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Localization of a fluorescent source without numerical fitting

2008; Optica Publishing Group; Volume: 16; Issue: 23 Linguagem: Inglês

10.1364/oe.16.018714

1094-4087

Sean B. Andersson,

Digital Holography and Microscopy

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.