Biomedical Imaging Group
Logo EPFL
    • Splines Tutorials
    • Splines Art Gallery
    • Wavelets Tutorials
    • Image denoising
    • ERC project: FUN-SP
    • Sparse Processes - Book Preview
    • ERC project: GlobalBioIm
    • The colored revolution of bioimaging
    • Deconvolution
    • SMLM
    • One-World Seminars: Representer theorems
    • A Unifying Representer Theorem
Follow us on Twitter.
Join our Github.
Masquer le formulaire de recherche
Menu
BIOMEDICAL IMAGING GROUP (BIG)
Laboratoire d'imagerie biomédicale (LIB)
  1. School of Engineering STI
  2. Institute IEM
  3.  LIB
  4.  Nanoparticle Localization
  • Laboratory
    • Laboratory
    • Laboratory
    • People
    • Jobs and Trainees
    • News
    • Events
    • Seminars
    • Resources (intranet)
    • Twitter
  • Research
    • Research
    • Researchs
    • Research Topics
    • Talks, Tutorials, and Reviews
  • Publications
    • Publications
    • Publications
    • Database of Publications
    • Talks, Tutorials, and Reviews
    • EPFL Infoscience
  • Code
    • Code
    • Code
    • Demos
    • Download Algorithms
    • Github
  • Teaching
    • Teaching
    • Teaching
    • Courses
    • Student projects
  • Splines
    • Teaching
    • Teaching
    • Splines Tutorials
    • Splines Art Gallery
    • Wavelets Tutorials
    • Image denoising
  • Sparsity
    • Teaching
    • Teaching
    • ERC project: FUN-SP
    • Sparse Processes - Book Preview
  • Imaging
    • Teaching
    • Teaching
    • ERC project: GlobalBioIm
    • The colored revolution of bioimaging
    • Deconvolution
    • SMLM
  • Machine Learning
    • Teaching
    • Teaching
    • One-World Seminars: Representer theorems
    • A Unifying Representer Theorem

Sub-Resolution Axial Localization of Nanoparticles in Fluorescence Microscopy

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

Proceedings of the SPIE European Conference on Biomedical Optics (ECBO'05), Münich, Federal Republic of Germany, June 12-16, 2005, vol. 5860, pp. 103-106.


In recent years single particle tracking has significantly contributed to the study of intracellular molecular dynamics [1]. Particle tracking primarily consists of localizing particles in three dimensions (3-D) from a number of images acquired by fluorescence microscopy. While many approaches have been proposed for the lateral (i.e., x-y) localization of particles from acquisitions that are (almost) in focus [2], far fewer methods currently exist for the precise determination of a particle's axial (i.e., z) position. Since the acquisition of high-resolution z-stacks is inefficient and impractical, axial localization methods try to estimate the position using few acquisitions of the particle. The difficulty of this task comes from the complexity of the diffraction patterns that appear as the particle moves out of focus. To avoid complex models, current axial localization methods often rely on empirical approaches [3].

Under optimal conditions, the spot of light that one observes when viewing a sub-resolution fluorescent particle with a microscope corresponds to the diffraction-limited point spread function (PSF) of the microscope. While most methods for tracking and localizing single particles try to exploit this property, they often make use of a simplified diffraction model. The principal difficulty with respect to axial localization is the non-stationarity of the PSF along the optical axis; if there is a difference in the refractive indices of the specimen and immersion medium, the PSF may vary depending on the particle's depth within the specimen, which is a frequently occurring situation in practice. In this paper we propose an algorithm based on maximum-likelihood estimation that employs an accurate PSF model to iteratively determine a particle's axial position. We describe an image formation model, propose a theoretical lower bound on the attainable localization precision, and show how our algorithm is derived. Finally, we demonstrate its efficiency by showing the correspondence with the theoretical lower bound.

References

  1. U. Kubitscheck, "Single Protein Molecules Visualized and Tracked in the Interior of Eukaryotic Cells," Single Molecules, vol. 3, no. 5-6, pp. 267-274, November 2002.

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

  3. M. Speidel, A. Jon, E.-L. Florin, "Three-Dimensional Tracking of Fluorescent Nanoparticles with Subnanometer Precision by Use of Off-Focus Imaging," Optics Letters, vol. 28, no. 2, pp. 69-71, January 2003.

@INPROCEEDINGS(http://bigwww.epfl.ch/publications/aguet0503.html,
AUTHOR="Aguet, F. and Van De Ville, D. and Unser, M.",
TITLE="Sub-Resolution Axial Localization of Nanoparticles in
	Fluorescence Microscopy",
BOOKTITLE="Proceedings of the {SPIE} {E}uropean Conference on Biomedical
	Optics: {C}onfocal, Multiphoton, and Nonlinear Microscopic Imaging
	{II} ({ECBO'05})",
YEAR="2005",
editor="Wilson, T.",
volume="5860",
series="",
pages="103--106",
address="M{\"{u}}nich, Federal Republic of Germany",
month="June 12-16,",
organization="",
publisher="",
note="")
© 2005 SPIE. Personal use of this material is permitted. However, permission to reprint/republish this material for advertising or promotional purposes or for creating new collective works for resale or redistribution to servers or lists, or to reuse any copyrighted component of this work in other works must be obtained from SPIE. This material is presented to ensure timely dissemination of scholarly and technical work. Copyright and all rights therein are retained by authors or by other copyright holders. All persons copying this information are expected to adhere to the terms and constraints invoked by each author's copyright. In most cases, these works may not be reposted without the explicit permission of the copyright holder.
  • Laboratory
  • Research
  • Publications
    • Database of Publications
    • Talks, Tutorials, and Reviews
    • EPFL Infoscience
  • Code
  • Teaching
Logo EPFL, Ecole polytechnique fédérale de Lausanne
Emergencies: +41 21 693 3000 Services and resources Contact Map Webmaster email

Follow EPFL on social media

Follow us on Facebook. Follow us on Twitter. Follow us on Instagram. Follow us on Youtube. Follow us on LinkedIn.
Accessibility Disclaimer Privacy policy

© 2023 EPFL, all rights reserved