This study was designed to determine whether an interactive three-dimensional presentation depicting liver and biliary anatomy is more effective for teaching medical students than a traditional textbook format presentation of the same material. Forty-six medical students volunteered for participation in this study. Baseline demographic information, spatial ability, and knowledge of relevant anatomy were measured. Participants were randomized into two groups and presented with a computer-based interactive learning module comprised of animations and still images to highlight various anatomical structures (3D group), or a computer-based text document containing the same images and text without animation or interactive features (2D group). Following each teaching module, students completed a satisfaction survey and nine-item anatomic knowledge post-test. The 3D group scored higher on the post-test than the 2D group, with a mean score of 74% and 64%, respectively; however, when baseline differences in pretest scores were accounted for, this difference was not statistically significant (P = 0.33). Spatial ability did not statistically significantly correlate with post-test scores for the 3D group or the 2D group. In the post-test satisfaction survey the 3D group expressed a statistically significantly higher overall satisfaction rating compared to students in the 2D control group (4.5 versus 3.7 out of 5, P = 0.02). While the interactive 3D multimedia module received higher satisfaction ratings from students, it neither enhanced nor inhibited learning of complex hepatobiliary anatomy compared to an informationally equivalent traditional textbook style approach. (Contains 3 tables and 2 figures.)