In this study, the preparation of patterned-fluoropolymer substrate using ion irradiationinducedsurface grafting of poly(acrylic acid) (PAA) and collagen modification was investigated forthe selective cell adhesion. First, the surface of a polytetrafluoroethylene (PTFE) film was selectivelyirradiated with proton (H+) ions and was functionalized by graft polymerization of acrylic acid onto theirradiated regions. In turn, the collagen was immobilized covalently on the PAA regions using EDC/NHS coupling reaction. The results of the surface characterization (including peroxide concentrationand grafting degree, chemical structure, and wettability), clearly revealed that PTFE surface wasfunctionalized with PAA (fluence dependent manner) and collagen was successfully immobilized on thegrafted PAA chains. Moreover, the result of the in vitro cell culture revealed that cells selectively adheredto the collagen patterns (but not on the control PTFE surfaces) and produced well-arranged 100 μm cellmicropatterns.