We report the preparation and application of indium tin oxide (ITO) coated fluorine-containing polyimide/organoclay nanocomposite substrate. Fluorine-containing polyimide/organoclay nanocomposite films were prepared through thermal imidization of poly(amic acid)/organoclay mixture films, whilst on which ITO thin films were coated on the films using a radio-frequency planar magnetron sputtering by varying the substrate temperature and the ITO thickness. Finally the ITO coated fluorine-containing polyimide/organoclay nanocomposite substrate was employed to make flexible organic light-emitting devices (OLED). Results showed that the lower sheet resistance was achieved when the substrate temperature was high and the ITO film was thick even though the optical transmittance was slightly lowered as the thickness increased. approximately 10 nm width ITO nanorods were found for all samples but the size of clusters with the nanorods was generally increased with the substrate temperature and the thickness. The flexible OLED made using the present substrate was quite stable even when the device was extremely bended.