Introduction: Recognition of variations in root canal anatomy is an essential prerequisite for successful endodontic diagnosis and treatment. Most of the endodontic failures result from missed canals. The mandibular first molar has the most complex root canal system and a high percentage of treatment failures are due to the impossibility of locating, instrumenting and obturating the middle mesial canal (MM). There are very few studies available regarding the incidence of MM canal in mandibular first molar. The present study evaluated the prevalence of the middle mesial canals in mandibular first molars using cone beam computed tomography (CBCT). Materials and Methodology: CBCT images of mandibular first molars were collected from patients (n= 100) who had undergone CBCT imaging. The images were analyzed and the prevalence of MM canals were calculated. Results: The data were analysed using descriptive statistics. In total of 100 patients, 60 were women and 40 were men, with age range 11-60 years old. 21 MM canals were identified in this study. In this, 11 were female and 10 were male. The prevalence of the MM canal in mandibular first molar MM canals were identified in 21% (21/100). Conclusion: Within the limitations of the study, it can be concluded that though advanced techniques like troughing and magnification help in finding and negotiating MM canals, 3D radiographies like CBCT can be as used as valuable adjunct to the above mentioned methods. When these canals are treated, the success rate of the endodontic treatment would definitely increase. Clinical studies with long-term follow-ups are needed to determine the effect of preparation of MM canals on the outcome of nonsurgical endodontic treatment in mandibular first molars. Keywords: Cone Beam Computed Tomography, Mandibular First Molar, Middle Mesial Canal, Prevalence.