Polyimide (PI) nanofibers with an average diameter of 300 nm, possessing superior electrolyte wettability and thermal stability, are synthesized by electrospinning a poly(amic acid) (PAA) solution followed by an imidization process. The large pore volume of the PI nanofiber mat can facilitate faster Li+-ion transport and greater rate capability, but it also cause an irreversible increase in cell impedance during long term cycling. To overcome these problems, thin Al2O3 over-layers are coated on both sides of a PI separator via a dip-coating process. The Al2O3-coated PI separators exhibit enhanced capacity, cyclability (95.53% retention after 200 cycles at 1 C), and rate capabilities (78.91% at 10 C) compared to the bare PI separator (68.65% at 10 C) and a commercial polypropylene separator (18.25% at 10 C) with a limited increase of cell impedance.