Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

An enzyme kinetic model for quantitative interpretation of the role of nicotinamide nucleotide transhydrogenase (NNT) in cell physiology.

Free radical biology & medicine·2026
Same author

Optimizing macrophage-targeted intracellular delivery systems for safe and effective immunotherapies.

Advanced drug delivery reviews·2026
Same author

Kv7.2 loss-of-function causes early hyperexcitability and network remodelling.

Brain : a journal of neurology·2026
Same author

Expertise is the new infrastructure.

Journal of microscopy·2026
Same author

Unperturbed dye-based imaging of spontaneous synchronized calcium activity in iPSC-derived neuronal cultures.

iScience·2026
Same author

Calciprotein particles disrupt autophagy in vascular endothelial cells and smooth muscle cells.

Atherosclerosis plus·2026

Related Experiment Video

Updated: Mar 1, 2026

Cellular Redox Profiling Using High-content Microscopy
11:37

Cellular Redox Profiling Using High-content Microscopy

Published on: May 14, 2017

11.6K

Cellular Redox Profiling Using High-content Microscopy.

Tom Sieprath1, Tobias Corne1, Joke Robijns2

  • 1Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp; Cell Systems and Imaging Research Group (CSI), Department of Molecular Biotechnology, Ghent University.

Journal of Visualized Experiments : Jove
|June 2, 2017
PubMed
Summary

Quantify reactive oxygen species (ROS) and mitochondrial function simultaneously in living cells. This high-content microscopy method reveals cellular health and responses to chemical treatments with high resolution.

More Related Videos

Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein
06:10

Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein

Published on: June 18, 2020

7.9K
Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator
07:47

Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator

Published on: October 20, 2021

3.4K

Related Experiment Videos

Last Updated: Mar 1, 2026

Cellular Redox Profiling Using High-content Microscopy
11:37

Cellular Redox Profiling Using High-content Microscopy

Published on: May 14, 2017

11.6K
Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein
06:10

Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein

Published on: June 18, 2020

7.9K
Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator
07:47

Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator

Published on: October 20, 2021

3.4K

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • Reactive oxygen species (ROS) are crucial for cellular signaling but excessive levels cause oxidative stress and damage.
  • Mitochondria are key sources and targets of ROS, necessitating joint analysis of their function and ROS production.
  • Understanding the interplay between mitochondrial health and ROS is vital for studying pathophysiological conditions.

Purpose of the Study:

  • To develop a high-content microscopy-based strategy for simultaneous quantification of intracellular ROS, mitochondrial membrane potential (ΔΨm), and mitochondrial morphology.
  • To enable detailed analysis of cellular health and responses at the individual cell level.
  • To provide a method for discriminating between cellular states following experimental perturbations.

Main Methods:

  • Utilized automated widefield fluorescence microscopy and image analysis of living adherent cells in multi-well plates.
  • Employed fluorescent reporter molecules CM-H2DCFDA for ROS and TMRM for ΔΨm and mitochondrial morphology.
  • Integrated signal intensity and morphological data for multivariate analysis.

Main Results:

  • Achieved simultaneous quantification of ROS, ΔΨm, and mitochondrial morphology in individual living cells.
  • Demonstrated high spatiotemporal resolution, surpassing traditional fluorimetry or flow cytometry.
  • Successfully discriminated between cellular states after chemical perturbation using combined feature analysis.

Conclusions:

  • The developed high-content microscopy assay provides a powerful tool for assessing cellular health by simultaneously measuring ROS and mitochondrial parameters.
  • This method allows for detailed, high-resolution analysis of cellular responses and differences between cell populations or treatments.
  • The approach offers a significant advancement for research into oxidative stress and mitochondrial dysfunction in various biological contexts.