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

24 Citations (Scopus)

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
Pages45-59
Number of pages15
Volume526
DOIs
StatePublished - 2013
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume526
ISSN (Print)00766879
ISSN (Electronic)15577988

Fingerprint

Cytophagocytosis
Fluorescent Dyes
Hydrogen Peroxide
Visualization
Imaging techniques
Sensitivity and Specificity
Microscopes
Monitoring
Sensors
Experiments

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

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

Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe. / Mishina, Nataliya M.; Markvicheva, Kseniya N.; Bilan, Dmitry S.; Matlashov, Mikhail E.; Shirmanova, Marina V.; Liebl, David; Schultz, Carsten; Lukyanov, Sergey; Belousov, Vsevolod V.

Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526 2013. p. 45-59 (Methods in Enzymology; Vol. 526).

Research output: Chapter in Book/Report/Conference proceedingChapter

Mishina, NM, Markvicheva, KN, Bilan, DS, Matlashov, ME, Shirmanova, MV, Liebl, D, Schultz, C, Lukyanov, S & Belousov, VV 2013, Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe. in Hydrogen Peroxide and Cell Signaling, Part A. vol. 526, Methods in Enzymology, vol. 526, pp. 45-59. https://doi.org/10.1016/B978-0-12-405883-5.00003-X
Mishina NM, Markvicheva KN, Bilan DS, Matlashov ME, Shirmanova MV, Liebl D et al. Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe. In Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526. 2013. p. 45-59. (Methods in Enzymology). https://doi.org/10.1016/B978-0-12-405883-5.00003-X
Mishina, Nataliya M. ; Markvicheva, Kseniya N. ; Bilan, Dmitry S. ; Matlashov, Mikhail E. ; Shirmanova, Marina V. ; Liebl, David ; Schultz, Carsten ; Lukyanov, Sergey ; Belousov, Vsevolod V. / Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe. Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526 2013. pp. 45-59 (Methods in Enzymology).
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