Imaging Intracellular H2O2 with the Genetically Encoded PerFRET and OxyFRET Probes

Balázs Enyedi; Miklós Geiszt

doi:10.1007/978-1-4939-9424-3_16

Imaging Intracellular H₂O₂ with the Genetically Encoded PerFRET and OxyFRET Probes

Methods Mol Biol. 2019:1982:275-282. doi: 10.1007/978-1-4939-9424-3_16.

Authors

Balázs Enyedi^{1

2}, Miklós Geiszt³

Affiliations

¹ Faculty of Medicine, Department of Physiology, Semmelweis University, Budapest, Hungary. [email protected].
² MTA-SE Lendﺲlet Tissue Damage Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary. [email protected].
³ Faculty of Medicine, Department of Physiology, Semmelweis University, Budapest, Hungary. [email protected].

PMID: 31172478
DOI: 10.1007/978-1-4939-9424-3_16

Abstract

Hydrogen peroxide (H₂O₂) is an important signaling intermediate with various regulatory and effector functions. Despite its significance, the subcellular organization of H₂O₂ signals is poorly understood. Introducing novel techniques for the intracellular detection of H₂O₂ would be essential for a more complete understanding of its role in cellular signaling. We previously reported the development of two novel fluorescence resonance energy transfer (FRET)-based protein sensors that showed opposite emission ratio changes upon reaction with H₂O₂. In this chapter, we detail the methods for using OxyFRET and PerFRET for the assessment of changes in subcellular H₂O₂ levels.

Keywords: Fluorescent biosensor imaging; Genetically encoded sensor; Hydrogen peroxide; OxyFRET; PerFRET.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biosensing Techniques
Data Analysis
Fluorescence Resonance Energy Transfer
Gene Expression*
Genes, Reporter*
HeLa Cells
Humans
Hydrogen Peroxide / metabolism*
Image Processing, Computer-Assisted
Intracellular Space
Microscopy, Fluorescence
Molecular Imaging* / methods

Substances

Hydrogen Peroxide