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Nat Methods. 2014 Mar;11(3):253-66. doi: 10.1038/nmeth.2843.

Precisely and accurately localizing single emitters in fluorescence microscopy.

Author information

1
1] Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ghent, Belgium. [2] Center for Nano- and Biophotonics, Ghent University, Ghent, Belgium.
2
1] Department of Nanophysics, Italian Institute of Technology, Genoa, Italy. [2] Department of Physics, University of Genoa, Genoa, Italy.
3
Department of Physics and Astronomy, University of Maine, Orono, Maine, USA.
4
1] Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA. [2] Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.

Abstract

Methods based on single-molecule localization and photophysics have brought nanoscale imaging with visible light into reach. This has enabled single-particle tracking applications for studying the dynamics of molecules and nanoparticles and contributed to the recent revolution in super-resolution localization microscopy techniques. Crucial to the optimization of such methods are the precision and accuracy with which single fluorophores and nanoparticles can be localized. We present a lucid synthesis of the developments on this localization precision and accuracy and their practical implications in order to guide the increasing number of researchers using single-particle tracking and super-resolution localization microscopy.

PMID:
24577276
DOI:
10.1038/nmeth.2843
[Indexed for MEDLINE]
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