Becton Dickinson LSR 2 Flow Cytometer

描述(由申请人提供)：宾夕法尼亚大学的研究人员越来越多地从事需要超过六个参数的流式细胞术的研究。此外，许多研究人员正在利用荧光共振能量转移(FRET)实验来描绘受体之间的相互作用。目前，这些类型的多参数研究必须在细胞分类器上进行(只有1个能够用首选的CFP/YFP对测量FRET)，因为没有能够进行复杂的多参数分析的分析仪器。由于这些仪器也用于预备性细胞分选，因此仅用于分析的等待时间通常为数周。缺乏高速6+参数分析流式细胞术已被证明是几个资金雄厚的研究人员的严重限制。 Becton-Dickinson LSR II(BD LSRII)是一种风冷四激光台式细胞仪，能够同时测量12个以上的参数。使用新的数字采集系统，它具有增强的探测和分辨率能力。由于配置了紫外光和紫外光激光器，它将支持钙离子通量(使用INDO-1)和FRET实验，这两种实验现在非常有限。这些扩展到流式细胞仪核心实验室的分析能力将使用户群体和机构中的许多研究人员特别受益，并将成为医学院现有仪器的重要补充。在这项提议中，我们提出了一个共享资源操作的模型，该模型将支持主要针对用户组的BD LSR II的有效使用。已经到位的是重要的机构支持，经验丰富的工作人员，以及备受推崇的流式细胞仪培训计划。这将使调查人员能够立即使用这项服务，并通过培训选定的学生和技术人员使用来进一步扩大使用范围。这将导致科学界更多地获得高水平分析细胞术。 通过将LSR II放置在最近翻新的现有空间中，并利用成熟的核心流式细胞仪设施运营模式，我们建议将这种细胞仪的好处最大化地惠及NIH资助的17名研究人员的用户群体(每年直接成本为9,007,559美元)。这将使研究人员能够从事以前有限的研究领域，这将导致生物医学科学的资金增加和进步。

DESCRIPTION (provided by applicant): Researchers at the University of Pennsylvania are increasingly engaged in investigations that require greater than six-parameter flow cytometry. In addition, many investigators are making use of fluorescent resonance energy transfer (FRET) experiments to map receptor interactions. Currently, these types of multiparameter studies must be performed on the cell sorters (only 1 capable of measuring FRET with the preferred CFP/YFP pair) as no analytical instrument capable of performing sophisticated multiparameter analyses is available. Since these instruments are also used for preparative cell sorting, the wait for analytical only applications is often weeks. Lack of access to high-speed 6+ parameter analytical flow cytometry has proven to be a serious limitation to several well-funded investigators. The Becton- Dickinson LSR II (BD LSRII) is an air-cooled four-laser bench top cytometer capable of measuring more than 12 parameters simultaneously. Using a new digital acquisition system, it has enhanced detection and resolution capabilities. As configured with UV and violet lasers, it will support both calcium flux (with Indo-1) and FRET experiments, which are now available on a very limited basis. These expanded analytical capabilities to the flow cytometry core laboratory will specifically benefit the user group and many researchers in the institution and be a significant addition to the available instrumentation in the School of Medicine. In this proposal, we present a model for shared resource operation that will support the efficient use of the BD LSR II for predominantly the user group. Already in place are significant institutional support, a highly experienced staff, and a well-respected training program in flow cytometry. This will allow investigators immediate access to the service and further extend access by training selected students and technical staff to use. This will result in increased availability of high-level analytical cytometry to the scientific community. By placing the LSR II in existing recently renovated space and utilizing a well-established model for core flow cytometry facility operation, we propose to maximize the benefit of this cytometer to the user group of 17 NIH-funded investigators ($9,007,559 direct costs/year). This will allow investigators to pursue many areas of research, previously with limited availability, which will result in increased funding and advancement of biomedical science.