Two-photon fluorescence microscopy imaging of cellular oxidative stress using profluorescent nitroxides.
Two-photon fluorescence microscopy imaging of cellular oxidative stress using profluorescent nitroxides.
复制标题
使用大量硝化氧化物对细胞氧化应激进行两光子荧光显微镜成像。
DOI：
10.1021/ja210315x
复制
发表时间：
2012-03-14
影响因子：
15
通讯作者：
Belfield, Kevin D.
中科院分区：
文献类型：
--
作者：
Ahn, Hyo-Yang;Fairfull-Smith, Kathryn E.;Morrow, Benjamin J.;Lussini, Vanessa;Kim, Bosung;Bondar, Mykhailo V.;Bottle, Steven E.;Belfield, Kevin D.
合成了一系列不同的发色团氮氧自由基和它们的非自由基甲氧基胺类似物，并研究了它们的线性光物理性质。邻近的自由基的存在掩盖了发色团通常的荧光发射，这些物质被描述为profluorescent。两个氮氧化物结合蒽和荧光素发色团（化合物7和19，分别）表现出约400 G. M的双光子吸收（2PA）截面。当在大于800 nm的波长下激发时。这两个profluorescent氮氧化物表现出对中国仓鼠卵巢（CHO）细胞的细胞毒性低。使用市售CellROX Deep Red氧化应激监测器的成像共定位实验证明了氮氧探针的良好细胞摄取。氮氧探针H2O2诱导的损伤的灵敏度也证明了单光子和双光子荧光显微镜。这些profluorescent氮氧化物探针是潜在的强大的工具，在生物系统中的氧化应激成像，它们基本上是“亮”在某些物种产生的氧化应激的存在下。荧光氮氧化物种类和它们的非自由基加合物之间的荧光量子产率的高比率提供了测量一系列细胞氧化还原环境所需的灵敏度。此外，它们合理的2PA横截面提供了使用双光子荧光显微镜的选择，这避免了通常遇到的与单光子成像相关的缺点，如光漂白和不良的组织穿透。
A range of varying chromophore nitroxide free radicals and their nonradical methoxyamine analogues were synthesized and their linear photophysical properties examined. The presence of the proximate free radical masks the chromophore’s usual fluorescence emission, and these species are described as profluorescent. Two nitroxides incorporating anthracene and fluorescein chromophores (compounds 7 and 19, respectively) exhibited two-photon absorption (2PA) cross sections of approximately 400 G.M. when excited at wavelengths greater than 800 nm. Both of these profluorescent nitroxides demonstrated low cytotoxicity toward Chinese hamster ovary (CHO) cells. Imaging colocalization experiments with the commercially available CellROX Deep Red oxidative stress monitor demonstrated good cellular uptake of the nitroxide probes. Sensitivity of the nitroxide probes to H2O2-induced damage was also demonstrated by both one- and two-photon fluorescence microscopy. These profluorescent nitroxide probes are potentially powerful tools for imaging oxidative stress in biological systems, and they essentially “light up” in the presence of certain species generated from oxidative stress. The high ratio of the fluorescence quantum yield between the profluorescent nitroxide species and their nonradical adducts provides the sensitivity required for measuring a range of cellular redox environments. Furthermore, their reasonable 2PA cross sections provide for the option of using two-photon fluorescence microscopy, which circumvents commonly encountered disadvantages associated with one-photon imaging such as photobleaching and poor tissue penetration.
登录
查看更多内容
影响因子：
3.2
作者：
Keddie, Daniel J.;Fairfull-Smith, Kathryn E.;Bottle, Steven E.
通讯作者：
Bottle, Steven E.
影响因子：
2.8
作者：
Fairfull-Smith, Kathryn E.;Bottle, Steven E.
通讯作者：
Bottle, Steven E.
影响因子：
2.9
作者：
MITCHELL, JB;SAMUNI, A;RUSSO, A
通讯作者：
RUSSO, A
影响因子：
5
作者：
Lipman, Tatiana;Tabakman, Rinat;Lazarovici, Philip
通讯作者：
Lazarovici, Philip
影响因子：
3.8
作者：
Makarov, Nikolay S.;Drobizhev, Mikhail;Rebane, Aleksander
通讯作者：
Rebane, Aleksander