Cross-cumulant enhanced radiality nanoscopy for multicolor superresolution subcellular imaging

Abstract

Fluorescence fluctuation-based superresolution techniques can achieve fast superresolution imaging on a cost-effective wide-field platform at a low light level with reduced phototoxicity. However, the current methods exhibit certain imaging deficiencies that misinterpret nanoscale features reconstructed from fluctuating image sequences, thus degrading the superresolution imaging quality and performance. Here we propose cross-cumulant enhanced radiality nanoscopy (CERN), which employs cross-cumulant analysis in tandem with radiality processing. We demonstrated that CERN can significantly improve the spatial resolution at a low light level while eliminating the misinterpretations of nanoscale features of the existing fluctuation-based superresolution methods. In the experiment, we further verified the superior performance of CERN over the current methods through performing multicolor superresolution imaging of subcellular microtubule networks and clathrin-coated pits as well as the high-precision reconstruction of densely packed RNA transcripts.