Development of fully integrated microfluidic system for detection of intracellular constituents and its applications for infectious viral disease = 세포내 성분 분석을 위한 통합 집적 미세유체 시스템의 개발 및 감염성 바이러스 검출로의 응용연구

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Cell lysis, electro-desalting, and sample clean-up by affinity-capturing are used to develop the sample preparation techniques for the embodiment of μ-TAS. And solid phase extraction (SPE) and two phase extraction are demonstrated as the sample separation steps in the microfluidic chip. On the basis of the micro sample preparation steps and separation steps, the fully integrated microfluidic system is developed for detection of intracellular constituents and infectious viral disease. As a first result of sample preparation steps, a microfluidic cell lysis chip equipped with a micro-mixer and solid phase extraction (SPE) was developed and used for quantitative analysis of intracellular proteins. This miniaturized sample preparation system can be employed for any purpose, in which cell disruption is needed to obtain intracellular constituents for the subsequent analysis. This system is comprised of a magnetically actuated micro-mixer to disrupt cells, a hydrophobic valve to manipulate the cell lysate, and a packed porous polymerized monolith chamber to filter debris from the cell lysate. Using recombinant Escherichia coli expressing intracellular enhanced green fluorescent protein (EGFP) as a model bacterium, we optimized the cell disruption condition with respect to the lysis buffer composition, mixing time, and the frequency of the diaphragm in the micro-mixer, which was magnetically actuated by an external magnetic stirrer in the micro-mixer chamber. The lysed sample prepared under the optimal condition was purified by the SPE packed 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 each by 90% and 94%. Thus, this microfluidic cell disruption chip can be used for the efficient lysis of cells for further analysis of intracellular contents in many applications. As another application of sample preparation steps, the simple and rapid ele...
Advisors
Hong, Won-Hiresearcher홍원희researcher
Description
한국과학기술원 : 생명화학공학과,
Publisher
한국과학기술원
Issue Date
2007
Identifier
268705/325007  / 020035895
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생명화학공학과, 2007. 8, [ xiii, 139 ]

Keywords

Lab on a chip; sensor; micro-mixer; cell lysis; viral disease; micro sample preparation; micro sample separation; 랩온어칩; 센서; 미세혼합기; 세포파쇄; 바이러스질환; 미세 시료전처리; 미세 시료분리; Lab on a chip; sensor; micro-mixer; cell lysis; viral disease; micro sample preparation; micro sample separation; 랩온어칩; 센서; 미세혼합기; 세포파쇄; 바이러스질환; 미세 시료전처리; 미세 시료분리

URI
http://hdl.handle.net/10203/29016
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=268705&flag=dissertation
Appears in Collection
CBE-Theses_Ph.D.(박사논문)
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