ABA-triblock copolymers (PHCPMFs) were prepared via atom transfer radical polymerization (ATRP), using modified perfluoropolyether (PFPE) as a macroinitiator (Br-PFPE-Br) to form the B block, and 2-hydroxy-3-cardanylpropyl methacrylate (HCPM) as a monomer to form the A blocks. Physically stable and transparent cross-linked PHCPMFs (C-PHCPMFs) films could be prepared on a silicon wafer using spin coating, followed by UV irradiation to form the cross-linked structure. C-PHCPMFs showed bactericidal properties and bacterial adhesion resistance. The bacterial adhesion resistance of the PHCPMFs films was found to closely correlate with the low surface energy and water contact angle hysteresis. In particular, the C-PHCPMF5 film, containing about two HCPM units in the A block, shows excellent biocidal/bacterial adhesion resistance and cell viability because it has an optimum amount of biocidal HCPM units, and a sufficient amount of biocompatible PFPE units to impart a low surface energy.