Fluorescence to measure light intensity.

Autor: Lahlou A; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France. Alienor.Lahlou@sony.com.; Sony Computer Science Laboratories, Paris, France. Alienor.Lahlou@sony.com., Tehrani HS; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Coghill I; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Shpinov Y; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Mandal M; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Plamont MA; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Aujard I; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France., Niu Y; Institute of Bio- and Geosciences/Plant Sciences, Forschungszentrum Jülich, Jülich, Germany., Nedbal L; Institute of Bio- and Geosciences/Plant Sciences, Forschungszentrum Jülich, Jülich, Germany.; Department of Biophysics, Faculty of Science, Palacký University, Olomouc, Czech Republic., Lazár D; Department of Biophysics, Faculty of Science, Palacký University, Olomouc, Czech Republic., Mahou P; Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, IP Paris, Palaiseau, France., Supatto W; Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, IP Paris, Palaiseau, France., Beaurepaire E; Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, IP Paris, Palaiseau, France., Eisenmann I; Laboratory of Physics of the École Normale Supérieure, University of PSL, CNRS, Sorbonne University, University of Paris City, Paris, France.; Institute of Biology of ENS (IBENS), École Normale Supérieure, CNRS, INSERM, University of PSL, Paris, France., Desprat N; Laboratory of Physics of the École Normale Supérieure, University of PSL, CNRS, Sorbonne University, University of Paris City, Paris, France.; Institute of Biology of ENS (IBENS), École Normale Supérieure, CNRS, INSERM, University of PSL, Paris, France., Croquette V; Laboratory of Physics of the École Normale Supérieure, University of PSL, CNRS, Sorbonne University, University of Paris City, Paris, France.; Institute of Biology of ENS (IBENS), École Normale Supérieure, CNRS, INSERM, University of PSL, Paris, France., Jeanneret R; Laboratory of Physics of the École Normale Supérieure, University of PSL, CNRS, Sorbonne University, University of Paris City, Paris, France.; Institute of Biology of ENS (IBENS), École Normale Supérieure, CNRS, INSERM, University of PSL, Paris, France., Le Saux T; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France. Thomas.Lesaux@ens.psl.eu., Jullien L; PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France. Ludovic.Jullien@ens.psl.eu.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2023 Dec; Vol. 20 (12), pp. 1930-1938. Date of Electronic Publication: 2023 Nov 23.
DOI: 10.1038/s41592-023-02063-y
Abstrakt: Despite the need for quantitative measurements of light intensity across many scientific disciplines, existing technologies for measuring light dose at the sample of a fluorescence microscope cannot simultaneously retrieve light intensity along with spatial distribution over a wide range of wavelengths and intensities. To address this limitation, we developed two rapid and straightforward protocols that use organic dyes and fluorescent proteins as actinometers. The first protocol relies on molecular systems whose fluorescence intensity decays and/or rises in a monoexponential fashion when constant light is applied. The second protocol relies on a broad-absorbing photochemically inert fluorophore to back-calculate the light intensity from one wavelength to another. As a demonstration of their use, the protocols are applied to quantitatively characterize the spatial distribution of light of various fluorescence imaging systems, and to calibrate illumination of commercially available instruments and light sources.
(© 2023. The Author(s).)
Databáze: MEDLINE
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