CELL SORTER/FLOW CYTOMETRY CORE FACILITY

细胞分选仪/流式细胞术核心设施

• 批准号: 8167638
• 负责人: NICOLA M KOUTTAB
• 金额: $ 12.37万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167638
• 关键词: Aerosols; Air; American; Area; Argon; Biohazardous Substance; Cell Separation; Cells; Centers of Research Excellence; Color; Computer Retrieval of Information on Scientific Projects Database; Computer software; Core Facility; Deposition; Ensure; Equipment; Event; Flow Cytometry; Fluorescein-5-isothiocyanate; Fluorochrome; Funding; Grant; Helium; Hospitals; Housing; Human Resources; Immunophenotyping; Institution; Ions; Laboratories; Lasers; Medicine; Neon; Pathologist; Pathology; Research; Research Personnel; Resources; Safety; Sampling; Sorting - Cell Movement; Source; Speed; System; Time; Training; United States National Institutes of Health; Universal Precautions; Water; college; design; detector; instrument; programs; square foot

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Modular Flow (MoFlo) 高速细胞分选仪（DakoCytomation；科罗拉多州柯林斯堡）设计用于以 70,000 个事件/秒的速度进行分选，纯度为 99%。 MoFlo 配置有三个激光器，能够将细胞分选到微量滴定板中。还包括水冷式 Coherent Innova 90-C 氩离子 350nm（紫外线）激光器、风冷式 Spectraphysicals 氩离子 488nm（蓝色）激光器和 Melles-Griot 633nm（红色）氦氖激光器。该配置提供了检测多达六种颜色的能力，并且软件配置允许使用以下荧光染料同时进行多达四种颜色的免疫表型分析：FITC、PE、PE-Cy-7 和 APC。此外，该仪器还配备了 CyClone 自动细胞沉积装置，可将细胞快速准确地沉积到各种多孔板格式（每孔单细胞到多个细胞）中。 MoFlo 是可公开重新配置的，并且如果我们的研究需求发生变化，还具有额外的扩展功能，可容纳最多四个额外的探测器。作为附加的安全功能，安装了 E-Vac（气溶胶排出系统）来容纳分拣过程中产生的气溶胶。 流式细胞术设施：该设施是罗杰威廉姆斯医院病理学和实验医学部的组成部分。因此，它已获得美国病理学家学会 (CAP) 认证，并且符合 CLIA 88 标准。该设施占地 800 平方英尺，并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。有两名全职技术人员可以提供帮助。 FACSCalibur：FACSCalibur（Becton Dickinson）配备了用于自动采集样品的装载机（FACS 装载机）。它还具有用于分析的 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 细胞分选仪相比，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。