Nanowire Enhanced Substrates for Microarrays

描述（由申请人提供）：用于荧光微阵列应用的现有衬底有许多局限性，包括灵敏度差，动态范围低，光斑均匀性可变以及机械点阵的大特征尺寸。尽管存在这些限制，荧光微阵列已经成为大规模基因组分析和新兴蛋白质组学产业的主要工具。到目前为止，引入新基板的尝试一直没有成功，主要是因为动力学性能降低以及对基本阵列制造和分析基础设施进行重大更改的要求。在这个项目中，我们将开发一种新型的纳米微阵列衬底，它将克服现有微阵列衬底的所有主要限制，但与现有的杂交协议、阵列制造和分析基础设施完全不兼容。这项技术是基于我们的能力，以控制和模式生长的SiO2涂层，纳米直径线在平面基底表面。这种新型材料提供了有效表面积的显着增加，但保留了表面功能化和分析发展所需的基本化学特性。在第一阶段，我们将优化材料，并开发与传统阵列制造和扫描仪器兼容的平面上沉积和图案的方法。我们将使用常规化学方法将寡核苷酸探针连接到增强表面，并使用标准方案将荧光靶标与这些探针杂交。我们将优化纳米线增强基板的性能，使单位面积的信号强度增加100倍，同时动态范围也随之增加。此外，我们将把斑点阵列的特征尺寸减小到远低于目前可实现的水平，同时增加斑点探针的均匀性。最后，我们将通过在纳米线增强表面上开发蛋白质结合试验来展示该底物的广泛效用。在非优化材料上的初步实验表明，这些里程碑是可以实现的。我们的长期目标是证明这种性能增强底物与所有阵列制造方法的兼容性，将斑点阵列的密度提高到现在通过光刻合成格式实现的密度，并最终制造出具有良好特征的高密度DNA和蛋白质阵列，以便在该底物上进行大规模基因组（蛋白质组学）分析。

DESCRIPTION (provided by applicant): Existing substrates for fluorescent microarray applications have many limitations including poor sensitivity, low dynamic range, variable spot uniformity and large feature sizes on mechanically spotted arrays. Despite these limitations the fluorescent microarray has become a major tool for large-scale genomic analyses and the emerging proteomic industry. Thus far, attempts to introduce new substrates have been unsuccessful, largely because of reduced kinetic performance and the requirements for major changes to the basic array fabrication and analysis infrastructure. In this program we will develop a novel, nano-enabled microarray substrate that will overcome all the major limitations of existing microarray substrates and yet will be entirely incompatible with existing hybridization protocols, array fabrication and analysis infrastructure. This technology is based upon our ability to control and pattern the growth of SiO2 coated, nanometer diameter wires on the surface of a planar substrate. This novel material provides dramatic increases in effective surface area and yet retains the basic chemical characteristics required for surface functionalization and assay development. In Phase I we will optimize the material and develop methods for depositing and patterning it on planar surfaces compatible with conventional array fabrication and scanning instrumentation. We will link oligonucleotide probes to the enhanced surface using conventional chemistries and hybridize fluorescent targets to these probes using standard protocols. We will optimize the performance of the nanowire enhanced substrates to achieve a 100-fold increase in signal intensity per unit area with a concomitant increase in dynamic range. Furthermore, we will decrease feature sizes on spotted arrays to well below currently achievable levels and at the same time increase the uniformity of the spotted probe. Finally we will demonstrate the broad utility of this substrate by developing a protein binding assay on the nanowire enhanced surface. Preliminary experiments on non-optimized materials indicate that these milestones will be achievable. Our long term aims are to demonstrate the compatibility of this performance enhancing substrate with all array fabrication approaches, to increase the density of spotted arrays to those now achieved by lithographically synthesized formats and finally to manufacture well characterized high density DNA and protein arrays to carry out large scale genomic (proteomic) analyses on this substrate.