Flow Cytometry CORE

流式细胞术核心

• 批准号: 10930585
• 负责人: Rachel R. Caspi
• 金额: $ 75.87万
• 依托单位: NATIONAL EYE INSTITUTE
• 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/10930585
• 关键词: 6 year old; Affect; Age; Aging; Apoptosis; Blood; Budgets; COVID-19 pandemic; Cells; Collaborations; Communities; Computer software; Contracts; DNA; Data Analyses; Detection; Education; Equipment; Eye; Flow Cytometry; Foundations; Future; Gene Expression; Genetic Transcription; Genotype; Grant; Hour; Human; Information Technology; Intramural N.I.H. Research Support; Intramural Research Program; Investments; Laboratories; Leukocytes; Liquid substance; Maintenance; Measurement; Membrane; Methods; Monitor; National Eye Institute; Peripheral Blood Mononuclear Cell; Phenotype; Ploidies; Postdoctoral Fellow; Preparation; Principal Investigator; Procedures; Production; Proliferating; Proteins; Protocols documentation; Publications; Recommendation; Reporter Genes; Research; Research Personnel; Research Project Grants; Sampling; Science; Services; Sorting; Source; Specimen; Stains; Students; Techniques; Tissues; Training; United States National Institutes of Health; Work; cytate; cytokine; data acquisition; equipment acquisition; human tissue; instrument; instrumentation; operation; recruit; square foot; volunteer

GENERAL OVERVIEW: The Flow Cytometry Core at NEI provides flow cytometry analytical and sorting equipment and services to the NEI Intramural community. It utilizes and develops state-of-the-art sample preparation, data acquisition and analysis, and sorting procedures in collaborative research projects. Provides training to students, fellows, and principal investigators on sample preparation, staining, and post-sort handling. Assesses technical research needs and recommends recruitment of the appropriate staff and acquisition of the equipment needed to meet those needs. The core also supports NIH intramural research outside NEI by processing samples for laboratories without access to flow cytometry instrumentation and collaborations with the Foundation for Advanced Education in the Sciences. The instrumentation of in the core is aging. This is the last year that the FACSAria II will have an option for a maintenance contract. There is newer instrumentation that is interesting to the NEI Intramural Research Program. The COVID19 crisis still affects the operation of the core. The future of the core is also pending the direction of the incoming Scientific Director for the NEI IRP. SERVICES PROVIDED BY THE CORE: This year, fifty-fivindividuals from twentyfour different laboratories used the facility. These services and collaborative services were performed for Principal Investigators (PIs) from 4 NEI organizations (NNRL, LI, DIR, OGVFB), plus other NIH investigators outside NEI. This year the core performed 500 hours of sorting and over 10,000 samples were analyzed. Among the techniques now in use within the core are methods for phenotyping live cells, gene expression detection through fluorescent reporter genes, monitoring membrane and DNA content changes due to apoptosis or proliferation, measurement of intracellular proteins and quantification of soluble proteins. The work involving human tissues includes the sorting of peripheral blood mononuclear cells to study their cytokine production, genotype and DNA or RNA expression. The sources are blood, buffy coat, and white cells. Some analytical work had been done with eye fluids, eye tissue specimen, protein, and tears. The National Eye Institute made a great investment in biosafety with the addition BD FACSAria Fusion flow cytometer equipped with a fully integrated biosafety cabinet. This sorter meets the recent NIH's operator and sample protection requirements as well as global standards for bioprotection for processing human samples. No human tissues were stored by the core. The Core encourages, but does not require, users to acknowledge the Core contribution in their publications. TRAINING: Several formal training sessions were offered by the Core to the NEI community. These courses included: Introduction to Flow Cytometry, Advanced Techniques (Ex. Apoptosis applications), Analytical Instrument Operation and Data Analysis with FloJo Software. The Core stuff received about 100 hrs. or training a year in flow cytometry. The Core stuff provided about 50 hrs of training to the NEI community and 60 hrs of training through FAES to the NIH community.

一般概述： NEI的流式细胞仪核心为NEI室内社区提供流式细胞仪分析和分类设备和服务。它利用并开发了最新的样本准备，数据获取和分析以及协作研究项目中的分类程序。向学生，研究员和主要调查人员提供有关样本准备，染色和后处理后处理的培训。评估技术研究需求，并建议招募适当的员工，并收购满足这些需求所需的设备。该核心还通过处理实验室的样本，而无需获得流式细胞仪仪器和与科学高级教育基金会的合作，从而支持NEI外的NIH内部研究。 核心中的仪器正在衰老。这是Facsaria II可以选择维护合同的去年。 NEI壁内研究计划有新的仪器。 COVID19危机仍然影响核心的运行。核心的未来还在等待NEI IRP即将上任的科学总监的方向。 核心提供的服务： 今年，来自二十四个不同实验室的五十三个个人使用了该设施。这些服务和协作服务是为4个NEI组织（NNRL，Li，Dir，Dir，OGVFB）的首席研究人员（PIS）以及NEI以外的其他NIH调查人员进行的。今年，分析了核心进行500小时的分类和10,000多个样品。 在核心中使用的技术中，在核心表型的方法中，通过荧光报告基因的基因表达检测，监测膜和DNA含量因细胞凋亡或增殖而导致的DNA含量变化，测量细胞内蛋白质以及可溶性蛋白质的定量。 涉及人体组织的工作包括分类外周血单核细胞，以研究其细胞因子产生，基因型和DNA或RNA表达。来源是血液，巴菲大衣和白细胞。一些分析工作是用眼睛流体，眼组织标本，蛋白质和眼泪进行的。国家眼科研究所（National Eye Institute）通过配备完全集成的生物安全柜的BD Facsaria Facusion流式细胞仪进行了巨大的投资。该分类器符合最近的NIH的运营商和样品保护要求，以及用于处理人类样品的全球生物保护标准。没有人体组织由核心存储。 核心鼓励但不需要用户承认其出版物中的核心贡献。 训练： 核心向NEI社区提供了几次正式的培训课程。这些课程包括：流式细胞仪概论，高级技术（例如，凋亡应用），分析仪器操作和FLOJO软件的数据分析。 核心的东西大约100小时。或培训一年的流式细胞仪。 核心内容为NEI社区提供了约50小时的培训，并通过FAES向NIH社区提供了60小时的培训。

项目成果

Unlike Th1, Th17 cells mediate sustained autoimmune inflammation and are highly resistant to restimulation-induced cell death.

• DOI: 10.4049/jimmunol.0900519
• 发表时间: 2009-12-01
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Shi G;Ramaswamy M;Vistica BP;Cox CA;Tan C;Wawrousek EF;Siegel RM;Gery I
• 通讯作者: Gery I

Triphasic developmentally guided protocol to generate retinal pigment epithelium from induced pluripotent stem cells.

• DOI: 10.1016/j.xpro.2022.101582
• 发表时间: 2022-09-16
• 期刊: STAR PROTOCOLS
• 作者: Sharma, Ruchi;Bose, Devika;Montford, Jair;Ortolan, Davide;Bharti, Kapil
• 通讯作者: Bharti, Kapil

Gene expression changes in aging retinal microglia: relationship to microglial support functions and regulation of activation.

• DOI: 10.1016/j.neurobiolaging.2013.03.022
• 发表时间: 2013-10
• 期刊: Neurobiology of aging
• 影响因子: 4.2
• 作者: Ma W;Cojocaru R;Gotoh N;Gieser L;Villasmil R;Cogliati T;Swaroop A;Wong WT
• 通讯作者: Wong WT