Cholangioscopy is reportedly useful for selective guidewire placement across difficult biliary strictures, but few methods are available for complete stricture of biliary anastomosis. This study aimed to propose a guidewire puncture technique to recanalize totally obstructed anastomosis and discuss its safety and feasibility. From January 2015 to December 2021, a total of 11 patients with complete biliary anastomotic stricture after liver transplantation were enrolled. These patients underwent peroral single operator cholangioscopy (SpyGlass), whereas two failed cases on SpyGlass finally underwent percutaneous transhepatic cholangioscopy (PTCS). The steps of the recanalization technique were as follows: the stricture was viewed carefully to detect the closure point (CP) of the scar endoscopically, then the CP was targeted by the hard tip of the guidewire and broke through under guidance of the cholangioscope and fluoroscope. Complete occlusions were confirmed by SpyGlass in all cases. A total of 13 hard-tip guidewire punctures were performed under cholangioscopy, and ten punctures were successful (technical success rate, 76.9% [10/13]). After recanalization of the occluded anastomosis, plastic stent or metallic stent was deployed in three and seven patients, respectively. No procedure-related complications occurred during or after the cholangioscopy-assisted guidewire puncture. After a mean follow-up of 12 months, stents had been removed in five patients. The other six patients were still receiving stent treatment. This study demonstrated that the guidewire puncture technique under cholangioscopy is safe and feasible for complete stricture of biliary anastomosis, and the success rate is satisfactory.