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Aim and objective: This study aims to compare the efficacy of root canal cleanliness using hand instrumentation and ultrasonic handpiece under the scanning electron microscope. Materials and Methods: Forty five central incisor were collected and access opening was done. cleaning and shaping of all the samples were done with three different groups 1- Ultrasonic file. group-2 with Hand files and Group-3 were prepared with Ultrasonic + hand files and debris and smear layer were evaluated with SEM . Results: Group 1 (ultrasonic) removed smear layer superiorly, followed by Group 3 (ultrasonic/hand instrumentation) and Group 2 (hand instrumentation), and Group 3 (ultrasonic/hand instrumentation) showed superior cleanliness of debris followed by Group 1 (ultrasonic) and Group 2 (hand instrumentation). Conclusion: (1) At the apical, middle, and coronal third level for removal of debris, Group 3 showed superior cleanliness followed by Group 1 and Group 2. (2) At the apical, middle, and coronal third level for removal of smear layer, Group 1 showed superior cleanliness followed by Group 3 and Group 2.