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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 17, Iss. 10 — May. 11, 2009
  • pp: 8264–8277

Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy

Michael J. Mlodzianoski, Manuel F. Juette, Glen L. Beane, and Joerg Bewersdorf  »View Author Affiliations

Optics Express, Vol. 17, Issue 10, pp. 8264-8277 (2009)

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Three-dimensional (3D) particle localization at the nanometer scale plays a central role in 3D particle tracking and 3D localization-based super-resolution microscopy. Here we introduce a localization algorithm that is independent of theoretical models and therefore generally applicable to a large number of experimental realizations. Applying this algorithm and a convertible experimental setup we compare the performance of the two major 3D techniques based on astigmatic distortions and on multiplane detection. In both methods we obtain experimental 3D localization accuracies in agreement with theoretical predictions and characterize the depth dependence of the localization accuracy in detail.

© 2009 Optical Society of America

OCIS Codes
(110.6880) Imaging systems : Three-dimensional image acquisition
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Imaging Systems

Original Manuscript: March 6, 2009
Revised Manuscript: April 20, 2009
Manuscript Accepted: April 24, 2009
Published: April 30, 2009

Virtual Issues
Vol. 4, Iss. 7 Virtual Journal for Biomedical Optics

Michael J. Mlodzianoski, Manuel F. Juette, Glen L. Beane, and Joerg Bewersdorf, "Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy," Opt. Express 17, 8264-8277 (2009)

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