Surface-Micromachined Microfiltration Membranes for Efficient Isolation and Functional Immunophenotyping of Subpopulations of Immune Cells

Authors

  • Weiqiang Chen,

    1. Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
    Current affiliation:
    1. These authors contributed equally to this work.
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  • Nien-Tsu Huang,

    1. Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
    Current affiliation:
    1. These authors contributed equally to this work.
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  • Boram Oh,

    1. Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
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  • Raymond H. W. Lam,

    1. Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Hong Kong, China
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  • Rong Fan,

    1. Department of Biomedical Engineering, Yale University, New Haven, CT 06511, USA
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  • Timothy T. Cornell,

    1. Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, MI 48109, USA
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  • Thomas P. Shanley,

    1. Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, MI 48109, USA
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  • Katsuo Kurabayashi,

    Corresponding author
    1. Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
    2. Department of Electrical Engineering and Computer Science, University of Michigan, Ann Arbor, MI 48109 USA
    • Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA.
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  • Jianping Fu

    Corresponding author
    1. Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
    2. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 USA
    • Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109 USA.
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Abstract

An accurate measurement of the immune status in patients with immune system disorders is critical in evaluating the stage of diseases and tailoring drug treatments. The functional cellular immunity test is a promising method to establish the diagnosis of immune dysfunctions. The conventional functional cellular immunity test involves measurements of the capacity of peripheral blood mononuclear cells to produce pro-inflammatory cytokines when stimulated ex vivo. However, this “bulk” assay measures the overall reactivity of a population of lymphocytes and monocytes, making it difficult to pinpoint the phenotype or real identity of the reactive immune cells involved. In this research, we develop a large surface micromachined poly-dimethylsiloxane (PDMS) microfiltration membrane (PMM) with high porosity, which is integrated in a microfluidic microfiltration platform. Using the PMM with functionalized microbeads conjugated with antibodies against specific cell surface proteins, we demonstrated rapid, efficient and high-throughput on-chip isolation, enrichment, and stimulation of subpopulations of immune cells from blood specimens. Furthermore, the PMM-integrated microfiltration platform, coupled with a no-wash homogeneous chemiluminescence assay (“AlphaLISA”), enables us to demonstrate rapid and sensitive on-chip immunophenotyping assays for subpopulations of immune cells isolated directly from minute quantities of blood samples.

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