FLUORESCENCE ACTIVATED CELL SORTER: INFECTIOUS DISEASE

这个子项目是利用由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。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：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器(FACS)，具有两路或四路分选选项，能够分选到试管或微孔板中，可以选择单细胞沉积，用于正向和侧向散射和十个荧光通道的检测器，收集样本的温度控制，以及防止气溶胶扩散。这项申请是为资金购买Becton-Dickinson FACSAria FACS，它是满足这些需求的理想选择。目前，UTHSC校园中只有一个细胞分选机在运行。目前运行的细胞分选机已有14年的历史，由于其年代久远，经常无法使用。许多机械和电子部件的新更换部件不再可用，只能通过从其他已停止使用的类似老化的机器中回收来获得。虽然现有的细胞分选机进行了改装，以实现高速分选，但它不能满足上述最低要求。现有的分选机只允许四个荧光通道，不能分选成微孔板，也不能进行单细胞沉积。自六十年代末和七十年代问世以来，流式细胞术已经成为几乎所有涉及细胞和细胞功能分析的研究的支柱。针对过多的细胞膜标志物、分化标志物或CD抗原的单抗，以及针对细胞质细胞器(如内体、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。研究人员在这一应用中使用的细胞分类包括对吞噬细菌的巨噬细胞进行分类，将白细胞分成淋巴细胞和单核细胞亚群，基于II类四聚体结合对特定的T细胞亚群进行分类，根据GFP报告基因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11肽的昆虫细胞，以及基于转基因表面Ig表达水平分离不同的B细胞亚群。

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.