title: Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells.
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.
authors.affiliation: School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Jaime J
authors.initials: JJ
authors.lastname: Benítez
authors.forename: Juraj
authors.initials: J
authors.lastname: Topolancik
authors.forename: Harvey C
authors.initials: HC
authors.lastname: Tian
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Wallin
authors.forename: David R
authors.initials: DR
authors.lastname: Latulippe
authors.forename: Kylan
authors.initials: K
authors.lastname: Szeto
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Murphy
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Cipriany
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Levy
authors.forename: Paul D
authors.initials: PD
authors.lastname: Soloway
authors.forename: Harold G
authors.initials: HG
authors.lastname: Craighead
mesh_terms: D045744:Cell Line, Tumor; D005591:Chemical Fractionation / Q000295:instrumentation*; D002877:Chromosomes, Human / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D059010:Single-Cell Analysis / Q000295:instrumentation*
chemical_list: D004247:DNA