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

CDK9 interacts with a RanGTP-importin-β complex to regulate erythroid enucleation.

Journal of cell science·2026
Same author

Protocol on utilizing murine gastric cancer organoids for modeling subcutaneous, orthotopic primary, and liver metastatic disease in mice.

STAR protocols·2025
Same author

DeepKymoTracker: A tool for accurate construction of cell lineage trees for highly motile cells.

PloS one·2025
Same author

Mutant TP53 switches therapeutic vulnerability during gastric cancer progression within interleukin-6 family cytokines.

Cell reports·2024
Same author

A Pipeline for Dynamic Analysis of Mitochondrial Content in Developing T Cells: Bridging the Gap Between High-Throughput Flow Cytometry and Single-Cell Microscopy Analysis.

Methods in molecular biology (Clifton, N.J.)·2024
Same author

Mitochondrial fusion and altered beta-oxidation drive muscle wasting in a Drosophila cachexia model.

EMBO reports·2024

Related Experiment Video

Updated: Sep 6, 2025

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy
15:00

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy

Published on: January 25, 2008

11.6K

Establishing a multiplex imaging panel to study T cell development in the thymus in mouse.

Amr H Allam1, Sarah M Russell2

  • 1Optical Sciences Centre, School of Science Computing and Engineering Technologies, Swinburne University of Technology, Hawthorn, VIC 3122, Australia; Olivia Newton-John Cancer Research Institute and La Trobe University School of Cancer Medicine, Heidelberg, VIC, 3084, Australia; Immune Signalling Laboratory, Peter MacCallum Cancer Centre, Melbourne, VIC 3000 Australia.

STAR Protocols
|June 30, 2022
PubMed
Summary

This study details multiplexed immunohistochemistry protocols for the mouse thymus. These methods allow for precise identification of beta-selection stage cells and monitoring of pre-T-cell receptor signaling.

Keywords:
Cell BiologyImmunologyMicroscopy

More Related Videos

3D Organotypic Co-culture Model Supporting Medullary Thymic Epithelial Cell Proliferation, Differentiation and Promiscuous Gene Expression
06:47

3D Organotypic Co-culture Model Supporting Medullary Thymic Epithelial Cell Proliferation, Differentiation and Promiscuous Gene Expression

Published on: July 30, 2015

10.9K
Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy
10:08

Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy

Published on: January 7, 2012

17.1K

Related Experiment Videos

Last Updated: Sep 6, 2025

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy
15:00

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy

Published on: January 25, 2008

11.6K
3D Organotypic Co-culture Model Supporting Medullary Thymic Epithelial Cell Proliferation, Differentiation and Promiscuous Gene Expression
06:47

3D Organotypic Co-culture Model Supporting Medullary Thymic Epithelial Cell Proliferation, Differentiation and Promiscuous Gene Expression

Published on: July 30, 2015

10.9K
Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy
10:08

Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy

Published on: January 7, 2012

17.1K

Area of Science:

  • Immunology
  • Cell Biology
  • Histology

Background:

  • Multiplexed immunohistochemistry (mIHC) is a powerful technique for analyzing cellular interactions within intact tissues.
  • The mouse thymus is a critical organ for T-cell development, involving specific differentiation stages.

Purpose of the Study:

  • To establish and describe protocols for applying mIHC to the mouse thymus.
  • To enable the identification of cells at the crucial beta-selection stage.
  • To facilitate monitoring of pre-T-cell receptor (pre-TCR) signaling and cellular responses during T-cell development.

Main Methods:

  • Development and optimization of multiplexed immunohistochemistry protocols.
  • Application of mIHC to mouse thymus tissue sections.
  • Specific staining strategies to identify beta-selection stage thymocytes.
  • Analysis of pre-TCR signaling pathway activation markers.

Main Results:

  • Successfully applied mIHC to mouse thymus, preserving tissue architecture.
  • Developed methods to accurately identify thymocytes at the beta-selection stage.
  • Demonstrated the ability to visualize and quantify pre-TCR signaling events in situ.

Conclusions:

  • Multiplexed immunohistochemistry is a viable and effective method for studying T-cell development in the mouse thymus.
  • These protocols provide a valuable tool for researchers investigating thymocyte differentiation and signaling pathways.
  • The described techniques advance the understanding of cellular events during T-cell maturation.