Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/1652
標題: 利用微環境操控、培養與即時觀測單一A549細胞之生長
Manipulation, culturing and real-time investigation of a single A549 cell by microenvironment
作者: 洪彥哲
Hung, Yen-Che
關鍵字: microchannels;微流道;microwells;single cell trapping;microwells;single cell trapping
出版社: 機械工程學系所
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摘要: 
現今,操控單一細胞技術中已有多種方法,可有效將細胞抓取於微環境內。這些技術包含光鑷子、介電泳、磁場與微孔洞設計等。其中微孔洞設計的方法,僅利用流體傳輸系統並可降低對細胞的傷害。然而現今微孔洞的設計,目的皆在抓取單一細胞,而忽略細胞於微孔內後續的生長與分裂等情形。本研究將建立一套單一細胞抓取與培養裝置,採用二進二出的雙主流道,多個分支流道型式與20個微孔洞。本實驗所採用的細胞為A549、MDCK和HeLa,其細胞貼附直徑約50μm
,故在細胞培養區域設計為(50μm×50μm×50μm),利用此微結構設計可成功的將單一細胞抓取於微孔洞內。主流道流入分支流道之流量將影響單一細胞是否被抓取之機率,而分支流量將依據抓取厚度s值決定,且s值與注入端流速和支流的流量有關。由模擬分析可知,於主流道後半段支流的s值較大,亦表示分支流量較大,故細胞較易被抓取於後段區域之微孔洞。實驗結果發現,細胞被抓取的孔洞較集中於13~20之間。於後續培養實驗,當細胞於培養區域內經過36、48小時的培養,可發現細胞有明顯貼附之情形與培養皿內細胞之生長情形一致。

Currently, there are various approaches related to trapping a single cell technology which can effectively capture a single cell. These trapping technologies include optical tweezers, electrophoresis, magnetic field, microwells. Among them, microwells, by fluid transmission system, might less affect the statistical nature of many cellular events. In the literature review of microwells, several studies presented how to trap cells in the wells, but lack of studies reported how to culture cells in the wells. This thesis developed a single cell trapping and culturing devices. The cells studies in this thesis were A549, MDCK and HeLa. The diameters of those cells are less than fifty micrometer. In order to culture those cells in the microwells, the size of microwells were set as to 50μm×50μm×50μm. In this thesis, a microfluidic device was developed which was able to capture a single cell and in the feature of two primary channels, multi sub-channels and twenty microwells. Experimental results show that the single cells were much easier to be trapped in 13th to 20th microwells. We found that the amount of the flow from primary channel into sub-channel affects the trapping the percentage of single cell seized. The virtual width of the flow entering a sub-channel is defined as “s” and can be simulated in the finite element analysis. Large s will increase the probability of single cell trapping. At the same flow rate, microwells near outlet have larger s. The s can be determined by flow rate and fluid quantity of sub-channels. After capturing single cells in the microwells, cells were cultured for a few days. After 36 and 48 hours, Cells showed similar morphology as the cells growth in the petri dishes.
URI: http://hdl.handle.net/11455/1652
其他識別: U0005-2208201115553800
Appears in Collections:機械工程學系所

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