The SPM could image the most detailed microstructure of a sample in a wet and dry state by measuring the interaction between the atoms on the sample surface and the extremely sharp probe tip. The refined fibre exhibited large wrinkles formed by fibrillar bundles, the disintegrated fibres extensively showed “scale-like features”. By using the Non-Contact Atomic Force Microscopy (NC-AFM) and Contact Atomic Force Microscopy (C-AFM) including Phase Detection Microscopy (PDM) and Force Modulation Microscopy (FMM), it was possible to investigate surface topography, surface roughness and mechanical property (hardness or visco-elasticity) of fibre surface in detail. The PDM and FMM images showed that the disintegrated only fibre displayed uniform mechanical properties, whereas the refined one did not. The surface roughness of pulp fibres was higher in refined fibres than in disintegrated fibres due to the presence of external fibrils. These SPM images would be used to provide visual evidence of morphological change of a single fibre created during mechanical treatments such as refining, drying, calendering and so on.