Self-supervised denoising for multimodal structured illumination microscopy enables long-term super-resolution live-cell imaging

Detection noise significantly degrades the quality of structured illumination microscopy (SIM) images, especially under low-light conditions. Although supervised learning based denoising methods have shown prominent advances in eliminating the noise-induced artifacts, the requirement of a large amount of high-quality training data severely limits their applications. Here we developed a pixel-realignment-based self-supervised denoising framework for SIM (PRS-SIM) that trains an SIM image denoiser with only noisy data and substantially removes the reconstruction artifacts. We demonstrated that PRS-SIM generates artifact-free images with 20-fold less fluorescence than ordinary imaging conditions while achieving comparable super-resolution capability to the ground truth (GT). Moreover, we developed an easy-to-use plugin that enables both training and implementation of PRS-SIM for multimodal SIM platforms including 2D/3D and linear/nonlinear SIM. With PRS-SIM, we achieved long-term super-resolution live-cell imaging of various vulnerable bioprocesses, revealing the clustered distribution of Clathrin-coated pits and detailed interaction dynamics of multiple organelles and the cytoskeleton.