单细胞光电化学传感平台的构建及胞内过氧化氢检测应用

Cells are known as the basic unit of life. Single-cell analysis, which can reveal the diversity and differences of cellular components and physiological behavior, is the frontier technology in the field of life science. Because of the tiny volume, complex microenvironment, low component content and the significant individual differences of the cell, single-cell analysis techniques should be characterized by high sensitivity, high selectivity and high detection throughput. Photoelectrochemical (PEC) sensing have been proven to be an effective method for high-throughput detection due to its addressing feature of excitation light source and noteworthy advantages of low cost and low background. However, there are no related work reports about the PEC analysis applied in quantitative detection of intracellular substances in single cell and high-throughput single-cell analysis. In this project, we plan to develop single cell PEC analysis platform based on fabricated microwell electrode array by combining light-addressing detection strategy with direct redox sensing principle. On this basis, the intracellular second messenger hydrogen peroxide is selected as the target molecule. The hydrogen peroxide content in different single cells will be quickly analyzed by the developed platform to reveal the heterogeneity of the cells. In summary, the research of this project can not only explore the application of PEC analysis in the field of material analysis in single cells, but also promote the development of high-throughput single cell detection technology.

细胞是组成生命体的基本单元。单细胞分析可以揭示细胞物质组成、生理行为的多样性和差异性，是生命分析的前沿领域。由于单细胞体积极小、微环境复杂、被测组分含量低且不同单细胞个体之间存在着明显差异，因此单细胞分析技术应具备高灵敏度、高选择性和高检测通量等特点。光电化学分析因其激发光源的寻址特性和低成本、低背景的优势，被证明是一种高效的高通量检测方法。但是，光电化学分析在对单细胞内物质的定量检测和高通量传感方面，迄今尚无相关工作报道。本项目中，我们将构建微孔电极阵列并采用寻址检测策略，基于目标物直接氧化还原的传感原理，发展单细胞光电化学分析平台。在此基础上，选择细胞内第二信使过氧化氢为目标分子，对不同单细胞内的过氧化氢含量进行快速分析检测，揭示细胞间的异质性。通过本项目研究，探索光电化学传感在单细胞内物质分析领域的应用，促进高通量单细胞检测技术的发展。

单细胞分析可以揭示细胞物质组成、生理行为的多样性和差异性，是生命分析的前沿领域。由于单细胞体积极小、微环境复杂、被测组分含量低且不同。单细胞个体之间存在着明显差异，因此单细胞分析技术应具备高灵敏度、高选择性和高检测通量等特点。本项目旨在将光电化学传感技术应用于单细胞检测领域，突破单细胞分析在高通量、原位检测方面的一些瓶颈，促进单细胞检测技术的发展。围绕这一目标，利用光电化学方法具有的低背景、高灵敏度、高时空分辨率和原位检测的优势，以及光源的寻址高通量检测能力构建了一系列光电化学生物传感器。项目执行期间,制备了黑磷/碳纳米管、硫铟锌、硫铟银、钒酸铋/碳纳米管等高转换效率的光电活性材料；构建了无标记的光电化学传感器实现血液中microRNA的高灵敏检测；构建了抗污染光电化学超微传感器成功对单细胞内胞质多巴胺分子进行了自驱动原位检测；设计并构建了寻址夹心式光电化学微阵列传感器实现对单细胞释放硫化氢的高通量检测，揭示了单细胞异质性；构建了柔性超灵敏光电化学传感器成功对汗液中的葡萄糖进行了原位监测，促进光电化学传感器的生物应用。本项目主要创新性成果包括：.① 成功构建了光电化学超微传感器插入单个细胞内进行原位检测；.② 首次制备了夹心式光电化学微传感器单细胞进行高通量分析。