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荧光法测量光强度。

Fluorescence to measure light intensity.

机构信息

PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne University, CNRS, Paris, France.

Sony Computer Science Laboratories, Paris, France.

出版信息

Nat Methods. 2023 Dec;20(12):1930-1938. doi: 10.1038/s41592-023-02063-y. Epub 2023 Nov 23.

DOI:10.1038/s41592-023-02063-y
PMID:37996751
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10703675/
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.

摘要

尽管许多科学学科都需要对光强度进行定量测量，但用于测量荧光显微镜样本中光剂量的现有技术无法同时在广泛的波长和强度范围内检索光强度的空间分布。为了解决这一限制，我们开发了两种快速而简单的协议，使用有机染料和荧光蛋白作为光量计。第一个协议依赖于分子系统，当施加恒定光时，其荧光强度会呈指数衰减和/或增加。第二个协议依赖于宽吸收的光化学惰性荧光团，从一个波长到另一个波长反算光强度。作为其使用的演示，该协议应用于定量表征各种荧光成像系统的光空间分布，并对市售仪器和光源的照明进行校准。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/d22e7238c91e/41592_2023_2063_Fig12_ESM.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/d22e7238c91e/41592_2023_2063_Fig12_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/18278a7ca6f0/41592_2023_2063_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/ce9b68a21df3/41592_2023_2063_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/c0a0ff66bbf8/41592_2023_2063_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/24b85c418e5b/41592_2023_2063_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/2371aa5886dc/41592_2023_2063_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/467778ed4a74/41592_2023_2063_Fig6_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/74136ef7c946/41592_2023_2063_Fig7_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/96643831c687/41592_2023_2063_Fig8_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/89e53260bba1/41592_2023_2063_Fig9_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/fc26d7182980/41592_2023_2063_Fig10_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/640a79aca22a/41592_2023_2063_Fig11_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92e3/10703675/d22e7238c91e/41592_2023_2063_Fig12_ESM.jpg

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2
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3
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4
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Chemistry. 2022 May 16;28(28):e202200497. doi: 10.1002/chem.202200497. Epub 2022 Apr 5.
5
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Nat Methods. 2021 Dec;18(12):1423-1426. doi: 10.1038/s41592-021-01162-y.
6
The polyphasic chlorophyll a fluorescence rise measured under high intensity of exciting light.在高强度激发光下测量的多相叶绿素a荧光上升。
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7
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Sci Rep. 2018 Apr 3;8(1):5421. doi: 10.1038/s41598-018-23735-2.
8
Resonant out-of-phase fluorescence microscopy and remote imaging overcome spectral limitations.共振异相荧光显微镜和远程成像克服了光谱限制。
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9
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10
Applications of Continuous-Flow Photochemistry in Organic Synthesis, Material Science, and Water Treatment.连续流动光化学在有机合成、材料科学和水处理中的应用。
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