Purpose <p>Laparoscopic cholecystectomy (LC) in patients with situs inversus totalis (SIT) is technically challenging due to the reversed anatomical orientation of the thoracoabdominal organs, which can increase operative complexity and the risk of complications. Although the incidence of cholelithiasis and cholecystitis in SIT is similar to the general population, altered anatomy may lead to diagnostic and intraoperative uncertainty. We describe LC in a patient with SIT using intraoperative indocyanine green (ICG) fluorescence cholangiography to guide the identification of biliary structures.</p> Methods <p>Preoperative ultrasound, computed tomography, and magnetic resonance imaging are used to delineate anatomy and exclude biliary duct dilation or biliary anomalies. ICG (0.5 mL of 2.5&#xa0;mg/mL) is administered intravenously one hour before surgery. A mirror-image port configuration is employed centered on the left hemiabdomen, with the surgeon standing on the patient’s right and the assistant on the left. During surgery, ICG fluorescence cholangiography is utilized intraoperatively to enhance visualization of the biliary anatomy and facilitate attainment of the critical view of safety.</p> Results <p>LC was successfully performed without complications. ICG imaging enabled clear identification of the cystic duct and common bile duct, mitigating the risks associated with anatomical variation in SIT.</p> Conclusion <p>LC in patients with SIT is rarely described and technically demanding due to reversed intra-abdominal anatomy. ICG fluorescence cholangiography is an innovative tool and valuable adjunct that improves biliary visualization and may enhance safety in complex anatomical settings.</p>