Journal
ANNUAL REVIEW OF BIOPHYSICS
Volume 51, Issue -, Pages 301-326Publisher
ANNUAL REVIEWS
DOI: 10.1146/annurev-biophys-102521-112912
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This review introduces super-resolution microscopy techniques, focusing on SMLM and MINFLUX, and summarizes recent technical developments. It also outlines key experimental conditions and analysis methods, highlighting the application of SMLM in studying the structures of biologically relevant molecular machines.
Super-resolution microscopy techniques, and specifically single-molecule localization microscopy (SMLM), are approaching nanometer resolution inside cells and thus have great potential to complement structural biology techniques such as electron microscopy for structural cell biology. In this review, we introduce the different flavors of super-resolution microscopy, with a special emphasis on SMLM and MINFLUX (minimal photon flux). We summarize recent technical developments that pushed these localization-based techniques to structural scales and review the experimental conditions that are key to obtaining data of the highest quality. Furthermore, we give an overview of different analysis methods and highlight studies that used SMLM to gain structural insights into biologically relevant molecular machines. Ultimately, we give our perspective on what is needed to push the resolution of these techniques even further and to apply them to investigating dynamic structural rearrangements in living cells.
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