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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 23 — Nov. 10, 2008
  • pp: 18714–18724

Localization of a fluorescent source without numerical fitting

Sean B. Andersson  »View Author Affiliations

Optics Express, Vol. 16, Issue 23, pp. 18714-18724 (2008)

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We present an algebraic solution to the problem of localizing a single fluorescent particle with sub-diffraction-limit accuracy. The algorithm is derived and its performance studied experimentally. Isolated 20 nm fluorescent beads were imaged using a wide-field microscope at two different positions separated by 100 nm and at a range of signal-to-noise ratios (SNR). The data were analyzed using both the new algorithm and the standard approach of fitting the data to a Gaussian profile. Results indicate that the proposed approach is nearly as accurate as Gaussian fitting across a wide range of SNR while executing over 200 times faster. In addition, the new algorithm is able to localize at lower SNR than the fitting method.

© 2008 Optical Society of America

OCIS Codes
(100.2960) Image processing : Image analysis
(180.2520) Microscopy : Fluorescence microscopy
(300.6280) Spectroscopy : Spectroscopy, fluorescence and luminescence

ToC Category:

Original Manuscript: August 29, 2008
Revised Manuscript: October 9, 2008
Manuscript Accepted: October 24, 2008
Published: October 29, 2008

Virtual Issues
Vol. 4, Iss. 1 Virtual Journal for Biomedical Optics

Sean Andersson, "Localization of a fluorescent source without numerical fitting," Opt. Express 16, 18714-18724 (2008)

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