Optimized localization analysis for single-molecule tracking and super-resolution microscopy

Kim Mortensen, L. S. Churchman, J. A. Spudich, Henrik Flyvbjerg

    Research output: Contribution to journalJournal articleResearchpeer-review


    We optimally localized isolated fluorescent beads and molecules imaged as diffraction-limited spots, determined the orientation of molecules and present reliable formulas for the precision of various localization methods. Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula for its precision exaggerates beyond Fisher's information limit, and weighted least-squares may do worse, whereas maximum-likelihood fitting is practically optimal.
    Original languageEnglish
    JournalNature Methods
    Issue number5
    Pages (from-to)377-U59
    Publication statusPublished - 2010

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