In endodontic therapy, access and instrumentation are strongly affected by root canal curvature. However, the few studies that have actually measured curvature are mostly from two-dimensional radiographs. The purpose of this study was to measure the three-dimensional (3D) canal curvature in maxillary first molars using micro-computed tomography (microCT) and mathematical modeling. Extracted maxillary first molars (46) were scanned by microCT (502 image slices/tooth, 1024 X 1024 pixels, voxel size of 19.5 x 19.5 x 39.0 microm) and their canals reconstructed by 3D modeling software. The intersection of major and minor axes in the canal space of each image slice were connected to create an imaginary central axis for each canal. The radius of curvature of the tangential circle was measured and inverted as a measure of curvature using custom-made mathematical modeling software. Root canal curvature was greatest in the apical third and least in the middle third for all canals. The greatest curvatures were in the mesiobuccal (MB) canal (0.76 +/- 0.48 mm(-1)) with abrupt curves, and the least curvatures were in the palatal (P) canal (0.38 +/- 0.34 mm(-1)) with a gradual curve. This study has measured the 3D curvature of root canals in maxillary first molars and reinforced the value of microCT with mathematical modeling.