Computational Correction of Spatially-Variant Optical Aberrations in 3D Single Molecule Localization Microscopy

Abstract

3D single-molecule localization microscopy relies on fitting the shape of point-spread-functions (PSFs) recorded on a wide-field detector. However, optical aberrations distort those shapes, which compromise the accuracy and precision of single-molecule localization microscopy. Here we employ a computational phase retrieval based on a vectorial PSF model to quantify the spatially-variance of optical aberrations in a two-channel ultrawide-field single-molecule localization microscope. The use of a spatially-variant PSF model enables accurate and precise emitter localization in x, y- and z-directions throughout the entire field-of-view.\n\nOCIS Codes(100.6640) Superresolution; (180.2520) Fluorescence microscopy; (180.6900) Three-dimensional microscopy; (100.5070) Phase retrieval; (150.1135) Algorithms; (220.1010) Aberrations (global).