BD FACSAria Flow Cytometer Cell Sorter

描述（由申请人提供）：本共享仪器拨款提案是用于购买BD FACSAria流式细胞仪细胞分选仪。这是一个最先进的全数字，三激光（488nm， 633nm和407nm）流式细胞仪细胞分选仪。它能够以每秒高达7万个细胞的采集速率分选2到4个种群。这个请求包括几个选项。水循环器将为样品和分选馏分提供最佳的温度控制。自动细胞沉积单元（ACDU）选项是一个集成的细胞沉积系统，用于多孔板和载玻片分选。气溶胶管理系统选项将使生物有害样品的分类成为可能。HP XW4000工作站提供了一个强大的数据管理系统。新的BD FACSAria细胞分选仪将被安置在USUHS生物医学仪器中心（BIC），流式细胞仪设施G235室，并将由流式细胞仪设施主任Karen M. Wolcott操作和维护。首席研究员William C. Gause将与内部咨询委员会协商，提供行政/科学监督。目前使用的流式细胞仪细胞分选仪是一台使用了13年的Beckman Coulter EPICS ELITE ESP。由于这种过时的仪器无法满足许多研究方案的需求，特别是nih支持的研究人员的需求，许多分选请求都被拒绝了。优先考虑由美国国立卫生研究院支持的科学家，他们正在参与这项拨款提案。它们包括：1)；可溶性MHC II-肽预防1型糖尿病；2)。Ras效应因子在乳腺分化中的作用3)。Darling, Thomas N. -结节性硬化症的肿瘤发生；4)。体内Th2细胞承诺、分化和细胞周期进程的调控；5)。HTLV-1的分子生物学及发病机制6)。Pollard, Harvey B. -囊性纤维化炎症通路的调控；7)。含黑视素视网膜神经节细胞的分离与表征8)。Schaefer, Brian C. - NF-KB信号在淋巴细胞活化和肿瘤发生中的抗原受体调控；9)。树突状细胞和T细胞在抗菌Ig反应中的作用10)。系统性红斑狼疮（SLE）期间淋巴细胞功能的调控和11)。《红斑狼疮的免疫发病机制》预计这组调查人员将需要该仪器总使用量的95%。剩下的5%将以先到先得的方式分配给USUHS、美国国立卫生研究院支持的研究科学家或USUHS社区的其他个人。分拣时间可能会不时提供给USUHS社区以外的调查人员，否则仪器将不会被使用。

DESCRIPTION (provided by applicant): This Shared Instrumentation Grant proposal is for the purchase of the BD FACSAria Flow Cytometer Cell Sorter. This is a state-of-the-art fully digital, three laser (488nm, 633nm and 407nm) Flow Cytometer Cell Sorter. It is capable of sorting two to four populations at an acquisition rate of up to 70,000 cells per second. This request includes several options. The water recirculator will provide optimal temperature control for the sample and sorted fractions. The Automated Cell Deposition Unit (ACDU) option is an integrated cell deposition system for multi-well plate and slide sorting. The aerosol management system option will enable sorting of biologically hazardous samples. The HP XW4000 Workstation provides a powerful data management system. The new BD FACSAria cell sorter will be housed at USUHS in the Biomedical Instrumentation Center (BIC), Flow Cytometry Facility, Room G235, and will be operated and maintained by the Flow Cytometry Facility Director, Karen M. Wolcott. The Principal Investigator, William C. Gause, will provide administrative/scientific oversight in consultation with an internal advisory committee. The BD FACSAria is requested as a replacement for the current flow cytometer cell sorter, a 13-year old Beckman Coulter EPICS ELITE ESP. Many requests for sorting have been declined due to the inability of this antiquated instrument to support the needs of many research protocols, especially those of the NIH-supported investigators identified in this grant. First priority will be given to the NIH-supported scientists who are participating in this grant proposal. They include: 1.) Brumeanu, Teodor D. - Prevention of Type 1 Diabetes by soluble MHC II- peptides; 2.) Cutler, Mary Lou - Role of Ras Effectors in Mammary Differentiation; 3.) Darling, Thomas N. - Tumorigenesis in Tuberous Sclerosis; 4.) Gause, William C. - Regulation of Th2 cell commitment, differentiation, and cell cycle progression in vivo; 5.) Giam, Chou-Zen - Molecular Biology and Pathogenesis of HTLV-1; 6.) Pollard, Harvey B. - Regulation of Inflammatory Pathways in Cystic Fibrosis; 7.) Provencio, Ignacio Isolation and Characterization of Melanopsin-Containing Retinal Ganglion Cells; 8.) Schaefer, Brian C. - Antigen receptor regulation of NF-KB signaling in lymphocyte activation and tumorigenesis; 9.) Snapper, Clifford M. - Dendritic and T cells in antibacterial Ig responses; 10.) Tsokos, George C. - Regulation of lymphocyte function during Systemic Lupus Erythematosus (SLE); and 11.) Via, Charles S. - Immunopathogenesis of Lupus. It is anticipated that this group of investigators will require 95% of the total usage of the instrument. The remaining 5% will be allocated to USUHS, NIH-supported research scientists or other individuals in the USUHS community on a first come, first served basis. Sorting time may be made available to investigators outside of the USUHS community from time to time when the instrument would otherwise not be used.