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Summary: Localization of a fluorescent source
without numerical fitting
Sean B. Andersson
Department of Mechanical Engineering, Boston University,
Boston, MA 02215
sanderss@bu.edu
Abstract: We present an algebraic solution to the problem of localizing
a single fluorescent particle with sub-diffraction-limit accuracy. The algo-
rithm 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: (180.2520) Fluorescence microscopy; (300.6280); Spectroscopy, fluorescence
and luminescence; (100.2960) Image analysis
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