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? 2000 Nature America Inc. ? review Observing single biomolecules at work with the atomic force microscope Andreas Engel1 and Daniel J. Müller1,2 Progress in the application of the atomic force microscope (AFM) to imaging and manipulating biomolecules is the result of improved instrumentation, sample preparation methods and image acquisition conditions. Biological membranes can be imaged in their native state at a lateral resolution of 0.5–1 nm and a vertical resolution of 0.1–0.2 nm. Conformational changes that are related to functions can be resolved to a similar resolution, complementing atomic structure data acquired by other methods. The unique capability of the AFM to directly observe single proteins in their native environments provides insights into the interactions of proteins that form functional assemblies. In addition, single molecule force spectroscopy combined with single molecule imaging provides unprecedented possibilities for analyzing intramolecular and intermolecular forces. This review discusses recent examples that illustrate the power of AFM. .com To observe biomolecules at work they must reside in their native environments. For soluble proteins this is a physiological buffer. Membrane proteins, in addition, need to be embedded in a lipid bilayer. The only instrument that can image samples at sub- nanometer resolutions and be operated in solution is the atomic force microscope (AFM)1. This instrument uses a sharp stylus at the end of a flexible cantilever to scan over the sample surface. Cantilever deflections measured at a resolution of a few angstroms http://structbio.nature ? are used to determine the surface contour of the sample, exploiting a sensitive feedback to minimi