The invention relates to a nano-particle fluorescence spatial coding anti-fake identification method. The method includes the following steps that: S1, fluorescence nano-particles are prepared; S2, the fluorescence nano-particles are adopted to make an anti-fake pattern; S3, the anti-fake pattern is irradiated by light, and an anti-fake model is built; and S4, a picture to be authenticated is processed in the same manner with the step S3, and fluorescence intensity and spatial distribution characteristic information are extracted, and similarity measurement is carried out through using the anti-fake model, so that authenticity of the picture can be determined. According to the method of the invention, the anti-fake pattern is unique, the distribution of the nanoparticles has random uncontrollability, and therefore, it is impossible to produce the same fluorescence distribution pattern, and thus, compared with the prior art, the method has high anti-fake degree.