溶液中一部分NADH是“暗的”：对通过荧光寿命进行代谢传感的影响。
A fraction of NADH in solution is "dark": Implications for metabolic sensing via fluorescence lifetime.
作者信息
Cao Simin, Zhou Zhongneng, Li Haoyang, Jia Menghui, Liu Yangyi, Wang Mengyu, Zhang Mengjie, Zhang Sanjun, Chen Jinquan, Xu Jianhua, Knutson Jay R
机构信息
State Key Laboratory of Precision Spectroscopy, East China Normal University, Shanghai 200062, China.
Laboratory for Advanced Microscopy and Biophotonics, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, United States.
出版信息
Chem Phys Lett. 2019 Jul;726:18-21. doi: 10.1016/j.cplett.2019.04.014. Epub 2019 Apr 5.
Abstract
The metabolic cofactor and energy carrier NADH (nicotinamide adenine dinucleotide, reduced) has fluorescence yield and lifetime that depends strongly on conformation, a fact that has enabled metabolic monitoring of cells via FLIM (Fluorescence Lifetime Microscopy). Using femtosecond fluorescence upconversion, we show that this molecule in solution participates in ultrafast self-quenching along with both bulk solvent relaxation and spectral relaxation on 1.4 and 26 ps timescales. This, in effect, means up to a third of NADH is effectively "dark" for FLIM in the 400-500 nm observation window commonly employed. Methods to compensate for, avoid or measure dark species corrections are outlined.
摘要
代谢辅助因子和能量载体还原型烟酰胺腺嘌呤二核苷酸（NADH）具有荧光产率和寿命，其强烈依赖于构象，这一事实使得通过荧光寿命显微镜（FLIM）对细胞进行代谢监测成为可能。利用飞秒荧光上转换技术，我们表明溶液中的该分子在1.4皮秒和26皮秒时间尺度上参与超快自猝灭以及大量溶剂弛豫和光谱弛豫。实际上，这意味着在通常使用的400 - 500纳米观察窗口中，高达三分之一的NADH对于FLIM而言实际上是“暗的”。文中概述了补偿、避免或测量暗物质校正的方法。
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