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.

总体概述： NEI 的流式细胞术核心为 NEI 校内社区提供流式细胞术分析和分选设备及服务。它在合作研究项目中利用和开发最先进的样品制备、数据采集和分析以及分类程序。为学生、研究员和主要研究人员提供样品制备、染色和分选后处理方面的培训。评估技术研究需求并建议招聘适当的人员并采购满足这些需求所需的设备。该核心还支持 NEI 之外的 NIH 校内研究，为无法使用流式细胞仪的实验室处理样本，并与科学高级教育基金会合作。 核心仪器仪表老化。这是 FACSAria II 可选择维护合同的最后一年。 NEI 校内研究项目对更新的仪器很感兴趣。新冠危机仍然影响着核心的运行。核心的未来也取决于即将上任的 NEI IRP 科学主任的方向。 核心提供的服务： 今年，来自二十四个不同实验室的五十五个人使用了该设施。这些服务和协作服务是为来自 4 个 NEI 组织（NNRL、LI、DIR、OGVFB）的首席研究员 (PI) 以及 NEI 以外的其他 NIH 研究人员提供的。今年，核心进行了 500 个小时的分选，分析了 10,000 多个样本。 目前核心使用的技术包括活细胞表型分析方法、通过荧光报告基因检测基因表达、监测细胞凋亡或增殖导致的膜和 DNA 含量变化、测量细胞内蛋白质以及定量可溶性蛋白质。 涉及人体组织的工作包括分选外周血单核细胞，以研究其细胞因子的产生、基因型以及 DNA 或 RNA 表达。来源是血液、血沉棕黄层和白细胞。对眼液、眼组织样本、蛋白质和眼泪进行了一些分析工作。美国国家眼科研究所在生物安全方面进行了大量投资，新增了配备完全集成生物安全柜的 BD FACSAria Fusion 流式细胞仪。该分选机满足最近 NIH 的操作员和样品保护要求以及处理人体样品的生物保护全球标准。核心中没有储存任何人体组织。 核心鼓励但不要求用户在其出版物中承认核心的贡献。 训练： 核心向 NEI 社区提供了几次正式培训课程。这些课程包括：流式细胞术简介、先进技术（例如细胞凋亡应用）、分析仪器操作和使用 FloJo 软件进行数据分析。 核心内容花了大约 100 小时。或接受一年的流式细胞术培训。 核心人员为 NEI 社区提供了大约 50 小时的培训，并通过 FAES 为 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