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Maximum-Likelihood Based Tracking of Fluorescent Nanoparticles

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

Proceedings of the 2005 Annual Meeting of the Swiss Society for Biomedical Engineering (SSBE'05), Lausanne VD, Swiss Confederation, September 1-2, 2005, pp. F01.



Novel fluorescent labeling techniques developed over the past decade have made it possible to study the dynamics of individual proteins inside living cells using fluorescence microscopy. These studies typically involve the acquisition of a time series of z-stacks, which are sequences of images taken at different focal distances. Single particle tracking techniques, designed to localize particles in three dimensions, have significantly contributed to the interpretation of such data. Due to the limited axial resolution of the z-stacks, it is common that particles must be localized from a single acquisition. As a consequence, since particles are likely to appear out of focus in the acquisition, determining their axial position is the most difficult aspect of the localization process. This is reflected in the literature, where many approaches to lateral localization (i.e., in the x-y plane) have been investigated (see, e.g., [1]), but only few methods for axial localization, with limited practical applicability, have been proposed.

References

  1. R.J. Ober, S. Ram, E.S. Ward, "Localization Accuracy in Single-Molecule Microscopy," Biophysical Journal, vol. 86, no. 2, February 2004, pp. 1185-1200.


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