Publication

On the Feasibility of Axial Tracking of a Fluorescent Nano-Particle Using a Defocusing Model

Michaël Unser
2004
Conference Papers

Abstract

The image of a sub-resolution nano-particle in fluorescence microscopy corresponds to a slice of the 3D point spread function (PSF). This slice relates to the out-of-focus distance of the nano-particle. In this paper, we investigate to which extent it is possible to estimate the out-of-focus distance of the nano-particle from a 2D image based on the knowledge of the 3D PSF. To this end, we compute the Cramér-Rao bound (CRB) that provides a lower bound on the error of the best estimator of the axial position. The calculation of the CRB involves the specification of a 3D PSF model, the assumption of a signal-dependent Poisson noise, and some acquisition parameters. Our derivation shows that the CRB depends on the defocusing distance. Interestingly, nanometer precision can be attained over a range of defocus distances and for sufficiently high SNR levels. The theoretical results are confirmed with simulated experiments using estimators based on the least-squares (LS) and normalized cross-correlation (NCC) criterion. The results obtained are very close to the theoretical CRB.

Official source

https://infoscience.epfl.ch/entities/publication/86e0fd0f-8bd7-46a9-a26d-eaaf1eed1a03

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Michaël Unser
2004
Conference Papers

Abstract

Official source

https://infoscience.epfl.ch/entities/publication/86e0fd0f-8bd7-46a9-a26d-eaaf1eed1a03

About this result

Related concepts (32)

Point spread function

The point spread function (PSF) describes the response of a focused optical imaging system to a point source or point object. A more general term for the PSF is the system's impulse response; the PSF is the impulse response or impulse response function (IRF) of a focused optical imaging system. The PSF in many contexts can be thought of as the extended blob in an image that represents a single point object, that is considered as a spatial impulse. In functional terms, it is the spatial domain version (i.e.

Super-resolution microscopy

Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that use the Pendry Superlens and near field scanning optical microscopy) or on the far-field.

Nanoparticle

A nanoparticle or ultrafine particle is usually defined as a particle of matter that is between 1 and 100 nanometres (nm) in diameter. The term is sometimes used for larger particles, up to 500 nm, or fibers and tubes that are less than 100 nm in only two directions. At the lowest range, metal particles smaller than 1 nm are usually called atom clusters instead.

Related publications (2)

On the Feasibility of Axial Tracking of a Fluorescent Nano-Particle Using a Defocusing Model

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