Through chemomechanical debridement of the root canal is a primary requisite for successful endodontic therapy. Thus the aim of this study was to evaluate the effects of using QmiX alone, QmiX with EndoActivator and QmiX with Er,Cr:YSGG laser for final irrigation on sealer penetration into the dentinal tubules. 75 extracted human mandibular premolar teeth were treated with sodium hypochlorite (NaOCl) irrigation. The samples were divided into 5 groups according to the final irrigation solution used: (1) 17% EDTA and 2.5% NaOCl, (2) QmiX (3) QmiX with Er,Cr:YSGG laser and (4) QmiX with EndoActivator (5) 2.5%NaOCl. All teeth were obturated using cold lateral condensation technique with gutta percha and AH 26 sealer (Dentsply; DeTrey,Konstanz, Germany) labeled with Rhodamine B dye. The teeth were sectioned at distances of 2 and 5 from root apex. Total percentage and maximum depth of sealer penetration were measured using confocal laser scanning microscopy. Results of one way Anova analysis showed that there was a significant difference in the percentage and depth of sealer penetration among all groups at 3 and 5 mm level sections (P < .05). Within the groups maximum sealer penetration was recorded for Er,Cr:YSGG laser activated group. Greater depth of sealer penetration was recorded at 5mm as compared to 3mm in all the groups. Activation of QMix using EndoActivator and Er,Cr:YSGG laser enhanced the sealer penetration at apical and middle third. Thus Er,Cr:YSGG laser and EndoActivator may act as an appropriate adjunct during chemomechanical preparation of the root canal.