High‐throughput single‐molecule localization microscopy: Potential clinical applications
Published in Clinical and Translational Medicine, 2023
Single-molecule localization microscopy (SMLM) has improved optical resolution to the range of several to tens of nanometres with high contrast, making it possible to noninvasively resolve nanoscale biological structures using fluorescence microscopy. 1 In SMLM (Figure 1A), small subsets of fluorescent molecules are randomly activated to ‘blink’due to the photon-switching property of fluorophores. Due to the diffraction of light, each blinking event will form an image of point spread function (PSF), typically with a width of∼ 200-300 nm. After collecting several thousands of frames and computationally localizing all these sparsely activated molecules using a mathematical PSF model, a pointillistic super-resolution image can be achieved, whose resolution is determined by both the localization accuracy and labelling density. Typically,∼ 20 nm resolution can be achieved by SMLM, which is an order of magnitude …