CELL SORTER/FLOW CYTOMETRY CORE FACILITY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 模块化流(MoFlo)高速细胞分选器(达科细胞分选器；科林斯堡，CO)设计为每秒70,000个事件进行分选，纯度为99%。MoFlo配置了三个激光器，配备了将细胞分选成微滴定板的能力。还包括一台水冷相干Innova 90-C Ar离子350 nm(UV)激光器，一台风冷光谱物理Ar离子488 nm(蓝)激光器，以及一台Melle-Griot 633 nm(红)氦-霓虹激光器。这种配置提供了最多检测六种颜色的能力，并且软件配置允许使用以下荧光染料同时进行最多四种颜色的免疫表型鉴定：FITC、PE、PE-Cy-7和APC。此外，该仪器配备了旋风自动细胞沉积单元，可以快速准确地将细胞沉积成各种多孔板格式(每孔一个细胞到多个细胞)。MoFlo是可公开重新配置的，如果我们的研究需要改变，它还具有额外的扩展能力，最多可以额外增加四个探测器。作为一项额外的安全功能，安装了E-Vac(气溶胶排气系统)，以容纳分类过程中产生的气溶胶。 流式细胞仪设备：该设备是罗杰·威廉姆斯医院病理和实验室医学部的一个组成部分。因此，它通过了美国病理学家学会(CAP)的认证，并符合CLIA 88标准。该设施占地800平方英尺，根据医院的需要进行维护和翻新。该设施在二级生物危害状态下运行，并严格遵守通用预防措施。有两名全职技术专家可以提供帮助。 FACSCalibur：FACSCalibur(Becton Dickinson)配备了用于自动采集样本的加载器(FACS加载器)。它还有一个用于分析的FACStation。分析程序包括：FACScomp、SimulSet、MultiSet、Attractors、CELLQuest和ModFit LT。 LSRII流式细胞仪：LSRII流式细胞仪(Becton Dickinson)也可用于免疫表型分析。与拥有专门操作员的MoFlo细胞分选仪不同，Cobre鼓励人员接受LSRII培训，这将使他们能够独立使用该仪器。培训由设施人员提供，他们将确保他们熟练操作设施设备。

This subproject is one of many research subprojects utilizing the resources provided by a Center 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 Center, which is not necessarily the institution for the investigator. The Modular Flow (MoFlo) high-speed cell sorter (DakoCytomation; Fort Collins, CO) is designed to sort at 70,000 events/second, with 99% purity. The MoFlo is configured with three lasers, equipped with the capability of sorting cells into microtiter plates. Also included are a water-cooled Coherent Innova 90-C Argon ion 350nm (UV) laser, an air-cooled Spectraphysics Argon ion 488nm (blue) laser, and a Melles-Griot 633nm (red) Helium-Neon laser. This configuration provides the capability of detecting up to six colors, and the software configuration allows for up to four color immunophenotyping simultaneously using the following fluorochromes: FITC, PE, PE-Cy-7, and APC. Additionally, the instrument is equipped with a CyClone Automated Cell Deposition Unit, which allows for rapid and accurate cell deposition into various multi-well plate formats (single cell to multiple cells per well). The MoFlo is openly reconfigurable and has additional expansion capabilities for up to four additional detectors, should the needs of our research shift. As an added safety feature, an E-Vac (aerosol evacuation system) is installed to contain aerosols produced during sorting. Flow Cytometry Facility: This facility is a component of the Department of Pathology and Laboratory Medicine at Roger Williams Hospital. As such it is certified by the College of American Pathologists (CAP), and is CLIA 88 compliant. The facility is housed in an 800 square foot area and is maintained and renovated as needed by the hospital. The facility operates at a level two biohazard status, and Universal Precautions are strictly adhered to. There are two full time technologists available for assistance. FACSCalibur: The FACSCalibur (Becton Dickinson) is equipped with a loader (FACS loader) for automatic acquisition of samples. It also has a FACStation for analysis. Analysis programs include: FACScomp, SimulSET, MultiSET, Attractors, CELLQuest, and ModFit LT. LSRII flow cytometer: An LSRII flow cytometer (Becton Dickinson) is also available for immunophenotyping analysis. In contrast to the MoFlo cell sorter, which has a dedicated operator, COBRE personnel are encouraged to train on the LSRII, which will allow them independent use of the instrument. Training is provided by facility personnel, who will ensure that they are proficient in operating the facility equipment.