Microelectrode arrays of single cell biosensors

描述(申请人提供)：这项研究将开发设计细胞-材料“生物界面”的方法，用于将蛋白质和细胞量身定做地固定在电极表面，并构建高精度、选择性、灵敏度和重复性的基于单细胞的传感器微阵列。基于电池的传感器微阵列在电池与材料的接口方面提出了主要的设计挑战，特别是在电极-衬底形式下。基于细胞的传感器技术受到多重系统性问题的严重限制，包括(1)将细胞附着在设计图案的表面的方法；(2)细胞生长不受控制；(3)细胞连接到电极后失去细胞功能；(4)细胞选择性(影响传感器的信噪比)；以及(5)图案化细胞的长期生存能力。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(二氧化硅上的金色)上，以调节自然细胞的附着和生长。底物背景(二氧化硅)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微阵列随后将与微电子和光学系统集成，以展示其在药物筛选和生物治疗检测中的有效性。目标1将重点研究聚乙二醇共价结合到硅氧化物背景衬底上的表面修饰协议，以实现最大的蛋白质抗性。将研究表面化学对聚乙二醇密度、稳定性和蛋白质长期截留率的影响。目的2将研究不同的蛋白质/多肽和表面几何形状对单细胞图案的细胞结合的影响，以保持电极上的高细胞覆盖率。Aim 3将通过将表面工程与先进的微电子技术相结合，设计和制造用于高通量药物筛选的单细胞传感器集成微阵列(IMA)。我们将开发一套全面的传感方案，包括硬件实现、计算数据采集和分析，以实现快速、准确、高效的药物筛查。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：药物筛选和生物硫脲检测。为了展示开发的传感器检测和监控外部刺激的能力，从IMA和FTIR研究中提取的数据将进行关联，以便可以从生物传感器-IMA读数中单独提取实用信息。

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.