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

• Published in: ACS 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
• Language: English
• 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
• ISSN: 1554-8937
• DOI: 10.1021/cb300625g

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.