Bilan, D.S., Pase, L., Joosen, L., Gorokhovatsky, A.Y., Ermakova, Y.G., Gadella, T.W., Grabher, C., Schultz, C., Lukyanov, S., and Belousov, V.V. (2013) HyPer-3: a genetically encoded H2O2 probe with improved performance for ratiometric and fluorescence lifetime imaging. ACS Chemical Biology. 8(3):535-542.
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.