FLUORESCENCE ACTIVATED CELL SORTER

荧光激活细胞分选仪

• 批准号: 7335195
• 负责人: TONY N. MARION
• 金额: $ 24.3万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7335195
• 关键词: flow cytometry; fluorescence

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): NIH funded researchers at The University of Tennessee Health Science Center currently have critical need for a high-speed fluorescence activated cell sorter (FACS) with two or four-way sort option, the ability to sort into tubes or microplates, the option for single-cell deposition, detectors for forward and side scatter and ten fluorescence channels, temperature control for collected samples, and protection from aerosol dispersion. This application is for funds to purchase a Becton-Dickinson FACSAria FACS that is ideally suited to meet those demands. There is currently only one cell sorter in operation on the UTHSC campus. The cell sorter currently in operation is 14 years old and because of its age, is frequently out-of-service. New replacement parts for many mechanical and electronic components are no longer available and can only be acquired by scavenging from other similarly aged machines that are removed from service. Although the existing cell sorter was modified for high-speed sorting, it does not meet the minimum requirements indicated above. The existing sorter allows only four fluorescence channels, cannot sort into microwell plates, and cannot perform single-cell deposition. Since its inception in the late sixties and seventies, flow cytometry has become a mainstay for almost all research that involves the analysis of cells and cell functions. Monoclonal antibodies to the plethora of cell membrane markers, differentiation markers or CD antigens, and probes for cytoplasmic organelles, such as endosomes, nuclei, and mitochondria have made flow cytometry an even greater research resource. The uses of cell sorting by the investigators in this application include sorting macrophages that have ingested bacteria, sorting leukocytes into lymphoid and monocytic subsets, sorting specific T cell subsets based upon class II tetramer binding, isolating brain cells based upon level of expression of GFP reporters, isolation of lung endothelial cells before and after trauma, using recombinant, soluble T cell receptors to isolate insect cells expressing recombinant MHC class ll-peptide, and isolation of distinct B cell subpopulations based upon level of transgenic surface Ig expression.

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（申请人提供）：NIH资助的田纳西大学健康科学中心的研究人员目前迫切需要一种高速荧光激活细胞分选仪（FACS），该分选仪具有双向或四向分选选项、分选到试管或微孔板中的能力、单细胞沉积选项、前向和侧向散射检测器以及十个荧光通道、收集样品的温度控制、和防止气溶胶分散。此申请用于购买Becton-Dickinson FACSAria FACS的资金，非常适合满足这些需求。目前只有一台细胞分选机在UTHSC校园内运行。目前正在运行的细胞分选机已有14年的历史，由于其老化，经常无法使用。许多机械和电子部件的新替换零件不再可用，只能通过从其他类似老化的机器中回收来获得。虽然现有的细胞分选机被修改为高速分选，但它不满足上述最低要求。现有分选仪仅允许四个荧光通道，不能分选到微孔板中，也不能进行单细胞沉积。自60年代末和70年代初问世以来，流式细胞术已成为几乎所有涉及细胞和细胞功能分析的研究的支柱。针对过多细胞膜标记物、分化标记物或CD抗原的单克隆抗体，以及针对细胞质细胞器（如内体、细胞核和线粒体）的探针，使得流式细胞术成为更大的研究资源。研究者在本申请中使用的细胞分选包括分选已摄入细菌的巨噬细胞，将白细胞分选成淋巴和单核细胞亚群，基于II类四聚体结合分选特异性T细胞亚群，基于GFP报告基因的表达水平分离脑细胞，在创伤前后分离肺内皮细胞，使用重组，可溶性T细胞受体以分离表达重组MHC II类肽的昆虫细胞，并根据转基因表面IG表达水平分离不同的B细胞亚群。