基于微流控芯片的便携式恒温灌流细胞培养系统的研究

发布时间：2018-02-26 21:38

本文关键词： 微流控芯片便携式恒温灌流细胞培养温度仿真循环系统酵母菌细胞临界频率　出处：《哈尔滨工业大学》2015年硕士论文　论文类型：学位论文

【摘要】：细胞是各项生命活动的基本单元,也是研究各种生命现象的基础。观察细胞的生长状况,对于探究生命的规律及疾病的预防、诊断和治疗等具有非常重要的意义。微流控芯片由于其微型化和高度集成化等特点以及快速、高效的优势,通过集成其他功能部件可实现对微粒、细胞等微米级尺寸的物体控制,在单细胞和高通量的研究中较传统方法具有性价比高、样品消耗量小等优势,在生命科学和化学领域中具有广阔的应用前景。本文在比较并分析了常规细胞培养方法及微流控细胞培养系统的优缺点的基础上,自行设计并制作了基于微流控技术的便携式恒温细胞培养系统,开展了灌流细胞培养实验,验证了该系统的各项功能。选取薄膜热敏电阻温度传感器和PID智能温度控制器作为恒温控制系统;采用多物理场仿真软件COMSOL Multiphysics 4.4对细胞培养微流控芯片及恒温培养腔体进行温度场仿真以探究能够满足细胞正常生长的恒温控制条件。利用ITO导电玻璃经清洗、光刻、电极成型等工艺步骤制作出恒温箱加热器;选择可编程单轴步进电机及驱动器、控制器来搭建细胞培养液循环系统。利用数控铣床加工出以聚甲基丙烯酸甲酯作为材料的微流控芯片,并使用经固化的聚二甲基硅氧烷作为顶部材料,利用高频等离子辉光放电仪将二者结合。本文将恒温细胞培养系统按功能分为细胞培养微流控芯片,细胞培养液循环系统,细胞恒温培养箱体三个部分并分别搭建、组装起来。当温度设定为310.15±0.2K的范围内,开始进行稳定性测试,从而证实了这个系统的可行性。在微流控芯片细胞培养平台上进行了酵母菌连续96h的培养对照实验,经比较并分析实验结果,证实了该恒温细胞培养系统所提供的恒定且适宜的培养温度及密闭的培养环境对细胞培养具有重要的作用和意义;验证了本恒温细胞培养系统在细胞培养方面的高效性、优越性和实用性等特点。利用微电极进行了酵母菌细胞临界频率的测量实验,为以后在片细胞培养以及相关实验开展提供参考。
[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 卤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.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：Q813;TN492

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