Use of spin traps to detect superoxide production in living cells by electron paramagnetic resonance (EPR) spectroscopy
Use of spin traps to detect superoxide production in living cells by electron paramagnetic resonance (EPR) spectroscopy
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DOI：
10.1016/j.ymeth.2016.05.001
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发表时间：
2016-10-15
期刊：
影响因子：
4.8
通讯作者：
Peyrot, Fabienne
中科院分区：
文献类型：
--
作者：
Abbas, Kahina;Babic, Nikola;Peyrot, Fabienne
检测由活细胞产生的超氧化物已经是生物学中四十多年来的持续挑战。已经提出了各种方法来解决这个问题，其中自旋捕获与环硝酮耦合到EPR光谱，自由基检测的金标准。这种技术是基于超氧化物的亲核加成到抗磁性环状硝酮，称为自旋陷阱，和自旋加合物的形成，即具有特征EPR光谱的持久自由基。自旋捕获首次应用于活细胞可以追溯到1979年。从那时起，该方法已经取得了相当大的改进，无论是在结构的自旋陷阱，EPR方法，并设计的实验，包括适当的控制。在这里，我们将集中在自旋捕获/EPR技术的技术方面，描绘最近的突破，固有的局限性，和潜在的文物。(C)2016 Elsevier Inc. All rights reserved.
Detection of superoxide produced by living cells has been an on-going challenge in biology for over forty years. Various methods have been proposed to address this issue, among which spin trapping with cyclic nitrones coupled to EPR spectroscopy, the gold standard for detection of radicals. This technique is based on the nucleophilic addition of superoxide to a diamagnetic cyclic nitrone, referred to as the spin trap, and the formation of a spin adduct, i.e. a persistent radical with a characteristic EPR spectrum. The first application of spin trapping to living cells dates back 1979. Since then, considerable improvements of the method have been achieved both in the structures of the spin traps, the EPR methodology, and the design of the experiments including appropriate controls. Here, we will concentrate on technical aspects of the spin trapping/EPR technique, delineating recent breakthroughs, inherent limitations, and potential artifacts. (C) 2016 Elsevier Inc. All rights reserved.