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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 胡文聰 | |
| dc.contributor.author | Yu-Kai Hsu | en |
| dc.contributor.author | 徐郁凱 | zh_TW |
| dc.date.accessioned | 2021-06-15T11:48:11Z | - |
| dc.date.available | 2021-08-25 | |
| dc.date.copyright | 2016-08-25 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-12 | |
| dc.identifier.citation | 1. Nagrath, S., et al., Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature, 2007. 450(7173): p. 1235-1239.
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Beveridge, R., Circulating tumor cells in the management of metastatic breast cancer patients. Community oncology, 2007. 4(2): p. 79-82. 24. De Giorgi, U., et al., Circulating tumor cells and [18F] fluorodeoxyglucose positron emission tomography/computed tomography for outcome prediction in metastatic breast cancer. Journal of Clinical Oncology, 2009. 27(20): p. 3303-3311. 25. Eifler, R.L., et al., Enrichment of circulating tumor cells from a large blood volume using leukapheresis and elutriation: proof of concept. Cytometry Part B: Clinical Cytometry, 2011. 80(2): p. 100-111. 26. Lin, H.K., et al., Portable filter-based microdevice for detection and characterization of circulating tumor cells. Clinical Cancer Research, 2010. 16(20): p. 5011-5018. 27. Kaiser, J., Cancer's circulation problem. Science, 2010. 327(5969): p. 1072-1074. 28. Zheng, S., et al., 3D microfilter device for viable circulating tumor cell (CTC) enrichment from blood. Biomedical microdevices, 2011. 13(1): p. 203-213. 29. Dharmasiri, U., et al., Highly efficient capture and enumeration of low abundance prostate cancer cells using prostate‐specific membrane antigen aptamers immobilized to a polymeric microfluidic device. Electrophoresis, 2009. 30(18): p. 3289-3300. 30. Adams, A.A., et al., Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. Journal of the American Chemical Society, 2008. 130(27): p. 8633-8641. 31. Tan, S.J., et al., Microdevice for the isolation and enumeration of cancer cells from blood. Biomedical microdevices, 2009. 11(4): p. 883-892. 32. Tan, S.J., et al., Versatile label free biochip for the detection of circulating tumor cells from peripheral blood in cancer patients. Biosensors and Bioelectronics, 2010. 26(4): p. 1701-1705. 33. Wang, S., et al., Highly efficient capture of circulating tumor cells by using nanostructured silicon substrates with integrated chaotic micromixers. Angewandte Chemie International Edition, 2011. 50(13): p. 3084-3088. 34. Schiro, P.G., et al., Sensitive and High‐Throughput Isolation of Rare Cells from Peripheral Blood with Ensemble‐Decision Aliquot Ranking. Angewandte Chemie International Edition, 2012. 51(19): p. 4618-4622. 35. Punnoose, E.A., et al., Molecular biomarker analyses using circulating tumor cells. PloS one, 2010. 5(9): p. e12517. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49784 | - |
| dc.description.abstract | 血液中某些稀少細胞經提取檢測後，能提供作為臨床疾病診斷的重要指標。例如，癌症轉移是經由腫瘤釋放出循環腫瘤細胞 (circulating tumor cells (CTCs))進入病人的血液或淋巴管中，再經由血液或淋巴循環系統散佈到身體其他器官並在適應後形成新的腫瘤。目前的醫學影像和血液中癌症的相關抗體診斷於癌症轉移初期階段難以檢測，觀察在病人血液中循環腫瘤細胞轉移的及時情況提供醫生在臨床診斷上的幫助。觀察病人血液中的CTC不僅能作為治療療程效果評估及治療過程的監視，且CTC於病人血液中數目與患者之「總生存時間」(OS)及「無疾病進展」(PFS)之關係亦被證實。然而CTC在血液中的數量極為稀少，約十億個血球細胞中才有數顆CTC，因此快速地分離並偵測血液中的CTC仍舊是工程及醫學上的重要挑戰。
本研究提出一自動化微流多工處理離心碟盤系統，利用密度梯度離心方式從血液中分離出CTC再以免疫螢光標定之後，對抓取之目標細胞進行辨識。本研究分別針對不同的需求提供兩種不同設計之微流結構離心碟盤系統運用於多工處理複數血液樣本中稀少細胞之分離，並根據不同的需求使用不同的容器安裝在碟盤上進行後續的細胞收集與染色，以及一自動化操作機台來執行大部分流程。其中一種細胞收集容器得以直接在碟盤上進行螢光染色，並將收集到的細胞置於螢光顯微鏡下進行辨識計數，且搭配自動化系統將繁雜的傳統密度梯度離心及染色的步驟予以自動化來減少人為操作實驗之誤差。另一種細胞收集容器則為手動操作染色步驟，再進行後續之辨認和基因分析或藥物測試。為了瞭解兩種不同設計之碟盤之效率及執行性，於3.75毫升健康人血中混入人類乳腺癌細胞株(MCF-7)來模擬病人血液中稀少之CTC進行測試，而收集到的細胞進行anti-EpCAM、anti-cytokeratin、anti-CD45及Hoechst33342免疫螢光標定，並以螢光顯微鏡辨識計數細胞回收率(recovery rate)。 實驗結果顯示，細胞回收率不受不同的碟盤設計影響，平均的回收率為81.4%，並且在不同碟盤的各個分離層中皆得到高於80%的回收率並且相互間無明顯之差異。因此本系統展現出高血液處理速率、高回收率、高自動化的操作流程。可望在未來提供醫生於臨床診斷上一個方便且信賴之診斷工具。 | zh_TW |
| dc.description.abstract | Rare cells in blood possess much clinical significance for diagnosis. One type of rare cells is circulating tumor cells which has been demonstrated to have prognosis value in metastatic breast, prostate and colorectal cancer patients. Hence, these cells have the potential to detect and monitor metastatic events. However, population of CTCs is extremely rare compared to other peripheral blood cells causing great difficulty in their detection and isolation. This thesis presents a microfluidic disk platform for multi-throughput enrichment of rare cells. This platform automates several sequential process: density separation, enrichment of rare cells and immunofluorescence staining with multiple blood samples simultaneously. About 100 tumor cells (MCF-7) were labeled by epithelial cell adhesion molecule (EpCAM) and spiked into multiple blood samples to interrogate the protocol. Results show the multi-sample disk can detect over 80% of spiked cells from multiple blood samples regardless of MCF-7 EpCAM expression levels. Moreover, the multi-sample disk enables different immunofluorescence staining process for the target cells collected in different collecting reservoirs. The disk should be amenable for multi-sample processing of CTC with high recovery rate. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T11:48:11Z (GMT). No. of bitstreams: 1 ntu-105-R03543067-1.pdf: 1651262 bytes, checksum: 2833f132b223b926b2bf9e4300be9f5f (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 謝詞 ii
中文摘要 iii Abstract v List of figures viii List of tables ix Chapter 1. Introduction 1 1.1 Clinical importance of rare cells 1 1.2 Technologies of Circulating Tumor Cells Enrichment 3 1.3 Development of CTC technology in our lab 5 Chapter 2. Design Feature and Methodology of the Multi-sample Disk 8 2.1 Density separation method 8 2.2 System setup 10 2.3 Disk and chip technologies 12 2.4 High throughput multi-sample disk 14 Chapter 3. Material and Methods 18 3.1 Materials 18 3.1.1 Disk fabrication 18 3.1.2 Cells and Cell Culture 21 3.1.3 Reagents 21 3.2 Methods 22 3.2.2 CTC isolation from human peripheral blood 22 3.2.3 Immunofluorescence and staining 24 Chapter 4. Results and Discussion 26 4.1 Validation of disk performance 26 4.2 Efficiency of different separation reservoirs 27 4.3 Mutual interference of different layers 29 4.4 Disk performance 31 Chapter 5. Conclusions 35 Reference 37 | |
| dc.language.iso | en | |
| dc.subject | 多工處理 | zh_TW |
| dc.subject | 微流體 | zh_TW |
| dc.subject | 密度梯度 | zh_TW |
| dc.subject | 碟盤 | zh_TW |
| dc.subject | 循環腫瘤細胞 | zh_TW |
| dc.subject | Circulating tumor cells | en |
| dc.subject | density gradient | en |
| dc.subject | disk | en |
| dc.subject | microfluidics | en |
| dc.subject | multi-sample | en |
| dc.title | 新型微流碟盤應用於多工處理全血中稀少細胞之分離抓取之研究 | zh_TW |
| dc.title | A Novel Microfluidic Disk Enabling Multi-sample Isolation of Rare Cells from Peripheral Blood | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李雨,許聿翔 | |
| dc.subject.keyword | 循環腫瘤細胞,碟盤,密度梯度,微流體,多工處理, | zh_TW |
| dc.subject.keyword | Circulating tumor cells,density gradient,disk,microfluidics,multi-sample, | en |
| dc.relation.page | 39 | |
| dc.identifier.doi | 10.6342/NTU201602143 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-12 | |
| 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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