Becton Dickinson LSR II Analytical Flow Cytometer

描述（由申请人提供）：流式细胞术已经成为基于细胞的分析的基本工具。然而，这些多功能机器技术复杂，设备昂贵，需要共享使用和核心设施的开发。目前，医学院（25个不同的nih资助用户）占校园流式细胞术使用的大部分。来自医学部门和其他地方的需求增加，证明了内部资助的成像和细胞术核心的发展是合理的，由Clynes博士领导。这是一个更大的机构努力建立医学转化研究的一部分，包括新成立的转化免疫学中心。特别的是，该提案联合了8名多学科生物医学研究人员（NIH资助的研究总计1500万美元/年），并寻求将具有流式细胞术技术专业知识的免疫学家与研究癌症、肺部和动脉粥样硬化斑块炎症的研究人员结合起来。这将在整个医学院为多功能LSR II流式细胞仪的使用、培训和维护创造一个共享资源。该核心将支持的NIH资助研究的例子包括：癌症中的干细胞/骨髓抑制细胞，移植排斥和耐受性，动脉粥样硬化的解决以及I型糖尿病中免疫球蛋白受体的治疗性阻断。所要求的LSR II需要三个可选和昂贵的附件，以允许和加快这些转化研究。1. 紫外激光允许免疫受体信号（钙通量），侧人口检测和细胞周期分析。2. 黄绿色激光拓宽了细胞仪检测各种红色荧光色素的能力，包括红色荧光蛋白，这对细胞谱系分析、基因表达和蛋白质运输很重要，大大增加了多功能性。3. 96孔连接将允许高通量分析CFSE增殖试验，多种分析，加速评估组织细胞/血液和培养原代细胞的大样本形式的炎症细胞因子，消除了耗时的基于ELISA的分析的需要。总而言之，LSRII将为超过25名nih资助的医学系研究人员促进多功能和用户友好型细胞仪核心的发展，并将体现转化免疫学中心与医学院一般社区的整合。

DESCRIPTION (provided by applicant): Flow cytometry has become the essential tool for cell-based assays. Yet these versatile machines are technologically complex and the equipment expensive requiring shared use and the development of core facilities. Currently, the Department of Medicine (25 different NIH-funded users) accounts for the majority of the campus-wide use of flow cytometry. The increased demand from the Department of Medicine and elsewhere at CUMC has justified the development of an internally funded Imaging and Cytometry Core, headed by Dr. Clynes. This is part of a larger institutional effort to build translational research in medicine, including the newly created Center of Translational Immunology. This proposal, in particular, joins 8 multidisciplinary biomedical researchers (whose NIH funded research totals >$15,000,000/year), and seeks to integrate immunologists with specific expertise in flow cytometric techniques with investigators studying inflammation in cancer, the lung, and the atheromatous plaque. This will create a shared resource for use, training, maintenance and of the versatile LSR II flow cytometer throughout the School of Medicine at CUMC. Examples of NIH funded research that this Core will support include: stem cells/myeloid suppressor cells in cancer, transplantation rejection and tolerance, resolution of atherosclerosis and therapeutic blockade of immunoglobulin receptors in type I diabetes. The requested LSR II requires three optional and costly accessories to permit and expedite these translational studies. 1. The UV laser permits immunoreceptor signaling (calcium flux), side population detection, and cell cycle analysis. 2. The yellow-green laser broadens the cytometers capacity to detect a wide variety of red flourochromes, including Red Flourescence Protein, important for cell lineage analysis, gene expression and protein trafficking, dramatically increasing versatility. 3. The 96-well attachment will allow high throughput analysis of CFSE proliferation assays, multiple analytes, expediting assessment of inflammatory cytokines in large sample format of tissue cells/blood and cultured primary cells eliminating the need for time-consuming ELISA based assays. In sum the LSRII will catalyze the development of a versatile and user-friendly Cytometry Core for over 25 NIH-funded investigators in the Department of Medicine and will make manifest the integration of the Center of Translational Immunology within the general community of the School of Medicine.