Chasing Uptake: Super-Resolution Microscopy in Endocytosis and Phagocytosis. Issue 9 (September 2019)

• Record Type: Journal Article
• Authors: Baranov, Maksim V.; Olea, Rodica Alis; van den Bogaart, Geert
• Is Part Of: Trends in cell biology. Volume 29:Issue 9(2019)
• Page Start: 727
• Page End: 739
• Publication Date: 2019-09
• Subjects: super-resolution microscopy -- trafficking -- organelles -- endocytosis -- endosomes; Cytology -- Periodicals; Cytology -- Research -- Periodicals; 571.6
• DOI: 10.1016/j.tcb.2019.05.006
• Languages: English
• ISSNs: 0962-8924

Abstract

Abstract : Since their invention about two decades ago, super-resolution microscopes have become a method of choice in cell biology. Owing to a spatial resolution below 50 nm, smaller than the size of most organelles, and an order of magnitude better than the diffraction limit of conventional light microscopes, super-resolution microscopy is a powerful technique for resolving intracellular trafficking. In this review we discuss discoveries in endocytosis and phagocytosis that have been made possible by super-resolution microscopy – from uptake at the plasma membrane, endocytic coat formation, and cytoskeletal rearrangements to endosomal maturation. The detailed visualization of the diverse molecular assemblies that mediate endocytic uptake will provide a better understanding of how cells ingest extracellular material. Highlights: Super-resolution microscopy is widely available to most cell biologists and has become the method of choice for studying endocytic and phagocytic trafficking. Current live imaging techniques allow 3D super-resolution microscopy with a temporal resolution of seconds to visualize uptake and trafficking processes at the whole-cell level. Quantification strategies combined with super-resolution microscopy have enabled a quantitative understanding of receptor clustering, the appearance of signaling lipids, and the dynamics of F-actin assembly on endosomes and phagosomes. Super-resolution microscopy is often combined with techniques such as electron microscopy and TIRF to enable more accurate localization of endocytic proteins in relation to stage of uptake, membrane curvature, and protein–protein interactions. Although generally of lower spatial resolution compared with other super-resolution microscopy techniques, structured illumination microscopy has at present yielded most new insights in endocytic trafficking because it combines improved spatial resolution with a reasonably high temporal resolution and can be readily used for live cell imaging applications.