A two-step process for preparing polyimide nanofiber was used in this work, and the nanofibers of precursor polyamic acid (PAA) were prepared by electrospinning. The polyimide nanofibers were obtained through thermal imidization at different thermal imidization temperatures. The influences of different imidization temperatures on morphology, chemical structure, thermal stability, and mechanical properties of the polyimide nanofibers were investigated by Fourier Transform Infrared Spectroscope (FT-IR), Scanning Electronic Microscopy (SEM), thermal gravimetric analysis (TGA) and Electromechanical Universal Testing Machine. The experimental results show that the imidization of PAA nanofibers was not complete under 290°C but complete under 350°C. With the rise of imidization temperature from 290 °C to 350°C, the polyimide nanofibers became thinner, and the thermal stability of polyimide nanofibers was also improved. The breaking strength and elongation at break of the nanofiber membranes increased at higher imidization temperature.