IDBR: Collaborative Research: Real time secretion: single cell analyzer

IDBR：协作研究：实时分泌：单细胞分析仪

• 批准号: 1152030
• 负责人: James Glazier
• 金额: $ 58.02万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1152030&HistoricalAwards=false
• 关键词: IDBR; Collaborative; Research; Real; time

IDBR: Collaborative Research: Real time secretion: single cell analyzerIntellectual merit: The cells of every organ or organism release (secrete) a wide variety of molecules into their surroundings. Secretion changes over time as cells grow, change structure and function, interact with other cells, and when they become damaged or diseased. Molecules produced by cells play key roles in the growth of embryos, infection, immune response and healing, and in regulating internal body functions despite fluctuating environmental conditions, aging, injury and illness. A tool to measure the types and numbers of molecules secreted by individual cells and their changes in space and time would help researchers understand how cells and organisms function and malfunction, in biomedical, biological and bio-industrial contexts. The tool must be able to sort individual cells based on their secretion behaviors and recover them undamaged for further use, e.g. for tissue engineering or medical treatment. Currently, no technology is simultaneously: 1) Sensitive enough to quantitatively measure many important molecules secreted by single cells at the very low concentrations typical in living organisms, 2) Fast enough to measure changes in extracellular molecular concentrations over periods of minutes, 3) Able to sort and recover live cells for further use, 4) Able to use cells as they occur naturally, without needing genetic modification or an external supply of chemical labels, 5) Capable of quick extension to conduct many different measurements simultaneously on each cell or the same measurement on many cells (high-throughput), 6) Inexpensive enough for wide use in research laboratories, clinics, and industry. The proposed Real Time Secretion-Single Cell Analyzer (RTS-SCA) will be a significant breakthrough that will enable researchers to directly measure secretion dynamics, investigate the degree of variation in secretion behavior within populations of cells and the selection of cells with desired properties. RTS-SCA will find applications in biological and chemical engineering, developmental, immune and cell biology, computational biology, pharmacology and medicine. The technology to be developed is a critical first step towards developing micro- or nano-probes that could later be used as diagnostic tools in living tissues.Broader impact: Bioengineering, developmental and cell biology, biomedical research, and applied bioagriculture and bioindustry all require a currently unavailable capability to dynamically quantify secretion at the single cell or tissue level. This ability will advance basic understanding of cell functions and interactions, and aid tissue engineering, therapy development and drug discovery. The technology will allow improved versions of common research techniques, such as flow cytometry, polymerase chain reaction, gel electrophoresis, gene sequencing or microarray analysis.Immediate applications of the RTS-SCA include the analysis of the key signaling molecules in embryonic development, immunology, wound healing, tissue function regulation, and diseases like cancer, in which both cancer and host-tissue cells send complex chemical messages to each other. Bioengineering and pharmacology need to identify and select particular cells that produce molecules of interest at high levels, and to determine optimal stimulation and culture conditions for molecule production. The RTS-SCA could help in optimizing cell-secretion behaviors, essential for the development of tissue regeneration therapies. The RTS-SCA?s ability to quantify the complex kinetics of insulin release, which involves disrupted pulsatile secretion from pancreatic tissues, could accelerate research in treatments for type-2 diabetes, a disease which accounts for 10% of the U.S. health-care expenditure. Since periodic insulin secretion is an indicator of healthy pancreas function, RTS-SCA would help in selecting tissue fragments for transplant therapy for type-1 diabetes. Dynamic analysis of molecules is especially important in immune system responses and diseases, many of which are mediated by currently unmeasurable time-varying secreted molecules, examples include atherosclerosis, allergies, rheumatoid arthritis and multiple sclerosis. The RTS-SCA will help understand how immune system cells sense and respond to external challenges facilitating design of more effective treatments and vaccines.Dissemination Plan: The scientists from Indiana University-Bloomington (IUB) and University of California San Diego (UCSD) have established collaborations with two major research centers, The IUB Center for Genomics and Bioinformatics and The Moores UCSD Cancer Center Microscopy Shared Resource, covering research needs in the Midwest, West Coast, and across the USA. To encourage the RTS-SCA's wide use, each center will receive an operating RTS-SCA. These centers will train undergraduates, graduate students, postdoctoral researchers, faculty and external partners to use the RTS-SCA for their research, maintain the instrument, advertise the instrument to outside collaborators, and provide appropriate outreach support through their networks.

IDBR：协作研究：实时分泌：单细胞分析术的优点：每个器官或生物体释放的细胞（分泌）各种分子到周围的环境中。随着细胞的成长，改变结构和功能，与其他细胞的互动以及它们受损或患病时，分泌会随着时间的流逝而变化。细胞产生的分子在胚胎，感染，免疫反应和愈合的生长中起着关键作用，以及尽管环境条件，衰老，损伤和疾病的波动，但在调节体内功能方面起作用。一种测量单个细胞分泌的分子的类型和数量的工具及其在时空的变化将有助于研究人员了解细胞和生物在生物医学，生物学和生物工业环境中的功能和故障。该工具必须能够根据单个细胞的分泌行为对单个细胞进行分类，并将其恢复未损坏以进一步使用，例如用于组织工程或医疗。目前，没有技术是同时的：1）足够敏感以测量单个细胞在非常低的浓度下分泌的许多重要分子在典型的生物体中典型的浓度分泌在每个细胞上同时进行了许多不同的测量，或在许多细胞上进行相同的测量（高通量），6）足够的便宜，足以在研究实验室，诊所和行业中进行广泛使用。提出的实时分泌细胞分析仪（RTS-SCA）将是一个重大突破，将使研究人员能够直接测量分泌动力学，研究细胞种群中分泌行为的变化程度以及具有所需特性的细胞选择。 RTS-SCA将在生物学和化学工程，发育，免疫和细胞生物学，计算生物学，药理学和医学中找到应用。这项要开发的技术是开发微型或纳米探针的关键第一步，后来可以用作生存组织中的诊断工具。Boader的影响：生物工程，发育和细胞生物学，生物医学研究以及应用生物质学和生物INDUSTRITY都需要在单个细胞或组织级别动态量化单个细胞或组织的能力。这种能力将提高对细胞功能和相互作用的基本了解，并帮助组织工程，治疗开发和药物发现。该技术将允许改进的常见研究技术，例如流式细胞术，聚合酶链反应，凝胶电泳，基因测序或微阵列分析。对RTS-SCA的临时应用包括对胚胎发育，伤口愈合，组织的癌症和分类的癌症的关键信号分子的分析，包括胚胎发育，伤口愈合和疾病的癌症，并在癌症中进行癌症。生物工程和药理学需要识别和选择在高水平上产生感兴趣的分子的特定细胞，并确定分子产生的最佳刺激和培养条件。 RTS-SCA可以帮助优化细胞分泌行为，这对于组织再生疗法的发展至关重要。 RTS-SCA能够量化胰岛素释放的复杂动力学的能力，涉及胰腺组织中断的脉动分泌，可以加速对2型糖尿病治疗的研究，该疾病占美国医疗保健支出的10％。由于周期性胰岛素分泌是健康胰腺功能的指标，因此RTS-SCA将有助于选择用于1型糖尿病的移植治疗的组织片段。分子的动态分析在免疫系统反应和疾病中尤为重要，其中许多是由当前不可估量的分泌分子介导的，例如动脉粥样硬化，过敏，类风湿关节炎和多发性硬化症。 The RTS-SCA will help understand how immune system cells sense and respond to external challenges facilitating design of more effective treatments and vaccines.Dissemination Plan: The scientists from Indiana University-Bloomington (IUB) and University of California San Diego (UCSD) have established collaborations with two major research centers, The IUB Center for Genomics and Bioinformatics and The Moores UCSD Cancer Center Microscopy Shared Resource, covering research中西部，西海岸以及整个美国的需求。为了鼓励RTS-SCA的广泛使用，每个中心将获得操作的RTS-SCA。这些中心将培训本科生，研究生，博士后研究人员，教职员工和外部合作伙伴，以使用RTS-SCA进行研究，维护乐器，向外部合作者宣传乐器，并通过其网络提供适当的外展支持。