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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 7, Iss. 3 — Feb. 29, 2012

Single-image axial localization precision analysis for individual fluorophores

Michael C. DeSantis, Shannon Kian Zareh, Xianglu Li, Robert E. Blankenship, and Y. M. Wang  »View Author Affiliations


Optics Express, Vol. 20, Issue 3, pp. 3057-3065 (2012)
http://dx.doi.org/10.1364/OE.20.003057


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Abstract

Bio-mechanism investigations demand single particle tracking with high spatial and temporal resolutions which require single fluorophore 3D localization measurements with matching precision and speed. Although the precision for lateral-localization measurements is well described by an analytical expression, for the axial direction, it is often obtained by repeating location measurements or by estimating a lower bound. Here, we report a precision expression for an axial-localization method that analyzes the standard deviations of single fluorophores’ intensity profiles. Like the lateral-localization precision, this expression includes all relevant experimental effects measurable from a Gaussian intensity profile of the fluorophore. This expression completes the precision analysis for single-image 3D localization of individual fluorophores and lifts the temporal resolution to the typical exposure timescales of milliseconds.

© 2012 OSA

OCIS Codes
(100.6640) Image processing : Superresolution
(100.6890) Image processing : Three-dimensional image processing
(110.2960) Imaging systems : Image analysis
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Microscopy

History
Original Manuscript: September 6, 2011
Revised Manuscript: January 13, 2012
Manuscript Accepted: January 19, 2012
Published: January 25, 2012

Virtual Issues
Vol. 7, Iss. 3 Virtual Journal for Biomedical Optics

Citation
Michael C. DeSantis, Shannon Kian Zareh, Xianglu Li, Robert E. Blankenship, and Y. M. Wang, "Single-image axial localization precision analysis for individual fluorophores," Opt. Express 20, 3057-3065 (2012)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-20-3-3057


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