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

doi:10.1016/B978-0-12-405883-5.00003-X

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 proceeding › Chapter

38 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 language	English (US)
Title of host publication	Hydrogen Peroxide and Cell Signaling, Part A
Publisher	Academic Press Inc.
Pages	45-59
Number of pages	15
ISBN (Print)	9780124058835
DOIs	https://doi.org/10.1016/B978-0-12-405883-5.00003-X
State	Published - 2013
Externally published	Yes

Publication series

Name	Methods in Enzymology
Volume	526
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

Access to Document

10.1016/B978-0-12-405883-5.00003-X

Cite this

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

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. Methods in Enzymology, vol. 526, Academic Press Inc., pp. 45-59. https://doi.org/10.1016/B978-0-12-405883-5.00003-X

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

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.",

keywords = "HyPer, Hydrogen peroxide, Imaging, Phagocytosis, Reactive oxygen species, Sensor",

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

note = "Funding Information: This work was supported by the Russian foundation for basic research (RFBR) (10-04-01561-a, V. V. B.; 11-04-12187-ofi, S. L.), a joint EMBL-RFBR grant (12-04-92427 to C. S. and V. V. B.), Measures to Attract Leading Scientists to Russian Educational Institutions program (11.G34.31.0017 to S. L.), and the ESF EuroMembrane consortium TraPPs (to C. S.). ",

year = "2013",

doi = "10.1016/B978-0-12-405883-5.00003-X",

language = "English (US)",

isbn = "9780124058835",

series = "Methods in Enzymology",

publisher = "Academic Press Inc.",

pages = "45--59",

booktitle = "Hydrogen Peroxide and Cell Signaling, Part A",

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TY - CHAP

T1 - Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe

AU - Mishina, Nataliya M.

AU - Markvicheva, Kseniya N.

AU - Bilan, Dmitry S.

AU - Matlashov, Mikhail E.

AU - Shirmanova, Marina V.

AU - Liebl, David

AU - Schultz, Carsten

AU - Lukyanov, Sergey

AU - Belousov, Vsevolod V.

N1 - Funding Information: This work was supported by the Russian foundation for basic research (RFBR) (10-04-01561-a, V. V. B.; 11-04-12187-ofi, S. L.), a joint EMBL-RFBR grant (12-04-92427 to C. S. and V. V. B.), Measures to Attract Leading Scientists to Russian Educational Institutions program (11.G34.31.0017 to S. L.), and the ESF EuroMembrane consortium TraPPs (to C. S.).

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

KW - HyPer

KW - Hydrogen peroxide

KW - Imaging

KW - Phagocytosis

KW - Reactive oxygen species

KW - Sensor

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U2 - 10.1016/B978-0-12-405883-5.00003-X

DO - 10.1016/B978-0-12-405883-5.00003-X

M3 - Chapter

C2 - 23791093

AN - SCOPUS:84879765656

SN - 9780124058835

T3 - Methods in Enzymology

SP - 45

EP - 59

BT - Hydrogen Peroxide and Cell Signaling, Part A

PB - Academic Press Inc.

ER -

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

Abstract

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Keywords

ASJC Scopus subject areas

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