The facility currently supports the research of the majority of NIAMS IRP investigators. In addition, it is actively involved in instrument training and flow cytometric assay development for all IRP users unfamiliar with this technique. The Facility currently supports the NIAMS IRP community with the following instrumentation: (One) BD Biosciences FACSAria Fusion cell sorter (5-laser excitation, 19-color detection, automated cell deposition unit, Biosafety Cabinet) new purchase to replace MoFlo cell sorter (One) BD Biosciences FACSAria IIIu cell sorter (5-laser excitation, 19-color detection, automated cell deposition unit) (One) BD Biosciences Influx cell sorter (7-laser excitation, 20-color detection, automated cell deposition unit, spectral analyzer (One) Beckman Coulter MoFlo cell sorter (3-laser excitation, 10-color detection, automated cell deposition unit) retired after 10+ years of service prior to laboratory relocation in June 2014 (One) BD Biosciences FACSCanto SORP analyzer w/HTS (4-laser excitation, 14-color detection, high-throughput sampler) (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) (One) Compucyte iCys research laser scanning cytometer (3-laser excitation, 5-parameter detection) (One) Amnis ImageStreamX Mark II high resolution microscope/cytometer (7-laser excitation, 3 imaging objectives, analysis software) 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. Usage of the facility to sort patient specimens and human-derived cells can now be done with a higher degree of safety with the recent addition of the FACSAria Fusion cell sorter, which features a laminar flow biosafety cabinet. The core facility relocation within Bldg. 10 from 6D to 10C also provided the opportunity to house the BD InFlux cell sorter in a Baker biosafety cabinet, as well. These sorters now provide better opportunities for NIAMS investigators to further the Bench to Bedside initiative promoted by NIH Director, Dr. Francis Collins. Equipped with seven lasers and 20+ fluorescence detectors, the Influx cell sorter and LSR Fortessa analyzers continue to provide state-of-the-art tools, enabling 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 utilize the Section's laser scanning cytometer, the Thorlabs (Compucyte) iCys, as well as the Amnis ImageStreamx Mark II. These microscope-based instruments provide fluorescent and morphological data on cells or tissue fixed to slides or multi-well plates (iCys) or cells in suspension as in traditional flow cytometry (ImageStream). Along with typical immunophenotyping, apoptosis assays and co-localization studies, the iCys can also re-analyze specific cells over time for kinetics-based assays. In contrast, the design of the ImageStream Mark II allows for higher throughput for more robust statistics, a characteristic of standard flow cytometers, while providing image analysis of a microscope-based system. The FCS has three dedicated personnel to support the need of NIAMS IRP investigators: Mr. Jeffrey Lay has been a part of the section since 2010. He has been recently trained to use the Amnis ImageStream and will continue to promote and support usage of the instrument. Kevin Tinsley, PhD, joined the core facility in early January 2013 as a contractor. Having completed his doctoral degree and postdoctoral fellowship in the field of immunology, he joined the Core with a substantial familiarity with flow cytometry. He completed his FACSAria training at BD Biosciences in 2013 and is now the primary operator of the FACSAria IIIu. While fulfilling his duties as Section Leader, Mr. James Simone continues to be involved in the promotion of flow cytometry at the NIH and within the Mid-Atlantic Region. As a 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 longtime membership in 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. More than two hundred NIAMS flow cytometry users have been trained to date on the different platforms available in the facility. We continue to use successfully the multi-level training scheme that was introduced in 2011. Providing a more logical training structure, a progressive group and individual-based training system is used to provide investigators and their staff better access to the Flow Cytometry Sections 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 instruments most appropriate for their particular research needs. After approval by the section leader, users can then operate the instruments unassisted. In all cases, however, the FCS staff is available for troubleshooting and consultation.

Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Arthritis, Musculoskeletal, Skin Dis
Department
Type
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