due to abdominal pain. After admission, she completed a chest X-ray, electrocardiogram, B-ultrasound, and magnetic resonance cholangiopancreatography. Imaging examination revealed SIT and cholecystolithiasis. The laboratory examination results were normal. All contraindications to surgery were ruled out, and then SILC was performed under general anesthesia on January 22, 2024. The surgeon and assistant stood on the right side of the patient, and the abdominal SILC port with 3 operation channels was placed through a 2-cm traumatic incision. We used electric hooks and separating forceps to dissect the Calot triangle and gallbladder of the patient, and the gallbladder was successfully removed. The patient recovered well after the surgery. CONCLUSIONS For SIT patients with cholecystolithiasis, SILC is a safe and effective minimally invasive treatment option which has fewer surgical incisions and less postoperative pain. However, it demands experienced SILC surgeons, thorough preoperative examinations, and close attention to incision conditions after surgery.