Introduction: A new method for the approximation of the root canal's cross-sectional shape and its working width using cone-beam computed tomographic (CBCT) or micro–computed tomographic (micro-CT) imaging was introduced. Methods: Scanned data from 29 extracted human mandibular first and second molar distal root canals without instrumentation were reconstructed and analyzed with a self-developed measurement algorithm. The 3-dimensional volume models were sliced perpendicular to the vertical axis. Using different 2-dimensional parametric models, the contour of each root canal slice was approximated and used to determine the canal's cross-sectional dimensions. The measurements of minor width, major width, and the root canal's conicity were statistically analyzed using analysis of variance. Results: The measured minor and major widths of the investigated root canals were significantly higher (probability value P < .05) when evaluated by CBCT images than the results obtained from micro-CT data. Both dimensions increased starting from the apical foramen (P < .01). The narrowest measured canal widths were 0.19–0.24 mm for CBCT imaging and 0.09–0.21 mm for micro-CT imaging in the apical part. The maximum values for conicity were between 13% and 17% in the cervical third. Conclusions: The 3-dimensional imaging data from CBCT and micro-CT imaging enabled a valuable anatomic assessment of the root canal's cross-sectional working width along the canal up to the physiological foramen in order to determine an adequate apical diameter as well as the correct measured taper in the cervical and medial part.