Background and Aims Fluorescence cholangiography by intrabiliary indocyanine green (ICG) injection during laparoscopic cholecystectomy (LC) has the advantage of immediate display with zero background liver fluorescence. It is more suitable for complex LC. This study was conducted to investigate the efficacy of fluorescent LC after percutaneous transhepatic gallbladder drainage (PTGBD) or endoscopic nasobiliary drainage (ENBD).Methods The clinical data of 120 patients with gallbladder stones undergoing LC from January 2019 to April 2021 were selected. Of the patients, 60 cases underwent fluorescence-navigated LC by injection of ICG through the PTGBD tube or ENBD tube (observation group), and 60 cases underwent conventional LC (control group). The display rates of the extrahepatic bile duct structures and the main clinical variables were compared between the two groups.Results The preoperative general data showed no significant difference between the two groups of patients (all P>0.05). Before dissection of the Calot's triangle, the display rates of the common hepatic duct, the common bile duct, the cystic duct, the junction of cystic duct and common bile duct and the neck of the gallbladder were significantly higher in observation group than those in control group (all P<0.05), but there was no significant difference in display rate of the right hepatic duct between the two groups (P>0.05); After dissection of the Calot's triangle, the display rates of the common hepatic duct, the common bile duct and the junction of cystic duct and common bile duct were significantly higher in observation group than those in control group (all P<0.05). In observation group compared with control group, the operative time, the intraoperative blood loss, the rate of subhepatic drainage tube placement, and length of postoperative hospital stay were all significantly reduced (all P<0.05). There was no significant difference in hospitalization cost and incidence rate of postoperative complications between the two groups (all P>0.05).Conclusions The application of fluorescence-navigated LC is beneficial to improve the display rate of the extrahepatic bile duct structures, decrease the degree of difficulty of complex LC after PTGBD and ENBD, shorten the operative time and postoperative hospital stay, and reduce the intraoperative blood loss and subhepatic drainage tube placement.