Objective:To evaluate the application value of fluorescent cholangiography via gallbladder injection of ICG in laparoscopic cholecystectomy (LC) for patients with gallstones and cholecystitis.Methods:Clinical data of 60 patients with gallstones and cholecystitis who underwent LC in the Second Affiliated Hospital of Kunming Medical University from September 2020 to February 2022 were retrospectively analyzed. Among them, 31 patients were male and 29 female, aged from 22 to 70 years, with a median age of 50 years. The informed consents of all patients were obtained and the local ethical committee approval was received. All patients were divided into the ICG group (n=30) and control group (n=30) according to whether ICG fluorescent cholangiography was applied during LC. Perioperative conditions of all patients were observed. The identification rate of extrahepatic bile duct between two groups was compared by Chi-square test. The operation time between two groups was compared by t test. Intraoperative blood loss between two groups was compared by Mann-Whitney U test.Results:The surgeries were successfully performed in both groups, and no perioperative death occurred. The identification rates of cystic duct and common bile duct in the ICG group were 77%(23/30) and 87%(26/30), significantly higher than 27%(8/30) and 33%(10/30) in the control group (χ2=15.017, 17.778; P<0.05). In the ICG group, the average time of dissecting Calot's triangle, separating gallbladder bed and operation time were (8.4±2.5), (10.9±3.4) and (49.8±9.9) min, significantly shorter than (10.6±3.5), (15.7±5.5) and (57.9±13.6) min in the control group (t=-2.769, -3.973, -2.655; P<0.05). The median intraoperative blood loss in the ICG group was 15(6) ml, significantly less than 23(11) ml in the control group (Z=-4.172, P<0.05). In the ICG group, no extrahepatic bile duct injury and conversion to open surgery were reported, whereas there was one case of conversion to open surgery and 2 cases of common bile duct injury in the control group, who were treated symptomatic treatments, and all cases were restored to normal in postoperative 1-month follow-up.Conclusions:For patients with gallstones and cholecystitis, fluorescent cholangiography via gallbladder injection of ICG during LC can accurately identify cystic duct from common bile duct, effectively avoid intraoperative iatrogenic bile duct injury, shorten the operation time and reduce the intraoperative blood loss.