Fluorinated lipids and surfactants are attractive biomimetic materials for the extraction and reorganization of membrane proteins because of the biological inertness of fluorocarbons. We investigated the fundamental physical properties of a partially fluorinated phospholipid (F4-DMPC), such as phase transition, area thermal expansion, and lateral lipid diffusion, to evaluate the intermolecular interaction of F4-DMPC in the hydrophobic region quantitatively on the basis of free-volume theory. Fluorescence microscope observation of the supported lipid bilayer (SLB) of F4-DMPC showed that the phase transition between the liquid crystalline and gel phases occurred at 5 degrees C and that the area thermal expansion coefficient was independent of the temperature near the phase transition temperature. We performed a single particle tracking of the F4-DMPC-SLB on a SiO2/Si substrate, to measure the diffusion coefficient and its temperature dependence. The apparent activation energy (E-a') of lateral lipid diffusion, which is an indicator of intermolecular interaction, was 39.1 kJ/mol for F4-DMPC, and 48.2 kJ/mol for a nonfluorinated 1,2-dioleoyl-sn-glycero-3-phosphocholine as a control. The difference of 9 kJ/mol in E-a' was significant compared with the difference due to the acyl chain species among nonfluorinated phosphatidylcholine and also that caused by the addition of cholesterol and alcohol in the bilayer membranes. We quantitatively evaluated the attenuation of intermolecular interaction, which results from the competition between the dipole-induced packing effect and steric effect at the fluorocarbon segment in F4-DMPC.