Modern Methods of Fluorescence Nanoscopy in Biology

Abstract

Optical microscopy has undergone significant changes in recent decades due to the breaking of the diffraction limit of optical resolution and the development of high-resolution imaging techniques, which are collectively known as fluorescence nanoscopy. These techniques allow researchers to observe biological structures and processes at a nanoscale level of detail, revealing previously hidden features and aiding in answering fundamental biological questions. Among the advanced methods of fluorescent nanoscopy are: STED (Stimulated Emission Depletion Microscopy), STORM (STochastic Optical Reconstruction Microscopy), PALM (Photo-activated Localization Microscopy), TIRF (Total Internal Reflection Fluorescence), SIM (Structured Illumination Microscopy), MINFLUX (Minimal Photon Fluxes), PAINT (Points Accumulation for Imaging in Nanoscale Topography) и RESOLFT (REversible Saturable Optical Fluorescence Transitions) and others. In addition, most of these methods make it possible to obtain volumetric (3D) images of the objects under study. In this review, we will look at the principles of these methods, their advantages and disadvantages, and their application in biological researches.