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

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

14.1K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
14.1K

You might also read

Related Articles

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

Sort by
Same author

The reproducibility gap in graph neural network workflows for cell dynamics: A checklist-driven case study.

Journal of microscopy·2026
Same author

GloBIAS: strengthening the foundations of bioimage analysis.

Nature methods·2026
Same author

Steroidal hormones and neurosteroids - novel therapeutic strategies in bacterial infections: Design, synthesis, and biological evaluation.

European journal of medicinal chemistry·2026
Same author

From Corrosion Control to Cell Adhesion: Parascholzite as a Functional Interface for Biodegradable Zinc Alloys.

Materials (Basel, Switzerland)·2026
Same author

Estimation of Covid-19 lungs damage based on computer tomography images analysis.

F1000Research·2025
Same author

Impact of Species, Growth Conditions, and Plant Processing on the Phytochemistry and Antimicrobial Activity of Agrimonia Extracts.

Chemistry & biodiversity·2025
Same journal

Chemical, Biological, and Ecological Evidence for Aerobic Deoxynivalenol Detoxification in Agronomic Soil-Derived Bacterial Communities.

Toxins·2026
Same journal

Botulinum Toxin Treatment for Uncommon Phenotypes of Laryngeal Adductor Breathing Dystonia.

Toxins·2026
Same journal

Enhancing Neuronal Networks with <i>Rhinella schneideri</i> Skin Secretion Molecules: Implications for Neurodegenerative Disorders.

Toxins·2026
Same journal

Dangerous Measures: A Case Report and Review of Motoro Ray Envenomation.

Toxins·2026
Same journal

The Impact of OnabotulinumtoxinA on Oral Pain Medication Prescription Fills and Low-Value Care in Patients with Cervical Dystonia in the United States: A Retrospective Claims Analysis.

Toxins·2026
Same journal

Broad-Spectrum Antiviral and Antibacterial Activity of the Scorpion Venom Peptide HP1090.

Toxins·2026
See all related articles

Related Experiment Video

Updated: Aug 1, 2025

An Automated Microscopic Scoring Method for the &#947;-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
08:23

An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes

Published on: December 25, 2021

4.9K

In Vitro High-Throughput Genotoxicity Testing Using γH2AX Biomarker, Microscopy and Reproducible Automatic Image

Bára Křížkovská1, Martin Schätz2, Jan Lipov1

  • 1Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic.

Toxins
|April 27, 2023
PubMed
Summary
This summary is machine-generated.

Quantifying phosphorylated histone (γH2AX) via microscopy offers a reproducible method for genotoxicity testing. Sharing workflows and data is key to advancing this sensitive, high-throughput bioimage analysis technique.

Keywords:
ImageJbioimage analysisgenotoxicityhigh-throughputin vitro testing

More Related Videos

Quantitation of &#947;H2AX Foci in Tissue Samples
08:48

Quantitation of γH2AX Foci in Tissue Samples

Published on: June 28, 2010

14.5K
Immunofluorescence Microscopy of &#947;H2AX and 53BP1 for Analyzing the Formation and Repair of DNA Double-strand Breaks
10:47

Immunofluorescence Microscopy of γH2AX and 53BP1 for Analyzing the Formation and Repair of DNA Double-strand Breaks

Published on: November 3, 2017

16.4K

Related Experiment Videos

Last Updated: Aug 1, 2025

An Automated Microscopic Scoring Method for the &#947;-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
08:23

An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes

Published on: December 25, 2021

4.9K
Quantitation of &#947;H2AX Foci in Tissue Samples
08:48

Quantitation of γH2AX Foci in Tissue Samples

Published on: June 28, 2010

14.5K
Immunofluorescence Microscopy of &#947;H2AX and 53BP1 for Analyzing the Formation and Repair of DNA Double-strand Breaks
10:47

Immunofluorescence Microscopy of γH2AX and 53BP1 for Analyzing the Formation and Repair of DNA Double-strand Breaks

Published on: November 3, 2017

16.4K

Area of Science:

  • Biomarker detection
  • Genotoxicity testing
  • Cellular assays

Background:

  • Phosphorylated histone (γH2AX) is a sensitive biomarker for detecting DNA double-strand breaks in vitro.
  • Microscopy offers an accessible method for γH2AX detection, complementing flow cytometry for genotoxicity assessment.
  • Lack of published details on fluorescence quantification hinders reproducibility in γH2AX-based assays.

Purpose of the Study:

  • To establish and validate a reproducible bioimage analysis workflow for quantifying γH2AX immunofluorescence.
  • To demonstrate the utility of open-source software for high-throughput genotoxicity screening.
  • To provide shareable data and scripts to enhance the reproducibility of γH2AX assays.

Main Methods:

  • Utilized valinomycin as a model genotoxin across HeLa and CHO-K1 cell lines.
  • Employed a commercial kit for γH2AX immunofluorescence detection.
  • Performed bioimage analysis using ImageJ, quantifying mean fluorescence intensity and nuclear area.

Main Results:

  • The developed method confirmed valinomycin's genotoxic and cytotoxic effects on both cell lines after 24 hours.
  • Area-scaled relative fold change in γH2AX fluorescence successfully indicated genotoxicity.
  • Relative nuclear area served as a reliable indicator of cytotoxicity.

Conclusions:

  • Bioimage analysis of overall γH2AX fluorescence intensity presents a viable alternative to flow cytometry for genotoxicity assessment.
  • Sharing workflows, data, and scripts is essential for advancing and standardizing bioimage analysis methods.
  • This approach supports sensitive, specific, and reproducible in vitro genotoxicity testing.