In the diagnosis of dental occlusion, it is necessary to quantitatively measure interocclusal contacts and transfer them to a computer model. In this aspect, three-dimensional computer models of upper and lower dental casts play a significant role. In this study, we proposed a new method to measure occlusal interaction by using a microfocus X-ray CT technique. Measurement accuracy was determined as ±0.03 mm in comparison with a coordinate measuring machine. A superimposition procedure for two sets of three-dimensional dental cast models was also established. Using the same dental cast, the standard deviation between the two sets of models was ±0.015 mm -which was defined as measurement precision. Between an optical laser scanner and the microfocus X-ray CT system, the standard deviation measured between the two models was ±0.05 mm. Data were acquired when upper and lower dental casts mounted on the bite impression were scanned, and then occlusal interaction, contacts, and distance distribution between the casts were visualized by a colored map on the cast models. Within the limitations of the current study, it was successfully demonstrated that microfocus Xray CT was well poised for quantitative measurement of occlusal interaction.