BD Biosciences LSR Flow Cytometer

描述（由申请人提供）：一组九个NIH资助的主要用户和 Fox Chase癌症中心的四名NIH资助的二级用户申请资金， BD Biosciences LSR流式细胞仪。目前可用的仪器， 能够提供>3色分析，BD FACStar' plus细胞分选仪位于 特别是由于对分拣的需求不断增加。另 仪器BDFACScan广泛用于简单的免疫表型分型（2色， P1）和细胞周期分析，但不能进行更复杂的多色 通过使用各种荧光报告物， 系统（例如，IRES-GFP逆转录病毒）。此外，许多用户在此 应用程序提出的实验，目前不能与目前的 仪器，特别是测定细胞内钙水平，或 DNA含量以及细胞表面标记。拟议 仪器是一个由用户操作的三激光分析仪， 最大限度地灵活安排时间，并提供更多的时间分析 这些新的能力。虽然研究计划的 不同的用户是相当多样化的，包括免疫生物学，病毒学， 细胞生物学，他们都有一个最终目标， 疾病过程。这是通过研究的机制， 淋巴细胞发育，免疫球蛋白重排，功能性 效应细胞亚群（如记忆B细胞，辅助T细胞， 细胞毒性T细胞和NK细胞），细胞周期的调节，信号转导 转导和调节细胞形态。拟议的一些 实验分析感染中枢神经系统的病毒。其他人则寻求 来增加我们对免疫反应的理解，包括体液和细胞， 可以调节，这对疫苗接种和自身免疫都有影响。 还有一些人更关注临床，寻求开发疫苗疗法 治疗某些癌症所有这些项目都有一个共同的需求， 单个细胞的多个精确测量，通常在复杂的 混合物，拟议的文书可以有效满足这一要求。 要求的仪器将在流式细胞仪中保存和维护 FCCC的设施。用户将接受机构工作人员的操作培训。 该共享设施是该系统的理想位置，因为它提供了流动 细胞计数技术专业知识和仪器访问NIH资助的实验室 在气候变化框架公约，是一个中心的合作努力，杰出的，训练有素的 技术支持.

DESCRIPTION (provided by applicant): A group of nine NIH-funded major users and four secondary NIH-funded users at Fox Chase Cancer Center request funds for a BD Biosciences LSR flow cytometer. The currently available instrumentation that is capable of providing >3 color analysis, a BD FACStar' plus cell sorter is at capacity, particularly due to increasing demands for sorting. The other instrument, a BD FACScan, is widely used for simple immunophenotyping (2 color, P1) and for cell cycle analysis, but cannot perform more complex multi-color analyses that are increasingly required by use of various fluorescent reporter systems (e.g., IRES-GFP retroviruses). Furthermore, many of the users in this application propose experiments that currently cannot be done with the present instrumentation, specifically determination of intracellular calcium levels or DNA content together with multicolor cell surface markers. The proposed instrument is a three-laser analyzer that will be user-operated, providing for maximum flexibility in scheduling and providing increased multicolor analysis capacity together with these new capabilities. While the research programs of the different users are quite diverse, including immunobiology, virology, and cell biology, they all have as an ultimate goal the understanding of human disease processes. This is being accomplished by studies of the mechanisms of lymphocyte development, imrnunoglobulin rearrangement, generation of functional subpopulations of effector cells (such as memory B cells, helper T cells, cytotoxic T cells, and NK cells), regulation of the cell cycle, signal transduction, and regulation of cell morphology. Some of the proposed experiments analyze viruses that infect the central nervous system. Others seek to increase our understanding how immune responses, both humoral and cellular, can be regulated, with implications for vaccination and also autoimmunity. Still others have a more clinical focus, seeking to develop a vaccine therapy for certain cancers. All of these projects have a common need for making multiple precise measurements of individual cells, typically in complex mixtures, a requirement that can be met effectively by the proposed instrument. The requested instrument will be housed and maintained in the Flow Cytometry Facility at FCCC. Users will be trained in its operation by Facility personnel. This shared Facility is an ideal location for this system as it provides Flow Cytometry technical expertise and instrument access to NIH funded laboratories at FCCC and is a hub of collaborative efforts with outstanding, highly trained technical support.