Microelectrode arrays of single cell biosensors

单细胞生物传感器微电极阵列

• 批准号: 7094925
• 负责人: Miqin Zhang
• 金额: $ 27.25万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-10 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7094925
• 关键词: biomaterial interface interaction; biosensor device; bioterrorism /chemical warfare; cell adhesion; chemical stability; computational biology; drug screening /evaluation; electrical impedance; high throughput technology; infrared spectrometry; interferometry; method development; microarray technology; microelectrodes; polyethylene glycols; protein binding; silicon compounds; surface coating; tissue /cell culture

DESCRIPTION (provided by applicant): This research will develop methods to engineer cell-material "bio-interfaces" for tailored immobilization of proteins and cells on electrode surfaces and for construction of micro-arrays of single cell-based sensors of high precision, selectivity, sensitivity, and reproducibility. Micro-arrays of cell-based sensors present major design challenges in interfacing cells with materials, especially in an electrode-substrate format. Cell-based sensor technology is severely limited by multiple systematic problems, including (1) methods for attaching cells onto surfaces of designed patterns; (2) unregulated cell growth; (3) loss of cell functionality after cells are attached to electrodes; (4) cell selectivity (affects sensor's signal/noise ratio); and (5) long-term viability of patterned cells. In this work, adhesive proteins or peptides will be patterned onto an electrode array (gold on SiO2) to mediate natural cell attachment and growth. The substrate background (SiO2) will be passivated to resist protein adsorption and cell attachment. This microarray of single-cell biosensors will then be integrated with microelectronic and optical systems to demonstrate its efficacy in drug screening and biothreat detection. Aim 1 will focus on the study of surface modification protocols for covalent binding of PEG onto a silicon oxide background substrate to achieve maximum protein resistance. The effects of surface chemistry on PEG density, stability, and long-term protein rejection will be investigated. Aim 2 will investigate the influence of different proteins/peptides and surface geometry on cell binding for single-cell patterning in order to maintain high cell coverage on the electrodes. Aim 3 will design and fabricate an integrated microarray (IMA) of single-cell based sensors for high-throughput drug screening by integrating surface engineering with the advanced technology of microelectronics. We will develop a comprehensive sensing scheme including hardware implementation, and computational data acquisition and analysis for fast, accurate, and efficient drug screening. Aim 4 will apply the developed IMA sensing system, and complementary FTIR analysis techniques, in two model applications: drug screening and biothreat detection. To demonstrate the developed sensor's ability to detect and monitor external stimuli, data extracted from the IMA and FTIR studies will be correlated so that practical information can be drawn from biosensor-IMA readings, singly.

描述（由申请人提供）：本研究将开发方法来设计细胞材料的“生物互化”，以量身定制电极表面上的蛋白质和细胞的固定，并构建具有高精度，选择性，敏感性和可重复性的单细胞传感器的微阵列。基于细胞的传感器的微阵列在将细胞与材料接口，尤其是电极 - 基板格式中提出了主要的设计挑战。基于细胞的传感器技术受到多种系统问题的严重限制，包括（1）将细胞连接到设计模式表面上的方法； （2）细胞生长不受管制； （3）细胞后连接到电极上的细胞功能； （4）细胞选择性（影响传感器的信号/噪声比）； （5）图案化细胞的长期生存能力。在这项工作中，将粘合蛋白或肽将图案化在电极阵列上（SIO2上的金），以介导天然细胞的附着和生长。底物背景（SIO2）将被钝化以抵抗蛋白质的吸附和细胞附着。然后，这种单细胞生物传感器的微阵列将与微电源和光学系统集成，以证明其在药物筛查和生物治疗检测中的功效。 AIM 1将集中于研究表面修饰方案，以使PEG共价结合到氧化硅背景底物上，以实现最大的蛋白质耐药性。将研究表面化学对PEG密度，稳定性和长期蛋白质排斥的影响。 AIM 2将研究不同蛋白/肽和表面几何形状对单细胞模式的细胞结合的影响，以维持电极上的高细胞覆盖率。 AIM 3将通过将表面工程与微电子的先进技术集成，设计和制造单细胞传感器的集成微阵列（IMA），以用于高通量药物筛选。我们将制定一个全面的感应方案，包括硬件实施，以及快速，准确和有效的药物筛查的计算数据采集和分析。 AIM 4将在两个模型应用中应用开发的IMA传感系统和互补的FTIR分析技术：药物筛查和生物治疗检测。为了证明开发的传感器检测和监测外部刺激的能力，将从IMA和FTIR研究中提取的数据进行关联，以便单独从BioSensor-IMA读取中汲取实际信息。