title: Microfluidic cell disruption system employing a magnetically actuated diaphragm.
abstract: A microfluidic cell lysis chip equipped with a micromixer and SPE unit was developed and used for quantitative analysis of intracellular proteins. This miniaturized sample preparation system can be employed for any purpose where cell disruption is needed to obtain intracellular constituents for the subsequent analysis. This system comprises a magnetically actuated micromixer to disrupt cells, a hydrophobic valve to manipulate the cell lysate, and a packed porous polymerized monolith chamber for SPE and filtering debris from the cell lysate. Using recombinant Escherichia coli expressing intracellular enhanced green fluorescent protein (EGFP) and lipase as model bacteria, we optimized the cell disruption condition with respect to the lysis buffer composition, mixing time, and the frequency of the diaphragm in the micromixer, which was magnetically actuated by an external magnetic stirrer in the micromixer chamber. The lysed sample prepared under the optimal condition was purified by the packed SPE in the microfluidic chip. At a frequency of 1.96 Hz, the final cell lysis efficiency and relative fluorescence intensity of EGFP after the cell disruption process were greater than 90 and 94%, respectively. Thus, this microfluidic cell disruption chip can be used for the efficient lysis of cells for further analysis of intracellular contents in many applications.
authors.affiliation: Separation Process Laboratory, Department of Chemical and Biomolecular Engineering (BK21 program), Korea Advanced Institute of Science and Technology, Daejeon, Korea.
authors.forename: Yun Suk
authors.initials: YS
authors.lastname: Huh
authors.forename: Jong Hyun
authors.initials: JH
authors.lastname: Choi
authors.forename: Kyoung Ae Kim
authors.initials: KA
authors.lastname: Huh
authors.forename: Kyoung Ae
authors.initials: KA
authors.lastname: Kim
authors.forename: Tae Jung
authors.initials: TJ
authors.lastname: Park
authors.forename: Yeon Ki
authors.initials: YK
authors.lastname: Hong
authors.forename: Do Hyun
authors.initials: DH
authors.lastname: Kim
authors.forename: Won Hi
authors.initials: WH
authors.lastname: Hong
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D008280:Magnetics; D044085:Microfluidics / Q000295:instrumentation*; D016133:Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D017931:DNA Primers