Abstract

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

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

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Author Affiliations

Sean Andersson

Boston University

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