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目的设计一种用于玻璃-聚二甲基硅氧烷(PDMS)灌注式细胞培养芯片的温度控制系统,代替传统的细胞培养箱,以提高与细胞灌注培养装置的可集成性,同时便于细胞培养过程中显微镜下动态观察。方法系统采用透光性良好的铟锡氧化物(ITO)镀膜导电玻璃作为加热元件,以Pt100传感器作为测温元件,与PID微控制器构成闭环温度控制系统;玻璃-PDMS细胞培养芯片采用微丝模塑软刻工艺加工而成;利用红外热成像仪对ITO加热元件表面上温度场均匀性进行验证;有限元数值分析对ITO加热元件上细胞培养芯片空间温度场分布进行模拟。结果与结论设计的温度控制系统结构简单,控制精度达±0.2℃;加热元件表面及玻璃-PDMS灌注式细胞培养芯片上培养腔内温度场空间分布均匀(1℃内),可满足实际细胞培养条件;在芯片上进行人肺癌细胞A549灌注培养实验显示,系统为细胞培养提供合适的温度环境且适合于细胞生物学行为的动态表征。
Objective To design a temperature control system for a glass-polydimethylsiloxane (PDMS) perfusion cell culture chip instead of a traditional cell incubator to improve the integration with a cell perfusion culture device while facilitating cell Dynamic observation under microscope during culture. Method The system uses indium tin oxide (ITO) coated conductive glass with good translucency as heating element and Pt100 sensor as temperature measuring element to form closed-loop temperature control system with PID microcontroller. The glass-PDMS cell culture chip adopts microfilament Molded soft-etching process. The uniformity of temperature field on the surface of ITO heating element was verified by infrared thermal imager. The temperature field distribution in the cell culture chip on ITO heating element was simulated by finite element analysis. RESULTS AND CONCLUSIONS The temperature control system was simple in structure with a control accuracy of ± 0.2 ℃. The spatial distribution of the temperature field in the culture chamber on the surface of the heating element and in the glass-PDM perfusion cell culture chip was uniform (within 1 ℃) The results of perfusion experiments on human lung cancer cells A549 showed that the system provided a suitable temperature environment for cell culture and was suitable for the dynamic characterization of cell biological behavior.