Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe

Nataliya M. Mishina, Kseniya N. Markvicheva, Dmitry S. Bilan, Mikhail E. Matlashov, Marina V. Shirmanova, David Liebl, Carsten Schultz, Sergey Lukyanov, Vsevolod V. Belousov

Research output: Chapter in Book/Report/Conference proceedingChapter

26 Scopus citations

Abstract

The fluorescent sensor HyPer allows monitoring of intracellular H 2O2 levels with a high degree of sensitivity and specificity. Here, we provide a detailed protocol of ratiometric imaging of H2O2 produced by cells during phagocytosis, including instructions for experiments on different commercial confocal systems, namely, Leica SP2, Leica SP5, and Carl Zeiss LSM, as well as wide-field Leica 6000 microscope. The general experimental scheme is easily adaptable for imaging H2O2 production by various cell types under a variety of conditions.

Original languageEnglish (US)
Title of host publicationHydrogen Peroxide and Cell Signaling, Part A
PublisherAcademic Press Inc.
Pages45-59
Number of pages15
ISBN (Print)9780124058835
DOIs
StatePublished - 2013

Publication series

NameMethods in Enzymology
Volume526
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • HyPer
  • Hydrogen peroxide
  • Imaging
  • Phagocytosis
  • Reactive oxygen species
  • Sensor

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Mishina, N. M., Markvicheva, K. N., Bilan, D. S., Matlashov, M. E., Shirmanova, M. V., Liebl, D., Schultz, C., Lukyanov, S., & Belousov, V. V. (2013). Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe. In Hydrogen Peroxide and Cell Signaling, Part A (pp. 45-59). (Methods in Enzymology; Vol. 526). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-405883-5.00003-X