Novel poly(ethylene-vinyl acetate) film with fluorinated surface structure was prepared via sequential surface reactions, which are alcoholysis reaction with sodium ethoxide and located fluorination reaction with 2,3,3,3-tetrafluoro-2-[1,1,2,3,3,3-hexafluoro-2-(heptafluoropropoxy)prop oxy] propionyl fluoride, respectively. The obtained novel film possesses ordered perfluoroalkyl ether surface layer proved by detailed ATR-FTIR, XPS, and contact angle measurement, which substantiates the success of this new route to get better-tailored surface meeting the biological needs. Hemocompatibility of the resultant films was evaluated by hemolysis test and platelet adhesion test. The results indicated that the poly(ethylene-vinyl acetate) film surface displayed low hemolytic activity and no platelet adhesion due to its extremely low surface tension, which results from the special micro-electric field of perfluoroalkyl ether layer. The yielded transparent film with good properties exhibits a promising application prospect in biology and medical science in view of its flexible substrate and unique biocompatible surface.