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Introduction: The study aimed to evaluate, through in vivo tomographic analysis, the prevalence of C-shaped canals in mandibular first and second molars of Brazilian individuals, analyzing its frequency by thirds of the roots, and in contralateral teeth. Methods: Images of 801 mandibular molars (379 first molars and 422 second molars) from 334 Brazilian individuals (142 men and 192 women) were identified through 1544 cone beam computed tomography (CBCT) exams, obtained from a private oral radiologic clinic. The cross-sectional configurations were analyzed to determine the frequency of C-shaped canals at three different axial levels and classified in categories by three experienced endodontists independently. Results: The incidence of C-shaped canals was 181 (23%). Considering the type of tooth, 91 (24.01%) were identified in the first molars, and 90 (21.32%) were found in the second molars. The incidence was significantly higher in female individuals (P < 0.05) for both first and second molars. The most common C-shaped canal configurations were: C1 (89.01% for first molars and 90% second molars), followed by C2 (8.79% for first molars and 6.66% for second molars) and C4 (2.19% for the first molars and 3.33% for the second molars). Bilateral C-shaped canals were significantly higher than unilateral for both first and second molars (P < 0.01). Conclusions: The prevalence of C-shaped canals in mandibular molars of the Brazilian individuals was higher than previously reported for both mandibular first (24.01%) and second molars (21.32%). The incidence was significantly higher in female individuals and the coronal portion of the roots. The classic C-shaped format “C1” was the most frequent anatomical configuration. Furthermore, the prevalence of bilateral C-shaped canals was higher for the first molar (61.70%) and lower for the second molar (38.29%). [ABSTRACT FROM AUTHOR]