The aim of this study was to evaluate morphological features of 70 single-rooted mandibular first premolars with radicular grooves (RG) using micro-CT technology. Teeth were scanned and evaluated regarding the morphology of the roots and root canals as well as length, depth and percentage frequency location of the RG. Volume, surface area and Structure Model Index (SMI) of the canals were measured for the full root length. Two-dimensional parameters and frequency of canal orifices were evaluated at 1, 2, and 3 mm levels from the apical foramen. The number of accessory canals, the dentinal thickness, and cross-sectional appearance of the canal at different root levels were also recorded. Expression of deep grooves was observed in 21.42% of the sample. Mean lengths of root and RG were 13.43 mm and 8.5 mm, respectively, while depth of the RG ranged from 0.75 to 1.13 mm. Mean canal volume, surface area and SMI were 10.78 mm 3 , 58.51 mm 2 , and 2.84, respectively. Apical delta was present in 4.35% of the sample and accessory canals were observed mostly at the middle and apical thirds. Two-dimensional parameters indicated an oval-shaped cross-sectional appearance of the root canal with a high percentage frequency of canal divisions (87.15%). Canal configuration type V (58.57%) was the most prevalent. C-shaped configuration was observed in 13 premolars (18.57%), whereas dentinal thickness ranged from 1.0 to 1.31 mm. Radicular grooves in mandibular first premolars was associated with the occurrence of several anatomical complexities, including C-shaped canals and divisions of the main root canal.