Biomimetic nanoparticles are hybrid nanostructures in which the uppermost layer is similar to a cell membrane. This report provides instructions for the preparation and physical characterization of three different types of biomimetic nanoparticles: (1) polystyrene sulfate nanoparticles covered with one cationic dioctadecyldimethylammonium bilayer; (2) silica nanoparticles covered with one neutral phosphatidylcholine bilayer; (3) miconazole particles covered with one anionic dihexadecylphosphate (DHP) bilayer. These examples provide the basis for the preparation and characterization of novel nanoparticles from hydrophobic or hydrophilic and organic or inorganic nanoparticle cores covered with functional outer layers. The major concepts and technical details for obtaining the optimal lipid coverage of supporting cores and for nanoparticle characterization are discussed.