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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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 -