Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells

Research output: Contribution to journalArticleAcademicpeer-review

18 Citations (Scopus)

Abstract

Self-seeding microwell chips can sort single cells into 6400 wells based on cell size and their identity verified by immunofluorescence staining. Here, we developed a microfluidic device in which these single cells can be placed, lysed and their DNA amplified for further interrogation. Whole blood spiked with MCF7 tumor cells was passed through the microwell chips after leukocyte depletion and 37% of the MCF7 cells were identified by epithelial cell adhesion molecule (EpCAM) staining in the microwells. Identified single cells were punched into the reaction chamber of the microfluidic device and reagents for cell lysis and DNA amplification introduced sequentially by peristaltic pumping of micro-valves. On-chip lysis and amplification was performed in 8 parallel chambers yielding a 10000 fold amplification of DNA. Accessibility of the sample through the reaction chamber allowed for easy retrieval and interrogation of target-specific genes to characterize the tumor cells.
Original languageEnglish
Pages (from-to)4331-4337
JournalLab on a chip
Volume15
Issue number22
DOIs
Publication statusPublished - 10 Sep 2015

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Lab-On-A-Chip Devices
Microfluidics
Amplification
DNA
Blood
Cells
Tumors
MCF-7 Cells
Neoplasms
Cell adhesion
Cell Adhesion Molecules
Staining and Labeling
Genes
Molecules
Cell Size
Fluorescent Antibody Technique
Leukocytes

Keywords

  • METIS-311901
  • IR-97494

Cite this

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title = "Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells",
abstract = "Self-seeding microwell chips can sort single cells into 6400 wells based on cell size and their identity verified by immunofluorescence staining. Here, we developed a microfluidic device in which these single cells can be placed, lysed and their DNA amplified for further interrogation. Whole blood spiked with MCF7 tumor cells was passed through the microwell chips after leukocyte depletion and 37{\%} of the MCF7 cells were identified by epithelial cell adhesion molecule (EpCAM) staining in the microwells. Identified single cells were punched into the reaction chamber of the microfluidic device and reagents for cell lysis and DNA amplification introduced sequentially by peristaltic pumping of micro-valves. On-chip lysis and amplification was performed in 8 parallel chambers yielding a 10000 fold amplification of DNA. Accessibility of the sample through the reaction chamber allowed for easy retrieval and interrogation of target-specific genes to characterize the tumor cells.",
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author = "Yang, {Yoon Sun} and Rho, {Hoon Suk} and Michiel Stevens and Tibbe, {Arjan G.J.} and Gardeniers, {Johannes G.E.} and Terstappen, {Leonardus Wendelinus Mathias Marie}",
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Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells. / Yang, Yoon Sun; Rho, Hoon Suk; Stevens, Michiel; Tibbe, Arjan G.J.; Gardeniers, Johannes G.E.; Terstappen, Leonardus Wendelinus Mathias Marie.

In: Lab on a chip, Vol. 15, No. 22, 10.09.2015, p. 4331-4337.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells

AU - Yang, Yoon Sun

AU - Rho, Hoon Suk

AU - Stevens, Michiel

AU - Tibbe, Arjan G.J.

AU - Gardeniers, Johannes G.E.

AU - Terstappen, Leonardus Wendelinus Mathias Marie

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AB - Self-seeding microwell chips can sort single cells into 6400 wells based on cell size and their identity verified by immunofluorescence staining. Here, we developed a microfluidic device in which these single cells can be placed, lysed and their DNA amplified for further interrogation. Whole blood spiked with MCF7 tumor cells was passed through the microwell chips after leukocyte depletion and 37% of the MCF7 cells were identified by epithelial cell adhesion molecule (EpCAM) staining in the microwells. Identified single cells were punched into the reaction chamber of the microfluidic device and reagents for cell lysis and DNA amplification introduced sequentially by peristaltic pumping of micro-valves. On-chip lysis and amplification was performed in 8 parallel chambers yielding a 10000 fold amplification of DNA. Accessibility of the sample through the reaction chamber allowed for easy retrieval and interrogation of target-specific genes to characterize the tumor cells.

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