test_moissonnage_selune
type: Agent

Continuous photobleaching in vesicles and living cells: a measure of diffusion and compartmentation.
type: Dataset
contact point: Organization
  fn: CCSD
keyword: mesh-models-statistical
keyword: mesh-dextrans
keyword: mesh-computer-simulation
keyword: mesh-diffusion
keyword: sdvibimalife-sciences-q-biobioengineeringimaging
keyword: journal-articles
keyword: mesh-reproducibility-of-results
keyword: mesh-microscopy-fluorescence
keyword: mesh-photons
keyword: mesh-software
keyword: mesh-fluorescence-recovery-after-photobleaching
keyword: mesh-hela-cells
keyword: mesh-permeability
keyword: mesh-humans
keyword: infoeu-reposemanticsarticle
keyword: mesh-biophysics
keyword: mesh-green-fluorescent-proteins
keyword: mesh-microscopy-confocal
keyword: mesh-photobleaching
keyword: mesh-models-theoretical
keyword: mesh-fluorescein-5-isothiocyanate
keyword: mesh-programming-languages
keyword: mesh-image-processing-computer-assisted
keyword: mesh-time-factors
keyword: mesh-light
landing page: 0006-3495
description: We present a comprehensive and analytical treatment of continuous photobleaching in a compartment, under single photon excitation. In the very short time regime (t<0.1 ms), the diffusion does not play any role. After a transition (or short time regime), one enters in the long time regime (t>0.1-5 s), for which the diffusion and the photobleaching balance each other. In this long time regime, the diffusion is either fast (i.e., the photobleaching probability of a molecule diffusing through the laser beam is low) so that the photobleaching rate is independent of the diffusion constant and dependent only of the laser power, or the diffusion is slow (i.e., the photobleaching probability is high) and the photobleaching rate is mainly dependent on the diffusion constant. We illustrate our theory by using giant unilamellar vesicles ranging from approximately 10 to 100 microm in diameter, loaded with molecules of various diffusion constants (from 20 to 300 microm2/s) and various photobleaching cross sections, illuminated under laser powers between 3 and 100 microW. We also demonstrated that information about compartmentation can be obtained by this method in living cells expressing enhanced green fluorescent proteins or that were loaded with small FITC-dextrans. Our quantitative approach shows that molecules freely diffusing in a cellular compartment do experience a continuous photobleaching. We provide a generic theoretical framework that should be taken into account when studying, under confocal microscopy, molecular interactions, permeability, etc.
issued: 2026-05-14T17:00:10.635083
language: en
modified: 2026-05-14T17:00:10.635089

Continuous photobleaching in vesicles and living cells: a measure of diffusion and compartmentation.
type: Distribution
access URL: https://hal.science/hal-00078470
format: HTML
issued: 2026-05-14T17:00:10.640465
modified: 2026-05-14T17:00:10.562773