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Updated: Mar 11, 2026
Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
Published on: March 25, 2020
Phenotyping Multiple Subsets of Immune Cells In Situ in FFPE Tissue Sections: An Overview of Methodologies
1Andor Technology, 300 Baker St 150, Concord, MA, 01720, USA. jim@jmansfield.com.
Insights
New methods analyze immune cells in tumor tissues. This allows researchers to understand immune cell roles and spatial relationships within solid tumors, advancing cancer immunotherapy research.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Cancer immunotherapy success highlights the need to study immune cells in solid tumors.
- Current methods like flow cytometry lack spatial resolution for in situ tissue analysis.
- Understanding immune cell organization in the tumor microenvironment is critical.
Purpose of the Study:
- To review methods for phenotyping immune cells in formalin-fixed, paraffin-embedded (FFPE) tissue sections.
- To discuss techniques for analyzing immune cell organization and interrelationships within tumors.
- To bridge the gap between immunological insights and spatial tissue analysis.
Main Methods:
- Review of staining techniques for immune cell identification in FFPE sections.
- Analysis of multiplex imaging approaches for detailed cellular phenotyping.
- Examination of image analysis strategies for assessing spatial relationships.
Main Results:
- Development of methodologies to phenotype immune cells directly within tissue sections.
- Enabling in situ analysis of immune cell subsets and their tumor microenvironment interactions.
- Providing spatially resolved data crucial for understanding tumor immunology.
Conclusions:
- Multiplex imaging and advanced analysis offer powerful tools for immune cell phenotyping in FFPE tissues.
- These methods are essential for understanding immune cell roles in solid tumors and guiding cancer immunotherapy.
- The reviewed techniques facilitate the study of immune cell organization and its impact on tumor progression.
Abstract:
The recent clinical success of new cancer immunotherapy agents and methods is driving the need to understand the role of immune cells in solid tissues, especially tumors. Immune cell phenotyping via flow cytometry, while a cornerstone of immunology, is not spatially resolved and cannot analyze immune cell subsets in situ in clinical biopsy sections or to determine their interrelationships. To address this problem, a number of methodologies have been developed in attempts to phenotype immune and other cells in images acquired from tissue sections and to assess their organization in the tumor and its microenvironment. This chapter review the staining and multiplex image analysis methods that have been developed for phenotyping immune and other cells in formalin-fixed, paraffin-embedded (FFPE) tissue sections.
