FLOW CYTOMETRY & HIGH SPEED CELL SORTING: GENETICS

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（由申请人提供）：UAB流式细胞术和高速细胞分选设备提供专家协助和仪器，用于分析UAB所有NIH资助的研究人员的颗粒和细胞。核心中的当前设备包括FACS Vantage（已升级为DiVa）、FACS Calibur和FACScan，这些设备是在NIH和该机构的支持下购买的。这些细胞仪在它们可以检测的荧光染料和荧光颗粒的数量和类型方面具有有限的能力。在该提案中，23名NIH资助的UAB研究者申请资金购买BD LSR II血细胞计数器。LSR II将配备固态488 nm蓝色激光器（20 mw）、JDS单相633 nm红色激光器（18 mw）和405 nm紫色激光器（25 mw）以及532 nm绿色激光器（可变0-150 mw）。该系统将能够利用三角形光学阵列检测来自蓝色激光器的2种颜色加上侧向和前向散射，利用三角形光学阵列检测来自红色激光器的3种颜色，利用八边形光学阵列检测来自紫色激光器的8种颜色，以及利用八边形光学阵列检测来自绿色激光器的5种颜色。该系统能够收集多达20个参数。收集的发射光将通过光纤技术传送到光学阵列。包括适当的带通滤波器和分色镜。LSR II细胞仪将提供UAB研究人员目前无法获得的三种技术：宽光谱荧光染料的多参数分析;多个报告结构的激发和检测发射;量子点荧光发射的激发和检测。该提案记录了共同研究者使用扩展的技术能力的实验计划，缺乏可获得的替代品，以及对该细胞仪的机构行政和物质支持。结合该基金在研究者教育和推广方面的努力，该提案将显著改善流式细胞术服务，以支持UAB的NIH资助项目。

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.