Flow Cytometry

流式细胞术

• 批准号: 7695941
• 负责人: MICHAEL ANDREEFF
• 金额: $ 35.96万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-28 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7695941
• 关键词: 5-(6)-carboxyfluorescein diacetate succinimidyl ester; Antigens; Apoptosis; Arts; BIRC4 gene; Biological Assay; Bromodeoxyuridine; Cancer Center; Cancer Center Support Grant; Cancer Immunology Science; Caspase; Cell division; Cells; Clinical Research; Communities; Confocal Microscopy; Core Facility; Cyclins; DNA; Detection; Development; Employee; Environment; Event; Faculty; Flow Cytometry; Fluorescence-Activated Cell Sorting; Funding; Future; Galactosidase; Goals; Grant; Green Fluorescent Proteins; Growth Factor; Hour; Image Analysis; Immunology; Laboratories; Laser Microscopy; Laser Scanning Cytometry; Lasers; Life; Malignant - descriptor; Membrane Potentials; Methods; Microscopy; Mission; Mitochondria; Molecular; Molecular Cytogenetics; Molecular and Cellular Biology; Multiparametric Analysis; NCI Center for Cancer Research; NGFR Protein; Nerve Growth Factor Receptors; Normal Cell; Numbers; Operative Surgical Procedures; PCNA gene; Paper; Peer Review; Peer Review Grants; Phenotype; Principal Investigator; Proteins; Publishing; Recording of previous events; Regulation; Reproduction spores; Research; Research Activity; Research Personnel; Residual Neoplasm; Scientist; Services; Signal Transduction; Site; Sorting - Cell Movement; Stem cell transplant; Stem cells; System; TP53 gene; Techniques; Technology; Time; Travel; cancer cell; cancer immunology function; cancer prevention; cell analyzer; cellular imaging; digital; fluorescence activated cell sorter device; instrument; leukemia; malignant breast neoplasm; melanoma; member; peer; progenitor; programs; single cell analysis; transgene expression; tumorigenesis

The Flow Cytometry and Cellular Imaging Core Facility (FCCICF) provides cellular analysis to investigators with peer-reviewed grants. The FCCICF has two sites, on the north campus and on the recently-opened south campus. It occupies 1900 sq. ft. and is directed by Dr. Michael Andreeff. The FCCICF develops and provides techniques for single-cell analysis. Cell phenotyping, proliferation, signaling and apoptosis assays have been established and modified for multiparametric analysis. Immunophenotypic analysis was combined with assays of intracellular proteins related to apoptosis (Bcl-2, Bcl-XL, BAG-1, p53, Rb, caspase activation, mitochondria! membrane potential and Fas), cell signaling (MARK), proliferation (Ki67, cyclins, BrDU, PCNA, DNA) and cell division history (CFSE). Quantitation of cellular antigens allows determination of the antibodybinding capacity per cell. Rare events and progenitor/stem cell subpopulations can be detected and isolated by three-laser excitation/eight-parameter fluorescence-activated cell sorting (FACS) for subsequent analysis by molecular cytogenetic and other molecular techniques including quantitation of intracellular PKCoc, Bax, Bcl-2, ERK, pERK, XIAP by laser scanning cytometry. FISH has been combined with apoptosis assays to discriminate apoptosis in normal and malignant cells. The number, phenotype and proliferation of minimal residual disease cells can be determined at levels of one malignant in 30,000 normal cells. Methods for detection of transgene expression in cells are in place using p-galactosidase (p-gal), nerve growth factor receptor (NGF-R), and green fluorescent protein (EGFP). Acquisition of 3 new FACS Aria cell sorters and a four-laser flow cytometer (B&D LSRII) has upgraded the facility to provide state-of-the-art isolation and analysis. Laser confocal microscopy has been used extensively and was upgraded by acquisition of an Olympus FV-500 multi-user instrument and a DSU spinning disc confocal system. High-impact studies in cancer prevention, growth factor signaling in breast cancer, apoptosis regulation in leukemias and multistep tumorigenesis all utilized confocal microscopy. The FCCICF now utilizes 17 major instrument systems. The Core supports numerous R01, P01, R21, and SPORE grants. Since the previous review, the number of users has increased 145% (169 investigators with peer-reviewed grants from 19 different programs). 67% of users have peer-reviewed funding. During the previous funding period, 11,668 hours of service was provided. Use has tripled to greater than 7,000 hours estimated for 2007. Future plans are focused on the continued development and application of cutting-edge technologies in cell analysis.

流式细胞术和细胞成像核心设施（FCCICF）为研究者提供细胞分析 同行评审的赠款FCCICF有两个网站，在北校区和最近开放的 南校区。占地1900平方米。英尺由Michael Andreeff博士指导。FCCICF开发和 提供单细胞分析技术。细胞表型、增殖、信号传导和凋亡测定 已建立和修改的多参数分析。结合免疫表型分析 用与凋亡相关的细胞内蛋白（Bcl-2，Bcl-XL，BAG-1，p53，Rb，半胱天冬酶激活， 线粒体！膜电位和Fas），细胞信号传导（MARK），增殖（Ki 67，细胞周期蛋白，BrDU，PCNA， DNA）和细胞分裂史（CFSE）。细胞抗原的定量允许确定抗体结合 每个电池的容量。可以检测和分离罕见事件和祖细胞/干细胞亚群 通过三激光激发/八参数荧光激活细胞分选（FACS）进行后续分析 通过分子细胞遗传学和其他分子技术，包括定量细胞内PKCoc，Bax， 激光扫描细胞仪检测Bcl-2、ERK、pERK、XIAP。FISH已与细胞凋亡测定相结合， 区分正常和恶性细胞的凋亡。微小型的数量、表型和增殖 残留的疾病细胞可以被确定为30，000个正常细胞中有一个恶性细胞的水平。方法 使用β-半乳糖苷酶（p-gal）、神经生长因子 受体（NGF-R）和绿色荧光蛋白（EGFP）。购置3台新的FACS Aria细胞分选仪和1台 四激光流式细胞仪（B&D LSRII）已经升级了设备，以提供最先进的分离和 分析.激光共聚焦显微镜已被广泛使用，并通过收购一个 Olympus FV-500多用户仪器和DSU旋转盘共焦系统。高影响力研究 癌症预防，乳腺癌中的生长因子信号传导，白血病中的凋亡调节和多步骤 肿瘤发生都使用共聚焦显微镜。FCCICF现在使用17个主要仪器系统。的 Core支持众多R 01、P01、R21和SPORE赠款。自上次审查以来， 用户增加了145%（169名研究人员，来自19个不同的项目，获得了同行评审的赠款）。67%的 用户有同行评审的资金。在上一个供资期间， 提供了估计2007年的使用量增加了两倍，超过7 000小时。未来计划的重点是 继续开发和应用细胞分析的尖端技术。