High-throughput single-cell manipulation system for a large number of target cells

Takahiro Arakawa, Masao Noguchi, Keiko Sumitomo, Yoshinori Yamaguchi, Shuichi Shoji

    Research output: Contribution to journalArticle

    26 Citations (Scopus)

    Abstract

    A sequential and high-throughput single-cell manipulation system for a large volume of cells was developed and the successive manipulation for single cell involving single-cell isolation, individual labeling, and individual rupture was realized in a microhydrodynamic flow channel fabricated by using two-dimensional simple flow channels. This microfluidic system consisted of the successive single-cell handlings of single-cell isolation from a large number of cells in cell suspension, labeling each isolated single cell and the lysate extraction from each labeled single cell. This microfluidic system was composed of main channels, cell-trapping pockets, drain channels, and single-cell content collection channels which were fabricated by polydimethylsiloxane. We demonstrated two kinds of prototypes for sequential single-cell manipulations, one was equipped with 16 single-cell isolation pockets in microchannel and the other was constructed of 512 single-cell isolation pockets. In this study, we demonstrated high-throughput and high-volume single-cell isolation with 512 pocket type device. The total number of isolated single cells in each isolation pocket from the cell suspension at a time was 426 for the cell line of African green monkey kidney, COS-1, and 360 for the rat primary brown preadipocytes, BAT. All isolated cells were stained with fluorescence dye injected into the same microchannel successfully. In addition, the extraction and collection of the cell contents was demonstrated using isolated stained COS-1 cells. The cell contents extracted from each captured cell were individually collected within each collection channel by local hydrodynamic flow. The sequential trapping, labeling, and content extraction with 512 pocket type devices realized high-throughput single-cell manipulations for innovative single-cell handling, feasible staining, and accurate cell rupture.

    Original languageEnglish
    Article number014114
    JournalBiomicrofluidics
    Volume5
    Issue number1
    DOIs
    Publication statusPublished - 2011 Mar 23

    Fingerprint

    Labeling
    manipulators
    Cell Count
    Throughput
    Channel flow
    Microchannels
    Microfluidics
    Suspensions
    cells
    Cell Separation
    Polydimethylsiloxane
    Rats
    Coloring Agents
    Hydrodynamics
    Dyes
    Fluorescence
    Cells
    isolation
    marking
    Rupture

    ASJC Scopus subject areas

    • Genetics
    • Molecular Biology
    • Condensed Matter Physics
    • Materials Science(all)
    • Physical and Theoretical Chemistry

    Cite this

    High-throughput single-cell manipulation system for a large number of target cells. / Arakawa, Takahiro; Noguchi, Masao; Sumitomo, Keiko; Yamaguchi, Yoshinori; Shoji, Shuichi.

    In: Biomicrofluidics, Vol. 5, No. 1, 014114, 23.03.2011.

    Research output: Contribution to journalArticle

    Arakawa, Takahiro ; Noguchi, Masao ; Sumitomo, Keiko ; Yamaguchi, Yoshinori ; Shoji, Shuichi. / High-throughput single-cell manipulation system for a large number of target cells. In: Biomicrofluidics. 2011 ; Vol. 5, No. 1.
    @article{6a2f64fbea514d089a3b926561ee91b8,
    title = "High-throughput single-cell manipulation system for a large number of target cells",
    abstract = "A sequential and high-throughput single-cell manipulation system for a large volume of cells was developed and the successive manipulation for single cell involving single-cell isolation, individual labeling, and individual rupture was realized in a microhydrodynamic flow channel fabricated by using two-dimensional simple flow channels. This microfluidic system consisted of the successive single-cell handlings of single-cell isolation from a large number of cells in cell suspension, labeling each isolated single cell and the lysate extraction from each labeled single cell. This microfluidic system was composed of main channels, cell-trapping pockets, drain channels, and single-cell content collection channels which were fabricated by polydimethylsiloxane. We demonstrated two kinds of prototypes for sequential single-cell manipulations, one was equipped with 16 single-cell isolation pockets in microchannel and the other was constructed of 512 single-cell isolation pockets. In this study, we demonstrated high-throughput and high-volume single-cell isolation with 512 pocket type device. The total number of isolated single cells in each isolation pocket from the cell suspension at a time was 426 for the cell line of African green monkey kidney, COS-1, and 360 for the rat primary brown preadipocytes, BAT. All isolated cells were stained with fluorescence dye injected into the same microchannel successfully. In addition, the extraction and collection of the cell contents was demonstrated using isolated stained COS-1 cells. The cell contents extracted from each captured cell were individually collected within each collection channel by local hydrodynamic flow. The sequential trapping, labeling, and content extraction with 512 pocket type devices realized high-throughput single-cell manipulations for innovative single-cell handling, feasible staining, and accurate cell rupture.",
    author = "Takahiro Arakawa and Masao Noguchi and Keiko Sumitomo and Yoshinori Yamaguchi and Shuichi Shoji",
    year = "2011",
    month = "3",
    day = "23",
    doi = "10.1063/1.3567101",
    language = "English",
    volume = "5",
    journal = "Biomicrofluidics",
    issn = "1932-1058",
    publisher = "American Institute of Physics Publising LLC",
    number = "1",

    }

    TY - JOUR

    T1 - High-throughput single-cell manipulation system for a large number of target cells

    AU - Arakawa, Takahiro

    AU - Noguchi, Masao

    AU - Sumitomo, Keiko

    AU - Yamaguchi, Yoshinori

    AU - Shoji, Shuichi

    PY - 2011/3/23

    Y1 - 2011/3/23

    N2 - A sequential and high-throughput single-cell manipulation system for a large volume of cells was developed and the successive manipulation for single cell involving single-cell isolation, individual labeling, and individual rupture was realized in a microhydrodynamic flow channel fabricated by using two-dimensional simple flow channels. This microfluidic system consisted of the successive single-cell handlings of single-cell isolation from a large number of cells in cell suspension, labeling each isolated single cell and the lysate extraction from each labeled single cell. This microfluidic system was composed of main channels, cell-trapping pockets, drain channels, and single-cell content collection channels which were fabricated by polydimethylsiloxane. We demonstrated two kinds of prototypes for sequential single-cell manipulations, one was equipped with 16 single-cell isolation pockets in microchannel and the other was constructed of 512 single-cell isolation pockets. In this study, we demonstrated high-throughput and high-volume single-cell isolation with 512 pocket type device. The total number of isolated single cells in each isolation pocket from the cell suspension at a time was 426 for the cell line of African green monkey kidney, COS-1, and 360 for the rat primary brown preadipocytes, BAT. All isolated cells were stained with fluorescence dye injected into the same microchannel successfully. In addition, the extraction and collection of the cell contents was demonstrated using isolated stained COS-1 cells. The cell contents extracted from each captured cell were individually collected within each collection channel by local hydrodynamic flow. The sequential trapping, labeling, and content extraction with 512 pocket type devices realized high-throughput single-cell manipulations for innovative single-cell handling, feasible staining, and accurate cell rupture.

    AB - A sequential and high-throughput single-cell manipulation system for a large volume of cells was developed and the successive manipulation for single cell involving single-cell isolation, individual labeling, and individual rupture was realized in a microhydrodynamic flow channel fabricated by using two-dimensional simple flow channels. This microfluidic system consisted of the successive single-cell handlings of single-cell isolation from a large number of cells in cell suspension, labeling each isolated single cell and the lysate extraction from each labeled single cell. This microfluidic system was composed of main channels, cell-trapping pockets, drain channels, and single-cell content collection channels which were fabricated by polydimethylsiloxane. We demonstrated two kinds of prototypes for sequential single-cell manipulations, one was equipped with 16 single-cell isolation pockets in microchannel and the other was constructed of 512 single-cell isolation pockets. In this study, we demonstrated high-throughput and high-volume single-cell isolation with 512 pocket type device. The total number of isolated single cells in each isolation pocket from the cell suspension at a time was 426 for the cell line of African green monkey kidney, COS-1, and 360 for the rat primary brown preadipocytes, BAT. All isolated cells were stained with fluorescence dye injected into the same microchannel successfully. In addition, the extraction and collection of the cell contents was demonstrated using isolated stained COS-1 cells. The cell contents extracted from each captured cell were individually collected within each collection channel by local hydrodynamic flow. The sequential trapping, labeling, and content extraction with 512 pocket type devices realized high-throughput single-cell manipulations for innovative single-cell handling, feasible staining, and accurate cell rupture.

    UR - http://www.scopus.com/inward/record.url?scp=79953271858&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=79953271858&partnerID=8YFLogxK

    U2 - 10.1063/1.3567101

    DO - 10.1063/1.3567101

    M3 - Article

    VL - 5

    JO - Biomicrofluidics

    JF - Biomicrofluidics

    SN - 1932-1058

    IS - 1

    M1 - 014114

    ER -