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

IDBR：合作研究：真实的时间分泌：单细胞分析仪智能优点：每个器官或生物体的细胞都会释放（分泌）各种各样的分子到周围环境中。随着细胞生长，结构和功能的改变，与其他细胞的相互作用，以及当它们受损或患病时，分泌会随着时间的推移而改变。细胞产生的分子在胚胎的生长、感染、免疫反应和愈合中起着关键作用，尽管环境条件波动、衰老、受伤和疾病，但在调节体内功能方面也起着关键作用。测量单个细胞分泌的分子的类型和数量及其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生物体在生物医学，生物学和生物工业背景下的功能和故障。该工具必须能够根据细胞的分泌行为对单个细胞进行分类，并将其无损地恢复以供进一步使用，例如用于组织工程或医学治疗。目前，没有技术可以同时：1）足够灵敏以定量测量由活生物体中典型的非常低浓度的单细胞分泌的许多重要分子，2）足够快以测量细胞外分子浓度在数分钟内的变化，3）能够分选和回收活细胞以供进一步使用，4）能够使用天然存在的细胞，不需要遗传修饰或化学标记的外部供应，5）能够快速延伸以在每个细胞上同时进行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）足够便宜以广泛用于研究实验室、诊所和工业。拟议的真实的时间分泌单细胞分析仪（RTS-SCA）将是一个重大的突破，将使研究人员能够直接测量分泌动力学，研究细胞群体内分泌行为的变化程度和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，药理学和医学中找到应用。该技术的开发是开发微或纳米探针的关键第一步，这些探针以后可用作活组织的诊断工具。更广泛的影响：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尚不具备的在单细胞或组织水平上动态定量分泌的能力。这种能力将促进对细胞功能和相互作用的基本理解，并有助于组织工程，治疗开发和药物发现。RTS-SCA的直接应用包括分析胚胎发育、免疫学、伤口愈合、组织功能调节和癌症等疾病中的关键信号分子。其中癌细胞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鉴定和选择高水平产生感兴趣分子的特定细胞，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，这对组织再生疗法的发展至关重要。RTS-SCA？研究人员能够量化胰岛素释放的复杂动力学，包括胰腺组织的脉动分泌，这可能会加速2型糖尿病治疗的研究，这种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，RTS-SCA将有助于选择用于1型糖尿病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特别重要，其中许多是由目前不可测量的时变分泌分子介导的，例如包括动脉粥样硬化、过敏、类风湿性关节炎和多发性硬化。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，从而促进设计更有效的治疗方法和疫苗。来自印第安纳州布卢明顿大学（IUB）和加州圣地亚哥大学（UCSD）的科学家与两个主要研究中心建立了合作关系，IUB基因组学和生物信息学中心和Moores UCSD癌症中心显微镜共享资源，涵盖中西部、西海岸和美国各地的研究需求。为了鼓励RTS-SCA的广泛使用，每个中心将收到一个运行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.