Preparation and use of MitoPY1 for imaging hydrogen peroxide in mitochondria of live cells

被引：142

作者：

Dickinson, Bryan C. ^{[1
]}

Lin, Vivian S. ^{[1
]}

Chang, Christopher J. ^{[1
]}

机构：

[1] Univ Calif Berkeley, Howard Hughes Med Inst, Dept Chem & Mol & Cell Biol, Berkeley, CA 94720 USA

来源：

NATURE PROTOCOLS | 2013年 / 8卷 / 06期

基金：

美国国家卫生研究院;

关键词：

FLUORESCENT-PROBES; OXIDATIVE STRESS; H2O2; CHEMISTRY; BIOLOGY; SPECIFICITY; ROS;

D O I：

10.1038/nprot.2013.064

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Mitochondria peroxy yellow 1 (MitoPY1) is a small-molecule fluorescent probe that selectively tracks to the mitochondria of live biological specimens and responds to local fluxes of hydrogen peroxide (H2O2) by a turn-on fluorescence enhancement. This bifunctional dye uses a triphenylphosphonium targeting group and a boronate-based molecular switch to selectively respond to H2O2 over competing reactive oxygen species (ROS) within the mitochondria. MitoPY1 can be used to measure mitochondrial H2O2 levels in both cell culture and tissue models. In this protocol, we describe the synthesis of MitoPY1 and how to use this chemical tool to visualize mitochondrial H2O2 in live cells. The preparation of MitoPY1 is anticipated to take 7-10 d, and assays involving microscopy of cultured mammalian cells can be performed in 1-2 d.

引用

页码：1249 / 1259

页数：11

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