Introduction: The aim of the study was to compare laser activated irrigation using Photon-induced photoacoustic streaming (PIPS) to passive ultrasonic activation (PUI) and in terms of the ability to disinfect the root canal. Methods: 45 single rooted upper anteriors were inoculated with E. faecalis and candida albicans to form a biofilm, then randomly divided into four groups that differ in the activation method (PIPS group, PUI group, and a control group that employs syringe irrigation without activation). Confocal scanning electron microscope (CLSM) was then used to evaluate and compare groups in terms of bacterial reduction. Results: There was a significant difference between different groups (p<0.001). The mean value and standard deviation (SD) of PIPS was (57.34±3.70), followed, while the lowest value was found in PUI group (41.40±6.38). Post hoc pairwise comparisons showed PIPS group to have a significantly higher value than other groups (p<0.001). Conclusions: Activation of Sodium hypochlorite irrigant using Er:YAG laser utilizing PIPS technique increased its biofilm eradication capability, showing the best biofilm eradication in the apical third. However, none of the root canal disinfection methods resulted in complete eradication.