Microelectrode arrays of single cell biosensors

这项研究将开发方法，工程细胞材料的“生物界面”，为定制的固定化， 蛋白质和细胞，并用于构建基于单细胞的传感器的微阵列， 具有高精密度、高选择性、高灵敏度和高重现性。基于细胞的传感器的微阵列目前主要 在将电池与材料，特别是电极-衬底形式的材料连接中的设计挑战。基于细胞 传感器技术受到多个系统问题的严重限制，包括（1）用于连接的方法 细胞在设计图案的表面上;（2）不受调节的细胞生长;（3）细胞生长后细胞功能丧失 附着在电极上;（4）细胞选择性（影响传感器的信噪比）;（5）长期生存能力 有图案的细胞。在这项工作中，粘附蛋白质或肽将被图案化到电极阵列（金 在SiO2上）介导天然细胞附着和生长。基板背景（SiO2）将被钝化 以抵抗蛋白质吸附和细胞附着。这种单细胞生物传感器的微阵列， 与微电子和光学系统集成，以证明其在药物筛选中的功效， 生物威胁检测目标1将集中于研究用于共价结合的表面改性方案， PEG到氧化硅背景基底上，以实现最大的蛋白质抗性。的影响 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。目标2将 研究不同蛋白质/肽和表面几何形状对单细胞细胞结合的影响 图案化以保持电极上的高细胞覆盖率。目标3将设计和制造一个 用于高通量药物筛选的基于单细胞的传感器的集成微阵列（IMA） 表面工程与先进的微电子技术。我们将制定一个全面的 传感方案，包括硬件实现，以及计算数据采集和分析， 快速、准确、高效的药物筛选。目标4将应用开发的IMA传感系统， 在两个模型应用中，使用互补的FTIR分析技术：药物筛选和生物威胁检测。 为了证明所开发的传感器检测和监测外部刺激的能力， IMA和FTIR研究将相互关联，以便从生物传感器IMA中获得实用信息 阅读，单。

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.