Breaking the Diffraction Barrier: Super-Resolution Imaging of Cells

Abstract

Anyone who has used a light microscope has wished that its resolution could be a little better. Now, after centuries of gradual improvements, fluorescence microscopy has made a quantum leap in its resolving power due, in large part, to advancements over the past several years in a new area of research called super-resolution fluorescence microscopy. In this Primer, we explain the principles of various super-resolution approaches, such as STED, (S)SIM, and STORM/(F)PALM. Then, we describe recent applications of super-resolution microscopy in cells, which demonstrate how these approaches are beginning to provide new insights into cell biology, microbiology, and neurobiology.

Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate

“…These probes can be activated in sparse numbers over time such that their images are optically resolvable and can be precisely localized 4 . These techniques have been exploited to image sub-cellular structures [5][6][7] , visualize protein (co)-organization [8][9][10] and quantify protein stoichiometry [11][12][13][14][15] .…”

Variation in Carbohydrates between Cancer and Normal Cell Membranes Revealed by Super‐Resolution Fluorescence Imaging

“…Herein, in addition to common monosaccharides mentioned above, we also selected one common oligosaccharide (pentasaccharide sequence Galβ1‐4GlcNAcβ1‐2 (Galβ1‐4GlcNAcβ1‐6) Manα1‐R) as the investigated target, which is a complement and comparison to monosaccharides, as well as confirms that the results of monosaccharides can reflect ones of carbohydrate‐chains. Through using super‐resolution microscopy with a nanometer‐level resolution,17, 18, 19 we studied the changed distributions of these selected carbohydrates on cell membranes, showing an overall alterant distribution of carbohydrate‐related molecules (glycoproteins, glycolipids, and glycosaminoglycans) containing the same carbohydrate residues, as well as suggesting the changed organization of cell membrane. Herein, we revealed the distributed alterations of carbohydrates on cancer cell (both cultured and primary cells) membranes compared with normal cell (both cultured and primary cells) membranes.…”

Label-free super-resolution imaging of adenoviruses by submerged microsphere optical nanoscopy

“…Fluorescence optical microscopy is a recently established method for the imaging of cellular structures, bacteria and viruses beyond the optical diffraction limit, down to a resolution of 6 nm. [1][2][3][4] This technique is based on the detection of light emitted by the fluorescing specimen when it is excited by light of a specific wavelength. Structured illumination, such as stimulated emission depletion and saturated structured illumination microscopy, which activate florescent light emission from a group of molecules simultaneously, are typically used.…”