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

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity06:08

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity

25.0K
A flow cytometry-based method to quantitatively determine the cytotoxic activity of human natural killer cells is shown...
25.0K
Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity10:14

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

18.6K
This protocol describes a sensitive, cell-based cytotoxicity assay. By enumerating the decrease in frequency of live target CD4+ T cells in the presence of an increasing number of effector CD8+ T cells, this assay allows for the direct assessment of cytolytic activity of antigen-specific CD8+ T...
18.6K
Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis02:40

Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis

690
This video demonstrates the lytic activity of natural killer cells against fluorescently-stained target cells. The method enables NK cell-induced cytotoxicity measurement by staining dead cells and using a flow cytometer to quantify live and dead target...
690
Assessing Antibody-dependent, Cell-mediated Cytotoxicity in Cancer Cells using Antibody-Dependent Cell-Mediated Cytotoxicity Reporter Bioassay05:21

Assessing Antibody-dependent, Cell-mediated Cytotoxicity in Cancer Cells using Antibody-Dependent Cell-Mediated Cytotoxicity Reporter Bioassay

849
Here, we present a protocol for an antibody-dependent, cell-mediated cytotoxicity (ADCC) assay using an ADCC bioassay kit. This method offers a valuable tool for elucidating the ADCC mechanism and evaluating the therapeutic potential of antibodies in cancer immunotherapy.
849
An Assay for Bacterial Extracellular Protein-Mediated Cytotoxicity on PMNs03:14

An Assay for Bacterial Extracellular Protein-Mediated Cytotoxicity on PMNs

449
This video demonstrates an assay to investigate the cytotoxicity of proteins produced by Staphylococcus aureus, a pathogenic bacterium, on isolated human polymorphonuclear leukocytes (PMNs). The secreted cytotoxic proteins form pores on the PMN cell membrane, initiating cell death. Upon staining the dead cells using propidium iodide, flow cytometry is performed to quantify the dead cells and assess the cytotoxicity of the...
449
Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes12:12

Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes

14.0K
Cell-mediated lymphocytotoxicity (CML) assays can be used to test autoreactive responses and study mechanisms of cell death in vitro. However, using live-cell confocal microscopic imaging techniques with fluorescent dyes, the type and kinetics of cell death as well as the pathways utilized can be studied in greater detail.
14.0K

You might also read

Related Articles

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

Sort by
Same author

Integrated multi-omics analysis reveals distinct microbiota-metabolite signatures and a novel HCN2-2-hydroxybutyric acid interaction in inflammatory bowel disease.

Frontiers in nutrition·2026
Same author

Traditional Chinese medicine syndrome patterns and associated factors in adults with type 2 diabetes and metabolic syndrome: a data-driven analysis.

Frontiers in endocrinology·2026
Same author

[Research advances in the biosynthesis and delivery of RNAi-based biopesticides].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology·2026
Same author

Mucosal Involvement in Bullous Pemphigoid: Severity Indicator and Associated Clinical Features from a Retrospective Cohort Study.

Journal of inflammation research·2026
Same author

Correction: KAP1-associated transcriptional inhibitory complex regulates C2C12 myoblasts differentiation and mitochondrial biogenesis via miR-133a repression.

Cell death & disease·2026
Same author

Estimated Plasma Volume Status May Modify the Association Between Estimated Glomerular Filtration Rate and Atrial Fibrillation in HFpEF: A Post Hoc Analysis of the TOPCAT Trial.

Cardiorenal medicine·2026

Related Experiment Video

Updated: Jan 19, 2026

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity
06:08

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity

Published on: August 9, 2017

25.0K

A versatile flow-based assay for immunocyte-mediated cytotoxicity.

Peter M Rabinovich1, Jialing Zhang1, Samuel R Kerr1

  • 1Department of Pathology, Yale School of Medicine, New Haven, CT 06525, USA.

Journal of Immunological Methods
|September 17, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a novel, rapid assay for measuring cell-mediated cytotoxicity by analyzing stained cell nuclei. This method accurately quantifies target and effector cell viability, improving immunotherapy assessment.

Keywords:
CancerChimeric Antigen Receptor (CAR)Cytotoxicity assayNK cellsNucleiStainingT cellsTumor Infiltrating Lymphocytes (TIL)

More Related Videos

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
10:14

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Published on: December 17, 2013

18.6K
Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis
02:40

Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis

690

Related Experiment Videos

Last Updated: Jan 19, 2026

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity
06:08

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity

Published on: August 9, 2017

25.0K
Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
10:14

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Published on: December 17, 2013

18.6K
Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis
02:40

Natural Killer Cell-Mediated Cytotoxicity Assay Sample Preparation for Flow Cytometric Analysis

690

Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Cell-mediated cytotoxicity is crucial for immune defense against pathogens and cancers.
  • Existing methods for assessing cytotoxicity have significant limitations.
  • Accurate measurement of both target and effector cell viability is essential.

Purpose of the Study:

  • To develop a novel, direct, sensitive, rapid, and adaptable method for measuring cell-mediated cytotoxicity.
  • To enable accurate assessment of both target and effector cell viability.
  • To overcome drawbacks of current cytotoxicity assays.

Main Methods:

  • Fluorescently labeling target cells with a protein-binding dye.
  • Incubating labeled target cells with effector cells to initiate killing.
  • Staining dead cells with a specific dye and releasing nuclei using Triton X-100.
  • Analyzing stained nuclei via flow cytometry to differentiate cell types and viability.

Main Results:

  • A novel assay provides four distinct nuclear staining patterns for target/effector and live/dead cell identification.
  • Analyzing nuclei significantly reduces flow cytometry errors caused by cell aggregates.
  • Assay completion time is reduced to 2 hours, enabling high-throughput analysis.
  • Successfully validated across NK-92 cells, CAR-T cells, and TILs.

Conclusions:

  • The developed nuclear analysis method is broadly applicable, highly sensitive, and easy to perform.
  • This technique serves as a powerful tool for evaluating immunotherapy-based cell-mediated cytotoxicity.
  • It offers a significant advancement over existing methods for cytotoxicity assessment.