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 pointspread-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 singlemolecule 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.