CCR VB FACS CORE

项目概要已建立疫苗分支FACS（流式细胞术）核心设施 满足NCI贝塞斯达校区对流式细胞术核心设施的未满足需求， 安全分选感染逆转录病毒的活细胞所需的BL 2/BL 3遏制，例如 HIV/SIV，以及血液和组织样本中已知或疑似的其他感染因子。的 设施具有目前用于标准和感染性分选的细胞分选机。在 此外，该设施还具有从五个流式细胞仪分析能力， CCR的三名研究者捐赠给该机构的仪器， 机构间援助方案供资。2008年5月完成了实验室空间的翻修， 细胞仪于2008年6月搬迁到该设施。该设施购买了4激光高 末端分析仪和3激光高速细胞分选仪。这些是在年初购买的。 2009财年，并于2009财年第2季度安装。该设施目前有两台运行2激光器4 彩色模拟流式细胞仪，一台闪烁式2激光6色数字流式细胞仪，一台 3台激光12色数字高端流式细胞仪和1台4台荧光分析仪 激光18色高端数字流式细胞仪。3激光数字化高端流式细胞仪 经过广泛的硬件和软件升级，增加了额外的荧光， 样本处理能力，2台激光模拟流式细胞仪均已升级 一个新的计算机系统。该设施有一个3激光12色数字细胞分选机配备 其具有用作感染性细胞分选器的容器。分拣机已完全投入使用， 自2009财年第3季度起，已用于感染性分选。该设施目前支持50多个 来自疫苗分支、受体生物学和基因表达实验室的研究人员， 和细胞肿瘤学实验室。由于该设施只开放了4个月， 预计调查员人数将大幅增加。实验目前 在该设施中进行的包括来自人的细胞的多参数单细胞分析， 非人灵长类动物和小鼠的主要来源。此外，还分析了 体外培养细胞用于这些实验的试剂包括单克隆和多克隆试剂。 具有荧光标记和荧光染料的抗体，用于对多种 细胞成分。该设施目前支持的应用包括多色 表型分析、细胞周期分析、凋亡分析、BrdU增殖分析 和CFSE、细胞内细胞因子分析、罕见事件分析和钙通量分析。 目前支持的项目包括但不限于艾滋病毒、SIV、 HTLV-1和HTLV-2疫苗在非人灵长类动物中的研究，并监测基因检测结果 表达对细胞增殖和凋亡的影响。

Project Summary The Vaccine Branch FACS (Flow Cytometry) Core Facility has been created to meet an unmet need on the Bethesda Campus of NCI for a Flow Cytometry Core Facility with BL2/BL3 containment necessary to safely sort live cells infected with retroviruses, such as HIV/SIV, and other infectious agents known or suspected in blood and tissue samples. The facility has a multicolor cell sorter currently in use for standard and infectious sorting. In addition, this facility has analysis capabilities from five multicolor flow cytometry instruments that were donated to the facility from three Investigators in the CCR, and through IATAP funding. Laboratory space renovations were completed in May 2008, and the donated flow cytometers were relocated to the facility in June 2008. The facility purchased a 4 laser high end analyzer and a 3 laser high speed cell sorter. These were purchased at the beginning of FY09 and installed in Q2 FY09. The facility currently maintains two investigator-run 2 laser 4 color analog flow cytometers, one investigator-run 2 laser 6 color digital flow cytometer, one investigator-run 3 laser 12 color digital high end flow cytometer, and one investigator-run 4 laser 18 color high end digital flow cytometer. The 3 laser digital high end flow cytometer has undergone extensive hardware and software upgrades to add additional fluorescent and sample processing abilities, and both of the 2 laser analog flow cytometers have been upgraded to a new computer system. The facility has a 3 laser 12 color digital cell sorter equipped with containment for use as an infectious cell sorter. The sorter is fully operational and has been used for infectious sorting as of Q3 FY09. The facility currently supports more than 50 Investigators from the Vaccine Branch, the Laboratory of Receptor Biology and Gene Expression, and the Laboratory of Cellular Oncology. Since the facility has only been open for 4 months, the number of investigators is expected to increase significantly. Experiments currently conducted in the facility involve multi-parameter single cell analysis of cells from human, non-human primate and mouse primary sources. Additionally analysis is also conducted on in vitro cultured cells. Reagents for these experiments include monoclonal and polyclonal antibodies with fluorescent labels and fluorescent dyes used for staining a variety of cellular components. Current applications being supported by the facility include multi-color phenotypic analysis, cell cycle analysis, apoptosis analysis, proliferation analysis with BrdU and CFSE, intracellular cytokine analysis, rare event analysis and calcium flux analysis. Currently supported projects include, but are not limited to, immune monitoring for HIV, SIV, HTLV-1 and HTLV-2 vaccine studies on non-human primates, and monitoring the results of gene expression on cell proliferation and apoptosis.