Cell Sorting

细胞分选

基本信息

批准号：
8335545
负责人：
MICHAEL V. SEIDEN
金额：
$ 2.33万
依托单位：
FOX CHASE CANCER CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-21 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8335545
关键词：
Advisory Committees Archives Bioinformatics Biomedical Research Biometry Blood Cells Calendar Cancer Center Support Grant Cell Cycle Cell Line Cell Separation Cell surface Cells Clone Cells Collaborations Color Comprehensive Cancer Center Computer software Computers Data Deposition Detection Development Discrimination Experimental Designs Flow Cytometry Fluorescence Fluorochrome Fostering Fox Chase Cancer Center Foxes Funding Host Defense Hour Human Resources Immune Immunofluorescence Immunologic Immunophenotyping Instruction Laboratories Lasers Measurement Parents Peer Review Ploidies Policies Population Preparation Reagent Recruitment Activity Research Research Personnel Resources Sampling Sorting - Cell Movement Speed Stream Suspension substance Suspensions System Variant base cell analyzer data management design digital instrument light scattering operation programs release of sequestered calcium ion into cytoplasm software development tool tumor

项目摘要

PROJECT SUMMARY (See instructions): The Cell Sorting Facility (CSF) offers Fox Chase Cancer Center (FCCC) investigators the capability of analyzing single cell suspensions by up to 14 independent measurements, 12 of which are based on immunofluorescence and two on light scattering. In calendar year 2009 the Facility was used by 31 peer-reviewed, funded investigators across four of the five CCSG Research Programs. The large majority of Facility usage (98.5%) was by peer-reviewed, funded investigators. Major applications of flow cytometry include discrimination and isolation (sorting) of cell subpopulations (at up to 20,000 cells/sec), isolation of rare variant (1/10'') cell subsets, characterization of tumors or cell lines, and single cell cloning of cell subpopulations by direct deposition into 96-well plates. The current sorter provides high-speed 4-stream sorting and digital detection. A UV-capable laser enables measurement of calcium flux and viable cell DNA content. In addition to the 3-laser 12-color cell sorter (FACS-VantageSE/DiVa), the Facility also includes an analysis-only 10-color LSR-II and a 3-color FACScan. In 2009 the Center provided funds ($240,000) for a new Bay Biosystems JSAN desktop cell sorter. The user-operated JSAN is capable of up to 8-color 2-stream sorting and utilizes 3-beam excitation with flow-cell detection, including a UV laser. The Facility aids users with advice on experimental design (including availability of reagents and appropriate fluorochromes), sample preparation, and data interpretation. Software developed with the FCCC Biostatistics and Bioinformatics Facility (BBF), the Flow-LIMS, is used for data annotation, organization, and archiving. Analysis is done on investigators' laboratory computers using a highly interactive, graphical software program, FlowJo. Hours of usage, 2,778/year (currently estimated at over 64% of theoretical capacity), has increased 34% since the last review and is projected to increase over the next five years with planned recruits across all five Programs, Hardy (Immune Cell Development and Host Defense (ICDHD) Program), has directed this resource since 1987 and has responsibility for Facility operations. Equitable access to the Facility is assured by Facility policy, oversight from a Facility Advisory Committee (FAC) and a governing Facilities Parent Oversight Committee (FPOC).

项目摘要（参见说明）：细胞分选设施 (CSF) 为福克斯蔡斯癌症中心 (FCCC) 的研究人员提供了通过多达 14 项独立测量来分析单细胞悬浮液的能力，其中 12 项基于免疫荧光，两项基于光散射。 2009 日历年，该设施被 5 个 CCSG 研究计划中的 4 个的 31 名经过同行评审、资助的研究人员使用。大的大多数设施的使用（98.5%）是由经过同行评审、资助的研究人员进行的。流式细胞术的主要应用包括细胞亚群的辨别和分离（分选）（速度高达 20,000 个细胞/秒）、稀有变异 (1/10'') 细胞亚群的分离、肿瘤或细胞系的表征以及单细胞克隆通过直接沉积到 96 孔板中来分析细胞亚群。目前的分拣机提供高速4流分选和数字检测。具有紫外线功能的激光器可以测量钙通量和活细胞 DNA 含量。除了 3 激光 12 色细胞分选仪 (FACS-VantageSE/DiVa) 外，该设施还包括仅供分析的 10 色 LSR-II 和 3 色 FACScan。 2009 年，该中心为新型 Bay Biosystems JSAN 台式细胞分选机提供资金（240,000 美元）。用户操作的 JSAN 能够进行多达 8 色 2 流分选，并利用 3 光束激发和流动池检测，包括 UV 激光器。该设施为用户提供实验设计（包括试剂和适当荧光染料的可用性）、样品制备和数据解释方面的建议。使用 FCCC 生物统计和生物信息学设施 (BBF) 开发的软件 Flow-LIMS，用于数据注释、组织和分析归档。分析是在研究人员的实验室计算机上使用高度交互的图形软件程序 FlowJo 进行的。使用小时数为 2,778 小时/年（目前估计超过理论容量的 64%），自上次审查以来增加了 34%，预计在未来五年内将增加，计划在所有五个项目、Hardy（免疫细胞开发）中进行招募和主机防御 (ICDHD) 计划），自 1987 年以来一直领导该资源并负责设施运营。设施政策、设施咨询委员会 (FAC) 和管理设施家长监督委员会 (FPOC) 的监督确保公平使用设施。