Flow cytometry support to research in immune, skin, muscle and bone diseases

流式细胞术支持免疫、皮肤、肌肉和骨骼疾病的研究

基本信息

批准号：
8344973
负责人：
Massimo Gadina
金额：
$ 63.42万
依托单位：
NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8344973
关键词：
Autoimmune Diseases Biological Assay Biology Bone Diseases Cell Count Cell Separation Cells Communities Consultations Cytometry Data Data Analyses Deposition Detection Development Discipline Disease Educational Status Equipment Flow Cytometry Fluorescence Fluorochrome Functional disorder Genes Goals Hypersensitivity Immune Immune System Diseases Immune System and Related Disorders Immunophenotyping Individual Instruction Interest Group International Kinetics Laboratories Lasers Measurement Microscope Muscle satellite cell Myopathy National Institute of Arthritis and Musculoskeletal and Skin Diseases Policies Proteins Regenerative Medicine Relative (related person)Reporter Research Research Personnel Research Support Safety Scanning Scheme Services Skin Slide Societies Sorting - Cell Movement Specimen Speed Structure System Techniques Time Training United States National Institutes of Health Vendor atopy base bone color detection detector experience instrument instrumentation meetings member novel peer skin disorder tissue fixing tool

项目摘要

The facility currently supports the research of numerous NIAMS IRP investigators. In addition, it is actively involved in training and assay support for all the IRP users who are unfamiliar with this technique. The Facility currently supports the NIAMS IRP community with the following instrumentation: (One) Beckman Coulter MoFlo cell sorter (3-laser excitation, 10-color detection, automated cell deposition unit) (One) BD Biosciences FACSAria IIIu high-speed sorter (5-laser excitation, 19-color detection, automated cell deposition unit) (One) Beckman Coulter CyAn analyzer (3-laser excitation, 9-color detection) (One) BD Biosciences FACSCanto II analyzer w/HTS (3-laser excitation, 8-color detection, high-throughput sampler) (Two) BD Biosciences FACSCalibur analyzers (2-laser excitation, 4-color detection) (One) Compucyte iCys Research Laser Scanning Cytometer (3-laser excitation, 5-parameter detection) And we expect to be installing four more cytometers by the end of September 2011. (One) BD Biosciences Influx cell sorter (7-laser excitation, 20-color detection, automated cell deposition unit, fully replaceable fluidics pack (to support the NCRM), spectral analyzer (One) BD Biosciences LSR Fortessa analyzer (7-laser excitation, 20-color detection, high-throughput sampler) (Two) BD Biosciences FACSVerse analyzers (3-laser excitation, 8-color detection, universal high-throughput sampler, volumetric measurement (for absolute cell counts) With the addition of the FACSVerse analyzers, the FACSCaliburs will be retired. More than one hundred fifty NIAMS flow cytometry users have been trained to date on the different platforms available in the facility. In 2011, we continue to use the revised, multi-level training scheme that was introduced in 2011. To provide a more logical training structure, a progressive group and individual-based training system is used to provide investigators and their staff better access to Flow Cytometry Section equipment and services. All new users attend a general introduction and orientation to facility equipment and policies. Then, according to needs and previous experience, they progress to individualized or group instruction on the instrument most appropriate for their particular research needs. After approval by the section leader, users can then operate the instruments unassisted. However, the FCS staff is always available for troubleshooting and consultation. In addition to the projects from those laboratories focused on mechanisms and dysfunctions of the immune system, the facility has continued to provide flow cytometry and sorting services to NIAMS laboratories engaged in disciplines not often served by flow cytometry: the Developmental Skin Biology Unit and the Laboratory of Muscle Stem Cells and Gene..The projects in these laboratories have been and continue to be quite encouraging. Usage of the facility has increased steadily over the last few years requiring the upgrade of our high-speed cell sorters and analyzers. This year, to deal with increasing demand, provide better tools for our investigators and support the newly-established National Center for Regenerative Medicine (NCRM), we are adding four new cytometers (listed above). Equipped with seven lasers and 20+ fluorescence detectors, both the Influx cell sorter and LSR Fortessa analyzer will provide state-of-the-art instrumentation that will allow NIAMS investigators to take advantage of the latest fluorochromes and fluorescent protein reporters. Those investigators whose research involves specimens that are not well suited to flow cytometry can continue to take advantage of the Sections laser scanning cytometer, the Compucyte iCys. This microscope-based instrument can provide fluorescent and morphological data on cells or tissue fixed to slides or multi-well plates. Along with typical immunophenotyping and co-localization studies, the iCys can also re-analyze specific cells over time for kinetics-based assays. While fulfilling his duties as the core Section Leader, Mr. James Simone, continues to be involved in the promotion of flow cytometry at NIH-wide level. As a member and co-chair of the Flow Cytometry Interest Group, he assists in organizing and promoting quarterly meetings that showcase leading researchers in the field and promote relationships with vendors of flow cytometry-related products. His long-time membership of the International Society for the Advancement of Cytometry (ISAC) and the ISAC Bio-Safety Committee helps facilitate the exchange of new ideas and information with peers from around the world.

该设施目前支持众多 NIAMS IRP 研究人员的研究。此外，它还积极参与对所有不熟悉该技术的 IRP 用户的培训和分析支持。该设施目前通过以下工具支持 NIAMS IRP 社区：（一）Beckman Coulter MoFlo细胞分选仪（3激光激发、10色检测、自动细胞沉积装置）（一）BD Biosciences FACSAria IIIu高速分选机（5激光激发、19色检测、自动细胞沉积装置）（一）Beckman Coulter CyAn分析仪（3激光激发，9色检测）（一）BD Biosciences FACSCanto II 分析仪带 HTS（3 激光激发、8 色检测、高通量进样器）（两台）BD Biosciences FACSCalibur分析仪（2激光激发，4色检测）（一）Compucyte iCys Research激光扫描细胞仪（3激光激发，5参数检测）我们预计到 2011 年 9 月底将再安装四台细胞仪。 (一) BD Biosciences Influx 细胞分选仪（7 激光激发、20 色检测、自动细胞沉积装置、完全可更换的流体包（支持 NCRM）、光谱分析仪（一）BD Biosciences LSR Fortessa分析仪（7激光激发、20色检测、高通量进样器）（两台）BD Biosciences FACSVerse 分析仪（3 激光激发、8 色检测、通用高通量进样器、体积测量（用于绝对细胞计数）随着 FACSVerse 分析仪的加入，FACSCaliburs 将退役。迄今为止，超过 150 名 NIAMS 流式细胞术用户已接受过关于该设施中可用的不同平台的培训。 2011年，我们继续沿用2011年引入的修订后的多层次培训计划。为了提供更合理的培训结构，采用渐进式小组和个人培训体系，使研究者及其工作人员能够更好地接触Flow细胞计数科设备和服务。所有新用户都会参加设施设备和政策的一般介绍和介绍。然后，根据需要和以前的经验，他们将继续使用最适合其特定研究需求的仪器进行个性化或小组指导。经部门领导批准后，用户即可独立操作仪器。不过，FCS 工作人员随时可以提供故障排除和咨询服务。除了那些专注于免疫系统机制和功能障碍的实验室的项目外，该设施还继续向从事流式细胞术不常服务的学科的 NIAMS 实验室提供流式细胞术和分选服务：发育皮肤生物学单位和实验室肌肉干细胞和基因的研究......这些实验室的项目已经并将继续令人鼓舞。在过去几年中，该设施的使用量稳步增加，需要升级我们的高速细胞分选仪和分析仪。今年，为了应对不断增长的需求、为我们的研究人员提供更好的工具并支持新成立的国家再生医学中心 (NCRM)，我们添加了四台新的细胞仪（如上所列）。 Influx 细胞分选仪和 LSR Fortessa 分析仪配备了 7 个激光器和 20 多个荧光检测器，将提供最先进的仪器，使 NIAMS 研究人员能够利用最新的荧光染料和荧光蛋白报告基因。那些研究涉及不太适合流式细胞术的标本的研究人员可以继续利用Sections 激光扫描细胞仪 Compucyte iCys。这种基于显微镜的仪器可以提供固定在载玻片或多孔板上的细胞或组织的荧光和形态学数据。除了典型的免疫表型分析和共定位研究之外，iCys 还可以随着时间的推移重新分析特定细胞，以进行基于动力学的测定。 James Simone 先生在履行核心科组长职责的同时，继续参与在 NIH 范围内推广流式细胞术。作为流式细胞术兴趣小组的成员和联合主席，他协助组织和推动季度会议，展示该领域的领先研究人员，并促进与流式细胞术相关产品供应商的关系。他是国际细胞计数促进会 (ISAC) 和 ISAC 生物安全委员会的长期会员，有助于促进与世界各地同行的新思想和信息交流。