Application of Flow Cytometry to Cell Biology

实验免疫学分支（EIB）流式细胞术核心设施目前 支持欧洲投资银行50多名研究人员的多个研究项目， 在癌症研究中心（CCR）的其他地方。这些调查涉及 多参数定量单细胞分析和基于电子细胞分离 根据与从不同物种和/或组织新鲜制备的细胞相关的参数， 以及一系列体外培养细胞。提供基础研究支持， EIB的成员和NCI癌症研究中心的其他研究人员。 目前获得资助的项目包括但不限于以下研究领域： 通过细胞表面分子的细胞内信号传导的体内和体外分析; B） 分析动物和/或细胞中的细胞过程和/或缺陷， 修改; c）免疫发病机制和后果的研究; d） 分析多种分子的配位细胞表面表达; e） T细胞库产生的研究; g）移植物表达的分析 抗原; h）研究涉及T细胞谱系发育的机制; i） 细胞死亡机制;和j）干细胞分析。该设施运营和维护2 操作员运行3激光流式细胞仪，具有细胞分选功能和3个用户-操作员流程 仅具有分析能力的细胞仪。该设施最近获得了一个 最先进的5激光，仅用于分析，流式细胞仪将用于扩展和扩展 可用的应用程序。机构工作人员为研究者提供以下方面的咨询： 实验设计、问题解决、试剂选择和数据分析与解释。 该设备支持多种流式细胞仪应用，包括： 分析（包括干细胞分析）和细胞分选;多色表型分析， 细胞周期分析、增殖分析、代谢分析，包括钙通量 分析、无菌细胞分选和细胞内细胞因子分析。该设施还作为 节约成本的措施，维持一个超过150种常用流式细胞仪的试剂库 由机构工作人员预先滴定和等分供多个EIB使用的试剂 investigators.试剂库通过批量购买和最大限度地减少劳动力来最大限度地降低成本， 表征单个批次试剂的工作量。该设施正在开发 用于流式细胞术分析的基于WINDOWS的PC软件， 目前可在仪器制造商提供的软件或第三方软件中使用 源

The Experimental Immunology Branch (EIB) Flow Cytometry Core Facility currently supports multiple research projects for more than 50 investigators from within the EIB and elsewhere in the Center for Cancer Research (CCR). These investigations involve multiparametric quantitative single cell analysis of, and electronic cell separation based upon, parameters associated with cells freshly prepared from different species and/or tissues, as well as a spectrum of in vitro cultured cells. Basic research support is provided to members of the EIB and to other investigators within the Center for Cancer Research, NCI. Currently supported projects include, but are not limited to, the following areas of study: a) in vivo and in vitro analyses of intra-cellular signaling via cell surface molecules; b) analyses of cellular processes and/or defects in animals and/or cells with genetic modifications; c) studies of the mechanisms and consequences of immune pathogenesis; d) analyses of the coordinate cell surface expression of a variety of molecules; e) investigations of T cell repertoire generation; g) analyses of expression of transplantation antigens; h) investigations of mechanisms involved in T cell lineage development; i) mechanisms of cell death; and j) stem cell analyses. The facility operates and maintains 2 operator run 3-laser flow cytometers with cell sorting capabilities and 3 user-operator flow cytometers with analysis only capabilities. The facility has recently acquired a state-of-the-art 5-laser, analysis only, flow cytometer that will be used to extend and expand available applications. Facility staff provide consultation to investigators in the areas of: experimental design, problem-solving, reagent selection and data analysis and interpretation. The facility supports a wide variety of flow cytometric applications including: rare event analysis (including stem cell analysis) and cell sorting; multi-color phenotypic analyses, cell cycle analysis, proliferation analysis, metabolic analyses including calcium flux analysis, sterile cell sorting, and intra-cellular cytokine analyses. The facility also, as a cost-savings measure, maintains a reagent bank of over 150 commonly used flow cytometry reagents that are pre-titred and aliquoted by facility personnel for use by multiple EIB investigators. The reagent bank minimizes costs by buying in bulk and minimizing labor and effort involved in characterizing individual batches of reagents. The facility is developing WINDOWS-based pc software for flow cytometry analysis that will provide capabilities not currently available in software available from instrument manufacturers or 3rd party software sources.