The aim of this study was to assess the microleakage underneath a pit and fissure sealant bonded to occlusal surfaces treated by Er:YAG laser To determine the most effective energy density of laser, fourteen specimens were irradiated from 50mJ to 300mJ at 3Hz. After irradiation, the lased specimens were observed under the scanning electron microscope. Thirty six non-carious extracted premolars were randomly assigned to four groups of nine teeth: group 1, no treatment on the occlusal surface; group 2, acid etching for 15 seconds; group 3, Er:YAG laser irradiation; group 4, acid etching followed by Er:YAG laser irradiation. The pits and fissures were sealed with unfilled sealant(Helioseal F) and the specimen teeth were thermo-cycled, immersed in 2% Rhodamine B solution, longitudinally sectioned and analyzed for microleakage with fluorescent microscope. The results were as follows: 1. Er:YAG lased surfaces with 50mJ, 3Hz showed a similar pattern of irregularity with acid etched enamel surfaces 2. The mean microleakage score increased in the order of group 2, 4, 3 and 1. There was no significant difference among group 1, 3 and 4(p>0.05), however group 2 showed significantly less microleakage compared with group 1 and 3. Conclusively, the laser irradiation seemed not enough to replace the acid etching for proper retention of pit and fissure sealants.