Fluorescence to measure light intensity

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 waveleng...

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Published in:Nature methods Vol. 20; no. 12; pp. 1930 - 1938
Main Authors: Lahlou, Aliénor, Tehrani, Hessam Sepasi, Coghill, Ian, Shpinov, Yuriy, Mandal, Mrinal, Plamont, Marie-Aude, Aujard, Isabelle, Niu, Yuxi, Nedbal, Ladislav, Lazár, Dusan, Mahou, Pierre, Supatto, Willy, Beaurepaire, Emmanuel, Eisenmann, Isabelle, Desprat, Nicolas, Croquette, Vincent, Jeanneret, Raphaël, Le Saux, Thomas, Jullien, Ludovic
Format: Journal Article
Language:English
Published: New York Nature Publishing Group US 01.12.2023
Nature Publishing Group
Subjects:
631/1647/245/2225
631/92/96
Actinometry
Bioengineering
Bioinformatics
Biological Microscopy
Biological Physics
Biological Techniques
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Chemical Sciences
Dyes
Fluorescence
Fluorescent Dyes - chemistry
Imaging
Instrumentation and Detectors
Life Sciences
Light
Light intensity
Light sources
Luminous intensity
Measuring instruments
Microscopes
Microscopy, Fluorescence - methods
Optics
Physics
Proteins
Proteomics
Spatial distribution
Spectrometry, Fluorescence
Wavelengths
Chemical tools
Fluorescence imaging
ISSN:1548-7091, 1548-7105, 1548-7105
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Abstract 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. Two methods for fluorescence-based actinometry using organic dyes and photoconvertible fluorescent proteins enable rapid and precise measurement of light intensity at the sample in fluorescence microscopes.
AbstractList 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.
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.Two methods for fluorescence-based actinometry using organic dyes and photoconvertible fluorescent proteins enable rapid and precise measurement of light intensity at the sample in fluorescence microscopes.
Abstract 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.
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.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.
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. Two methods for fluorescence-based actinometry using organic dyes and photoconvertible fluorescent proteins enable rapid and precise measurement of light intensity at the sample in fluorescence microscopes.
Author Coghill, Ian
Nedbal, Ladislav
Jeanneret, Raphaël
Supatto, Willy
Niu, Yuxi
Desprat, Nicolas
Mandal, Mrinal
Jullien, Ludovic
Mahou, Pierre
Shpinov, Yuriy
Plamont, Marie-Aude
Croquette, Vincent
Eisenmann, Isabelle
Lazár, Dusan
Le Saux, Thomas
Aujard, Isabelle
Lahlou, Aliénor
Tehrani, Hessam Sepasi
Beaurepaire, Emmanuel
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– notice: The Author(s) 2023. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
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ISSN 1548-7091
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Issue 12
Keywords Chemical tools
Fluorescence imaging
Language English
License 2023. The Author(s).
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PublicationSubtitle Techniques for life scientists and chemists
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Snippet Despite the need for quantitative measurements of light intensity across many scientific disciplines, existing technologies for measuring light dose at the...
Abstract Despite the need for quantitative measurements of light intensity across many scientific disciplines, existing technologies for measuring light dose...
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StartPage 1930
SubjectTerms 631/1647/245/2225
631/92/96
Actinometry
Bioengineering
Bioinformatics
Biological Microscopy
Biological Physics
Biological Techniques
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Chemical Sciences
Dyes
Fluorescence
Fluorescent Dyes - chemistry
Imaging
Instrumentation and Detectors
Life Sciences
Light
Light intensity
Light sources
Luminous intensity
Measuring instruments
Microscopes
Microscopy, Fluorescence - methods
Optics
Physics
Proteins
Proteomics
Spatial distribution
Spectrometry, Fluorescence
Wavelengths
Title Fluorescence to measure light intensity
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Volume 20
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