Shared Resource: Cell Sorting Facility

细胞/分子分析共享资源：细胞分选设备(CSF) 项目摘要/摘要 细胞分选设施(CSF)为癌症中心成员提供访问和技术支持 流式细胞术技术，一种分析和分离(分类)复杂混合物中细胞的方法，基于 各种参数，包括光散射和荧光。理查德领导下的中国证监会 Hardy，PhD(BCDF)和1.3 FTE员工的目标是提供最高质量的分析和分类 癌症中心成员的能力。随着流式细胞仪技术的成熟，它已经成为必不可少的 研究的组成部分，用于各种不同的实验室。31名癌症中心成员使用了脑脊液 公历年2014年。2014年，94%的服务小时记录在CSF中，用于支持同行审查 资助研究。来自所有5个研究项目的48名癌症中心成员在2011年期间使用了CSF- 2014年。由于仪器的复杂性、成本和口径，服务是最方便的 集中式脑脊液。在每个实验室都有单独的流式细胞仪，这将是一种成本高昂的做法。 流式细胞仪分析和/或分类的性质并不真正适合外部(商业) 服务。也就是说，样本通常是活细胞，在分析之前不久就准备好了(因此很容易腐烂) 许多实验涉及使用不同的新试剂组合(荧光标记试剂 单抗)。哈代28年的经验和专业知识是实施 先进的技术和创新的方法。哈代的领导力和机构承诺导致了 购买一辆高度定制的FACSAria(509,500美元)。这种新仪器最大限度地激发了体育运动和体育运动 串列与绿色激光共轭，相对于蓝色488 nm激发提供更大的信噪比。这个 绿色激光还能够检测到某些荧光蛋白质，而这些蛋白质不能用以前的 优势分拣机。Aria还运行当前版本的仪器控制软件，带有编程的自动- 每天早上在设置时进行校准。LSR-II也进行了升级，以增强其能力，增加了绿色 激光(能最有效地激发PE和PE通道)，取代红色激光以增加激光 功率，并增加荧光探测器的数量。此仪器还可以运行当前的仪器 每天早上自动校准的控制软件。这款在Aria和LSR轨道上运行的自动校准软件 每个通道的敏感度等提供了额外的质量保证，标记了之前的潜在问题 他们会逐渐走向失败。内部开发的数据管理系统Flow-LIMS，允许所有癌症 中心成员对本地计算机上的流式细胞仪数据进行注释、访问和管理 实验室，旨在促进实验数据的共享，并有助于促进合作 在实验室之间。证监会由一个专门的设施咨询委员会(FAC)提供建议，该委员会将召开 每年一次。FAC的建议直接提交给管理机构家长监督委员会，该委员会负责 确保证监会继续拥有所需的资源，以提供最优质的服务。

CELLULAR/MOLECULAR ANALYSES SHARED RESOURCE: CELL SORTING FACILITY (CSF) PROJECT SUMMARY/ABSTRACT The Cell Sorting Facility (CSF) provides Cancer Center members access to and technical support for the technology of flow cytometry, a method for analyzing and isolating (sorting) cells in a complex mixture, based on a variety of parameters, including light scatter and fluorescence. The CSF under the direction of Richard Hardy, PhD (BCDF) and a staff of 1.3 FTEs has a goal to provide the highest quality analytic and sorting capabilities to Cancer Center members. As flow cytometry technology has matured, it has become an essential component of research, utilized in a diverse array of laboratories. 31 Cancer Center members used the CSF in calendar year 2014. 94% of service hours logged in the CSF during 2014 were in support of peer-review funded research. 48 Cancer Center members from all 5 Research Programs used the CSF in the period 2011- 2014. Due to the complexity, cost and caliber of the instrumentation, the services are best facilitated though the centralized CSF. It would be cost prohibitive to have individual flow cytometers available in each laboratory. The nature of flow cytometry analysis and/or sorting is not really appropriate for an external (commercial) service. That is, samples are often live cells, prepared shortly before analysis (therefore quite perishable) and many experiments involve the use of diverse combinations of novel reagents (fluorochrome-labeled monoclonal antibodies). The 28 years of experience and expertise of Hardy is key in implementing state-of- the-art technology and innovative approaches. Hardy's leadership coupled with institutional commitment led to the purchase of a highly customized FACSAria ($509,500). This new instrument optimally excites PE and PE tandem conjugates with a green laser, providing greater signal-to-noise relative to blue 488nm excitation. The green laser also enables detection of certain fluorescent proteins that could not be detected using the previous Vantage sorter. The Aria also runs the current version of instrument control software with a programmed auto- calibration at setup each morning. The LSR-II was also upgraded to enhance its capabilities, by adding a green laser (enabling most efficient excitation of PE and PE tandems), replacing the red laser to increase laser power, and increasing the number of fluorescence detectors. This instrument also runs the current instrument control software with auto-calibration each morning. This auto-calibration software on the Aria and LSR tracks the sensitivity in each channel and so provides added quality assurance, flagging potential problems before they progress to failure. The in-house developed data management system, Flow-LIMS, that allows all Cancer Center members to annotate, access, and manage flow cytometry data on computers in their local laboratories, has been designed to facilitate sharing of experimental data, and it helps to foster collaboration between laboratories. The CSF is advised by a dedicated Facility Advisory Committee (FAC) that meets annually. FAC recommendations are directed to a governing Facility Parent Oversight Committee, that serves to ensure that the CSF continues to have the necesssary resources to provide the highest quality services.