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On the Fundamental Limits of Nano-Particle Tracking Along the Optical Axis

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

Proceedings of the 2004 Annual Meeting of the Swiss Society of Biomedical Engineering (SSBE'04), Zürich ZH, Swiss Confederation, September 2-3, 2004, poster 30.



Tracking fluorescent nanoparticles is of particular interest for many applications in biology. Most tracking procedures try to reconstruct the particle's trajectory in an XY plane, or eventually in an XYZ volume when multiple acquisitions at different focal distances are available. In this work, we present the fundamental limits of recovering a nano-particle's position along the optical axis, from single or multiple plane z-stack acquisitions. In microscopy imaging, nano-particles can be considered as point sources with respect to the microscope's resolution. Therefore, the observed image at a certain out-of-focus distance corresponds to the microscope's defocused point spread function (PSF). Given such an image, it is possible to determine the axial position of a defocused particle. In this work, we show the restrictions and limitations on the precision of this estimation procedure as a function of common imaging parameters and noise statistics. We also show how taking into account the influence of using multiple acquisitions at different focal distances can increase the precision.


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