Parallel single-cell analysis microfluidic platform

Floris Teunis Gerardus van den Brink, Elmar Gool, Jean-Philippe Frimat, Johan G. Bomer, Albert van den Berg, Severine le Gac

    Research output: Contribution to journalArticleAcademicpeer-review

    28 Citations (Scopus)

    Abstract

    We report a PDMS microfluidic platform for parallel single-cell analysis (PaSCAl) as a powerful tool to decipher the heterogeneity found in cell populations. Cells are trapped individually in dedicated pockets, and thereafter, a number of invasive or non-invasive analysis schemes are performed. First, we report single-cell trapping in a fast (2–5 min) and reproducible manner with a single-cell capture yield of 85% using two cell lines (P3x63Ag8 and MCF-7), employing a protocol which is scalable and easily amenable to automation. Following this, a mixed population of P3x63Ag8 and MCF-7cells is stained in situ using the nucleic acid probe (Hoechst) and a phycoerythrin-labeled monoclonal antibody directed at EpCAM present on the surface of the breast cancer cells MCF-7 and absent on the myeloma cells P3x63Ag8 to illustrate the potential of the device to analyze cell population heterogeneity. Next, cells are porated in situ using chemicals in a reversible (digitonin) or irreversible way (lithium dodecyl sulfate). This is visualized by the transportation of fluorescent dyes through the membrane (propidium iodide and calcein). Finally, an electrical protocol is developed for combined cell permeabilization and electroosmotic flow (EOF)-based extraction of the cell content. It is validated here using calcein-loaded cells and visualized through the progressive recovery of calcein in the side channels, indicating successful retrieval of individual cell content.
    Original languageUndefined
    Pages (from-to)3094-3100
    Number of pages7
    JournalElectrophoresis
    Volume32
    Issue number22
    DOIs
    Publication statusPublished - Nov 2011

    Keywords

    • Cancer cell / Microfluidics / Parallelization / Population heterogeneity / Singlecell analysis
    • IR-78992
    • EWI-21018
    • METIS-281652

    Cite this

    van den Brink, Floris Teunis Gerardus ; Gool, Elmar ; Frimat, Jean-Philippe ; Bomer, Johan G. ; van den Berg, Albert ; le Gac, Severine. / Parallel single-cell analysis microfluidic platform. In: Electrophoresis. 2011 ; Vol. 32, No. 22. pp. 3094-3100.
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    abstract = "We report a PDMS microfluidic platform for parallel single-cell analysis (PaSCAl) as a powerful tool to decipher the heterogeneity found in cell populations. Cells are trapped individually in dedicated pockets, and thereafter, a number of invasive or non-invasive analysis schemes are performed. First, we report single-cell trapping in a fast (2–5 min) and reproducible manner with a single-cell capture yield of 85{\%} using two cell lines (P3x63Ag8 and MCF-7), employing a protocol which is scalable and easily amenable to automation. Following this, a mixed population of P3x63Ag8 and MCF-7cells is stained in situ using the nucleic acid probe (Hoechst) and a phycoerythrin-labeled monoclonal antibody directed at EpCAM present on the surface of the breast cancer cells MCF-7 and absent on the myeloma cells P3x63Ag8 to illustrate the potential of the device to analyze cell population heterogeneity. Next, cells are porated in situ using chemicals in a reversible (digitonin) or irreversible way (lithium dodecyl sulfate). This is visualized by the transportation of fluorescent dyes through the membrane (propidium iodide and calcein). Finally, an electrical protocol is developed for combined cell permeabilization and electroosmotic flow (EOF)-based extraction of the cell content. It is validated here using calcein-loaded cells and visualized through the progressive recovery of calcein in the side channels, indicating successful retrieval of individual cell content.",
    keywords = "Cancer cell / Microfluidics / Parallelization / Population heterogeneity / Singlecell analysis, IR-78992, EWI-21018, METIS-281652",
    author = "{van den Brink}, {Floris Teunis Gerardus} and Elmar Gool and Jean-Philippe Frimat and Bomer, {Johan G.} and {van den Berg}, Albert and {le Gac}, Severine",
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    doi = "10.1002/elps.201100413",
    language = "Undefined",
    volume = "32",
    pages = "3094--3100",
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    Parallel single-cell analysis microfluidic platform. / van den Brink, Floris Teunis Gerardus; Gool, Elmar; Frimat, Jean-Philippe; Bomer, Johan G.; van den Berg, Albert; le Gac, Severine.

    In: Electrophoresis, Vol. 32, No. 22, 11.2011, p. 3094-3100.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Parallel single-cell analysis microfluidic platform

    AU - van den Brink, Floris Teunis Gerardus

    AU - Gool, Elmar

    AU - Frimat, Jean-Philippe

    AU - Bomer, Johan G.

    AU - van den Berg, Albert

    AU - le Gac, Severine

    N1 - 10.1002/elps.201100413

    PY - 2011/11

    Y1 - 2011/11

    N2 - We report a PDMS microfluidic platform for parallel single-cell analysis (PaSCAl) as a powerful tool to decipher the heterogeneity found in cell populations. Cells are trapped individually in dedicated pockets, and thereafter, a number of invasive or non-invasive analysis schemes are performed. First, we report single-cell trapping in a fast (2–5 min) and reproducible manner with a single-cell capture yield of 85% using two cell lines (P3x63Ag8 and MCF-7), employing a protocol which is scalable and easily amenable to automation. Following this, a mixed population of P3x63Ag8 and MCF-7cells is stained in situ using the nucleic acid probe (Hoechst) and a phycoerythrin-labeled monoclonal antibody directed at EpCAM present on the surface of the breast cancer cells MCF-7 and absent on the myeloma cells P3x63Ag8 to illustrate the potential of the device to analyze cell population heterogeneity. Next, cells are porated in situ using chemicals in a reversible (digitonin) or irreversible way (lithium dodecyl sulfate). This is visualized by the transportation of fluorescent dyes through the membrane (propidium iodide and calcein). Finally, an electrical protocol is developed for combined cell permeabilization and electroosmotic flow (EOF)-based extraction of the cell content. It is validated here using calcein-loaded cells and visualized through the progressive recovery of calcein in the side channels, indicating successful retrieval of individual cell content.

    AB - We report a PDMS microfluidic platform for parallel single-cell analysis (PaSCAl) as a powerful tool to decipher the heterogeneity found in cell populations. Cells are trapped individually in dedicated pockets, and thereafter, a number of invasive or non-invasive analysis schemes are performed. First, we report single-cell trapping in a fast (2–5 min) and reproducible manner with a single-cell capture yield of 85% using two cell lines (P3x63Ag8 and MCF-7), employing a protocol which is scalable and easily amenable to automation. Following this, a mixed population of P3x63Ag8 and MCF-7cells is stained in situ using the nucleic acid probe (Hoechst) and a phycoerythrin-labeled monoclonal antibody directed at EpCAM present on the surface of the breast cancer cells MCF-7 and absent on the myeloma cells P3x63Ag8 to illustrate the potential of the device to analyze cell population heterogeneity. Next, cells are porated in situ using chemicals in a reversible (digitonin) or irreversible way (lithium dodecyl sulfate). This is visualized by the transportation of fluorescent dyes through the membrane (propidium iodide and calcein). Finally, an electrical protocol is developed for combined cell permeabilization and electroosmotic flow (EOF)-based extraction of the cell content. It is validated here using calcein-loaded cells and visualized through the progressive recovery of calcein in the side channels, indicating successful retrieval of individual cell content.

    KW - Cancer cell / Microfluidics / Parallelization / Population heterogeneity / Singlecell analysis

    KW - IR-78992

    KW - EWI-21018

    KW - METIS-281652

    U2 - 10.1002/elps.201100413

    DO - 10.1002/elps.201100413

    M3 - Article

    VL - 32

    SP - 3094

    EP - 3100

    JO - Electrophoresis

    JF - Electrophoresis

    SN - 0173-0835

    IS - 22

    ER -