Objective To evaluate the clinical application value of SpyGlass DS digital cholangioscopy in diagnosis and therapy of patients with biliary tract diseases. Methods Seven patients with biliary tract diseases were examined and treated with SpyGlass DS system. Three patients with large choledocholithiasis underwent laser lithotripsy, and then the broken stones were removed by ERCP. Four patients with indeterminate bile duct stricture underwent SpyGlass DS examination and SpyGlass DS-guided biopsies if necessary. Results SpyGlass DS cholangioscopy was successfully performed in all patients with a mean procedure time of 12.6 min. Complete stone clearance was achieved in all 3 patients with large biliary stones. Of the 4 patients with indeterminate biliary stricture, 2 underwent SpyGlass DS-guided biopsies and were diagnosed as malignant biliary tumor; the other 2 patients were diagnosed as inflammatory bile duct stenosis after visualizing the target lesions. Two patients developed hyperamylasemia after the procedure, and recovered spontaneously. Conclusion SpyGlass DS cholangioscopy can provide an accurate diagnosis in indeterminate biliary strictures and facilitate therapy in patients with challenging biliary stones.