In this paper laser ablation of polyethersulfone (PES) films regarding to the change in biocompatibility of the surface is investigated at 3 different wavelengths of 193nm (ArF), 248 nm (KrF) and 308 nm (XeCl). The optimum laser fluence and number of pulses for the improvement of the surface biocompatibility is found by examination of the surface behavior in contact with platelets and fibroblasts cells at 3 wavelengths. These biological modifications are explained by alteration of the surface morphology and chemistry following irradiation. The results show that the KrF laser is the best choice for treatment of PES in biological applications.