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 microscopes. 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.