Towards Mega-Throughput, Label-free Genomics and Proteomics: Revolutionizing Micr

描述(由申请人提供) 摘要：微阵列为功能基因组学、蛋白质组学、表观遗传学、医学诊断学甚至组织工程等领域的各种关键研究提供了高通量的平台。与先进的生化检测、成像和生物信息学技术相结合，现在可以经济高效地监测基因、蛋白质或其他生物标志物的表达行为，以及筛选各种细胞系的基因组和蛋白质组内容，芯片上的药物分析，甚至单核苷酸多态性的检测。因此，微阵列技术为进行高通量筛选实验提供了一个重要的平台，这些实验有助于我们了解发生在纳米尺度上的细胞、基因组和蛋白质组过程。在这项提议中，我们的目标是创造下一代微阵列技术，以实现前所未有的超大吞吐量，即每秒可以对数百万个DNA/蛋白质微点进行无标记成像。我们将这些革命性技术的总称称为纳米等离子体卢卡斯。具体地说，我们的目标是实现&gt；120 cm2/秒或&gt；450万点/秒的高灵敏度和无标记DNA/蛋白质微阵列成像，与最先进的技术相比，这构成了&gt；3个数量级的速度提升。为了不干扰原始目标分子的自然生化、物理和结构特性，无标记成像尤其重要。它还使测量更加定量，显著提高了数据质量；消除了不方便的标记步骤，进一步降低了成本；并避免了二次探针之间的交叉反应问题，这可以显著提高对弱或过渡性分子相互作用的检测。这种超大吞吐能力将使蛋白质组学/遗传学研究的进展速度发生重大变化，最终可能导致改进的战略/疗法的开发，以对抗以前难以解决的生物医学问题和包括癌症在内的各种疾病。此外，纳米等离子卢卡斯平台不需要任何透镜、显微镜物镜或其他大宗光学部件，因此提供了一个非常紧凑的芯片上平台，该平台可以很容易地与微流体系统合并，从而实现点式操作。 与公共健康相关：在这项高风险、高回报的提案中，我们的目标是创造下一代微阵列技术，使蛋白质组学和遗传学研究的进展速度发生重大变化，最终可能导致开发出更好的战略和疗法，以对抗以前难以解决的生物医学问题和包括癌症在内的各种疾病。

DESCRIPTION (Provided by the applicant) Abstract: Micro-arrays provide a high-throughput platform for various key studies in functional-genomics, proteomics, epigenetics, medical-diagnostics and even tissue-engineering. Together with advanced biochemical detection, imaging and bioinformatics technologies, it is now possible to cost-effectively monitor the expression behavior of genes, proteins or other biomarkers, as well as screening the genome and proteome content of various cell lines, on-chip drug profiling or even detection of single-nucleotide-polymorphism. Therefore, micro-array technologies provide a vital platform for performing high-throughput screening experiments that shed light on our understanding of cellular, genomic, and proteomic processes occurring at the nano-scale. In this proposal, we aim to create the next-generation of micro-array technologies to achieve an unprecedented mega-throughput, i.e., label-free imaging of millions of DNA/protein microspots would be feasible per second. We term the broad-umbrella of these revolutionary technologies as Nano-plasmonic LUCAS. Specifically, we aim achieve a throughput of >120 cm2/second or >4.5 million spots/second for highlysensitive and label-free imaging of DNA/protein micro-arrays, which constitutes a speed improvement of >3 orders-of-magnitude when compared to the state-of-the-art. Label-free imaging is especially important not to perturb the natural bio-chemical, physical and structural properties of the original molecule-of-interest. It also makes the measurements much more quantitative, significantly improving the data quality; eliminates inconvenient labeling steps which further reduces the cost; and avoids cross-reactivity issues among secondary-probes which can significantly improve the detection of weak or transitional molecular interactions. This mega-throughput capability will revolutionize the speed of progress that is taken in proteomics/genetics research by orders-of-magnitude that could eventually lead to the development of improved strategies/therapies for combating previously intractable bio-medical problems and various diseases including cancer. Furthermore, the Nano-plasmonic LUCAS platform does not require any lenses, microscope-objectives or other bulk optical components, and therefore offers an extremely compact on-chip platform that can easily be merged with micro-fluidic systems to permit point-of-care operation. Public Health Relevance: In this high-risk high pay-off proposal, we aim to create the next generation of micro-array technologies to revolutionize the speed of progress that is taken in proteomics and genetics research by orders of magnitude that could eventually lead to the development of improved strategies and therapies for combating previously intractable bio-medical problems and various diseases including cancer.