A non-destructive method to inverse-calculate a fiber probe aperture size, and a prediction method of the simulation and fabrication profile of near field photolithography are provided. The error between an experimental result of the photolithography and a simulation result of the theoretical model of near field photolithography is set as an objective function to inverse-calculate a fiber probe aperture size that can match with the photolithography experiment and the theoretical model of near field photolithography. Finally, by comparing the fabrication profile of the photolithography experiment and that of the simulation result of the inverse-calculated fiber probe aperture size, it is verified that the inverse-calculated fiber probe aperture size is reasonable and acceptable. By combining with other parameter control of near field photolithography, the present invention also proposes a prediction method of the fabrication profile of near field photolithography by means of inversely calculating the fiber probe aperture size.