本文關(guān)鍵詞： 微流控芯片 便攜式 恒溫 灌流細(xì)胞培養(yǎng) 溫度仿真 循環(huán)系統(tǒng) 酵母菌 細(xì)胞臨界頻率　出處：《哈爾濱工業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：細(xì)胞是各項(xiàng)生命活動(dòng)的基本單元,也是研究各種生命現(xiàn)象的基礎(chǔ)。觀察細(xì)胞的生長(zhǎng)狀況,對(duì)于探究生命的規(guī)律及疾病的預(yù)防、診斷和治療等具有非常重要的意義。微流控芯片由于其微型化和高度集成化等特點(diǎn)以及快速、高效的優(yōu)勢(shì),通過集成其他功能部件可實(shí)現(xiàn)對(duì)微粒、細(xì)胞等微米級(jí)尺寸的物體控制,在單細(xì)胞和高通量的研究中較傳統(tǒng)方法具有性價(jià)比高、樣品消耗量小等優(yōu)勢(shì),在生命科學(xué)和化學(xué)領(lǐng)域中具有廣闊的應(yīng)用前景。本文在比較并分析了常規(guī)細(xì)胞培養(yǎng)方法及微流控細(xì)胞培養(yǎng)系統(tǒng)的優(yōu)缺點(diǎn)的基礎(chǔ)上,自行設(shè)計(jì)并制作了基于微流控技術(shù)的便攜式恒溫細(xì)胞培養(yǎng)系統(tǒng),開展了灌流細(xì)胞培養(yǎng)實(shí)驗(yàn),驗(yàn)證了該系統(tǒng)的各項(xiàng)功能。選取薄膜熱敏電阻溫度傳感器和PID智能溫度控制器作為恒溫控制系統(tǒng);采用多物理場(chǎng)仿真軟件COMSOL Multiphysics 4.4對(duì)細(xì)胞培養(yǎng)微流控芯片及恒溫培養(yǎng)腔體進(jìn)行溫度場(chǎng)仿真以探究能夠滿足細(xì)胞正常生長(zhǎng)的恒溫控制條件。利用ITO導(dǎo)電玻璃經(jīng)清洗、光刻、電極成型等工藝步驟制作出恒溫箱加熱器;選擇可編程單軸步進(jìn)電機(jī)及驅(qū)動(dòng)器、控制器來搭建細(xì)胞培養(yǎng)液循環(huán)系統(tǒng)。利用數(shù)控銑床加工出以聚甲基丙烯酸甲酯作為材料的微流控芯片,并使用經(jīng)固化的聚二甲基硅氧烷作為頂部材料,利用高頻等離子輝光放電儀將二者結(jié)合。本文將恒溫細(xì)胞培養(yǎng)系統(tǒng)按功能分為細(xì)胞培養(yǎng)微流控芯片,細(xì)胞培養(yǎng)液循環(huán)系統(tǒng),細(xì)胞恒溫培養(yǎng)箱體三個(gè)部分并分別搭建、組裝起來。當(dāng)溫度設(shè)定為310.15±0.2K的范圍內(nèi),開始進(jìn)行穩(wěn)定性測(cè)試,從而證實(shí)了這個(gè)系統(tǒng)的可行性。在微流控芯片細(xì)胞培養(yǎng)平臺(tái)上進(jìn)行了酵母菌連續(xù)96h的培養(yǎng)對(duì)照實(shí)驗(yàn),經(jīng)比較并分析實(shí)驗(yàn)結(jié)果,證實(shí)了該恒溫細(xì)胞培養(yǎng)系統(tǒng)所提供的恒定且適宜的培養(yǎng)溫度及密閉的培養(yǎng)環(huán)境對(duì)細(xì)胞培養(yǎng)具有重要的作用和意義;驗(yàn)證了本恒溫細(xì)胞培養(yǎng)系統(tǒng)在細(xì)胞培養(yǎng)方面的高效性、優(yōu)越性和實(shí)用性等特點(diǎn)。利用微電極進(jìn)行了酵母菌細(xì)胞臨界頻率的測(cè)量實(shí)驗(yàn),為以后在片細(xì)胞培養(yǎng)以及相關(guān)實(shí)驗(yàn)開展提供參考。
[Abstract]:Cells are the basic units of all life activities and the basis for studying various life phenomena. Observing the growth of cells, exploring the laws of life and the prevention of diseases, Microfluidic chips have the advantages of miniaturization and high integration, as well as the advantages of fast and high efficiency. By integrating other functional components, microfluidic chips can realize the implementation of microparticles. In single-cell and high-throughput research, object control with cell size of micron size has the advantages of high performance-to-price ratio and small sample consumption. In this paper, the advantages and disadvantages of conventional cell culture methods and microfluidic cell culture systems are compared and analyzed. A portable constant temperature cell culture system based on microfluidic technology was designed and manufactured, and the experiment of perfusion cell culture was carried out. The functions of the system are verified. The thin film thermistor temperature sensor and PID intelligent temperature controller are selected as the constant temperature control system. The multi-physical field simulation software COMSOL Multiphysics 4.4 was used to simulate the temperature field of cell culture microfluidic chip and thermostat chamber in order to explore the constant-temperature control conditions which could satisfy the normal growth of cells. The ITO conductive glass was cleaned and lithography. The thermostat heater is made by the electrode forming process, and the programmable single-axis stepping motor and driver are selected. The microfluidic chip with polymethyl methacrylate as the material and the cured polydimethylsiloxane as the top material was fabricated by the numerical control milling machine. The thermostatic cell culture system was divided into three parts according to its function: cell culture microfluidic chip, cell culture medium circulatory system, cell constant temperature incubator. When the temperature was set to 310.15 鹵0.2K, the stability test was carried out, which proved the feasibility of the system. The control experiment of yeast culture for 96 hours was carried out on the microfluidic chip cell culture platform. By comparing and analyzing the experimental results, it is proved that the constant and suitable culture temperature and the closed culture environment provided by the thermostatic cell culture system play an important role in cell culture. The high efficiency, superiority and practicability of the thermostatic cell culture system in cell culture were verified. The cell critical frequency of yeast was measured by microelectrode. It provides a reference for cell culture and related experiments in the future.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：Q813;TN492

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本文編號(hào)：1539772