Single‐molecule force spectroscopy of biomolecules using AFM. A multigenerational perspective

Single-molecule force spectroscopy (SMFS) using atomic force microscopy (AFM) allows the direct application and measurement of forces on individual molecular systems; revolutionizing our understanding of biomolecular mechanics and receptor-ligand interactions and opening the field of single-molecule mechanobiology. This review traces the evolution of the field, from foundational landmark experiments to recent developments that allowed revisiting those seminal works. We discuss the critical technological advancements, functionalisation strategies and theoretical developments that have shaped the discipline. Finally, we suggest future directions to further elucidate the intricate relationship between structure and mechanical properties of single biomolecules.