Prospects for Fluorescence Nanoscopy

Abstract

Overcoming Abbe's diffraction limit has been a challenging task and one of great interest for biological investigations. The emergence of fluorescence nanoscopy circumvents the diffraction barrier with nearly limitless power for optical microscopy, which enables investigations of the microscopic world in the 1-100 nm range. Proposed variants, such as expansion microscopy (ExM), stimulated emission depletion microscopy (STED), and Airyscan, are innovative in three aspects: sampling, illumination, and detection. These techniques show increasing strength in bioimaging subcellular structures. In this Perspective, we highlight advances in and prospects of fluorescence nanoscopy.

Resolution Enhancement and Background Suppression in Optical Super‐Resolution Imaging for Biological Applications

“…In our perspective, based on subtraction technique, detector array microscopy will be a fashion in subcellular research field because of its versatile setup, high acquisition speed, and relatively high spatial 3D resolution. [ 4 ] For example, MINFLUX is a promising approach for achieving nanoscale localization with minimal photon flux; however, for each molecular localization, it requires four illuminations, which greatly decreases the acquisition speed. [ 140 ] Using a detector array scheme, with only one illumination, the subtraction of fluorescence signals from an inner‐ring detector and an outer‐ring detector could achieve molecular localization.…”

“…Because the dimensions of sub‐cellular structures (like microfilament diameter), organelles (like lysosomes), virus, and proteins are normally beyond the resolving ability of conventional optics, the demand for a higher spatial resolution in optical microscopy has prompted the development of optical super‐resolution imaging techniques that are capable of breaking this optical barrier. [ 4 ]…”

Testing Anti-Biofilm Polymeric Surfaces: Where to Start?

“…The result is a resolution increased of two times in comparison to the optical microscopies [320]. Compared to other super-resolution microscopies, SIM is quite popular as standard fluorophores can be used, does not require special mounting and preparation techniques [321]. The technical details of these methods have been extensively reviewed by Li et al [321] and Sydor et al [322].…”

Optical Spectroscopy of Surfaces, Interfaces, and Thin Films: A Status Report

“…Reviews of other optical microscopy and imaging methods have also appeared. Li, Kuang, and Liu wrote a concise perspective describing novel innovations and prospects for fluorescence nanoscopy; notable is the ability of these imaging modes to overcome the diffraction limit that has plagued the spatial resolution of optical microscopy in the past …”