Background and Objectives: A reliable method to eradicate the bacteria of residual carious dentin has not yet been developed. The aim of this study was to evaluate the antibacterial effect of a diode laser on Streptococcus mutans through different thickness (500, 1,000, and 2,000 mm) of human dentin. The thermal effect of laser irradiation was also investigated. Study Design/Materials and Methods: Dentin specimens were inoculated with 2 ml of S. mutans on one side and irradiated by a diode laser on the other side with a power output ranging from 0.5 to 7 W. The laser tip was swept with the whole irradiation area of 7 mmÂ3 mm at a speed of about 10 mm/second with a total irradiation time of 30 seconds. Cooling with distilled water (30 ml/minute) was applied simultaneously during laser irradiation. After laser irradiation, the bacteria was removed from the dentin surfaces and cultured for 48 hours at 378C anaerobically to assess the colony forming units (CFU) per ml. The morphology of the lased bacteria and the temperature rise during laser irradiation were observed by scanning electron microscope (SEM) and measured by thermocouple, respectively. Results: The results revealed that 7 W of laser power could kill 97.7% of CFU through 500 mm thickness of dentin. However, the bactericidal efficiency was significantly reduced as the dentin thickness was increased. The morphological changes of lased bacteria ranged from less affected such as loss of their wall bands and existence of minicells to more severely degenerated, such as disintegration and fusion of cells with pores on the cell wall. Only the dentin specimens with a thickness of 500 mm exhibited a temperature rise greater than 5.58C after receiving 5 or 7 W of laser irradiation. Conclusions: A diode laser can eliminate the Streptococcus mutans of the residual carious dentin without inducing high pulpal temperature rise when the remaining dentin thickness is greater than 1 mm.