For the past twenty years, significant progress has been made in both developing cryogenic transmission electron microscopy (cryo-TEM) technology and understanding assembled behavior of amphiphilic molecules. Cryo-TEM can provide high-resolution images of complex fluids in a near in situ state. Samples embedded in a thin layer of vitrified solvent do not exhibit artifacts that would normally occur when using chemical fixation or staining-and-drying techniques. Cryo-TEM has been useful in imaging biological molecules in aqueous solutions. Cryo-TEM has become a powerful tool in the study of in situ-assembled structures of amphiphiles in solution as a complementary tool to small-angle X-ray and neutron scattering, light scattering, rheology measurements, and nuclear magnetic resonance. The application of cryo-TEM in the study of assembled behavior of amphiphilic block copolymers, hydrogels, and other complex soft systems continues to emerge. In this context, the usage of cryo-TEM in the field of amphiphilic complex fluids and self-assembled nano-materials is briefly reviewed, and its unique role in exploring the nature of assembled structure in liquid suspension is highlighted.