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.  Photobleaching Kinetics
  • 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

Photobleaching Kinetics and Time-Integrated Emission of Fluorescent Probes in Cellular Membranes

D. Wüstner, T. Christensen, L.M. Solanko, D. Sage

Molecules, vol. 19, no. 8, pp. 11096-11130, August 2014.


Since the pioneering work of Hirschfeld, it is known that time-integrated emission (TiEm) of a fluorophore is independent of fluorescence quantum yield and illumination intensity. Practical implementation of this important result for determining exact probe distribution in living cells is often hampered by the presence of autofluorescence. Using kinetic modelling of photobleaching combined with pixel-wise bleach rate fitting of decay models with an updated plugin to the ImageJ program, it is shown that the TiEm of a fluorophore in living cells can be determined exactly from the product of bleaching amplitude and time constant. This applies to mono-exponential bleaching from the first excited singlet and/or triplet state and to multi-exponential combinations of such processes. The TiEm can be used to correct for illumination shading and background autofluorescence without the need for fluorescent test layers or separate imaging of non-stained cells. We apply the method to simulated images and to images of cells, whose membranes were labelled with fluorescent sterols and sphingolipids. Our bleaching model can be extended to include a probability density function (PDF) of intrinsic bleach rate constants with a memory kernel. This approach results in a time-dependent bleach rate coefficient and is exemplified for fluorescent sterols in restricted intracellular environments, like lipid droplets. We show that for small deviations from the classical exponential bleaching, the TiEm of decay functions with rate coefficients remains largely independent of fluorescence lifetime and illumination, and thereby represents a faithful measure of probe distribution.

@ARTICLE(http://bigwww.epfl.ch/publications/wuestner1401.html,
AUTHOR="W{\"{u}}stner, D. and Christensen, T. and Solanko, L.M. and
	Sage, D.",
TITLE="Photobleaching Kinetics and Time-Integrated Emission of
	Fluorescent Probes in Cellular Membranes",
JOURNAL="Molecules",
YEAR="2014",
volume="19",
number="8",
pages="11096--11130",
month="August",
note="")

© 2014 MDPI AG. 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 MDPI AG. 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