Objective To assess the feasibility of three-dimensional contrast-enhanced ultrasonic cholangiography(3D-CEUSC) in cadaver liver.Methods The 3D-CEUSC was performed in 6 cases of cadaver liver.Image quality of 3D-CEUSC was evaluated.The visualization of branching orders,the degree of visibility and coincidence of morphous were compared with those of cholangiagraphy using fluoroscopy.Results The imaging quality of 3D-CEUSC was inferior to that of cholangiography with significant difference.The three-dimensional biliary tree structures were visualized in all 6 3D-CEUSC.The maximum visualization of branching orders in 3D-CEUSC was (3.67±0.52),which was equal to the results(4.00±0.63)by cholangiography (P=0.465).The degrees of visibility of biliary tree were equivalent with those by cholangiography in the first and second order with significant difference.The coincidence of morphous was excellent compared the images of 3D-CEUSC with direct X-ray cholangiography.Conclusion 3D-CEUSC is a new technique as a useful supplement to cholangiography in evaluation of biliary anatomy and variation before graft harvesting in LDLT.