FLOW CYTOMETRY Flow cytometry - Jianke Zhang, PhD, KCC, TJU. The Flow Cytometry Shared Resource (FCSR) is an established KCC shared resource. It is located in a dedicated laboratory space of about 600 ft2. The FCSR is the only shared resource that provides access to all TJU investigators. The mission of FCSR is to provide high quality, efficient, and affordable analytical cytometry services to the KCC members in order to serve the basic science and the translational programs. Cytomation MoFlo high-speed cell sorter: This instrument was purchased 2 years ago using an NIH equipment grant supplemented by institutional funds. It is capable of sorting up to 70,000 particles per second, and has an air-cooled 488 argon laser and a 633 Helium-Neon laser. It allows analysis and sorting based on up to 7-color immunofluorescence detection combined with analysis of forward- and right angle-light scatter. Coulter EliteESP Cell Sorter: The Enhanced Sensitivity Performance (ESP) Flow Cytometer is capable of sorting between 5,000 and 15,000 particles per second (defined as high speed). The flow cytometer is equipped with 3 lasers, including a Coherent Innova 305 water-cooled Argon laser, an air-cooled Argon laser (488nm), and an air-cooled Helium-Neon laser (633nm). Coulter XL-MCL Automated Flow Cytometer: The Coulter XL-MCL is a cytometry system designed for high-throughput qualitative and quantitative measurements of biological and physical properties of cells and other particles. Becton-Dickinson FACSCalibur Benchtop Analyzer: The FACSCalibur bench top analyzer is equipped with an air-cooled Argon laser, an air-cooled Helium-Neon laser, and 5 photomultiplier tubes. The computer workstation is connected to the network and is a Becton Dickinson FACStation. Becton-Dickinson FACScan Benchtop Analyzer has an air-cooled Argon laser, and a temperature regulation unit. Analysis is based on up to 3-color immunofluorescence and light scatter characteristics. Workstation and software for extended data analysis: The shared resource has a network-connected computer workstation for off-line reanalysis of stored data. This system is a Celeron 400Mhz/64MB RAM/13GB HD/CD-ROM, loaded with various cytometry programs including FloJo, EXPO2, MulticycleAV, Multitime, MFI, and WinMDI. Flow cytometry - Bruce Boman, MD, PhD, CCHS. The Christiana Care Flow Cytometry Facility is located in the new Helen F. Graham Cancer Center Pavilion. The flow cytometry facility provides state-of-the-art fluorescence-activated cell sorting and analysis, including multi-color cell surface phenotyping, detection of intracellular and cell surface molecules, studies of cell proliferation, such as cell cycle analysis and apoptosis, and isolation of cell subpopulations expressing specific markers such as stem-cell markers. The shared resource allows for reliable and accurate characterization of a wide spectrum of biological samples and provides services that can be tailored to experimental designs in a variety of fields toward the understanding of the biology and treatment of cancer and other disease processes. The facility will house a BD FACSAria™ cell sorter for high performance flow cytometry. This easy-to-use benchtop system delivers high-speed sorting and multicolor analysis. BD FACS™ Accudrop technology is integrated into the system for quick and accurate drop-delay determination. Flow cytometry - Robert Mason, PhD, Nemours/AIDHC. The Flow Cytometry/FACS facility provides tools to study, analyze, select and purify cells using endogenously produced fluorescent tags or antibodies to specific cell surface or intracellular markers. The facility operates the Beckman-Coulter Altra Fluorescence Activated Cell Sorter (FACS). An advantage of this cell sorter is that it has 3 lasers with multiple variation of wavelengths and the capability of high-speed sorting, which results in higher yields or viable cells. The Flow Cytometry/FACS Core staff is available to advise on study design, perform analyses, and sort cells into specific populations for additional study or culture.