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Sub-Resolution Maximum-Likelihood Based Localization of Fluorescent Nanoparticles in Three Dimensions

F. Aguet, D. Van De Ville, M. Unser

Proceedings of the Fourth IEEE International Symposium on Biomedical Imaging: From Nano to Macro (ISBI'07), Arlington VA, USA, April 12-15, 2007, pp. 932-935.

Several recent studies have shown that fluorescent particles can be localized with an accuracy that is well beyond traditional resolution limits. Using a theoretical model of the image formation process that accounts for possible sources of noise, Cramér-Rao bounds have been used to define the theoretical limits. A crucial influence on these bounds is the mismatch of refractive indices that is usually present between immersion medium and specimen. This results in an axially shift-variant point spread function, meaning that the bounds change as a function of the particle's position in the z-direction. We investigate the theoretical bounds for this shift-variant model, and propose a maximum-likelihood estimator for the particle position in 3D (XYZ position). Using this estimator, sub-resolution localization at the nanometer scale is demonstrated on experimental data. The results provide optimal conditions for particle tracking and localization experiments.

AUTHOR="Aguet, F. and Van De Ville, D. and Unser, M.",
TITLE="Sub-Resolution Maximum-Likelihood Based Localization of
        Fluorescent Nanoparticles in Three Dimensions",
BOOKTITLE="Proceedings of the Fourth {IEEE} International Symposium on
        Biomedical Imaging: {F}rom Nano to Macro ({ISBI'07})",
address="Arlington VA, USA",
month="April 12-15,",

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