CCR Vaccine Branch FACS Core

CCR 疫苗分支 FACS 核心

• 批准号: 10487215
• 负责人: Katherine McKinnon
• 金额: $ 66.77万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487215
• 关键词: Apoptosis; Biological; Blood specimen; Bromodeoxyuridine; CCR; COVID-19 vaccine; Cancer Biology; Cell Cycle; Cell Proliferation; Cells; Collaborations; Color; Containment; Core Facility; Cultured Cells; Event; Experimental Designs; Flow Cytometry; Fluorescent Dyes; Funding; Gene Expression; Goals; HIV; Human; Human T-lymphotropic virus 1; Human T-lymphotropic virus 2; Immunologic Monitoring; In Vitro; Infectious Agent; Label; Laboratories; Lasers; Monitor; Monoclonal Antibodies; Mus; NCI Center for Cancer Research; National Cancer Institute; National Institute of Allergy and Infectious Disease; Pediatric Oncology; Phenotype; Reagent; Receptor Biology and Gene Expression; Research Personnel; Research Project Grants; Retroviridae; Running; SIV; Safety; Sorting - Cell Movement; Source; Speed; Stains; Techniques; Technology; Tissue Sample; Training; Vaccines; analog; biological research; cancer genetics; cellular oncology; cytokine; digital; experimental study; instrument; instrumentation; nonhuman primate; polyclonal antibody; release of sequestered calcium ion into cytoplasm; single cell analysis; vaccine trial

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.

创建了疫苗分支FACS（流式细胞术）核心设施，以满足NCI Bethesda校区对具有BL 2/BL 3防护的流式细胞术核心设施的未满足需求，该防护是安全分选受逆转录病毒（如HIV/SIV）以及血液和组织样本（包括SAR）中已知或疑似的其他感染因子感染的活细胞所必需的。该设施有一台目前用于标准和感染性分选的细胞分选机。此外，该设施还拥有五台流式细胞仪的分析能力，这些仪器是CCR的三名研究人员捐赠给该设施的，并通过IATAP提供资金。该机构购买了一台4激光高端分析仪和一台3激光高速细胞分选仪。这些是在2009财年初购买的，并在2009财年第二季度安装。该工厂于2016年购买了28色BD FACSymphony A5。正在购买一台新的28色FACSymphony A5，以取代停产的12色BD LSRII流式细胞仪。该机构目前拥有一台2激光4色模拟流式细胞仪、两台4激光18色高端数字流式细胞仪和两台5激光28色高端数字流式细胞仪。该机构还拥有两台细胞分选仪：配备有容器的4激光16色数字细胞分选仪用作感染性细胞分选仪，以及生物安全柜中的5激光19色细胞分选仪。该设施目前支持来自疫苗分支、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实验室和儿科肿瘤学分支的70多名NCI/CCR研究人员。除了NCI调查员外，设施经理还与NEI和NIAID的一名调查员合作。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主要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。用于这些实验的试剂包括具有荧光标记的单克隆和多克隆抗体以及用于染色多种细胞组分的荧光染料。该设施目前支持的应用包括多色表型分析、细胞周期分析、凋亡分析、BrdU和CFSE增殖分析、细胞内细胞因子分析、罕见事件分析和钙通量分析。目前支持的项目包括但不限于对HIV、SIV、HTLV-1和HTLV-2的免疫监测，以及对非人灵长类动物的COVID-19疫苗研究，以及监测基因表达对细胞增殖和凋亡的结果。