Various methods and materials used in the treatment of dentin hypersensitivity are thought to achieve a therapeutic benefit by tubule occlusion. The aim of the present study was to evaluate the combined occluding effect of sodium fluoride varnish and Nd:YAG laser irradiation on human dentinal tubules. Thirty-six dentin specimens with exposed dentinal tubule orifices were used in this study. The samples were randomly divided into four groups. Groups A, B, and C were varnished by sodium fluoride, whereas group D served as a control. Then, group C was lased by 30 mJ of Nd:YAG laser, 10 pulses/s for 2 min by light painting. Three hours later, groups B and C were brushed by an electrical toothbrush for 30 min. Under SEM observation, the control group showed numerous exposed dentinal tubule orifices, and the sodium fluoride varnished specimens showed closure of exposed dentinal tubule orifices. After electrical toothbrushing, most of the sodium fluoride varnish was brushed away, except in the specimens that were irradiated by Nd:YAG laser. Over 90% of the dentinal tubule orifices were occluded by sodium fluoride varnish combined with Nd:YAG laser irradiation.