基于微流控芯片的细胞代谢指纹可视化方法研究-生物学专业论文.docxVIP

摘 要 本文立足于开发一种细胞代谢分析研究的新方法，利用卟啉等染料分子对生命物 质分子的敏感识别能力构建一种基于 PVDF（聚偏氟乙烯）膜的比色阵列，基于比色 阵列对多成分物质检测的交叉响应的能力及其对细胞代谢物变化响应所表现出的不同 的细胞代谢指纹图谱而用于细胞代谢物变化监测和细胞区分；然后在借鉴 Biolog 系统 对微生物代谢指纹图谱时建立的微生物标准化培养方法的基础上进行了细胞培养标准 化的探索，并在此基础上标准化培养了 6 种不同的动物细胞，同时利用所构建的比色 阵列进行细胞代谢指纹图谱研究，结合 HCA、PCA 等分析方法成功地将细胞代谢指 纹可视化的方法用于细胞代谢监测和细胞区分；基于微流控芯片及其检测系统在细胞 培养、流体操纵和实时检测方面所表现出的优点，设计了 4 块可用于比色阵列检测的 微流控分析芯片，并且在此基础上初步构建了微流控芯片-比色阵列-CCD 的微检测系 统用于芯片上细胞培养和实时比色阵列检测，并在对检测方法和技术充分了解的基础 上得出这种新的微流控芯片比色阵列检测方法具有简单、小型化、对细胞无损、对细 胞代谢物多成分物质响应的实时检测（监测）的优点，很有潜力用于细胞研究和药物 筛选。 关键词： 细胞标准化培养 比色阵列 微流控芯片 细胞代谢指纹图谱 可视化方法 ABSTRACT The study focus on developing an new method for cell metabolic fingerprinting analysis. First a PVDF (polyvinylidene difluoride) membrane colorimetric array are constructed utilizing the sensitive detection ability for bio-molecules of porphyrin and other dyes, the membrane colorimetric array has a cross-reaction property for multicomponent substance detecton and shows great color profile response to cell metabolites change. The color profile change of the colorimetric array could greatly translate the cell metabolic fingerprinting and make the cell metabolic fingerprinting visualized. Then the colorimetric method was successfully used for cell metabolites change monitoring and cell discriminant with cell standard culure. The cell standard cell culture method are explored with the conference of Biolog stystem for standard microorganism culture, six kinds of cells are cultured with the cell standard culture and used for colorimetric array detection, and the HCA and PCA analysis show that the colorimetric array cell metabolic fingerprinting method could be used for cell discriminant. Based on the cell culture, fluid control and real-time detection abilities of microfluidic chip, four microfluidic chip are designed for colorimetric array real-time detection, and further used for the microfluidic chip-colorimetric array-CCD contruction. The microsystem integrate on chip cell culture and colorimetric detection