Core D - BSL3/Flow Cytometry Core

核心 D - BSL3/流式细胞术核心

• 批准号: 8453384
• 负责人: JANICE E CLEMENTS
• 金额: $ 18.9万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8453384
• 关键词: Aliquot; Animals; Archives; Astrocytes; Autopsy; Biological Assay; Blood Cells; Body Fluids; Brain; Cell Line; Cell Separation; Cells; DNA; Data; Data Analyses; Databases; Documentation; Ensure; Equipment; Equipment and supply inventories; Flow Cytometry; HIV; Highly Active Antiretroviral Therapy; In Vitro; Laboratories; Lymphocyte; Macaca; Measurement; Microglia; Modeling; Monitor; Mononuclear; Peripheral; Plasma; Preparation; Primary Cell Cultures; Process; Property; RNA; Reagent; Reporting; Reproducibility; Research Project Grants; SIV; Sampling; Specimen; Spleen; Techniques; Time; Tissues; Viral; Viral Load result; Virus; Work; Workplace; in vivo; lymph nodes; macrophage; programs; research study; wasting

The SIV Biosafety Level 3 (BSL3) / Flow Cytometry Core Laboratory will be directed by Dr. M. Christine Zink. The BSL3 technical staff will be responsible for processing all samples from the infected and uninfected control macaques throughout the experiments described in this Program and will provide samples and reagents to all projects in the program and perform measurement of markers on cells by flow cytometry (FACS). This core laboratory will fulfill the following Specific Aims: 1. To provide a safe work environment to perform work with infectious materials (ALL) 2. To provide support in the preparation and quantitation of virus stocks (ALL) 3. To receive, process, aliquot and log into appropriate databases all samples from macaques (ALL) 4. To isolate PBMCs, pDCs and mononuclear cells from blood (and lymph nodes, spleen and brain at necropsy) from animals for quantitation and longitudinal characterization of cells (ALL) 5. To isolate macrophages/microglia, astrocytes and lymphocytes from the brains of all infected and uninfected macaques for analysis. (Project 2 and 3) 6. To aliquot and archive plasma and CSF samples for viral load and other assays. (ALL) 7. Organizing, monitoring, and maintaining a physical inventory of specimens derived from all experiments {in vivo and in vitro), along with associated documentation and analytical reports (ALL) 8. To prepare samples for Real Time PCR (extraction of RNA or DNA). (ALL) 9. To establish primary cell cultures, and maintain cell lines. (ALL) 10. To perform, FACS experiments, analyze data and submit data to the database (ALL) 11. To property dispose of all the waste generated within the BL3 laboratory (ALL) 12. Maintain all equipment central to the function of the core (ALL)

SIV生物安全级别3（BSL3） /流式细胞仪核心实验室将由M. Christine Zink博士执导。 BSL3技术人员将负责在本计划中描述的整个实验中处理来自感染和未感染的对照猕猴的所有样品，并将为程序中的所有项目提供样品和试剂，并通过流式细胞仪对细胞上的标记进行测量。 （FACS）。 该核心实验室将实现以下特定目标： 1。提供一个安全的工作环境来使用传染性材料（所有）进行工作 2。为病毒库存的准备和定量提供支持（所有） 3。接收，处理，等分并登录适当的数据库 4。从动物中分离出血液（和淋巴结，脾脏和大脑）的PBMC，PDC和单核细胞与动物的血液（淋巴结，脾脏和大脑），以定量和纵向表征细胞（所有） 5。为了分离巨噬细胞/小胶质细胞，来自所有感染和未感染的猕猴的大脑的星形胶质细胞和淋巴细胞进行分析。 （项目2和3） 6。到等分等分等离子体和CSF样品进行病毒载荷和其他测定法。 （全部） 7。组织，监测和维持从所有实验（体内和体外）得出的标本的物理清单，以及相关的文档和分析报告（全部） 8。为实时PCR准备样品（RNA或DNA的提取）。 （全部） 9。建立原代细胞培养并维持细胞系。 （全部） 10。要执行，FACS实验，分析数据并将数据提交到数据库（全部） 11。财产处置BL3实验室中产生的所有废物（全部） 12.维护核心功能（全部）功能的所有设备