The work aims to study the method and proper conditions of quick preparation of structural color of photonic crystal on the glass substrate. This could be acquired by combining ink-jet printing polystyrene microsphere (PS) emulsion and self-assembly of drops. First, PS emulsion was prepared by emulsion polymerization. Second, emulsion was prepared as proper ink-jet ink and some printing conditions (including the wetting and temperature of substrate and printed times) were optimized to regularly array PS particles. Finally, the printing structural colors were represented with fiber optic spectrometer and colorimeter. With the moisturizer composed of 10%PS ink and 30% ethylene glycol, the structural color could be rapidly prepared by only one-time printing when the substrate was at 45 ℃. The main reflection peak of curve was at 400 nm, the corresponding reflectivity was 83%, and the LCH was respectively 38.55, 25.16 and 272.8. Structural color can be quickly achieved via one-time printing of PS emulsion, and the disadvantages of traditional time-/material-consuming vertical deposition method were overcome. The preparation of structural color is featured by the such advantages as simplicity, unique coloration and excellent ageing resistance and it can be applied in packaging, anti-counterfeiting and other fields.