In this work, we applied total internal reflection microscopy (TIRM) to directly measure the interactions between three different kinds of macroscopic surfaces: namely bare polystyrene (PS) particle and bare silica surface (bare-PS/bare-silica), PS particle and silica surfaces both coated with bovine serum albumin (BSA) (BSA-PS/BSA-silica), and PS particle and silica surfaces both modified with polyethylene glycol (PEG) (PEG-PS/PEG-silica) polymers, in phosphate buffer solution (PBS) and fetal bovine serum (FBS). Our results showed that in PBS, all the bare-PS, BSA-PS, and PEG-PS particles were irreversibly deposited onto the bare silica surface or surfaces coated either with BSA or PEG. However, in FBS, the interaction potentials between the particle and surface exhibited both free-diffusing particle and stuck particle profiles. Dynamic light scattering (DLS) and elliposmeter measurements indicated that there was a layer of serum proteins adsorbed on the PS particle and silica surface. TIRM measurement revealed that such adsorbed serum proteins can mediate the surface-surface interactions by providing additional stabilization under certain conditions, but also promoting bridging effect between the two surfaces. The measured potential profile of the stuck particle in FBS thus was much wider than in PBS. These quantitative measurements provide insights that serum proteins adsorbed onto surfaces can regulate surface-surface interactions, thus leading to unique moving behavior and stability of colloidal particles in the serum environment.