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Updated: May 10, 2026
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Imaging of mtHyPer7, a Ratiometric Biosensor for Mitochondrial Peroxide, in Living Yeast Cells
Published on: June 2, 2023
Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe
Nataliya M Mishina1, Kseniya N Markvicheva, Dmitry S Bilan
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, RAS, Moscow, Russia.
Methods in Enzymology
|June 25, 2013
Summary
This study details a protocol for using the HyPer fluorescent sensor to measure hydrogen peroxide (H2O2) in cells during phagocytosis. The method is adaptable across various confocal and wide-field microscopes.
Area of Science:
- Cell biology
- Biochemistry
- Microscopy
Background:
- Intracellular hydrogen peroxide (H2O2) plays a critical role in cellular signaling and responses.
- Monitoring H2O2 production in real-time is essential for understanding cellular processes like phagocytosis.
- Existing methods may lack the sensitivity or specificity required for accurate H2O2 quantification.
Purpose of the Study:
- To provide a detailed, adaptable protocol for ratiometric imaging of intracellular H2O2.
- To enable sensitive and specific monitoring of H2O2 production during phagocytosis.
- To facilitate H2O2 imaging across different microscopy platforms.
Main Methods:
- Utilized the HyPer fluorescent sensor for ratiometric imaging of H2O2.
- Developed experimental schemes for Leica SP2, Leica SP5, Carl Zeiss LSM confocal systems, and Leica 6000 wide-field microscope.
- Focused on H2O2 production by cells during phagocytosis.
Main Results:
- Demonstrated the feasibility of ratiometric H2O2 imaging using HyPer.
- Provided specific instructions for adapting the protocol to various commercial microscopes.
- Established the general applicability of the experimental scheme for diverse cell types and conditions.
Conclusions:
- The presented protocol offers a sensitive and specific method for quantifying cellular H2O2.
- The adaptability of the method across multiple microscope systems enhances its utility for researchers.
- This protocol facilitates the study of H2O2's role in phagocytosis and other cellular processes.
