Background: preparation of root canals is an important step in root canal treatment. Mechanical instrumentation of root canals cause an irregular layer of debris, known as the smear layer. As a result, several studies reported that preferring the removal of the smear layer. Objective: To study the influence of the energy (100 mJ) of Erbium, Chromium: Yttrium Scandium Gallium Garnet (Er,Cr:YSGG) laser at short pulse duration (60 μs)  on smear layer removal of apical third after using Photon induced photoacoustic streaming technique. Materials and methods: Eighteen straight single-rooted mandibular premolars were used. The roots length were uniform to 14mm from the anatomic apex and instrumented using Protaper Gold Rotary NiTi file system to size 40 taper 0.06. Er,Cr:YSGG pulsed laser (waterlase iplus Biolase, CA, USA) 2780 nm was used at short pulse duration (60 μs, 5 Hz ,air and water off) delivered by MD/iplus Glass Tips (MZ6) 600 μm in diameter, length=6 mm, calibration factor= 1.00. Ultrasonic activator ENDO1 (Guilin Woodpecker Medical Instrument Co., Ltd.china) was used with Ultrasonic activator tip ED98 (25#, 2% taper, 18.5mm) (Guilin Woodpecker Medical Instrument Co., Ltd.china). The samples were arbitrarily divided into three groups (n=6) and used as follows: (A) conventional irrigation with 5 ml of 17 % EDTA, (B) passive ultrasonic irrigation with 5 ml of 17 % EDTA,  (C) Er,Cr:YSGG induced photoacoustic streaming  with 5 ml of 17 % EDTA. After final irrigation with normal saline and drying, the roots were painted with nail varnish externally, and 2% methylene blue dye injected into canal until the canal filled with dye. The tooth was splitted at the fourth millimeters from root apex with a diamond disc representing the apical third. Picture was taken by Professional Digital SLR camera (Nikon D7100, Nikon Corporation, Thailand) with 40X magnification. The dye penetration was measured by using analytical software (measure picture CAD-KAS Kessler Germany). ANOVA test was used to analyse repeated measure mean between tested concentration (5.25% NaOCL, 17% EDTA) and control. Data was expressed as mean±SD. LSD (Least Significant Difference) test was used to calculate the significant differences between tested mean. Results: The mean values of the percentage of dye penetration area in two experimental groups was higher than the control group, as the follow (20.79±1.15control, 34.5453±0.45 ultrasonic group, 85.9804±2.69 Er,Cr.YSGG+17%EDTA at 0.5 W). The Percentage of dye penetration in Er,Cr:YSGG laser induced photoacoustic streaming with 17% EDTA was significantly high (P value 0.001), followed by ultrasonic activated group, while control group result in less Percentage of dye penetration than both study groups.  Conclusion: In this study, we concluded that Er,Cr:YSGG  pulsed laser  (0.5 W,5 Hz , air and water off) at short pulse duration (60 μs) with 17% EDTA using PIPS technique for activation of irrigant in endodontic treatment give better result than ultrasonic activation  in smear layer removal