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Updated: May 7, 2026
Enrichment and Characterization of the Tumor Immune and Non-immune Microenvironments in Established Subcutaneous Murine Tumors
Published on: June 7, 2018
Multicolor flow cytometric analysis of immune cell subsets in tumor-bearing mice
Lauren J Bayne1, Robert H Vonderheide
1Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104;
Insights
This protocol details preparing single-cell suspensions from mouse tissues for multicolor flow cytometry. It enables identification of key immune cell populations like T lymphocytes and B lymphocytes.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Characterizing immune cell populations is crucial for understanding immune responses and disease pathogenesis.
- Standardized protocols are needed for reliable analysis of immune cells from solid tissues and spleens.
Purpose of the Study:
- To describe a reproducible protocol for generating single-cell suspensions from mouse spleen and solid tissues.
- To outline the application of multicolor flow cytometry for immune cell subset identification using these suspensions.
Main Methods:
- Single-cell suspension preparation from mouse spleen or solid tissues.
- Multicolor flow cytometry analysis.
- Immunophenotyping based on cell surface molecule expression.
Main Results:
- Successful generation of single-cell suspensions suitable for flow cytometry.
- Identification of distinct immune cell populations including leukocytes, T lymphocytes (CD4+, CD8+), B lymphocytes, NK cells, dendritic cells, and immature myeloid cells.
Conclusions:
- This protocol provides a robust method for immune cell analysis in mice.
- It facilitates comprehensive evaluation of immune cell subsets for research applications.
Abstract:
This protocol describes a procedure for obtaining a single-cell suspension from mouse spleen or solid tissue. Once a single-cell suspension is prepared, multicolor flow cytometry is used to identify subsets of immune cells in the samples of interest. This protocol can be used to evaluate the following immune cell populations, defined by the expression of certain cell surface molecules: total leukocytes, T lymphocytes and CD4(+) and CD8(+) T-cell subsets, B lymphocytes, natural killer cells, dendritic cells, and immature myeloid cells.
