Ta2O5 films were prepared with conventional electron beam evaporation and annealed in O2 at 673 K for 12 h. Laser-induced damage thresholds (LIDTs) of the films were performed at 532 and 1064 nm in 1-on-1 regime firstly, and then were performed at 532, 800, and 1064 nm in n-on-1 regime, respectively. The results showed that the LIDTs in n-on-1 regime were higher than that in 1-on-1 regime at 532 and 1064 nm. In addition, in n-on-1 regime, the LIDT increased with the increase of wavelength. Furthermore, both the optical property and LIDT of Ta2O5 films were influenced by annealing in O2.