Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors

Abstract In this paper, we propose a novel and sensitive ratiometric analysis method that uses the fractional intensities of time-resolved fluorescence of genetically encoded fluorescent NADH/NAD+ biosensors, Peredox, SoNar, and Frex. When the conformations of the biosensors change upon NADH/NAD+ bi...

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Autores principales:	Mengfang Chang, Lei Li, Hanyang Hu, Qingxun Hu, Aoxue Wang, Xiaodan Cao, Xiantong Yu, Sanjun Zhang, Yuzheng Zhao, Jinquan Chen, Yi Yang, Jianhua Xu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b1416b4418bf4c22bdaaa271fb4b9633
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spelling	oai:doaj.org-article:b1416b4418bf4c22bdaaa271fb4b96332021-12-02T16:06:00ZUsing Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors10.1038/s41598-017-04051-72045-2322https://doaj.org/article/b1416b4418bf4c22bdaaa271fb4b96332017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04051-7https://doaj.org/toc/2045-2322Abstract In this paper, we propose a novel and sensitive ratiometric analysis method that uses the fractional intensities of time-resolved fluorescence of genetically encoded fluorescent NADH/NAD+ biosensors, Peredox, SoNar, and Frex. When the conformations of the biosensors change upon NADH/NAD+ binding, the fractional intensities (α i τ i ) have opposite changing trends. Their ratios could be exploited to quantify NADH/NAD+ levels with a larger dynamic range and higher resolution versus commonly used fluorescence intensity and lifetime methods. Moreover, only one excitation and one emission wavelength are required for this ratiometric measurement. This eliminates problems of traditional excitation-ratiometric and emission-ratiometric methods. This method could be used to simplify the design and achieve highly sensitive analyte quantification of genetically encoded fluorescent biosensors. Wide potential applications could be developed for imaging live cell metabolism based on this new method.Mengfang ChangLei LiHanyang HuQingxun HuAoxue WangXiaodan CaoXiantong YuSanjun ZhangYuzheng ZhaoJinquan ChenYi YangJianhua XuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mengfang Chang Lei Li Hanyang Hu Qingxun Hu Aoxue Wang Xiaodan Cao Xiantong Yu Sanjun Zhang Yuzheng Zhao Jinquan Chen Yi Yang Jianhua Xu Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
description	Abstract In this paper, we propose a novel and sensitive ratiometric analysis method that uses the fractional intensities of time-resolved fluorescence of genetically encoded fluorescent NADH/NAD+ biosensors, Peredox, SoNar, and Frex. When the conformations of the biosensors change upon NADH/NAD+ binding, the fractional intensities (α i τ i ) have opposite changing trends. Their ratios could be exploited to quantify NADH/NAD+ levels with a larger dynamic range and higher resolution versus commonly used fluorescence intensity and lifetime methods. Moreover, only one excitation and one emission wavelength are required for this ratiometric measurement. This eliminates problems of traditional excitation-ratiometric and emission-ratiometric methods. This method could be used to simplify the design and achieve highly sensitive analyte quantification of genetically encoded fluorescent biosensors. Wide potential applications could be developed for imaging live cell metabolism based on this new method.
format	article
author	Mengfang Chang Lei Li Hanyang Hu Qingxun Hu Aoxue Wang Xiaodan Cao Xiantong Yu Sanjun Zhang Yuzheng Zhao Jinquan Chen Yi Yang Jianhua Xu
author_facet	Mengfang Chang Lei Li Hanyang Hu Qingxun Hu Aoxue Wang Xiaodan Cao Xiantong Yu Sanjun Zhang Yuzheng Zhao Jinquan Chen Yi Yang Jianhua Xu
author_sort	Mengfang Chang
title	Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
title_short	Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
title_full	Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
title_fullStr	Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
title_full_unstemmed	Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors
title_sort	using fractional intensities of time-resolved fluorescence to sensitively quantify nadh/nad+ with genetically encoded fluorescent biosensors
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/b1416b4418bf4c22bdaaa271fb4b9633
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Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors

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