title: Quantitative evaluation of software packages for single-molecule localization microscopy.
abstract: The quality of super-resolution images obtained by single-molecule localization microscopy (SMLM) depends largely on the software used to detect and accurately localize point sources. In this work, we focus on the computational aspects of super-resolution microscopy and present a comprehensive evaluation of localization software packages. Our philosophy is to evaluate each package as a whole, thus maintaining the integrity of the software. We prepared synthetic data that represent three-dimensional structures modeled after biological components, taking excitation parameters, noise sources, point-spread functions and pixelation into account. We then asked developers to run their software on our data; most responded favorably, allowing us to present a broad picture of the methods available. We evaluated their results using quantitative and user-interpretable criteria: detection rate, accuracy, quality of image reconstruction, resolution, software usability and computational resources. These metrics reflect the various tradeoffs of SMLM software packages and help users to choose the software that fits their needs.
authors.affiliation: Biomedical Imaging Group, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Sage
authors.affiliation: Biomedical Imaging Group, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
authors.forename: Hagai
authors.initials: H
authors.lastname: Kirshner
authors.affiliation: Center for Genomic Regulation, Barcelona, Spain.
authors.forename: Thomas
authors.identifier: 000000029632918X
authors.initials: T
authors.lastname: Pengo
authors.affiliation: 1] Howard Hughes Medical Institute, University of California (UCSF), San Francisco, California, USA. [2] Department of Cellular and Molecular Pharmacology, UCSF, San Francisco, California, USA.
authors.forename: Nico
authors.identifier: 0000000261798613
authors.initials: N
authors.lastname: Stuurman
authors.affiliation: Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.
authors.forename: Junhong
authors.initials: J
authors.lastname: Min
authors.affiliation: Laboratory of Experimental Biophysics, EPFL, Lausanne, Switzerland.
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
authors.affiliation: Biomedical Imaging Group, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Unser
mesh_terms: D000465:Algorithms; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D019295:Computational Biology / Q000379:methods; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000737:chemistry; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods; D008853:Microscopy / Q000379:methods*; D008870:Microtubules / Q000737:chemistry; D015203:Reproducibility of Results; D012984:Software; D013269:Stochastic Processes; D014404:Tubulin / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D014404:Tubulin