title: Single particle maximum likelihood reconstruction from superresolution microscopy images.
abstract: Point localization superresolution microscopy enables fluorescently tagged molecules to be imaged beyond the optical diffraction limit, reaching single molecule localization precisions down to a few nanometers. For small objects whose sizes are few times this precision, localization uncertainty prevents the straightforward extraction of a structural model from the reconstructed images. We demonstrate in the present work that this limitation can be overcome at the single particle level, requiring no particle averaging, by using a maximum likelihood reconstruction (MLR) method perfectly suited to the stochastic nature of such superresolution imaging. We validate this method by extracting structural information from both simulated and experimental PALM data of immature virus-like particles of the Human Immunodeficiency Virus (HIV-1). MLR allows us to measure the radii of individual viruses with precision of a few nanometers and confirms the incomplete closure of the viral protein lattice. The quantitative results of our analysis are consistent with previous cryoelectron microscopy characterizations. Our study establishes the framework for a method that can be broadly applied to PALM data to determine the structural parameters for an existing structural model, and is particularly well suited to heterogeneous features due to its single particle implementation.
authors.affiliation: Univ Lyon, Ens de Lyon, Univ Claude Bernard, CNRS, Laboratoire de Physique, F-69342 Lyon, France.
authors.forename: Timothée
authors.initials: T
authors.lastname: Verdier
authors.affiliation: Laboratory for Experimental Biophysics, School of Basic Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.
authors.forename: Julia
authors.initials: J
authors.lastname: Gunzenhauser
authors.affiliation: Laboratory for Experimental Biophysics, School of Basic Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
authors.affiliation: Univ Lyon, Ens de Lyon, Univ Claude Bernard, CNRS, Laboratoire de Physique, F-69342 Lyon, France.
authors.forename: Martin
authors.initials: M
authors.lastname: Castelnovo
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D015497:HIV-1 / Q000648:ultrastructure; D016013:Likelihood Functions; D008853:Microscopy / Q000379:methods*; D013269:Stochastic Processes; D014771:Virion / Q000648:ultrastructure