Manipulation, culturing and real-time investigation of a single A549 cell by microenvironment

• Main Authors: Yen-Che Hung, 洪彥哲
• Other Authors: 吳嘉哲
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/339e75

碩士 === 國立中興大學 === 機械工程學系所 === 99 === 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.