Literature regarding the influence of Er,Cr:YSGG laser pulse frequency with different output power levels on adhesion properties of adhesive resin to lased enamel is limited. Therefore, the aim of the present study was to evaluate the effects of laser pulse frequency (20, 35, and 50 Hz) at two different output power settings (3 and 6 W) of Er,Cr:YSGG on the microtensile bond strength (μTBS) of adhesive resin to enamel. Crowns of 35 intact bovine incisors were embedded into self-cure acrylic resin individually, and then flat enamel surfaces were prepared with 600-grit silicon carbide papers under water cooling. Teeth were divided randomly into seven groups. Enamel surfaces were irradiated with Er,Cr:YSGG laser operated at one of six output power-pulse frequency combinations (6 W20 Hz, 6... W-35 Hz, 6 W-50 Hz, 3 W-20 Hz, 3 W-35 Hz, and 3 W-50 Hz) in groups 1-6, respectively. Bur-treated surfaces served as a control in group 7. After surface treatments and bonding procedures, composite build-ups were done in three layers up to a height of 4 mm. Next, all bonded teeth were sectioned into the resin-enamel sticks to be tested in a μTBS testing machine. The μTBS data were analyzed with univariate analysis of variance under a general linear model with the factor 'tooth' added as a random effect to the design. Resin-enamel interfaces were evaluated with scanning electron microscopy (SEM). The μTBS to laser-irradiated enamel in group 1 (6 W-20 Hz) was significantly lower than those of bur-treated enamel (p<0.05). However, group 6 (3 W-50 Hz) showed significantly higher μTBS values than did bur-treated teeth (p<0.05). SEM evaluation revealed enormous morphological alterations of laser-irradiated specimens, such as extensive vertical and horizontal microcracks and gaps, with the exception of group 6. The bonding effectiveness of adhesive resin to laser-irradiated enamel was affected by the pulse frequency of the Er,Cr:YSGG laser. Although the parameters recommended by the manufacturer lowered μTBS, increasing the pulse rate may maintain optimum μTBS.