Ultimate Use of Two-Photon Fluorescence Microscopy to Map Orientational Behavior of Fluorophores
| Autor: | Alla Kress, Xiao Wang, Paulina Gasecka, Patrick Ferrand, Fatma-Zohra Bioud, Sophie Brasselet, Julien Duboisset |
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| Přispěvatelé: | MOSAIC (MOSAIC), Institut FRESNEL (FRESNEL), Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU), Chinese Scholarship Council, ANR-10-BLAN-1509,ReceptOrient,Observation de l'activation du récepteur des lymphocytes T (TCR) en temps réel: suivi des orientations dynamiques du domaine intracellulaire du complexe récepteur dans les cellules vivantes par imagerie en fluorescence polarisée résolue de temps(2010), ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010), ANR-11-IDEX-0001,Amidex,INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE(2011), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Předmět: |
Distribution (number theory)
[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging Gaussian Population Biophysics Pyridinium Compounds 02 engineering and technology Linear dichroism 01 natural sciences 010309 optics Quantitative Biology::Subcellular Processes symbols.namesake Optics 0103 physical sciences Microscopy Chlorocebus aethiops Animals education Fluorescent Dyes education.field_of_study [PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics] business.industry Chemistry Isotropy Cell Membrane 021001 nanoscience & nanotechnology Fluorescence [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics Distribution function Microscopy Fluorescence Multiphoton Models Chemical Cell Biophysics COS Cells Liposomes symbols [SPI.OPTI]Engineering Sciences [physics]/Optics / Photonic 0210 nano-technology business Biological system |
| Zdroj: | Biophysical Journal Biophysical Journal, Biophysical Society, 2014, 106 (11), pp.2330-2339. ⟨10.1016/j.bpj.2014.04.011⟩ Biophysical Journal, 2014, 106 (11), pp.2330-2339. ⟨10.1016/j.bpj.2014.04.011⟩ |
| ISSN: | 0006-3495 1542-0086 |
| DOI: | 10.1016/j.bpj.2014.04.011 |
| Popis: | International audience; The orientational distribution of fluorophores is an important reporter of the structure and function of their molecular environment. Although this distribution affects the fluorescence signal under polarized-light excitation, its retrieval is limited to a small number of parameters. Because of this limitation, the need for a geometrical model (cone, Gaussian, etc.) to effect such retrieval is often invoked. In this work, using a symmetry decomposition of the distribution function of the fluorescent molecules, we show that polarized two-photon fluorescence based on tunable linear dichroism allows for the retrieval of this distribution with reasonable fidelity and without invoking either an a priori knowledge of the system to be investigated or a geometrical model. We establish the optimal level of detail to which any distribution can be retrieved using this technique. As applied to artificial lipid vesicles and cell membranes, the ability of this method to identify and quantify specific structural properties that complement the more traditional molecular-order information is demonstrated. In particular, we analyze situations that give access to the sharpness of the angular constraint, and to the evidence of an isotropic population of fluorophores within the focal volume encompassing the membrane. Moreover, this technique has the potential to address complex situations such as the distribution of a tethered membrane protein label in an ordered environment. |
| Databáze: | OpenAIRE |
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