Work

Protocol for multi-organ isolation of murine leukocytes for flow cytometry

Year: 2026

Type: article

Abstract: The immune system comprises several specialized cells across multiple organs. Here, we present a protocol for the isolation of leukocytes from murine blood, spleen, mesenteric lymph nodes, liver, kidney, and skeletal muscle. We describe steps for cell isolation and gradient density separation and provide four antibody panels to identify adaptive immune cells (e.g., CD4 + and CD8 + T cell subsets) by flow cytometry. This protocol provides a basic framework for phenotyping immune cells from in vivo mouse models. • A streamlined protocol for murine leukocyte isolation from multiple organs • Guidelines for multi-organ processing for in vivo mouse models • Instructions for multiparameter flow cytometry analyses of immune T lymphocyte subsets Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The immune system comprises several specialized cells across multiple organs. Here, we present a protocol for the isolation of leukocytes from murine blood, spleen, mesenteric lymph nodes, liver, kidney, and skeletal muscle. We describe steps for cell isolation and gradient density separation and provide four antibody panels to identify adaptive immune cells (e.g., CD4 + and CD8 + T cell subsets) by flow cytometry. This protocol provides a basic framework for phenotyping immune cells from in vivo mouse models.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104318

Authors: Paul Jerard Layug, Agata Marcinow, David Sontag, Asher A. Mendelson, Janilyn Arsenio

Institution: University of Manitoba

Language: English

Topic: T-cell and B-cell Immunology

Subfield: Immunology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550