OBJECTIVE Several studies demonstrate variability of root canal morphology of anterior teeth. This in-vivo study was designed to analyze and characterize root canal morphology of maxillary and mandibular anterior teeth using three-dimensional imaging. METHODS Patients who required CBCT radiographic examinations as part of their routine examination, diagnosis, and treatment planning were enrolled on the study. A total of 1,200 healthy, untreated, fully developed maxillary and mandibular incisors and canines were included. The following observations were recorded: (i) number of roots and their morphology; (ii) number of canals per root; and (iii) primary variations in the morphology of the root canal systems. RESULTS Root canal morphology of maxillary teeth was consistent with a single canal extending from the pulp chamber to the apex in 98% of the cases. In contrast, in mandibular central and lateral incisors, the presence of a single canal represented 64.5% and 60.5% respectively. The second most common morphology was the presence of one canal leaving the pulp chamber, which divided into two within the root, and then merged to exit as one canal in 18% and 25.5% respectively. The presence of one canal leaving the pulp chamber and dividing short of the apex into two separate and distinct canals with separate apical foramina (variant 5) was recorded in 14.5% and 12% respectively. CONCLUSIONS There was a low incidence of anatomical variation in maxillary anterior teeth. In contrast, mandibular anterior teeth presented with a high prevalence of two canals, especially central and lateral incisors. The reported data may help clinicians understand the variations in root canal morphology of anterior teeth in order to overcome problems associated with shaping and cleaning procedures.