Introduction: Dentin hypersensitivity (DH) in one of the most common causes of patient discomfort in the general population and its prevalence. There is currently no known specialised therapy that can effectively eradicate dentinal hypersensitivity, according to the literature. Aim: The purpose of this study was to evaluate and contrast the effectiveness of diode laser alone against iontophoresis with 1.23% sodium fluoride (NaF) gel in occluding dentinal tubules. Materials and Methods: In this in vitro study, 20 teeth were included with intact root surfaces unaltered by extraction procedure for specimen preparation. Each tooth was cleaned, air dried and cut into three sections. Total 60 sections were prepared, which were acid etched. In control group, no treatment was carried. In iontophoresis treatment group, Samples were placed into a foam tray containing 1.23% NaF Gel and exposed to 1.5 mA output current for three minutes. In laser treatment group, specimens were lased with 940± 10nm diode laser at 0.5 W/PW in a noncontact mode for 30 seconds. Specimens were evaluated under Scanning Electron Microscope (SEM) at 10KV to 20KV under x5000 x1000 magnification for surface characteristics and patency of dentinal tubules. Total number of tubules visible, open, completely, and partially occluded were recorded in each microphotograph and compared. Results: On comparison, laser group showed maximum number of occluded dentinal tubules, 14.36% followed by iontophoresis group, 7.4% and control group 0.2% Conclusion: The blockage of dentinal tubules was treated more successfully with diode laser than with iontophoresis. As a result, it can be applied to the treatment of DH patients.