The invention discloses a method of producing polymer colloid crystal film on a hydrophobic substrate and belongs to the technical field of colloid crystal production. The method comprises the steps as follows: polymer colloid particles synthetized through emulsifier-free emulsion polymerization are self-assembled and grown on a hydrophobic substrate by utilizing traditional vertical deposition method; the two-dimension or the three-dimension polymer colloid crystal film of monodisperse polymer colloid particles on the surface of the substrate is obtained; the two-dimension or the three-dimension colloid crystal film is completely peeled off as a whole from the hydrophobic substrate by adopting a film peeling technology and floats on water surface; and the film can be freely transported to the surface of any hydrophobic substrate. The invention has low cost, high efficiency and simple technology, thereby greatly enlarging the application of the colloid crystal in the fields of photonic crystals, the micro-nano machining, micro optical lenses, the sensing, high density magnetic storage devices, etc.