Abstract: The dynamics of polystyrene (PS) microsphere in thin layers of meniscus self-assembly 2D colloidal crystals was studied systemically. The mechanism of void and multilayer flaw was analyzed based on the model of assembly. A large scale, ordered monolayer of polystyrene sub-microsphere with a diameter of 300 nm was obtained by dip coating on glass surface. The substrate withdrawal velocity and the colloidal suspension concentration were studied for large scale 2D colloidal crystals in this work. Large domain 2D colloidal crystals could be rapidly fabricated from high concentration, reducing influence of environment fluctuation. By the experimental result and theoretical relationship between withdrawal velocity and colloidal concentration to synthetize 2D colloidal crystals, the experiment result is consistent with Nagayama’s model when particles concentration is smaller than 8%.