A Perspective on Data Processing in Super-resolution Fluorescence Microscopy Imaging

With super-resolution microscopy,we attempt to visualize(biological)structures and processes at the sub-cellular level(i.e.,nanoscale).To obtain this information,the samples are labeled with fluorophores that have a stochastic on/off switching of their emissions,which help to overcome the optical diffraction limit of around 250 nm,related to the use of optical micro-scopes.However,nowadays,research focuses on the imaging of live cells and thicker samples.These investigations require a high amount of simultaneously active fluorophores(i.e.,high-density imaging)and are challenging due to the collapse of the single-molecule localization techniques and the increased background in the image.Therefore,recent efforts have shifted towards the development of new ways to process the data.This publication gives an introduction to wide-field super-resolution fluorescence microscopy,explaining the concepts of the technique,and then gives an overview of the recently developed methods to provide super-resolution images for high-density data of live cells and ways to overcome the issues related to the imaging of these samples.