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

ACS Chem Biol. 2013 Mar 15;8(3):535-42. doi: 10.1021/cb300625g. Epub 2013 Jan 7.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fluorescence*
  • Fluorescent Dyes / analysis*
  • Hydrogen Peroxide / analysis*
  • Hydrogen Peroxide / metabolism
  • Larva / metabolism
  • Luminescent Measurements / methods*
  • Oxidation-Reduction
  • Zebrafish / growth & development
  • Zebrafish / metabolism


  • Fluorescent Dyes
  • Hydrogen Peroxide