Imaging H2O2 microdomains in receptor tyrosine kinases signaling

Nataliya M. Mishina, Kseniya N. Markvicheva, Arkady F. Fradkov, Elena V. Zagaynova, Carsten Schultz, Sergey Lukyanov, Vsevolod V. Belousov

Research output: Chapter in Book/Report/Conference proceedingChapter

10 Citations (Scopus)

Abstract

HyPer, a ratiometric genetically encoded fluorescent sensor, is a popular tool for intracellular hydrogen peroxide detection. When expressed in cultured cells, the freely diffusing version of the sensor (HyPer-cyto) detects temporal patterns of H2O2 generation. However, rapid diffusion of the probe within the nucleocytoplasmic compartment averages the H 2O2 signal even in cases of local oxidant production. Consequently, we immobilized the sensor within specific subcellular compartments allowing it to monitor local increases in H2O2. Here, we provide a protocol of ratiometric imaging and ImageJ-based quantification of H2O2 microdomains produced by cells upon physiological stimulation.

Original languageEnglish (US)
Title of host publicationHydrogen Peroxide and Cell Signaling, Part A
Pages175-187
Number of pages13
Volume526
DOIs
StatePublished - 2013
Externally publishedYes

Publication series

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

Fingerprint

Receptor Protein-Tyrosine Kinases
Oxidants
Hydrogen Peroxide
Cultured Cells
Imaging techniques
Sensors
Cells

Keywords

  • Hydrogen peroxide microdomains
  • HyPer
  • Imaging
  • NADPH oxidases
  • Receptor tyrosine kinases
  • Sensor
  • Signaling

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Medicine(all)

Cite this

Mishina, N. M., Markvicheva, K. N., Fradkov, A. F., Zagaynova, E. V., Schultz, C., Lukyanov, S., & Belousov, V. V. (2013). Imaging H2O2 microdomains in receptor tyrosine kinases signaling. In Hydrogen Peroxide and Cell Signaling, Part A (Vol. 526, pp. 175-187). (Methods in Enzymology; Vol. 526). https://doi.org/10.1016/B978-0-12-405883-5.00011-9

Imaging H2O2 microdomains in receptor tyrosine kinases signaling. / Mishina, Nataliya M.; Markvicheva, Kseniya N.; Fradkov, Arkady F.; Zagaynova, Elena V.; Schultz, Carsten; Lukyanov, Sergey; Belousov, Vsevolod V.

Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526 2013. p. 175-187 (Methods in Enzymology; Vol. 526).

Research output: Chapter in Book/Report/Conference proceedingChapter

Mishina, NM, Markvicheva, KN, Fradkov, AF, Zagaynova, EV, Schultz, C, Lukyanov, S & Belousov, VV 2013, Imaging H2O2 microdomains in receptor tyrosine kinases signaling. in Hydrogen Peroxide and Cell Signaling, Part A. vol. 526, Methods in Enzymology, vol. 526, pp. 175-187. https://doi.org/10.1016/B978-0-12-405883-5.00011-9
Mishina NM, Markvicheva KN, Fradkov AF, Zagaynova EV, Schultz C, Lukyanov S et al. Imaging H2O2 microdomains in receptor tyrosine kinases signaling. In Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526. 2013. p. 175-187. (Methods in Enzymology). https://doi.org/10.1016/B978-0-12-405883-5.00011-9
Mishina, Nataliya M. ; Markvicheva, Kseniya N. ; Fradkov, Arkady F. ; Zagaynova, Elena V. ; Schultz, Carsten ; Lukyanov, Sergey ; Belousov, Vsevolod V. / Imaging H2O2 microdomains in receptor tyrosine kinases signaling. Hydrogen Peroxide and Cell Signaling, Part A. Vol. 526 2013. pp. 175-187 (Methods in Enzymology).
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