Knowledge of the segmental anatomy and intersegmental biliary connections is an essential prerequisite to the effective management of patients with complex biliary strictures. Three dimensional (3D) imaging has the ability to demonstrate complex anatomical relationships that are difficult to appreciate on simple noninvasive two dimensional (2D) imaging. Our aim was to develop a technique for accurate, non-invasive 3D computed tomography (CT) cholangiography. Contiguous 4 mm CT sections were obtained through the liver during a dynamic bolus of 200 ml IV contrast. 3D surface reconstructions were then performed, the biliary system was isolated from surrounding hepatic parenchyma using segmentation and contrast threshold algorithms. 14 patients (six females, eight males, median age 68 years (range 48-82)) were studied. 13/14 had malignant biliary obstruction and one had obstruction secondary to a pancreatic pseudocyst. Obstruction was at the liver hilum in eight, the common bile duct in five and the common hepatic duct in one. Four patients had biliary endoprostheses but were symptomatic from inadequate drainage. There was good demonstration of the biliary anatomy, obstructed segments and intersegmental biliary connections in 13/14; irregular biliary dilatation secondary to primary sclerosing cholangitis rendered interpretation difficult in one. 3D cholangiography provided a useful adjunct to other imaging techniques. In particular, in patients with complex hilar strictures it aided implementation of appropriate interventional drainage procedures.