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- Title
Optimized localization analysis for single-molecule tracking and super-resolution microscopy.
- Authors
Mortensen, Kim I.; Churchman, L. Stirling; Spudich, James A.; Flyvbjerg, Henrik
- 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.
- Subjects
MICROSCOPY; OPTICAL diffraction; LEAST squares; PHOTONS; POLAR coordinates (Mathematics); MAGNETIC dipoles; POLYSTYRENE
- Publication
Nature Methods, 2010, Vol 7, Issue 5, p377
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth.1447