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Nat Methods. 2010 May;7(5):377-81. doi: 10.1038/nmeth.1447. Epub 2010 Apr 4.

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

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  • 1Department of Micro- and Nanotechnology, Technical University of Denmark, Kongens Lyngby, Denmark.

Abstract

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.

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PMID:
20364147
[PubMed - indexed for MEDLINE]
PMCID:
PMC3127582
Free PMC Article

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