Thermal and microstructural events resulting from KTP laser use during root canal preparation were investigated in 30 extracted single-rooted human teeth. In the first section of this study, thermal events occurring on the root surfaces of 18 teeth during and after exposure of the root canal were measured using thermography. A variety of parameters were used to determine settings that would be effective without causing thermal damage to the periodontal ligament. In the second section of the study, root canals of 12 teeth exposed to KTP laser irradiation at parameters derived from section 1 were evaluated using Scanning electron microscopy. KTP laser application at a power setting of 3 W, an exposure time of 2 s, and a frequency of 5 Hz, applied five times, removed smear layer and debris from the root canal surface at temperatures below the thermal injury threshold for periodontal tissue.