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Related Experiment Video

Updated: Oct 26, 2025

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors
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Morpho-metabotyping the oxidative stress response.

Mate Rusz1,2, Giorgia Del Favero3,4, Yasin El Abiead1

  • 1Institute of Analytical Chemistry, University of Vienna, Währinger Str. 38, 1090, Vienna, Austria.

Scientific Reports
|July 30, 2021
PubMed
Summary

This study introduces a new workflow combining metabolomics and live cell imaging to comprehensively analyze cellular responses to oxidative stress. The method reveals how cancer cells adapt their metabolism and mitochondrial function when exposed to hydrogen peroxide.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Metabolomics

Background:

  • Oxidative stress and reactive oxygen species (ROS) are critical in numerous biological processes, yet comprehensive cellular analysis remains challenging.
  • Existing methods struggle to integrate biochemical and functional cellular status, limiting a holistic understanding of stress responses.

Purpose of the Study:

  • To develop and validate a fully parallelized workflow integrating metabolomics and live cell imaging microscopy.
  • To investigate the morphological and metabolic adaptations of cancer cells to short-term hydrogen peroxide (H2O2) exposure in vitro.

Main Methods:

  • A novel workflow combining high-throughput metabolomics (quantifying >100 metabolites) with live cell imaging microscopy.
  • Correlative imaging strategy to link metabolic profiles with mitochondrial morphology and function.
  • Analysis of cancer cells exposed to hydrogen peroxide (H2O2) in vitro.

Main Results:

  • Elevated superoxide concentrations with significant mitochondrial localization were observed.
  • Multi-parametric image analysis indicated a shift towards mitochondrial fragmentation.
  • Metabolic profiling revealed impaired mitochondrial function, shifts in cellular defense pathways, and energy compensation mechanisms.

Conclusions:

  • The developed workflow effectively captures comprehensive cellular readouts under oxidative stress conditions.
  • This integrated approach provides deep insights into cancer cell adaptation to short-term H2O2-induced oxidative stress.
  • The workflow is adaptable for studying various short-term cellular stress phenomena.