CCR Vaccine Branch FACS Core

疫苗分部FACS(流式细胞术)核心设施是为了满足NCI贝塞斯达校区对流式细胞术核心设施的未得到满足的需求而创建的，该核心设施具有必要的BL2/BL3容器，用于安全地分离感染了逆转录病毒的活细胞，如HIV/SIV，以及血液和组织样本中已知或怀疑的其他传染病病原体，包括SARS。该设施目前有一台多色细胞分选机，用于标准和传染性分选。此外，该设施还拥有五台多色流式细胞仪的分析能力，这些仪器是由CCR的三名研究人员捐赠给该设施的，并通过IATAP资助。该设施购买了一台4激光高端分析仪和一台3激光高速细胞分选机。这些设备是在09财年初购买的，并于2009财年第二季度安装。该工厂在2016年购买了一辆28色的BD FACSymphony A5。正在购买一台新的28色FACSymphony A5，以取代已停产的12色BD LSRII流式细胞仪。该设施目前拥有一台研究人员运行的2激光4色模拟流式细胞仪，两台研究人员运行的4激光18色高端数字流量细胞仪，两台研究人员运行的5激光28色高端数字流量细胞仪。该设施还拥有两台细胞分选机：一台配备封闭件的4激光16色数字细胞分选机，用于传染病细胞分选机；一台生物安全柜中的5激光19色细胞分选机。该设施目前支持来自疫苗分部、受体生物学和基因表达实验室、细胞肿瘤学实验室以及癌症生物学和遗传学和儿科肿瘤学分部的70多名NCI/CCR研究人员。除了NCI的调查人员外，设施经理还在与NEI和NIAID的一名调查人员合作。目前在该设施中进行的实验涉及对来自人类、非人类灵长类动物和小鼠原始来源的细胞进行多参数单细胞分析。此外，还对体外培养的细胞进行了分析。这些实验的试剂包括带有荧光标记的单克隆和多克隆抗体，以及用于对各种细胞成分进行染色的荧光染料。该设施目前支持的应用包括多色表型分析、细胞周期分析、细胞凋亡分析、BrdU和CFSE增殖分析、细胞内细胞因子分析、罕见事件分析和钙离子通量分析。目前支持的项目包括但不限于艾滋病毒、SIV、HTLV-1和HTLV-2的免疫监测，非人类灵长类动物的新冠肺炎疫苗研究，以及监测基因表达对细胞增殖和凋亡的结果。

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 including SARs. 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. 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 purchase a 28-color BD FACSymphony A5 in 2016. A new 28-color FACSymphony A5 is being purchase to replace a discontinued 12-color BD LSRII flow cytometer. The facility currently maintains one investigator-run 2 laser 4 color analog flow cytometer, two investigator-run 4 laser 18 color high end digital flow cytometers, two investigator run 5 laser 28 color high end digital flow cytometers.The facility also has two cell sorters: a 4 laser 16 color digital cell sorter equipped with containment for use as an infectious cell sorter and a 5 laser 19 color cell sorter in a biological safety cabinet. The facility currently supports more than 70 NCI/CCR Investigators from the Vaccine Branch, the Laboratory of Receptor Biology and Gene Expression, the Laboratory of Cellular Oncology, and the Laboratory of Cancer Biology and Genetics and Pediatric Oncology Branch. In addition to NCI investigators, the Facility Manager is working on a collaboration with an investigator from NEI and NIAID. 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, and COVID-19 vaccine studies on non-human primates, and monitoring the results of gene expression on cell proliferation and apoptosis.