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

Bryan C. Dickinson; Vivian S. Lin; Christopher J. Chang

doi:10.1038/nprot.2013.064

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

Bryan C. Dickinson, Vivian S. Lin, Christopher J. Chang

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146 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	1249-1259
Number of pages	11
Journal	Nature Protocols
Volume	8
Issue number	6
DOIs	https://doi.org/10.1038/nprot.2013.064
State	Published - Jun 2013
Externally published	Yes

All Science Journal Classification (ASJC) codes

General Biochemistry, Genetics and Molecular Biology

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10.1038/nprot.2013.064

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title = "Preparation and use of MitoPY1 for imaging hydrogen peroxide in mitochondria of live cells",

abstract = "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.",

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

Abstract

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