Simultaneous AFM topography and recognition imaging at the plasma membrane of mammalian cells

Elucidation the nano-organization of membrane proteins at/within the plasma membrane is probably themost demanding and still challenging task in cell biology since requires experimental approaches withnanoscale resolution. During last decade, atomic force microscopy (AFM)-based simultaneous topographyand recognition imaging (TREC) has become a powerful tool to quickly obtain local receptor nano-maps oncomplex heterogeneous biosurfaces such as cells and membranes. Here we emphasize the TREC techniqueand explain how to unravel the nano-landscape of mammalian cells. We describe the procedures for allsteps of the experiment including tip functionalization with ligand molecules, sample preparation, andlocalization of key molecules on the cell surface. We also discuss the current limitations and futureperspectives of this technique.