title: Fast and Precise 3D Fluorophore Localization based on Gradient Fitting.
abstract: Astigmatism imaging approach has been widely used to encode the fluorophore's 3D position in single-particle tracking and super-resolution localization microscopy. Here, we present a new high-speed localization algorithm based on gradient fitting to precisely decode the 3D subpixel position of the fluorophore. This algebraic algorithm determines the center of the fluorescent emitter by finding the position with the best-fit gradient direction distribution to the measured point spread function (PSF), and can retrieve the 3D subpixel position of the fluorophore in a single iteration. Through numerical simulation and experiments with mammalian cells, we demonstrate that our algorithm yields comparable localization precision to the traditional iterative Gaussian function fitting (GF) based method, while exhibits over two orders-of-magnitude faster execution speed. Our algorithm is a promising high-speed analyzing method for 3D particle tracking and super-resolution localization microscopy.
authors.affiliation: Biomedical and Optical Imaging Laboratory, Departments of Medicine and Bioengineering, University of Pittsburgh, Pittsburgh PA 15213, USA.
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Ma
authors.affiliation: Biomedical and Optical Imaging Laboratory, Departments of Medicine and Bioengineering, University of Pittsburgh, Pittsburgh PA 15213, USA.
authors.forename: Jianquan
authors.initials: J
authors.lastname: Xu
authors.affiliation: Biomedical and Optical Imaging Laboratory, Departments of Medicine and Bioengineering, University of Pittsburgh, Pittsburgh PA 15213, USA.|School of Medicine, Tsinghua University, China.
authors.forename: Jingyi
authors.initials: J
authors.lastname: Jin
authors.affiliation: School of Medicine, Tsinghua University, China.|Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh PA 15213, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh PA 15213, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Lan
authors.affiliation: Biomedical and Optical Imaging Laboratory, Departments of Medicine and Bioengineering, University of Pittsburgh, Pittsburgh PA 15213, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000465:Algorithms; D000818:Animals; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D008870:Microtubules / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes