HyPer-3: A Genetically Encoded H2O2 Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging
ACS Chemical Biology2012Vol. 8(3), pp. 535–542
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Dmitry S. Bilan, Luke Pase, Linda Joosen, Andrey Yu. Gorokhovatsky, Yulia G. Ermakova, Theodorus W. J. Gadella, Clemens Grabher, Carsten Schultz, Sergey Lukyanov, Vsevolod V. Belousov
Abstract
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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