FLUORESCENCE ACTIVATED CELL SORTER: AUTOIMMUNE

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：美国国立卫生研究院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器（FACS），该分选器具有两种或四种分选选项，分选到管或微孔板的能力，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，以及防止气溶胶分散。此申请用于购买Becton-Dickinson FACSAria FACS的资金，该FACS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，由于其年龄，经常停止使用。许多机械和电子部件的新替换部件不再可用，只能通过从停止使用的其他类似老化的机器上清除来获得。虽然对现有的细胞分选机进行了高速分选改造，但仍不能满足上述最低要求。现有的分选器只允许四个荧光通道，不能在微孔板上进行分选，也不能进行单细胞沉积。流式细胞术自六七十年代末问世以来，已成为几乎所有涉及细胞和细胞功能分析的研究的支柱。针对大量细胞膜标记物、分化标记物或CD抗原的单克隆抗体，以及针对胞质细胞器（如核内体、细胞核和线粒体）的探针，使流式细胞术成为更大的研究资源。研究人员在此应用中使用细胞分选，包括分选摄取细菌的巨噬细胞，将白细胞分选为淋巴细胞和单核细胞亚群，根据II类四聚体结合分选特定的T细胞亚群，根据GFP报告者的表达水平分离脑细胞，分离创伤前后的肺内皮细胞，使用重组的可溶性T细胞受体分离表达重组MHC类肽的昆虫细胞。以及基于转基因表面Ig表达水平的不同B细胞亚群的分离。

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.