CORE--FLOW CYTOMETRY

这一集中的核心资源的目的是提供（1）专门知识 （2）实验设计中的技术和设备支持 利用流式细胞术进行研究项目的实验 在调查与艾滋病毒有关的基础和临床问题方面， 发病机制主要目标是提供一个智力环境， 和物质资源，使初级调查员能够应用流程 他们的项目，并鼓励建立细胞计数技术， 研究人员启动创新试点研究。 为此，若干 不同的资金来源（UCSD，CFAR，VA艾滋病研究中心，VAMC 研究核心）已被整合，以支持和扩大我们的流程 细胞仪设施，以满足圣地亚哥学术研究的需要， 社区CFAR奖仍然是这一领域的一个组成部分， 奋进。 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 用双激光器加上紫外激光器来检测钙流和细胞DNA， 定量主要用于多参数细胞分析。一 Becton-Dickinson FACStar Plus分选机，带有第五个检测器，用于3色 分析，作为一个高效率的细胞分选机， 或固定的细胞样品。一个最近资助的高速双激光细胞 正在购置一台分拣机（FACS Vantage Turbo），并将其安置在一个受限制的 BSL-2空间;它将执行股骨柄所需的罕见事件恢复 细胞研究和HIV感染标本的活细胞分选。专用 技术人员操作这些流式细胞仪，并在 为个人研究者充电的基础。支助人员包括 一名经验丰富的高级技师和一名经过培训的初级技师 技术员;正在招聘一名中级经验丰富的技术员 目前正在操作新的细胞分选机， 材料核心总监，谁是经验丰富的流式细胞术和 研究应用，提供实验设计咨询， 向使用该设施的调查人员解释数据。 在过去的三年里，CFAR资源使流式细胞术 核心支持艾滋病毒研究项目在这样的不同领域：卡波西斯 肉瘤（CA 67394）、克隆性B细胞发育（AI 34001）、巨噬细胞 功能（AI 35258，NS 31060），细胞因子应答（AI 38858），治疗- 诱导免疫恢复（AI 27670），基因治疗（DK 49618，AI 36612）， 抗病毒药物开发（GM 24979）和儿科艾滋病（AI 39004）。与 流式细胞术在生物分子领域的应用 研究，持续的CFAR支持将是至关重要的， 利用艾滋病毒新的科学机会的基础设施 research.

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.