Biomolecules such as proteins, lipids, and other amphiphiles organise at soft interfaces to form functional biological assemblies, yet resolving this organisation across length scales remains methodologically challenging. This talk addresses that challenge at two complementary levels, first through mesoscale organization and subsequently at a molecular scale. The first part of the talk focuses on biomolecular interfaces constructed by stimuli-responsive liquid crystals (LCs). When confined in planar or spherical geometries in contact with an aqueous environment, the orientational order of thermotropic nematic LCs can amplify interfacial molecular events into easily measurable optical signals. The responsiveness of the LCs will be demonstrated to investigate biomolecular interactions ranging from protein aggregation to chemoresponsive drug release. The second part of the talk moves from mesoscale response to molecular level interrogation using vibrational sum frequency generation (SFG) spectroscopy applied to biologically relevant photoresponsive and membrane-mimetic interfaces. This approach resolves how molecular organization and conformation at these interfaces evolve during externally triggered processes. In particular, I will discuss the structural reorganisation of photoresponsive amphiphiles and polyelectrolyte–surfactant complexes accompanying light-triggered drug release at aqueous and lipid interfaces, establishing a correlation between interfacial molecular conformation and functional release behaviour. Together, combining functional soft materials with interface-specific spectroscopy can bridge molecular organization across length scales, providing mechanistic insight into emergent function.