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

该设施目前支持许多NIAMS IRP调查人员的研究。此外，它还积极参与对所有不熟悉这项技术的IRP用户的培训和化验支持。 该基金目前通过下列手段支持NIAMS IRP社区： (一)Beckman Coulter MoFlo细胞分类器(3激光激发、10色检测、自动细胞沉积装置) (一)BD Bioscience FACSAria IIIu高速分选机(5激光激发，19色检测，自动细胞沉积单元) (1)Beckman Coulter青色分析仪(3激光激发，9色检测) (1)BD Bioscience FACSCanto II分析仪，带HTS(3激光激发、8色检测、高通量采样器) (2)BD Bioscience FACSCalibur分析仪(2激光激发、4色检测) (一)复合式ICYS研究激光扫描细胞仪(3激光激发，5参数检测) 我们预计在2011年9月底之前再安装四个细胞仪。 (一)BD Bioscience Inflation细胞分选机(7激光激发，20色检测，自动细胞沉积单元，完全可更换的流体组件(支持NCRM)，光谱分析仪 (1)BD Bioscience LSR Fortessa分析仪(7路激光激发、20色检测、高通量采样器) (2)BD Bioscience FACSVerse分析仪(3激光激发、8色检测、通用高通量采样器、体积测量(用于绝对细胞计数)) 随着FACSVerse分析仪的增加，FACSCaliburs将被淘汰。 到目前为止，已有150多名NIAMS流式细胞仪用户接受了关于该设施可用不同平台的培训。2011年，我们继续使用2011年引入的经修订的多层次培训计划。为了提供更合理的培训结构，采用了以小组和个人为基础的循序渐进的培训制度，使调查员及其工作人员能够更好地获得流式细胞科的设备和服务。所有新用户都参加了设施设备和政策的一般介绍和培训。然后，根据需要和先前的经验，他们就最适合他们特定研究需要的仪器进行个性化或小组指导。在得到科长的批准后，用户可以在没有辅助的情况下操作仪器。然而，功能界别的工作人员随时可以进行故障排除和咨询。 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，该设施还继续为NIAMS实验室提供流式细胞仪和分类服务，这些实验室从事的学科通常不是流式细胞仪：发育皮肤生物学单元和肌肉干细胞和基因实验室。这些实验室的项目一直并将继续令人鼓舞。 在过去的几年里，该设备的使用量稳步增加，需要升级我们的高速细胞分选仪和分析仪。 今年，为了应对日益增长的需求，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)，我们将增加四个新的细胞仪(如上所示)。配备了7个激光器和20+个荧光探测器的Inumx细胞分选仪和LSR Fortessa分析仪都将提供最先进的仪器，使NIAMS的研究人员能够利用最新的荧光色素和荧光蛋白质报告。 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切片激光扫描细胞仪，即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数据。除了典型的免疫表型和共定位研究，ICYS还可以随着时间的推移重新分析特定细胞，进行基于动力学的分析。 詹姆斯·西蒙先生在履行其作为核心科长的职责的同时，继续参与在全美卫生研究院推广流式细胞术。作为流式细胞术兴趣小组的成员和联席主席，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。他长期担任国际细胞测量促进会(ISAC)和ISAC生物安全委员会的成员，有助于促进与来自世界各地的同行交流新的想法和信息。

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.