Star copolymer films were studied for confirming the role of film morphology in antimicrobial activity previously reported. With this scope, films of a two-branches copolymer, namely m-PEG-(MMA-ran-DMAEMA)2, containing ≈ 40 % in mol of non-quaternized 2-(dimethylamino)ethyl methacrylate, were produced by using spin-coating, drop-casting and casting deposition techniques, thus obtaining ultra-thin, thin and thick films, respectively. The morphology is generally flat for thin and thick films, but it becomes substrate dependent for ultra-thin films where the film planarization effect is not efficient. Mechanical properties of such films were investigated by Force Volume Maps in both air and liquid. In air, ultra-thin films are in the substrate-dominated zone and thus the elastic modulus E is overestimated, while E reaches its bulk value for thin and thick films. In liquid (water), E follows an exponential decay for all films with a minimum soaked time t0 of 0.37 and 2.65 h for ultra-thin and thin and thick films, respectively. After this time, E saturates to a value in average 92 % smaller than that measured in air due to films swelling. Such results confirm the role of morphology envisaged in literature, suggesting also an additional role of mechanical properties in the antimicrobial activity.