Acquisition of Shared Equipment - Cell Analyzer/Flow Cytometer

 描述（由申请人提供）： 这是一项共享设备提案，目的是在ORD共享设备方案的支持下，购置一台最先进的细胞分析仪/流式细胞仪。所提出的仪器BD LSRFortessa是一种先进的数字细胞分析仪，提供了广泛的关键，敏感的技术，以研究临床研究标本或动物模型样品中细胞内信号通路的表型，行为和激活。我们研究中心现有的流式细胞仪已有11年的历史。它已经失去了其科学用途，无法升级。BD LSRFortessa提供的技术更加先进，能够测量细胞行为和代谢的细微变化。因此，该仪器提供了高效，准确的新工具来回答有关人类疾病的关键问题。由于所提出的仪器以测量细胞行为及其随疾病进展而发生的变化为中心，因此它广泛适用于我们不同的研究人员群体。这包括神经科学家，糖尿病和代谢，癌症，血管生物学和创伤性脑损伤的研究人员。我们已经确定了12名研究人员将使用这一工具。其中，8个具有VA资助的研究项目（主要用户，占仪器时间的75%），而4个次要用户具有位于VA Puget Sound Health Care System的研究项目，并具有由NIH资助的研究项目。我们在本提案中提出了一个全面的计划，用于共享，管理和护理仪器，以及不同的VA调查人员将如何利用这项技术。在本提案的研究计划部分，我们为每个VA研究者提供了一个叙述，以及他们将如何利用这一共享工具。本文摘录总结了其中一些不同的用途：通过对活化依赖性表位的分析，测量接受血管手术的退伍军人患者的血小板和单核细胞活化;分析来自重要血液样本的细胞内信号传导途径;测量血小板和内皮衍生微粒。测量药物治疗后基因表达的变化;体内神经退行性变相关聚集体形成的定量以及细胞活力分析。分析STAT-1蛋白突变对动脉粥样硬化血管中细胞数量和类型的影响。阐明细胞内应激信号通路（如JNK信号通路）的激活和细胞凋亡的激活，介导淀粉样蛋白诱导的β细胞丢失，因为它与2型糖尿病发病机制有关。从阿尔茨海默氏症和对照死后脑组织中鉴定特定的脑细胞（例如神经元和小胶质细胞）。确定循环或组织驻留炎性细胞的细胞表型，用于随后的体内研究。总之，这种新仪器将提供一个最先进的科学平台，以支持许多VA研究人员。该仪器将位于我们主要研究大楼的地理中心，并由已建立的细胞和分子成像核心管理。我们拥有完善的行政基础设施，用于仪器的管理、调度和支持。

 DESCRIPTION (provided by applicant): This is a shared equipment proposal to acquire a state-of-the-art cell analyzer/flow cytometer, under the auspices of the ORD Shared Equipment Program. The instrument proposed, a BD LSRFortessa, is an advanced digital cell analyzer that offers a broad range of critical, sensitive techniques to study phenotypes, behavior and activation of intracellular signaling pathways in samples from clinical research specimens or animal models. The present flow cytometer at our research center is over 11 years old. It has outlived its scientific usefulness and cannot be upgraded. The technology offered by the BD LSRFortessa is significantly more advanced, enabling measurement of subtle changes in cellular behavior and metabolism. This instrument therefore offers efficient, accurate new tools to answer key questions about human disease. Because the proposed instrument is centered on the measurement of cellular behavior and changes therein that occur with disease progression, it is widely applicable to our diverse community of researchers. This includes neuroscientists, researchers in diabetes and metabolism, cancer, vascular biology, and traumatic brain injury. We have identified 12 investigators that will use this instrument. Of these, 8 have VA-funded research programs (Major users, accounting for 75% of instrument time) while 4 minor users have research programs geographically located at VA Puget Sound Health Care System, and have research programs funded by NIH. We present in this proposal a comprehensive plan for the sharing, administration, and care of the instrument, and how different VA investigators will take advantage of this technology. In the Research Plan section of this proposal, we present a narrative for each VA investigator, and how they would take advantage of this shared instrument. Herein are excerpts to summarize some of these different uses: ¿ Measurements of platelet and monocyte activation in Veteran patients undergoing vascular surgery, through multicolor analyses of activation-dependent epitopes; analyses of intracellular signaling pathways from vital blood samples; measurements of platelet- and endothelial-derived microparticles. ¿ Measuring gene expression changes in response to drug treatment; quantitation of neurodegeneration- related aggregate formation in vivo, and cell viability analysis. ¿ Analyzing the effect of mutations in the STAT-1 protein on the numbers and types of cells present in atherosclerotic vessels. ¿ Delineating activation of intracellular stress signalng pathways (e.g. JNK signaling) and activation of apoptosis, which mediate amyloid-induced ß-cell loss as it relates to type 2 diabetes pathogenesis. ¿ Identification of specific brain cells e.g. neurons and microglia) from Alzheimer's and control postmortem brain tissues. ¿ Determining cellular phenotype of circulating or tissue-resident inflammatory cells for use in subsequent in vivo studies. In summary, this new instrument would provide a state-of-the-art scientific platform to support many VA investigators. The instrument will be located at the geographic center of our main research building, and managed by the established Cellular and Molecular Imaging Core. We have a well-developed administrative infrastructure for the management, scheduling, and support of the instrument.