Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells

Jaime J. Benítez, Juraj Topolancik, Harvey C. Tian, Christopher B. Wallin, David R. Latulippe, Kylan Szeto, Patrick J. Murphy, Benjamin R. Cipriany, Stephen L. Levy, Paul D. Soloway, Harold G. Craighead

    Research output: Contribution to journalArticlepeer-review

    57 Scopus citations

    Abstract

    We describe a microfluidic device for the extraction, purification and stretching of human chromosomal DNA from single cells. A two-dimensional array of micropillars in a microfluidic polydimethylsiloxane channel was designed to capture a single human cell. Megabase-long DNA strands released from the cell upon lysis are trapped in the micropillar array and stretched under optimal hydrodynamic flow conditions. Intact chromosomal DNA is entangled in the array, while other cellular components are washed from the channel. To demonstrate the entrapment principle, a single chromosome was hybridized to whole chromosome paints, and imaged by fluorescence microscopy. DNA extracted from a single cell and small cell populations (less than 100) was released from the device by restriction endonuclease digestion under continuous flow and collected for off-chip analysis. Quantification of the extracted material reveals that the microdevice efficiently extracts essentially all chromosomal DNA. The device described represents a novel platform to perform a variety of analyses on chromosomal DNA at the single cell level.

    Original languageEnglish (US)
    Pages (from-to)4848-4854
    Number of pages7
    JournalLab on a Chip
    Volume12
    Issue number22
    DOIs
    StatePublished - Nov 21 2012

    ASJC Scopus subject areas

    • Bioengineering
    • Biochemistry
    • General Chemistry
    • Biomedical Engineering

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