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Measuring Calcium and ROS by Genetically Encoded Protein Sensors and Fluorescent Dyes

  • Christine S. Gibhardt
  • Adina Vultur
  • Ivan BogeskiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1925)

Abstract

Oxidative modifications of cellular building blocks such as proteins, lipids, and DNA have a major impact on cell behavior, fate, and clinical outcome. Reactive oxygen species (ROS) are important factors that influence these redox processes. Calcium ion (Ca2+) dynamics and signals are also essential regulators of key cellular processes. Therefore, the combined and precise monitoring of ROS and Ca2+ in single cells, with a high spatial and temporal resolution and in physiological environments, is essential to better understand their functional impact. Here, we describe protocols to detect one of the most prominent ROS (hydrogen peroxide, H2O2) using genetically encoded protein sensors and fluorescent dyes. We also provide guidelines on how to simultaneously detect Ca2+ and H2O2 and how to examine the influence of Ca2+ signals on cellular ROS production and vice versa.

Key words

Calcium ROS Redox Imaging Microscopy Sensor H2O2 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Christine S. Gibhardt
    • 1
  • Adina Vultur
    • 1
  • Ivan Bogeski
    • 1
    Email author
  1. 1.Molecular Physiology, Institute of Cardiovascular PhysiologyUniversity Medical Center, Georg-August-UniversityGöttingenGermany

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