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 Experiment Videos

Mitotic index determination by flow cytometry.

David A F Gillespie1, Mark Walker

  • 1CR-UK Beatson Laboratories, Beatson Institute for Cancer Research, Garscube Estate, Bearsden, Glasgow G61 1BD.

Sub-Cellular Biochemistry
|July 13, 2007
PubMed
Summary
This summary is machine-generated.

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

Large-scale exome array summary statistics resources for glycemic traits to aid effector gene prioritization.

Wellcome open research·2024
Same author

Pregnancies with 'double-positive' multiple marker screening results: a population-based study in Ontario, Canada.

BMC pregnancy and childbirth·2024
Same author

Elevations in plasma glucagon are associated with reduced insulin clearance after ingestion of a mixed-macronutrient meal in people with and without type 2 diabetes.

Diabetologia·2024
Same author

Interventions for intimate partner violence during the perinatal period: A scoping review: A systematic review.

Campbell systematic reviews·2024
Same author

Rare germline structural variants increase risk for pediatric solid tumors.

bioRxiv : the preprint server for biology·2024
Same author

Sustaining e-health innovations in a complex hospital environment: learning through evidence.

Frontiers in digital health·2024
Same journal

Future Directions in Biotechnological and Pharmacological Applications of CAIs.

Sub-cellular biochemistry·2026
Same journal

Industrial and Environmental Applications of Carbonic Anhydrases.

Sub-cellular biochemistry·2026
Same journal

Applications of Carbonic Anhydrase Inhibitors in Arthritis, Neuropathic Pain, Acute Mountain Sickness, and Cerebral Ischemia.

Sub-cellular biochemistry·2026
Same journal

Applications of Carbonic Anhydrase Inhibitors in Neurological Disorders, Mechanisms and Therapeutic Potential.

Sub-cellular biochemistry·2026
Same journal

Carbonic Anhydrase Inhibitors in Oncology.

Sub-cellular biochemistry·2026
Same journal

Therapeutic Applications of Carbonic Anhydrase Inhibitors in Ophthalmology.

Sub-cellular biochemistry·2026
See all related articles

This study introduces a method using histone H3 phosphorylation (pS10H3) and flow cytometry to quickly assess mitotic cells. It measures G2 checkpoint efficiency and identifies cells entering mitosis with unreplicated DNA.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Chromosome condensation during mitosis involves histone H3 phosphorylation at serine 10 (pS10H3).
  • Assessing mitotic cell populations is crucial for understanding cell cycle regulation and response to DNA damage.

Purpose of the Study:

  • To develop a rapid and accurate method for determining the percentage of mitotic cells.
  • To evaluate the efficiency of the G2 checkpoint in blocking mitosis after DNA damage.
  • To identify cells entering mitosis with incompletely replicated DNA due to checkpoint failure.

Main Methods:

  • Utilizing phospho-specific antibodies to detect pS10H3.
  • Combining pS10H3 detection with DNA content flow cytometry.
  • Employing nocodazole to arrest cells in mitosis for rate determination.

Related Experiment Videos

Main Results:

  • The pS10H3 and DNA content flow cytometry method accurately quantifies mitotic cells.
  • Comparing mitotic accumulation rates before and after DNA damage assesses G2 checkpoint function.
  • The method identifies cells with abnormal DNA content entering mitosis, indicating S-M checkpoint failure.

Conclusions:

  • Phosphorylation of histone H3 at serine 10 (pS10H3) detection coupled with flow cytometry is an efficient tool for cell cycle analysis.
  • This technique allows for the assessment of G2 checkpoint integrity and the detection of checkpoint failures.
  • It provides insights into cellular responses to DNA damage and replication stress.