Surfactants containing fluorocarbon chains have been drawing increasing attentions, because they self-assemble into a variety of microscopic and mesoscopic domains. They tend to form highly ordered patterns at the air/water interface, which are clearly different from those formed by their hydrocarbon analogues. Focusing on fluorinated surfactants possessing unique physical characteristics, this review aims to give an overview how the interplay of line tension and dipole interaction, the comprehensive principle, governs the pattern formation of two-dimensional selfassemblies. This review also covers several key experimental and analytical techniques that are useful for the characterization of the shape, size, correlation, and viscoelasticity of hierarchical self-assemblies on water surfaces. Finally, several biomedical applications, including biomimetic surface coating, multi-modal contrast agents in medical diagnostics, and controlled delivery of