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具有许多限制，包括低灵敏度、低动态范围、可变的斑点均匀度和机械斑点阵列上的大特征尺寸。尽管有这些局限性，荧光微阵列已经成为大规模基因组分析和新兴蛋白质组产业的主要工具。到目前为止，引入新衬底的尝试没有成功，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的重大改变的要求。在这个项目中，我们将开发一种新型的纳米微阵列基板，它将克服现有微阵列基板的所有主要限制，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。这项技术基于我们在平面基板表面控制和图案化生长二氧化硅涂层纳米直径线材的能力。这种新型材料显著增加了有效表面积，但保留了表面功能化和分析开发所需的基本化学特性。在第一阶段，我们将优化材料，开发与传统阵列制造和扫描仪器兼容的平面表面沉积和图案化方法。我们将使用传统的化学方法将寡核苷酸探针连接到增强的表面，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衬底的性能，以实现单位面积信号强度增加100倍，同时动态范围也随之增加。此外，我们将把斑点阵列上的特征尺寸降低到远低于当前可实现的水平，同时提高斑点探测器的均匀性。最后，我们将通过在纳米线增强的表面上开发蛋白质结合实验来展示这种底物的广泛用途。对非优化材料的初步实验表明，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型基板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，并最终制造具有良好特性的高密度DNA和蛋白质阵列，以便在这种基板上进行大规模基因组(蛋白质)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.