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

Bottom-Up Absorptive and Stretchable Plasmonic Tape for Field-Deployable In Vivo Fruit Safety Surveillance.

ACS sensors·2026
Same author

Temporal control of Ninj1 activation determines cell-to-cell heterogeneity in IL-33 release.

Communications biology·2026
Same author

Length-based separation of Arthrospira (Spirulina) platensis trichomes via the self-alignment effect of helical filaments in a straight microchannel.

Microsystems & nanoengineering·2026
Same author

High-speed fluorescence lifetime imaging microscopy: techniques, applications, and prospects.

Biophotonics discovery·2026
Same author

Intelligent image-activated sorting of large cells enabled by elasto-inertial focusing.

Lab on a chip·2026
Same author

cGAS-IFN-I responses by extracting nuclear DNA from dying cells via nucleocytosis.

Nature communications·2026
Same journal

A Modular High-Parameter Flow Cytometry Framework: Pre-Analytical Optimization and Validation for Clinical Research.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
Same journal

Quantitative Detection of Entotic Cell-In-Cell Structures Using Deformable Segmentation and Deep Learning.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
Same journal

Comparison of Tissue Preparations to Identify and Phenotype T Cells in Human Colorectal Tumor Tissue.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
Same journal

Refractive Index-Correlated Pseudocoloring for Adaptive Color Fusion in Holotomographic Cytology.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
Same journal

Ensembling Unets for Rare Chromosomal Aberration Detection in Metaphase Images, Uncertainty Quantification, and Ionizing Radiation Dose Estimation.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
Same journal

OMIP-121: Immune Phenotyping of Canine Peripheral Leukocytes by Mass Cytometry.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2026
See all related articles

Related Experiment Video

Updated: May 26, 2025

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
16:10

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

23.8K

Investigating T-Cell Receptor Dynamics Under In Vitro Antibody-Based Stimulation Using Imaging Flow Cytometry.

Akihiro Isozaki1,2, Kazuma Kita3, Natsumi Tiffany Ishii2

  • 1Department of Mechanical Engineering, Ritsumeikan University, Kusatsu, Shiga, Japan.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|February 21, 2025
PubMed
Summary
This summary is machine-generated.

T cell receptor dynamics were studied using antibody stimulation. CD3 and CD28 receptors showed distinct surface changes before cytokine production, indicating potential early activation markers.

Keywords:
in vitro stimulationT cellsimaging flow cytometryreceptor clustering

More Related Videos

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
09:04

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

13.4K
Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry
10:01

Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry

Published on: December 16, 2022

3.8K

Related Experiment Videos

Last Updated: May 26, 2025

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
16:10

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

23.8K
Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
09:04

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

13.4K
Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry
10:01

Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry

Published on: December 16, 2022

3.8K

Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • T cells are crucial immune responders activated by antigen-presenting cells (APCs) through T-cell receptors (TCRs) and immunological synapses.
  • Imaging flow cytometry allows detailed analysis of cellular interactions, but in vitro T-cell receptor dynamics post-stimulation lack comprehensive study.

Purpose of the Study:

  • To investigate the temporal dynamics of T-cell surface receptors (CD3 and CD28) following in vitro antibody-based stimulation without APCs.
  • To determine if receptor morphological changes precede downstream activation events like cytokine production.

Main Methods:

  • Established a Th1 cell clone for in vitro antibody-based stimulation using centrifugation, anti-human CD3, and anti-human CD28 antibodies.
  • Utilized imaging flow cytometry to capture and analyze bright-field and fluorescence images of single cells at various time points post-stimulation.

Main Results:

  • CD3 and CD28 receptors exhibited distinct temporal relocation patterns on the T-cell surface immediately after stimulation.
  • CD3 receptors dispersed by 3.5 hours, while CD28 receptors remained clustered for 7.5 hours post-stimulation.
  • Observed morphological changes in CD3 and CD28 receptor distribution preceded measurable cytokine production.

Conclusions:

  • The distinct temporal dynamics of CD3 and CD28 receptors following in vitro stimulation provide insights into T-cell activation mechanisms.
  • CD3 and CD28 receptor redistribution patterns may serve as early indicators of T-cell activation, preceding cytokine release.