Physicochemical properties such as cell morphology, cell elasticity and cell adhesion of live cells are different when cells are under different physiological conditions. Therefore, these physicochemical properties can reflect complex physiological processes occurred in cells. Atomic force microscope (AFM) has super spacious imaging resolution and high force resolution (pico-Newton) in mechanical measurements, and can work in near-physiological conditions to observe ultrastructure of cell surface and measure cytomechanics. Consequently, AFM is widely regarded as an ideal tool to investigate physicochemical properties of cells. In this review, principles, traditional operation modes and applications of traditional AFM in the study of cell physicochemical properties are briefly introduced. Particular emphasis is given on the applications of newly-developed AFM techniques including imaging and cytomechanics in the study of cell physicochemical properties. © 2016, Editorial Board of “Journal of Chemical Engineering of Chinese Universities”. All right reserved.