CORE--FLOW CYTOMETRY

该集中核心资源的目的是提供(1)专业知识 在实验设计方面，以及(2)在技术和设备支持方面 利用流式细胞术进行研究项目的实验表现 在有关艾滋病毒的基本和临床问题的调查中 发病机制。首要目标是提供智力环境 以及物质资源，使初级调查人员能够应用流动 细胞检测技术对他们的项目和鼓励建立 调查人员启动创新的先导研究。为此，有几个 不同的资金来源(UCSD、CFAR、退伍军人事务部艾滋病研究中心、VAMC Research Core)已集成，以支持和扩展我们的流程 满足圣地亚哥学院研究需要的细胞学设施 社区。CFAR奖仍然是这方面一个综合组成部分 努力吧。 流式细胞仪设施包含三个主要设备， 用于执行高度互补的功能。Coulter Elite分拣机 用双激光加紫外光激光测量钙离子通量和细胞DNA 定量主要用于多参数细胞分析。一个 Becton-Dickinson FACStar Plus分选器，带第五个三色探测器 分析，作为一种高效的细胞分选机进行非生物危害 或者固定的细胞样本。一种最近资助的高速双激光电池 分类器(FACS Vantage Turbo)正在被收购并安置在一个受限的 BSL-2空间；它将执行茎所需的两种罕见事件恢复 艾滋病毒感染标本的细胞研究和活细胞分选。专属的 技术人员操作这些流式细胞仪，工作在 个人调查员的充值依据。支持人员包括一名 经验丰富的高级技师和训练有素的初级技师 技术员；正在招聘一名经验丰富的中级技术员 目前正在操作新的细胞分选机，用于生物危险工作 材料。核心主管，在流式细胞仪和 研究应用，提供实验设计和实验设计咨询 向使用该设施的调查人员解释数据。 在过去的三年中，CFAR资源使流式细胞术 核心支持不同领域的艾滋病毒研究项目，如：卡波斯 肉瘤(CA67394)，克隆性B细胞发育(AI34001)，巨噬细胞 功能(AI35258，NS31060)，细胞因子反应(AI38858)，治疗- 诱导免疫修复(AI27670)、基因治疗(DK49618、AI36612)、 抗病毒药物开发(GM24979)和儿童艾滋病(AI39004)。使用 流式细胞术在生物分子领域的拓展应用 研究，持续的CFAR支持将是提供 基础设施将利用艾滋病毒领域的新科学机遇 研究。

The purpose of this centralized core resource is to provide (1) expertise in experimental design, and (2) technical and equipment support in the performance of experiments for research projects utilizing flow cytometry in the investigation of basic and clinical questions relating to HIV pathogenesis. The prime goal is to provide the intellectual environment and the material resources to enable junior investigators to apply flow cytometry technology to their projects and to encourage established investigators to initiate innovative pilot studies. To this end, several different funding sources (UCSD, CFAR, VA AIDS Research Center, VAMC Research Core) have been integrated to support and expand our flow cytometry facility to meet the research needs of the San Diego academic community. The CFAR award continues to be an integrate component in this endeavor. The flow cytometry facility contains three major pieces of equipment that are used to perform highly complementary functions. A Coulter Elite sorter with dual lasers plus a UV laser for calcium flux and cellular DNA is quantitation is utilized primarily for multi-parameter cell analyses. A Becton-Dickinson FACStar Plus sorter, with a fifth detector for 3-color analysis, operates as a high efficiency cell sorter for non-biohazardous or fixed cell samples. A recently funded, high speed, dual laser cell sorter (FACS Vantage Turbo) is being acquired and housed in a restricted BSL-2 space; it will perform both rare events recovery, needed for stem cell research and live cell sorting of HIV-infected specimens. Dedicated technicians operate these flow cytometers, and work is performed on a recharge basis for individual investigators. Support personnel include a highly experienced senior technician and a more junior, trained technician; a mid-level experienced technician is being recruited currently to operate the new cell sorter for work with biohazardous material. The core director, who is experienced in flow cytometry and research application, provides consultation on experimental design and data interpretation to investigators using the facility. In the past three years, CFAR resources have enabled the flow cytometry core to support HIV research projects in such diverse areas as: Kaposis sarcoma (CA67394), clonal B-cell development (AI34001), macrophages functions (AI35258, NS31060), cytokine responses (AI38858), therapy- induced immune restoration (AI27670), gene therapy (DK49618, AI36612), antiviral drug development (GM24979), and pediatric AIDS (AI39004). With the expanding application of flow cytometry to areas of biomolecular research, continued CFAR support will be crucial to providing the infrastructure to take advantage of new scientific opportunities in HIV research.