A thorough understanding of both common and uncommon root canal anatomies is crucial for the success of root canal treatments, as missing a canal can lead to treatment failure. Although the maxillary second molar typically features three canals, the occurrence of five canals, particularly a mesial buccal third canal (MB3) canal in the mesiobuccal root, is extremely rare. This case report documents such a rare occurrence in a maxillary second molar with five canals. With the assistance of a dental operating microscope, all canals were successfully located, and root canal preparation, irrigation, and filling were accomplished. This case report underscores the significance of in-depth knowledge of root canal anatomy and the invaluable aid of a dental operating microscope in achieving successful root canal treatments.