Porous silica films with various porosities were prepared via a sol-gel method using a nonionic amphiphilic triblock copolymer F127 as the structure-directing agent. Doppler broadening of positron annihilation radiation (DBAR) spectra were collected for the prepared films using a variable energy slow positron beam. Different linear relationships between positron annihilation line shape parameters S and W are found for the as-deposited films and calcined ones, indicative of the decomposition of the copolymer porogen in the as-deposited films upon calcination. This also reveals the variation of positron annihilation sites as a function of F127 loading or porosity. Strong correlations between positronium 3γ annihilation fraction, S parameter and porosity of the mesoporous silica films with isolated pores are obtained, which may provide a complementary method to determine closed porosities of mesoporous silica films by DBAR.