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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 19, Iss. 8 — Apr. 11, 2011
  • pp: 7020–7033

Measuring localization performance of super-resolution algorithms on very active samples

Steve Wolter, Ulrike Endesfelder, Sebastian van de Linde, Mike Heilemann, and Markus Sauer  »View Author Affiliations


Optics Express, Vol. 19, Issue 8, pp. 7020-7033 (2011)
http://dx.doi.org/10.1364/OE.19.007020


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Abstract

Super-resolution fluorescence imaging based on single-molecule localization relies critically on the availability of efficient processing algorithms to distinguish, identify, and localize emissions of single fluorophores. In multiple current applications, such as three-dimensional, time-resolved or cluster imaging, high densities of fluorophore emissions are common. Here, we provide an analytic tool to test the performance and quality of localization microscopy algorithms and demonstrate that common algorithms encounter difficulties for samples with high fluorophore density. We demonstrate that, for typical single-molecule localization microscopy methods such as dSTORM and the commonly used rapidSTORM scheme, computational precision limits the acceptable density of concurrently active fluorophores to 0.6 per square micrometer and that the number of successfully localized fluorophores per frame is limited to 0.2 per square micrometer.

© 2011 OSA

OCIS Codes
(100.6640) Image processing : Superresolution
(180.2520) Microscopy : Fluorescence microscopy
(350.4800) Other areas of optics : Optical standards and testing

ToC Category:
Image Processing

History
Original Manuscript: December 21, 2010
Revised Manuscript: February 23, 2011
Manuscript Accepted: February 27, 2011
Published: March 29, 2011

Virtual Issues
Vol. 6, Iss. 5 Virtual Journal for Biomedical Optics

Citation
Steve Wolter, Ulrike Endesfelder, Sebastian van de Linde, Mike Heilemann, and Markus Sauer, "Measuring localization performance of super-resolution algorithms on very active samples," Opt. Express 19, 7020-7033 (2011)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-19-8-7020


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