Two new ways of cellulose nanocrystal（CNC）film fabrication and characterization of the films by infrared reflection absorption spectroscopy（IRRAS）and atomic force microscopy（AFM）were discussed in this paper.CNC films can be formed via electrostatic driven physisorption to an amine-terminated self-assembled monolayer（SAM）on evaporated gold substrate either by immersion in CNC suspensions or through a spin-coating process.AFM can not only characterize the surface topography of non-conductive CNC films and analyze the roughness of the surface，but also reach accurate film thickness through scratching technique.AFM height images showed that CNC films are well packed with rod-like CNC nanoparticles.Through at least three times of experiments and accurate statistical analysis，it is known that spin-coated CNC films，with a roughness RMS of approximately 2.7 nm，are slightly smoother than immersed CNC films.According to the AFM scratching results，the thickness of the immersed CNC film is maximum 15 nm due to the repulsion between particles，while spin coating process can make much thicker CNC films，with a thickness of over 50 nm.The relationship between CNC suspension concentrations and spin-coated CNC film thickness was investigated.It was found that the thickness can be well controlled by the CNC solution concentration.The IRRAS results also confirmed that the spin-coated CNC film thickness increases with the increase of CNC suspension concentrations.The successful fabrication of CNC films provided a high-surface area substrate using green materials for studying surface chemistry，and could be used in pharmaceuticals，drug delivery，biosensors，solar energy battery，and other fields.