The influence of laser conditioning on defects of HfO2 monolayer films prepared by electron beam evaporation (EBE) is investigated utilizing the spot-size effect of the laser-induced damage. It is found that the laser-induced damage threshold of HfO2 monolayer films can be increased by a factor of 1.3-1.6. It is also found that the defects with low threshold can be removed by laser conditioning and defects with higher threshold may be removed partially.