The goal of the present article is to illustrate and analyze the applications and the potential of microcomputed tomography (micro-CT) in the analysis of tooth anatomy and root canal morphology. The authors performed a micro-CT analysis of the following different teeth: maxillary first molars with a second canal in the mesiobuccal (MB) root, mandibular first molars with complex anatomy in the mesial root, premolars with single and double roots and with complicated apical anatomy. The hardware device used in this study was a desktop X-ray microfocus CT scanner (SkyScan 1072, SkyScan bvba, Aartselaar, Belgium). A specific software ResolveRT Amira (Visage Imaging) was used for the 3D analysis and imaging. The authors obtained three-dimensional images from 15 teeth. It was possible to precisely visualize and analyze external and internal anatomy of teeth, showing the finest details. Among the 5 upper molars analyzed, in three cases, the MB canals joined into one canal, while in the other two molars the two mesial canals were separate. Among the lower molars two of the five samples exhibited a single canal in the mesial root, which had a broad, flat appearance in a mesiodistal dimension. In the five premolar teeth, the canals were independent; however, the apical delta and ramifications of the root canals were quite complex. Micro-CT offers a simple and reproducible technique for 3D noninvasive assessment of the anatomy of root canal systems.