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Fast temporal detection of intracellular hydrogen peroxide by HyPer

  • Yang, Yu-Mi (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry) ;
  • Lee, Sung Jun (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry) ;
  • Shin, Dong Min (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry)
  • Received : 2013.11.26
  • Accepted : 2013.12.16
  • Published : 2013.12.31

Abstract

HyPer is the genetically encoded biosensor of intracellular hydrogen peroxide ($H_2O_2$), the most stable of the reactive oxygen species (ROS) generated by living cells. HyPer has a high sensitivity and specificity for detecting intracellular $H_2O_2$ by confocal laser microscopy. However, it was not known whether high speed ratiometric imaging of $H_2O_2$ by HyPer is possible. We thus investigated the sensitivity of HyPer in detecting changes to the intracellular $H_2O_2$ levels in HEK293 and PC12 cells using a microfluorometer imaging system. Increase in the HyPer ratio were clearly evident on stimulations of more than $100{\mu}M$ $H_2O_2$ and fast changes in the HyPer ratio were observed on ratiometric fluorescent images after $H_2O_2$ treatment. These results suggest that HyPer is a potent biosensor of the fast temporal production of intracellular $H_2O_2$.

Keywords

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