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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃榮山(Long-Sun Huang) | |
dc.contributor.author | Tien-Ming Cheng | en |
dc.contributor.author | 鄭天民 | zh_TW |
dc.date.accessioned | 2021-06-13T04:27:03Z | - |
dc.date.available | 2006-07-29 | |
dc.date.copyright | 2006-07-29 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-20 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33153 | - |
dc.description.abstract | 在過往的數十年中,有關分子生物技術上的許多重大突破與發展,依序解開了許多生物組織的分子網絡與其交互作用,其成果也廣泛運用於製藥或是生命科學之基礎研究。然而,許多更基本的問題依舊懸而未解。比方說,單單一顆存在於人體組織眾多細胞內的幹細胞,是如何啟動增長或演化的機制,達到修補組織的目的?一顆癌化的細胞,是如何藉由人體的循環系統,代謝與擴散至身體不同的器官,並進行吸附與攻擊?很顯然的,單純倚靠現代的分生技術很難解答這些問題。而對於新研究領域的需求,也促成了各式新技術的發展與投入。
本篇論文在微製造的基礎之下,希望可以為系統細胞生物研究領域提供新的研究利器。一個以微流力為基礎的晶片,與光學系統以及微環境控制設備組合成一個多功能的細胞培養平台。本文的主題主要在各個不同科技領域的整合,包括微流體力學、微製造、基本電學、光學系統以及細胞生物學。這也是第一次,細胞培養、篩選、與觀測三大重要的功能同時被整合於同一晶片系統之中。在開始的章節中,論文的重心主要在闡述為微製程的設計,與應用於流體操作的封裝技術,而微流道的詳細測試結果則是另一個主軸。緊接著,有關介電泳技術的理論與實際應用於細胞的相關步驟也詳細的收錄於本文篇章之中。最後,光學系統與微環境的控制完全整合於系統之中,而完成了一個包含了往復式細胞篩選、長時間細胞觀測與培養的複雜系統。數種有關人體疾病的關鍵細胞,骨癌細胞、口腔內皮細胞、人類胚胎肝細胞,以及骨髓幹細胞也實際於系統中測試與培養。完成的系統,相信有潛力將對於人體臨床、基礎研究有一定程度的貢獻。 | zh_TW |
dc.description.abstract | In the past decades, extensive investments in basic biomolecular research have yielded about the many and complex processes involved in the development of an organism. But many questions still remain. How does a single rare cell, such as the stem cell embedded in the human tissue, give rise to a complex, multi-cellular organism? How does a rare tumor cell traveling through the human circuiting system and adhesive to a specifically targeted organ? Obviously, these questions represent a fundamental challenge in developmental biology. The demands of the new methodologies hasten the emerging technologies, such as the BioMEMS and Nanobiotechnology, trying to carry out the remained issues and obtaining the new scientific discovery.
The aim of work this thesis described is to accomplish a microfluidic based system, which is composed of a microfabricated core chip, optical instruments, and a microincubator. The subject matter is coordinated from the connection and integration of different research field, including microfabrication, microscopic optics, and cell biology. This is also the first time a microfabricated platform integrates the reciprocated sorting, in-situing culturing, long-term culture, and nanobiotechnology. In this thesis, early work is devoted on the microfabrication, and the related packaging. Reciprocated cell sorting is one of steps of a complete system. High speed sorting is achieved by the trail design and realizing of the fundamental theories of the hydrodynamics focusing and dielectrophoresis. On-chip culture is testified using several kinds of human living cells. Cytokinesises of the cancerous cells, primary cultured cells, and stem cells, are real-time monitored. The accomplished system is useful in fundamental of rare cell study, and potential in personal cell therapy. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T04:27:03Z (GMT). No. of bitstreams: 1 ntu-95-R93543028-1.pdf: 5990529 bytes, checksum: 46980a6a8ef7be3e8b19eb3f24432aa5 (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
中文摘要 .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .iii Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . .iv List of Symbols. . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .x 1 Introduction 1.1 Biological Application Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1 1.2 Integrated Technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.2.1 MEMS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.2.2 Lab-On-A-Chip . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 1.2.3 Nnaobiotechnology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 1.3 Microfluidic Device for Cellomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 1.3.1 Transport, Sorting, Manipulation and Characterization in Mechanical Way . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1.3.2 Electric Trapping and Sorting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1.4 Design Fundamentals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 1.4.1 Flow cytomrtry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.4.2 IACS (Image Activated Cell Sorting) . . . . . . . . . . . . . . . . . . . . . . . . . . 9 1.4.3 Design Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10 1.5 Outline of the Thesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14 1.5.1 Spiral Design model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 1.5.2 Chapter Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 2 Microfabrication of the Micro-Bioreactor . . . . . . . . . . . . . . . . . . . . . . . . . . . .20 2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20 2.2 Process Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20 2.2.1 Silicon Mold Fabrication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21 2.2.2 Detail Size Measurements of the Si Mold . . . . . . . . . . . . . . . . . . . . . .24 2.3 Electronics Fabrication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27 2.3.1 Fabricated Electrodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 2.4 PDMS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 2.4.1 Curing, Patterning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 2.4.2 Plasma Bonding (Activation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 2.5 Assembly of the chip . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 2.5.1 Alignments & Couplings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .31 2.5.2 SU8 Pillar as Aligned Adaptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33 2.5.3 Interconnection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 2.6 Chapter Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35 3 Flow Circuit 3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36 3.2 Technical Specification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36 3.3 Setup and Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .38 3.3.1 Flow Circuit Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 3.4 Hydrodynamic Focusing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .41 3.4.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 3.4.2 Definition of the Nozzle Geometry . . . . . . . . . . . . . . . . . . . . . . . . . . .43 3.4.3 PartⅠ: Theoretical Central Stream Width . . . . . . . . . . . . . . . . . . . . . .45 3.4.4 PartⅡ: Particle Velocity - Predicting Assay. . . . . . . . . . . . . . . . . . . . .47 3.5 Experimental Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . .50 3.5.1 Flow instabilities. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51 3.6 Chapter Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .57 4 Dielectrophoresis 4.1 Chapter Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58 4.1.1 Benefits of the DEP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59 4.2 Theory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59 4.2.1 Dielectrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .60 4.3 Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .64 4.3.1 2D Cross-sectional Simulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65 4.3.2 Parameter Specification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .66 4.3.3 Geometry and Mesh . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .66 4.4 Crossover Frequency Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 4.5 Electrode Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .72 4.5.1 Settling Speed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 4.5.2 High Speed Sorting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 4.5.3 Choice of Electrode Geometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74 4.6 Results and Discussions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79 4.7 Chapter Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .80 5 Screening of the System 5.1 Chapter Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 5.2 Optical System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .81 5.2.1 Optical Microscopy and High Speed Camera . . . . . . . . . . . . . . . . . . .81 5.2.2 Cytoviva . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .82 5.2.3 Fluorescent Emission and Fluorochome . . . . . . . . . . . . . . . . . . . . . . . 83 5.3 Microfluidic Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 5.3.1 Cell Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .85 5.3.2 Non-fouling Coating . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86 5.4 Results of Cell Screening . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87 5.4.1 Particle Tracking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87 5.4.2 Morphological Observation of the Rare Cells . . . . . . . . . . . . . . . . . . .88 5.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Summarized Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .92 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96 Appendix A Instrumental Specification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 Appendix B Fluorescent Spectrum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .105 Appendix C Detail Schematic Layout of the Microchannels . . . . . . . . . . . . . . . . .106 | |
dc.language.iso | en | |
dc.title | 微型罕見細胞反應晶片之製造與平台開發:
往復式細胞篩選、培養與即時觀測 | zh_TW |
dc.title | A Novel Rare Cell Micro-Bioreactor with Reciprocated Cell Sorting, In-Situ Culturing, and Real-time Investigation | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 賴信志(Hsin-Chih Lai),施文彬(Wen-Pin Shih),蔡睿哲(Jui-che Tsai) | |
dc.subject.keyword | 微機電,微製造,微流,介電泳,稀有細胞,骨髓幹細胞,細胞篩選,細胞培養,長時間監測,平台整合, | zh_TW |
dc.subject.keyword | MEMS,microfluidic,mFACS,hydrodynamic focusing,dielectrophoresis,cell sorting, rare cell,bone marrow stem cell,in-situing culturing,time-lapsed investigation,osteosarcoma,cytoviva, | en |
dc.relation.page | 116 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-22 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 應用力學研究所 | zh_TW |
顯示於系所單位: | 應用力學研究所 |
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