Microelectrode arrays of single cell biosensors

这项研究将开发设计细胞-材料“生物界面”的方法，以便量身定做固定化 用于电极表面的蛋白质和细胞以及构建基于单个细胞的传感器的微阵列 精密度高、选择性好、灵敏度高、重现性好。基于细胞的传感器微阵列是目前主要的 电池与材料接口的设计挑战，特别是在电极-衬底形式。基于单元格 传感器技术受到多个系统问题的严重限制，包括(1)连接方法 细胞在设计图案的表面上；(2)细胞生长不受调控；(3)细胞后丧失细胞功能 连接到电极上；(4)电池的选择性(影响传感器的信噪比)；以及(5)长期生存能力 图案化的细胞。在这项工作中，粘附性蛋白质或多肽将被图案化到电极阵列(金 在二氧化硅上)，以调节自然细胞的附着和生长。衬底背景(二氧化硅)将被钝化 抵抗蛋白质吸附和细胞附着。这个单细胞生物传感器的微阵列将被 与微电子和光学系统集成，以展示其在药物筛选和 生物活性检测。目标1将重点研究用于共价结合的表面修饰方案 钉在硅氧化物背景衬底上，以实现最大的蛋白质抗性。的影响 将研究表面化学对聚乙二醇密度、稳定性和蛋白质长期排斥的影响。目标2将 研究不同蛋白质/多肽和表面几何形状对单细胞结合的影响 为了在电极上保持较高的电池覆盖率而进行图案化。AIM 3将设计和制造一个 用于高通量药物筛选的单细胞传感器集成微阵列 表面工程与先进的微电子技术。我们将制定一项全面的 包括硬件实现和计算数据采集与分析的传感方案 快速、准确、高效的药物筛查。目标4将应用开发的IMA传感系统，以及 互补的FTIR分析技术，在两个模型应用中：药物筛选和生物治疗检测。 为了证明开发的传感器检测和监控外部刺激的能力，从 IMA和FTIR研究将相互关联，以便从生物传感器中提取实用信息-IMA 读数，单独的。

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 biothreatdetection. 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.