| Microfluidic cell disruption system employing a magnetically actuated diaphragm. | |
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MedLine Citation:
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PMID: 18008309 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Yun Suk Huh; Jong Hyun Choi; Kyoung Ae Kim Huh; Kyoung Ae Kim; Tae Jung Park; Yeon Ki Hong; Do Hyun Kim; Won Hi Hong; Sang Yup Lee |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Electrophoresis Volume: 28 ISSN: 0173-0835 ISO Abbreviation: Electrophoresis Publication Date: 2007 Dec |
Date Detail:
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Created Date: 2007-12-17 Completed Date: 2008-04-01 Revised Date: 2008-08-13 |
Medline Journal Info:
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Nlm Unique ID: 8204476 Medline TA: Electrophoresis Country: Germany |
Other Details:
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Languages: eng Pagination: 4748-57 Citation Subset: IM |
Affiliation:
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Separation Process Laboratory, Department of Chemical and Biomolecular Engineering (BK21 program), Korea Advanced Institute of Science and Technology, Daejeon, Korea. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Base Sequence DNA Primers Magnetics* Microfluidics / instrumentation* Polymerase Chain Reaction Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers |
| Comments/Corrections | |
Erratum In:
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Electrophoresis. 2008 May;29(9):1954 Note: Kim, Kyoung Ae [added]; Kim, Do Hyun [added]Electrophoresis. 2008 Jul;29(13):2927 Note: Kim, Kyoung Ae [corrected to Huh, Kyoung Ae Kim] |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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