High-speed atomic force microscopy of membrane and membrane protein dynamics

High-speed atomic force microscopy (HS-AFM) enables direct nanometer-resolution visualization of single molecules and molecular assemblies in real-time and under physiological conditions, providing unique insights into how membranes and membrane proteins move and interact within native lipid environments. Recent methodological advances and integration with complementary techniques have extended HS-AFM to increasingly complex, physiologically relevant systems, bridging gaps between high-resolution static structural methods and low-resolution functional dynamics. Here, we highlight how HS-AFM has changed our understanding of membrane organization, protein conformational dynamics, and lipid–protein coupling. By capturing transient events inaccessible to ensemble approaches, HS-AFM is transforming our ability to connect structural snapshots with functional behavior, advancing dynamic structural biology.