HyPer-3: a genetically encoded H(2)O(2) probe with improved performance for ratiometric and fluorescence lifetime imaging

High-performance sensors for reactive oxygen species are instrumental to monitor dynamic events in cells and organisms. Here, we present HyPer-3, a genetically encoded fluorescent indicator for intracellular H2O2 exhibiting improved performance with respect to response time and speed. HyPer-3 has an...

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Published inACS chemical biology Vol. 8; no. 3; pp. 535 - 542
Main Authors Bilan, Dmitry S, Pase, Luke, Joosen, Linda, Gorokhovatsky, Andrey Yu, Ermakova, Yulia G, Gadella, Theodorus W J, Grabher, Clemens, Schultz, Carsten, Lukyanov, Sergey, Belousov, Vsevolod V
Format Journal Article
LanguageEnglish
Published United States 15.03.2013
Subjects
Animals
Fluorescence
Fluorescent Dyes - analysis
Hydrogen Peroxide - analysis
Hydrogen Peroxide - metabolism
Larva - metabolism
Luminescent Measurements - methods
Oxidation-Reduction
Zebrafish - growth & development
Zebrafish - metabolism
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Summary:High-performance sensors for reactive oxygen species are instrumental to monitor dynamic events in cells and organisms. Here, we present HyPer-3, a genetically encoded fluorescent indicator for intracellular H2O2 exhibiting improved performance with respect to response time and speed. HyPer-3 has an expanded dynamic range compared to HyPer and significantly faster oxidation/reduction dynamics compared to HyPer-2. We demonstrate this performance by in vivo imaging of tissue-scale H2O2 gradients in zebrafish larvae. Moreover, HyPer-3 was successfully employed for single-wavelength fluorescent lifetime imaging of H2O2 levels both in vitro and in vivo.
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ISSN:1554-8937
DOI:10.1021/cb300625g