Cytomation MoFlo cytometer and high-speed cell sorter

描述(由申请者提供)：该建议书申请资金购买一台Cytomotion MoFlo高速、高纯度流式细胞仪细胞分选机，以支持北卡罗来纳州立大学九(9)名生物医学研究人员的研究，总共有11个NIH资助的项目。该仪器将被安置在NCSU的流式细胞仪核心设施中。其中一些项目侧重于T淋巴细胞的小亚群，这些T淋巴细胞调节艾滋病毒引起的艾滋病的FIV模型中的耐药性或免疫缺陷。两个项目研究了支气管上皮细胞群体及其功能。另一个项目包括利用基因敲除小鼠研究与慢性炎症性肠病相关的细胞因子；利用转基因的转录因子分析调节视网膜母细胞瘤易感性的特定内源性细胞周期基因的表达；以及基因打靶和动物克隆技术。在所有情况下，MoFIo技术都需要根据表面标记或细胞周期来分离细胞亚群，以分析特定的蛋白质和/或细胞功能。PI目前有可用于细胞分选的FACSCaliber，但其分选速度(500个细胞/秒)太慢。MoFIO的分选速度高达70,000个细胞/秒，并具有从混合种群中同时分选多达4个细胞子集的能力。北卡罗来纳大学教堂山分校有一个MoFIo分校，杜克大学有两个FACSVantage分校，这两个分校都距离NCSU 30英里。安排这些设备的难度和较长的通勤时间(最多单程最少45分钟)使得在常规基础上使用这项技术是不切实际的。该大学将为MoFIo操作员提供支持，并支付费用，使该仪器进入运行状态。MoFlo监督委员会制定的使用费将涵盖后续的服务合同。客户对MoFIo的使用将由监督委员会根据当前的流式细胞仪设施指南确定。我们会优先考虑这项建议的私人机构督导人员，但亦会鼓励其他教员在有需要时使用这项技术。

DESCRIPTION (provided by applicant): This proposal requests funds to purchase a Cytomotion MoFlo high speed, high purity flow cytometric cell sorter to support the research of nine (9) Biomedical research faculty with a total of 11 NIH supported projects at North Carolina State University. The instrument will be housed in the Flow Cytometry Core Facility at NCSU. Some of the projects focus on small subsets of T lymphocytes that regulate resistance or immunodeficiency in the FIV-model for HIV-induced AIDS. Two projects examine bronchial epithelial cell populations and their functions. Another project includes utilizing knock-out mice to study the factors of cells involved in chronic inflammatory bowel disease; utilizing transfected mutated transcription factors to analyze expression of specific endo cell-cycle genes regulating susceptibility to retinoblastoma; and gene targeting and animal cloning technology. In all cases, MoFIo technology is required to separate cell subsets based on either surface markers or cell cycle to analyze specific proteins and/or cell function. The PIs currently have available a FACSCaliber for cell sorting, but its sorting speed (500 cells/sec) to too slow. The MoFIo sorts at a rate up to 70,000 cells/sec, and has the capability of simultaneous sorting of up to 4 cell subsets from a mixed population. There is a MoFIo at the University of North Carolina-Chapel Hill and two FACSVantage sortesr at Duke University, both 30 miles from NCSU. The difficulty in scheduling these instruments and long commuting time (minimum of 45 minutes one way at best) makes it impractical to use this technology on a routine basis. The University will provide support for the MoFIo operator and costs to place the instrument in operational order. Users fees established by the MoFlo oversight committee will cover subsequent service contracts. Client use of the MoFIo will be determined by the oversight committee under current guidelines for the Flow Cytometry Facility. The PIs on this proposal will be given priority; however, other faculty will be encouraged to use the technology as the need arises.