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

IDBR：合作研究：实时分泌：单细胞分析仪智能优点：每个器官或有机体的细胞向其周围释放(分泌)各种分子。随着细胞的生长、结构和功能的改变、与其他细胞的相互作用，以及当细胞受损或患病时，分泌物会随着时间的推移而变化。尽管环境条件、衰老、损伤和疾病不断变化，但细胞产生的分子在胚胎的生长、感染、免疫反应和愈合以及调节体内功能方面发挥着关键作用。测量单个细胞分泌的分子的类型和数量以及它们在空间和时间上的变化的工具将有助于研究人员了解细胞和有机体在生物医学、生物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其进行分类，并将其原封不动地恢复以供进一步使用，例如用于组织工程或医疗。目前，没有一种技术同时存在：1)足够灵敏以定量测量单个细胞在活体中典型的极低浓度下分泌的许多重要分子；2)足够快以测量几分钟内细胞外分子浓度的变化；3)能够分拣和回收活细胞以供进一步使用；4)能够在自然发生时使用细胞，而不需要基因改造或外部提供化学标签；5)能够快速扩展以同时对每个细胞进行许多不同的测量或对多个细胞进行相同的测量(高通量)；6)足够便宜，可以在研究实验室、临床和工业中广泛使用。提出的实时分泌-单细胞分析仪(RTS-SCA)将是一项重大突破，将使研究人员能够直接测量分泌动力学，调查细胞群体内分泌行为的变异程度，并选择具有所需特性的细胞。RTS-SCA将在生物和化学工程、发育、免疫和细胞生物学、计算生物学、药理学和医学中得到应用。将要开发的技术是开发微或纳米探针的关键的第一步，这些探针以后可以用作活组织的诊断工具。广泛的影响：生物工程、发育和细胞生物学、生物医学研究以及应用生物农业和生物工业都需要目前无法在单细胞或组织水平上动态量化分泌物的能力。这种能力将促进对细胞功能和相互作用的基本了解，并有助于组织工程、治疗开发和药物发现。这项技术将使常见研究技术的改进版本成为可能，如流式细胞仪、聚合酶链式反应、凝胶电泳、基因测序或微阵列分析。RTS-SCA的即时应用包括分析胚胎发育、免疫学、伤口愈合、组织功能调节以及癌症等疾病中的关键信号分子，在这些疾病中，癌症和宿主组织细胞相互发送复杂的化学信息。生物工程和药理学需要识别和选择在高水平产生感兴趣分子的特定细胞，并确定产生分子的最佳刺激和培养条件。RTS-SCA可以帮助优化细胞分泌行为，这对组织再生疗法的发展至关重要。S能够量化复杂的胰岛素释放动力学，这涉及胰腺组织脉动性分泌的中断，这可能会加速2型糖尿病治疗的研究，2型糖尿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一个指标，RTS-SCA将有助于为1型糖尿病的移植治疗选择组织片段。分子的动态分析在免疫系统反应和疾病中特别重要，其中许多是由目前无法测量的时变分泌分子介导的，例如动脉粥样硬化、过敏、类风湿性关节炎和多发性硬化症。RTS-SCA将有助于了解免疫系统细胞如何感知和应对外部挑战，从而有助于设计更有效的治疗和疫苗。授精计划：来自印第安纳大学布鲁明顿分校(IUB)和加州大学圣地亚哥分校(UCSD)的科学家与两个主要研究中心--IUB基因组和生物信息学中心以及摩尔大学UCSD癌症中心显微镜共享资源--建立了合作关系，涵盖了中西部、西海岸和美国各地的研究需求。为了鼓励RTS-SCA的广泛使用，每个中心都将收到一份运行中的RTS-SCA。这些中心将培训本科生、研究生、博士后研究人员、教师和外部合作伙伴使用RTS-SCA进行研究，维护仪器，向外部合作者宣传仪器，并通过其网络提供适当的外展支持。

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.