The objective of this in vitro study was to evaluate the immediate and short-term effects of laser neodymium-doped yttrium aluminum garnet (Nd:YAG) irradiation and in-office desensitizing treatment on dentin tubule occlusion.Literature shows a lack of long-lasting treatments for dentin hypersensitivity.Forty-eight dentin slabs (4×4×2 mm) were ground flat, polished, and treated with 27% ethylenediaminetetraacetic acid (EDTA) to open the dentinal tubules. Specimens were randomly divided into the following experimental groups (n=12): Group 1: Control (no treatment); Group 2: Nd:YAG laser irradiation (100 mJ, 85 J/cm(2) per pulse with a quartz fiber of 400 μm, in scanning movements); Group 3: In-office prophylaxis with pumice; Group 4: In-office Colgate Sensitive Pro-Relief™ Desensitizing Paste. Treatments were performed according to the manufacturer's instructions. After treatment, the specimens were submitted to a sequence of erosive and abrasive challenges, twice a day for 5 days. The specimens were qualitatively and quantitatively evaluated by scanning electron microscopy immediately after treatment and after 4 and 5 days. The response variable was the amount of occluded dentin tubules per area, determined by three different examiners with the use of visual criteria, with a standardized grade created in the PowerPoint program. Data were compared with ANOVA and Tukey's test, considering a 5% significance level.Immediately after treatment, a reduction in the number of opened dentin tubules was observed for the laser group when compared with the control group (p0.05). After the experimental procedures, there were no quantitative differences between the amount of opened dentin tubules for all groups; however, micrographs showed some qualitative tubule occlusion for the laser group after the erosive/abrasive challenge.only laser irradiation was capable of immediately sealing the dentinal tubules; however, none of the treatments showed efficacy in maintaining tubule occlusion after the chemical and mechanical challenges.