High-speed atomic force microscopy of membrane proteins: From dynamic imaging to integrative structural biology

High-speed atomic force microscopy (HS-AFM) enables direct, label-free visualization of membrane proteins in lipid bilayers with nanometer spatial resolution and millisecond temporal resolution. Recent studies revealed transient conformational states, dynamic oligomerization changes, and membrane-mediated organization of membrane proteins that are not readily captured by conventional structural or biophysical approaches. Advances in experimental design-including membrane reconstitution strategies, stimulus-coupled imaging, high-temporal-resolution acquisition, and computational reconstruction-have further expanded the scope of HS-AFM. When integrated with complementary techniques such as cryo-electron microscopy, molecular dynamics simulations, and single-molecule methods, HS-AFM provides a powerful and versatile tool for linking membrane protein structure, dynamics, and function in native-like environments.