Fluorescence microscopy, especially confocal microscopy, has revolutionized the field of biological imaging. Breaking theoptical diffraction barrier of conventional light microscopy, through the advent of super-resolution microscopy, has usheredin the potential for a second revolution through unprecedented insight into nanoscale structure and dynamics in biologicalsystems. Stimulated emission depletion (STED) microscopy is one such super-resolution microscopy technique whichprovides real-time enhanced-resolution imaging capabilities. In addition, it can be easily integrated with well-establishedfluorescence-based techniques such as fluorescence correlation spectroscopy (FCS) in order to capture the structure ofcellular membranes at the nanoscale with high temporal resolution. In this review, we discuss the theory of STED anddifferent modalities of operation in order to achieve the best resolution. Various applications of this technique in cellimaging, especially that of neuronal cell imaging, are discussed as well as examples of application of STED imaging inunravelling structure formation on biological membranes. Finally, we have discussed examples from some of our recentstudies on nanoscale structure and dynamics of lipids in model membranes, due to interaction with proteins, as revealed bycombination of STED and FCS techniques.
Volume 48, 2023
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