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Localization of a fluorescent source without numerical fitting

Summary: Localization of a fluorescent source
without numerical fitting
Sean B. Andersson
Department of Mechanical Engineering, Boston University,
Boston, MA 02215
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


Source: Andersson, Sean B. - Department of Aerospace and Mechanical Engineering, Boston University
Boston University, Center for Nanoscience and Nanobiotechnology


Collections: Engineering; Materials Science