Background: Chemo mechanical debridement is an important part of endodontic treatment. Elimination of pulpal tissue, microbiota and their by-products, organic and inorganic debris removal by using instruments and intracanal irrigants are objectives of this important phase of treatment. The aim of this study was to ex-vivo evaluation of smear layer removal in conventional clinical methods, Passive Ultrasonic Irrigation, Laser Activated Irrigation.Material and Methods: 54 anterior mandibular teeth was used in this study. After preparation of samples, they were divided in to 3 groups of Conventional Irrigation +Smear layer removal, Passive Ultrasonic Irrigation, Laser Activated Irrigation. Then, the samples were cut longitudinally in mesiodistal direction for examination by SEM (1500X) for assessing smear layer removal.Results: In coronal, middle and apical one third, the best result was obtained from Conventional Irrigation + Smear layer removal group for smear layer removal.Conclusion: According to better smear layer removal in Conventional Irrigation + Smear layer, it is considered as gold standard protocol and other groups cannot be a suitable substitution for this method.