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Related Experiment Video

Updated: May 16, 2026

Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
09:32

Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells

Published on: February 8, 2018

Protocol for mapping tumor infiltration by murine T cells using multiplex immunofluorescence.

Raymond J Lim1, Camelia Dumitras2, Kostyantyn Krysan3

  • 1Division of Pulmonary and Critical Care, Department of Medicine, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, CA 90095, USA.

STAR Protocols
|May 14, 2026
PubMed
Summary

This study introduces a 7-plex immunofluorescence protocol for analyzing immune cells and the tumor microenvironment in mouse models. It enables spatial mapping of CD4+ and CD8+ T cells to understand tumor immunology.

Keywords:
CancerCell BiologyImmunology

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Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
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Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment

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Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
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Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment

Published on: March 25, 2020

Related Experiment Videos

Last Updated: May 16, 2026

Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
09:32

Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells

Published on: February 8, 2018

Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
06:05

Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment

Published on: June 2, 2023

Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
11:00

Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment

Published on: March 25, 2020

Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • The tumor microenvironment (TME) is complex, involving intricate interactions between tumor cells and immune cells.
  • Understanding immune cell infiltration and function within the TME is crucial for developing effective cancer immunotherapies.

Purpose of the Study:

  • To present a novel 7-plex immunofluorescence staining protocol for spatial analysis of murine tumor tissues.
  • To enable detailed characterization of immune cell populations, particularly CD4+ and CD8+ T cells, within the TME.

Main Methods:

  • Sequential antibody labeling strategy for multiplexed immunofluorescence staining.
  • Application of the protocol to murine tumor models for spatial tissue analysis.
  • Identification and characterization of T cell subsets (CD4+, CD8+) and their activation/regulatory states.

Main Results:

  • The protocol successfully visualizes and quantifies CD4+ and CD8+ T cells within the tumor microenvironment.
  • Spatial distribution and infiltration patterns of T cells are mapped, revealing their localization relative to tumor structures.
  • The method allows for the investigation of T cell activation and regulatory phenotypes in situ.

Conclusions:

  • This 7-plex immunofluorescence protocol provides a powerful tool for dissecting the spatial immune landscape of tumors in murine models.
  • It facilitates a deeper understanding of immune cell dynamics and interactions within the TME, crucial for cancer research.
  • The protocol supports the investigation of immune cell infiltration and function, aiding in the development of novel immunotherapeutic strategies.