FLOW CYTOMETRY & HIGH SPEED CELL SORTING: INFLAMMATORY DISEASES

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)：UAB流式细胞术和高速细胞分选设施为所有NIH资助的UAB研究人员提供分析颗粒和细胞的专家协助和仪器。目前的核心设备包括一台FACS Vantage(后来升级为DIVA)、一台FACS Calibur和一台FACScan，这些设备都是在NIH和该机构的支持下购买的。这些细胞仪在它们可以检测到的荧光染料和荧光颗粒的数量和类型方面能力有限。在这项提案中，23名由NIH资助的UAB调查人员申请资金购买BD LSR II细胞仪。LSR II将配备固态488 nm蓝光激光器(20 MW)、JDS单相633 nm红光激光器(18 MW)、405 nm紫光激光器(25 MW)和532 nm绿光激光器(0-150 mW可变)。该系统将具有利用三角光学阵列从蓝色激光正侧和前向散射检测2种颜色的能力，使用三角光学阵列检测来自红色激光的3种颜色，使用八角形光学阵列检测来自紫色激光的8种颜色，以及使用八角形光学阵列检测来自绿色激光的5种颜色。该系统能够采集多达20个参数。收集到的发射光将通过光纤技术传输到光学阵列。包括适当的带通滤光器和二色镜。LSR II细胞仪将提供UAB研究人员目前无法使用的三项技术：广谱荧光的多参数分析；多报告结构的激发和发射检测；量子点荧光发射的激发和检测。该提案记录了协查人员使用扩大的技术能力的实验计划、缺乏可利用的替代方案以及对该细胞仪的机构行政和物质支持。结合该基金在研究员教育和外展方面的努力，这项提议将显著改善流式细胞仪服务，以支持UAB由NIH资助的项目。

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): The UAB Flow Cytometry and High-Speed Cell Sorting Facility provides expert assistance and instruments for analysis of particles and cells that is available to all NIH-funded researchers at UAB. The current equipment in the core includes a FACS Vantage (since upgraded to DiVa), a FACS Calibur and a FACScan, which were purchased with support from both the NIH and the institution. These cytometers have limited capabilities in the number and type of fluorochromes and fluorescent particles they can detect. In this proposal, twenty-three NIH-funded, UAB investigators request funds for purchase of a BD LSR II cytometer. The LSR II will be equipped with a solid-state 488nm blue laser (20mw) a JDS Uniphase 633nm Red laser (18mw) and 405nm Violet laser (25mw) and a 532nm Green laser (Variable 0-150 mw). The system will have the ability to detect 2 colors from the blue laser plus side and forward scatter utilizing a trigon optical array, 3 colors from the red laser utilizing a trigon optical array, 8 colors off the violet laser utilizing an octagon optical array and 5 colors off the green laser utilizing an octagon optical array. The system is capable of collecting up to 20 parameters. The collected emission light will be delivered to the optical arrays by fiber optic technology. Appropriate bandpass filters and dichroic mirrors are included. The LSR II cytometer will provide access to three technologies not currently available to UAB researchers: Multi-parameter analysis of a wide spectrum of fluorochromes; Excitation and detection of emission for multiple reporter constructs; Excitation and detection of fluorescent emission from quantum dots. This proposal documents the experimental plans of the co-investigators for use of the expanded technical capabilities, the absence of accessible alternatives, and the institutional administrative and material support for this cytometer. Combined with efforts by the Facility in investigator education and outreach, this proposal will significantly improve the flow cytometry services in support of NIH funded projects at UAB.