Optimized localization analysis for single-molecule tracking and super-resolution microscopy (original) (raw)
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- Published: 04 April 2010
Nature Methods volume 7, pages 377–381 (2010)Cite this article
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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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References
- Born, M. & Wolf, E. Principles of Optics (Cambridge University Press, New York, 1999).
- Barak, L.S. & Webb, W.W. J. Cell Biol. 95, 846–852 (1982).
Article CAS Google Scholar - Yildiz, A. et al. Science 300, 2061–2065 (2003).
Article CAS Google Scholar - Okten, Z., Churchman, L.S., Rock, R.S. & Spudich, J.A. Nat. Struct. Mol. Biol. 11, 884–887 (2004).
Article Google Scholar - Betzig, E. et al. Science 313, 1642–1645 (2006).
Article CAS Google Scholar - Moerner, W.E. Proc. Natl. Acad. Sci. USA 104, 12596–12602 (2007).
Article CAS Google Scholar - Abraham, A.V., Ram, S., Chao, J., Ward, E.S. & Ober, R.J. Opt. Express 17, 23352–23373 (2010).
Article Google Scholar - Rao, C.R. Linear Statistical Inference and Its Applications (Wiley, New York, New York, 1973).
- Ober, R.J., Ram, S. & Ward, E.S. Biophys. J. 86, 1185–1200 (2004).
Article CAS Google Scholar - Robbins, M.S. & Hadwen, B.J. IEEE Trans. Electron. Dev. 50, 1227–1232 (2003).
Article Google Scholar - Enderlein, J., Toprak, E. & Selvin, P. Opt. Express 14, 8111–8120 (2006).
Article CAS Google Scholar - Forkey, J.N., Quinlan, M.E., Shaw, M.A., Corrie, J.E.T. & Goldman, Y.E. Nature 422, 399–404 (2003).
Article CAS Google Scholar - Toprak, E. et al. Proc. Natl. Acad. Sci. USA 103, 6495–6499 (2006).
Article CAS Google Scholar - Aguet, F., Geissbühler, S., Märki, I., Lasser, T. & Unser, M. Opt. Express 17, 6829–6848 (2009).
Article CAS Google Scholar - Toprak, E. & Selvin, P.R. Annu. Rev. Biophys. Biomol. Struct. 36, 349–369 (2007).
Article CAS Google Scholar - Thompson, R.E., Larson, D.R. & Webb, W.W. Biophys. J. 82, 2775–2783 (2002).
Article CAS Google Scholar - Bobroff, N. Rev. Sci. Instrum. 57, 1152–1157 (1986).
Article Google Scholar - Carter, A.R. et al. Appl. Opt. 46, 421–427 (2007).
Article Google Scholar - Berg-Sørensen, K. & Flyvbjerg, H. Rev. Sci. Instrum. 75, 594–612 (2004).
Article Google Scholar - Axelrod, D., Burghardt, T.P. & Thompson, N.L. Annu. Rev. Biophys. Bioeng. 13, 247–268 (1984).
Article CAS Google Scholar - Ulbrich, M.H. & Isacoff, E.Y. Nat. Methods 4, 319–321 (2007).
Article CAS Google Scholar - Churchman, L.S., Flyvbjerg, H. & Spudich, J.A. Biophys. J. 90, 668–671 (2006).
Article CAS Google Scholar
Acknowledgements
We thank S.M. Block, W.E. Moerner and R.S. Rock for discussions; Z.D. Bryant for allowing us to use his microscope for some of the data collection and M.W. Elting and J.M. Sung for assisting us. This work was supported by the European Union (FP7-HEALTH-F4-2008-201418, Revolutionary Approaches and Devices for Nucleic Acid Analysis to H.F.), by the US National Institutes of Health (GM33289 to L.S.C. and J.A.S.), by the Human Frontier Science Program (GP0054/2009-C to J.A.S. and H.F.) and the Damon Runyon Cancer Research Foundation (DRG-1997-08 to L.S.C.).
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Author notes
- Kim I Mortensen and L Stirling Churchman: These authors contributed equally to this work.
Authors and Affiliations
- Department of Micro- and Nanotechnology, Technical University of Denmark, Kongens Lyngby, Denmark
Kim I Mortensen & Henrik Flyvbjerg - Department of Biochemistry, Stanford University School of Medicine, Stanford, California, USA
L Stirling Churchman & James A Spudich - Department of Physics, Stanford University School of Medicine, Stanford, California, USA
L Stirling Churchman
Authors
- Kim I Mortensen
You can also search for this author inPubMed Google Scholar - L Stirling Churchman
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Contributions
H.F., K.I.M. and L.S.C. designed research; K.I.M. and H.F. performed the theoretical calculations and analyzed data; J.A.S. supervised the experiments; L.S.C. conducted experiments; K.I.M. did numerical simulations; H.F., K.I.M., L.S.C. and J.A.S. wrote the paper.
Corresponding author
Correspondence toHenrik Flyvbjerg.
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The authors declare no competing financial interests.
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Mortensen, K., Churchman, L., Spudich, J. et al. Optimized localization analysis for single-molecule tracking and super-resolution microscopy.Nat Methods 7, 377–381 (2010). https://doi.org/10.1038/nmeth.1447
- Received: 14 October 2009
- Accepted: 22 February 2010
- Published: 04 April 2010
- Issue Date: May 2010
- DOI: https://doi.org/10.1038/nmeth.1447