用于单细胞原位实时检测的超微阵列传感器研究

Cell is the basis of the physiological functions of the organism and all lives, and the secretion of the cells stimulated has a very important role in the life activities. The microelectrode sensor with integrated multi-array can be used for monitoring of exocytosis from single cell and in situ real-time quantitative detection of single cell, which is significant on research of cell physiological and pathological processes and drug screening..New methods will be developed for the preparation of the integrated ultra-microelectrode biosensor; new technologies will be developed for detection of an extremely small number secretion from single-cell to meet the needs of life science research of the nerves and other major diseases in this study. Deep exploration and research will be done on the electrode preparation process for the integration of advanced micro devices, design of integrated sensor, cell-anchored technology. Key technologies will be developed for multi-parameter detection of multi-array sensor to immobilize and character specific active substances on the electrode's surface; Biocompatibility and stability of the sensor will be studied for in situ, real-time, multi-channel and multi-component monitoring secretion of single cell. The feedback studies on electrode preparation process and modified methods will be done to improve biocompatibility and stability of the sensor. The study will facilitate the revelation of the mechanism on some major human diseases, and promote the joint development of biomedical electronics, micro-nano electronics and information science.

细胞是有机体的生理功能和一切生命现象的基础，细胞的受激释放在生命活动中具有十分重要的作用。通过集成多阵列微电极传感器研究单细胞动态变化，进行细胞分泌物原位实时定量检测分析，对细胞生理、病理过程和药物筛选等研究有重要意义。.本项目面向生命科学研究和重大疾病早期检测治疗等方面的需求，开展集成超微电极阵列生物传感器制备的新方法、新技术及单细胞痕量物质检测等研究。在集成超微多阵列电极制备工艺路线、传感器集成设计、细胞自锚定技术等方面进行深入探索研究；进行多阵列传感器的多参数检测选择修饰关键技术研究，实现不同特异活性物质在电极表面的固定和表征；通过单细胞的原位、实时、微量、多通道、多组分检测，对电极制备工艺和修饰方法等进行反馈研究，解决传感器的生物相容性和稳定性等关键问题。本研究将为人类神经等重大疾病的机理揭示与治疗提供研究基础，并促进生物医学电子学、微纳电子学、信息科学等多学科领域联合。

本课题主要围绕神经多递质检测新方法、微电极修饰和神经三维细胞培养及检测等关键科学问题开展研究，全面完成了研究任务和考核指标。在神经超微电极阵列器件制备和表面修饰研究、神经三维细胞培养及多种神经递质检测方面取得重要进展。制备出60通道和128通道的超微电极阵列和多种纳米材料修饰传感器，新型微电极阵列制造工艺和相关制备关键技术获得了国家发明专利。对电极进行了纳米修饰，增加了灵敏度和细胞锚定能力，细胞覆盖率达到60%。进行了单细胞的动作电位发放和单细胞及囊泡水平的多巴胺等痕量化学分泌物电化学信号检测，获得了µV量级电生理信号，实现了多巴胺、5-羟色胺、葡萄糖、谷氨酸等多种细胞分泌物的检测，获得了nM~µM的分泌物的pA量级信号。.课题实施过程中，与北大医学部、协和医科大学细胞中心、牛津大学工程系等单位开展了广泛交流合作，发表论文25篇，其中，SCI/EI收录12篇；申请发明专利12项；授权发明专利6项，授权实用新型专利2项，获得国家技术发明奖二等奖1项，获得中科院电子所菁英人才荣誉称号。

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