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Updated: Jun 10, 2026
Determination of Regulatory T Cell Subsets in Murine Thymus, Pancreatic Draining Lymph Node and Spleen Using Flow Cytometry
Published on: February 27, 2019
Protocol for identifying recirculating thymic regulatory T cells and characterizing the role of Eos in their function
Xuan Xie1, Tom Hill2, Francisco Otaizo-Carrasquero3
1Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
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Previous studies on the characterization of regulatory T cells (Tregs) in the thymus have proven to be simplistic. Here, we present a protocol for identifying recirculating thymic Tregs in mice and characterizing the role of Eos in their function. We describe steps for thymic dissection, magnetic cell separator (autoMACS) and fluorescence-activated cell sorting (FACS), single-cell sequencing, and downstream computational analysis. This protocol details a refined classification based on analysis of gene expression from single-cell RNA sequencing (scRNA-seq) and TCR sequencing data. For details on the use of this protocol, refer to Xie et al.1.
