Friction has been considered to mediate physiological activities of cells, however, the biological friction between a single cell and its ligand-bound surface has not been thoroughly explored. Herein, we established a friction model for single cells based on an atomic force microscopy (AFM) combined with an inverted fluorescence microscopy (IFM) to study the friction between a highly sensitive platelet and fibrinogen-coated surface. The study revealed that the friction between the platelet and fibrinogen-coated tip is mainly influenced by specific ligand–receptor interaction. Further, we modeled the biological friction, which consists of specific interaction, non-specific interaction, and mechanical effect. Besides, the results suggested that the velocity can also affect specific ligand–receptor interactions, resulting in the friction change and platelet adhesion to fibrinogen surfaces. The study built a friction model between a single cell and its ligand-bound surface and provided a potential method to study the biological friction by the combination of AFM and IFM.