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Related Experiment Video

Updated: May 30, 2025

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
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30-Color Longitudinal Full-Spectrum Immunophenotyping and Sorting of Human Circulating Immune Cell Subsets Implicated

Nikhil Jiwrajka, Florin Tuluc, Maria C Jaimes

    Biorxiv : the Preprint Server for Biology
    |January 27, 2025
    PubMed
    Summary
    This summary is machine-generated.

    This study introduces a 30-color flow cytometry panel for precise immune cell subset analysis in autoimmune diseases. It enables efficient sorting and high-parameter immunophenotyping of critical immune cells from limited samples.

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    Area of Science:

    • Immunology
    • Autoimmune Diseases
    • Flow Cytometry

    Background:

    • Systemic autoimmune diseases like rheumatoid arthritis and lupus involve complex immune cell dysregulation.
    • Accurate enumeration and purification of immune cell subsets are crucial for understanding pathogenesis.
    • Existing methods may lack the resolution or efficiency for comprehensive immune profiling.

    Purpose of the Study:

    • To develop and validate a 30-color flow cytometry panel for detailed immune cell subset analysis.
    • To facilitate the enumeration and purification of circulating immune cells implicated in autoimmune diseases.
    • To provide a robust tool for high-parameter immunophenotyping from limited sample quantities.

    Main Methods:

    • Development of a 30-color flow cytometry panel utilizing 30 markers.
    • Application to peripheral blood mononuclear cells and other tissue types using CD45 and autofluorescence spectral signatures.
    • Profiling of T cell subsets (18 markers), B cell subsets (>10 markers), and immune lineages (8 markers).

    Main Results:

    • The panel reproducibly identifies distinct immune cell populations with excellent resolution.
    • Minimal batch effects were observed over several months of data acquisition.
    • The panel enables efficient sorting of immune cell subsets for downstream applications.

    Conclusions:

    • This 30-color panel offers a practical and high-resolution approach for immune cell subset analysis in autoimmune disease research.
    • It allows for simultaneous high-parameter immunophenotyping and cell sorting, conserving valuable sample material.
    • The panel is suitable for investigating immune cell involvement in systemic autoimmune diseases and beyond.