Two kinds of polyaniline films with different microstructures were prepared and investigated. One of the films was prepared with polyaniline having nanofiber structure, which was synthesized by rapidly mixed reaction; and the other was prepared with polyaniline having granular particle structure, synthesized by conventional chemical oxidation method. The spectroscopic, electrochemical, and transport properties of the two kinds of films were studied with an interest in an influence of film microstructure on mobilities of charge carriers in polymer films. A clear difference was observed in the mobility versus oxidation level plot between the two kinds of polyaniline films. The results show that nanofiber structure of polyaniline film enhances mobilities, but affect little features of the mobility change induced by oxidation, which implies that although the nanofibers structure facilitates the charge transport in neutral or oxidized films, but it is irrelevant to the evolution of metallic conduction in polyaniline films.