Localized Surface Plasmon Resonance (LSPR) Biosensing Microarray for Multiplex, Real-time Single-Cell Immunophenotyping

用于多重、实时单细胞免疫表型分析的局域表面等离子共振 (LSPR) 生物传感微阵列

• 批准号: 1263889
• 负责人: Katsuo Kurabayashi
• 金额: $ 33万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1263889&HistoricalAwards=false
• 关键词: Localized; Surface; Plasmon; Resonance; LSPR

1263889 - KurabayashiThe immune system plays a critical role in protecting a living organism against invasions of viruses,bacteria, and parasitic worms and in distinguishing diseased tissue from healthy tissue. Leukocytes -white blood cells - are important blood constituents that support the immune system and haveseveral cellular subsets determined by their morphology and response to an antigen exposure. Comprehensive characterization of the immune cell functions is critical to diagnose, stratify, and monitor infections, malignancies, autoimmune disorders, and injuries or to study the efficacy of new drugs. In particular, quantitative analysis of the cytokine secretion profiles of individual cells is required for precise determination of the subtle variations in the cellular function and immune status across their sub-groups. However, there are some shortcomings in existing methods that need to be overcome for realizing rapid, efficient, dynamic analysis of leukocyte cytokine-secretion profiles. This research aims to establish a biosensing platform that enables high-throughput, multiplexed, dynamic measurement of cytokines secreted by target immune cells isolated from whole blood at thesingle-cell level. This single-cell functional assay efficiently permits on-chip isolation and stimulation of single cells and detection of cell-secreted proteins. Our approach will employ localized surface plasmon resonance (LSPR) imaging within microchamber arrays, each capturing a single cell. As a result of miniaturization of the whole system, both the sample volume and the total assay time canbe reduced. The microarray form of the microfluidic platform coupled with LSPR biosensing will allow for massively parallel label-free, real-time quantification and mapping of proteins secreted individual cells with a simple optics setup. The analysis performed for a large number of single cells will allow for immunophenotypical screening of leukocyte subsets while detecting subtle differences and time variations of their immune functions. If the isolation process only yields extraction of identical cell subsets, the large number of measurements per sample will lead to high statistical accuracy (i.e., reduced false positives). NontechnicalThis project will develop novel means to analyze immune cells present in blood. Specifically, the proposal will develop devices to capture individual immune cells and then monitor release of certain proteins from the captured cells. The levels and the types of proteins produced by the immune cells reflect on the state of health or disease of an individual. Therefore proposed devices will have significant diagnostic implications.

1263889 - Kurabayashi免疫系统在保护生物体免受病毒、细菌和寄生虫的侵袭以及区分患病组织和健康组织方面发挥着关键作用。白细胞 - 白细胞 - 是支持免疫系统的重要血液成分，并具有由其形态和对抗原暴露的反应决定的多个细胞亚群。免疫细胞功能的全面表征对于诊断、分层和监测感染、恶性肿瘤、自身免疫性疾病和损伤或研究新药的疗效至关重要。特别是，需要对单个细胞的细胞因子分泌谱进行定量分析，以精确确定其亚组中细胞功能和免疫状态的细微变化。然而，现有方法存在一些缺点需要克服，以实现白细胞细胞因子分泌谱的快速、高效、动态分析。本研究旨在建立一个生物传感平台，能够在单细胞水平上对从全血中分离的目标免疫细胞分泌的细胞因子进行高通量、多重、动态测量。这种单细胞功能测定有效地允许在芯片上分离和刺激单细胞以及检测细胞分泌的蛋白质。我们的方法将在微室阵列内采用局域表面等离子共振（LSPR）成像，每个微室阵列捕获一个细胞。由于整个系统的小型化，样品体积和总测定时间都可以减少。微流体平台的微阵列形式与 LSPR 生物传感相结合，将允许通过简单的光学设置对单个细胞分泌的蛋白质进行大规模并行、无标记、实时定量和绘图。对大量单细胞进行的分析将允许对白细胞亚群进行免疫表型筛选，同时检测其免疫功能的细微差异和时间变化。如果分离过程仅提取相同的细胞子集，则每个样本的大量测量将导致较高的统计准确性（即减少误报）。非技术性该项目将开发分析血液中存在的免疫细胞的新方法。 具体来说，该提案将开发捕获单个免疫细胞的设备，然后监测捕获细胞中某些蛋白质的释放。 免疫细胞产生的蛋白质的水平和类型反映了个体的健康或疾病状态。 因此，所提出的设备将具有重要的诊断意义。