Anode materials providing a high specific capacity with a high cycling performance are one of the key parameters for lithium ion batteries’ (LIBs) applications. Herein, a high-capacity NiFe2O4(NFO) film anode is prepared by E-beam evaporation, and the effect of the heat treatment is studied on the microstructure and electrochemical properties of LIBs. The NiFe2O4 film annealed at 800 °C (NFO-800) showed a highly crystallized structure and different surface morphologies when compared to the electrode annealed at a lower temperature (NFO-600, NFO-700). In the electrochemical measurements, the high specific capacity (1804 mA g−1) and capacity retention ratio (95%) after 100 cycles were also achieved by the NFO-800 electrode. The main reason for the good electrochemical performance of the NFO-800 electrode is a high structure integrity, which could improve the cycle stability with a high discharge capacity. The NiFe2O4 electrode with an annealing process could be further proposed as an alternative ferrite material.