Lateral diffusion of single chain related to the crystallization process of poly(ethylene oxide) (PEO) in its monolayers on silica surfaces is studied by single molecule fluorescence microscopy and single molecule tracking techniques. Diffusion of PEO chains is observed in the supercooled state before crystallization as well as in the noncrystalline regions alter crystallization. lithe postcrystallization monolayers, the diffusion coefficient of PEO chains is an order of magnitude lower than that in the supercooled state before crystallization. The origin is attributed to the change of polymer surface concentration due to the consumption of polymer molecules in the crystal formation. This is supported by the results showing a monotonous decrease of diffusion coefficient with the thickness decrease of the monolayer in its supercooled state. The PEO chains take a more flattened conformation under lower surface concentration and stick stronger to the surface. As a consequence, the diffusion rate is reduced. The results clearly demonstrate a strong mutual effect between the crystallization process and the mass transportation for the polymer crystallization under surface confinement.