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This lecture covers the principles and applications of Atomic Force Microscopy (AFM) in mechanobiology, focusing on how mechanics regulate life. It explains the essential components of AFM, including the cantilever, substrate, and piezo-based tip scanning setup. The lecture discusses the z-position feedback loop for imaging sample surfaces, cantilever properties, force curve-based sample characterization, force spectroscopy for cell-cell adhesion, imaging modes like contact and dynamic mode, high-resolution imaging techniques, key inventions in AFM, imaging examples, cantilever-based buoyant mass measurements, fluid force microscopy, biophysical aspects of cell separation, and long-term AFM imaging of mycobacteria. It concludes by highlighting the versatility of AFM in various biological applications.