ABSTRACTProtein adsorption is the first phenomenon which occurs at nanoscale level when a given surface came into contact with a living fluid cell such as blood. Investigation of this adsorption at nanoscale provides useful information about kinetics and mechanism of conformation of proteins on a given surface. The present study investigates the adsorption of proteins using tapping/intermittent mode atomic force microscopy (T-AFM). The approach taken here is that hydrogenated amorphous carbon coating (a-C:H) is used as a model surface because it is amorphous, smooth, inert and hydrophobic. Two proteins namely albumin and fibrinogen in phosphate buffer (PBS) and de-ionized water are drop casted to study the adsorption kinetics. First and second resonance AFM data was used to investigate the adsorbed layer of proteins. AFM force curve and scratch experiment were used to verify the adhesion and thickness of the adsorbed layer. Combination of height, phase images along with the AFM force curve and scratch experiment shows inhomogeneous distribution of albumin protein in phosphate buffer compared to other protein solutions.