Flow Cytometry CORE

流式细胞术核心

• 批准号: 10266926
• 负责人: Rachel R. Caspi
• 金额: $ 56.45万
• 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/10266926
• 关键词: 2 year old; Age; Aging; Apoptosis; Blood; Budgets; COVID-19 pandemic; Cells; Collaborations; Communities; Computer software; Cytometry; DNA; DNA Methylation; Data Analyses; Detection; Education; Energy Metabolism; Equipment; Eye; Flow Cytometry; Foundations; Gene Expression; Genetic Transcription; Genotype; Grant; Hour; Human; Information Technology; Interleukins; Intramural N.I.H. Research Support; Investments; Laboratories; Leukocytes; Liquid substance; Measurement; Membrane; Methods; Monitor; National Eye Institute; National Institute of Allergy and Infectious Disease; Peripheral Blood Mononuclear Cell; Phenotype; Ploidies; Postdoctoral Fellow; Preparation; Principal Investigator; Procedures; Production; Proteins; Protocols documentation; Publications; Publishing; Reporter Genes; Research; Research Personnel; Research Project Grants; Resources; Sampling; Science; Secure; Services; Sorting - Cell Movement; Source; Specimen; Stains; Students; Supervision; Techniques; Tissues; Training; United States National Institutes of Health; Work; autoimmune uveitis; coronavirus disease; cytokine; data acquisition; equipment acquisition; exosome; genome-wide; human tissue; instrument; instrumentation; neuroinflammation; operation; recruit; retinal rods; square foot; volunteer; 2 year old; Age; Aging; Apoptosis; Blood; Budgets; COVID-19 pandemic; Cells; Collaborations; Communities; Computer software; Cytometry; DNA; DNA Methylation; Data Analyses; Detection; Education; Energy Metabolism; Equipment; Eye; Flow Cytometry; Foundations; Gene Expression; Genetic Transcription; Genotype; Grant; Hour; Human; Information Technology; Interleukins; Intramural N.I.H. Research Support; Investments; Laboratories; Leukocytes; Liquid substance; Measurement; Membrane; Methods; Monitor; National Eye Institute; National Institute of Allergy and Infectious Disease; Peripheral Blood Mononuclear Cell; Phenotype; Ploidies; Postdoctoral Fellow; Preparation; Principal Investigator; Procedures; Production; Proteins; Protocols documentation; Publications; Publishing; Reporter Genes; Research; Research Personnel; Research Project Grants; Resources; Sampling; Science; Secure; Services; Sorting - Cell Movement; Source; Specimen; Stains; Students; Supervision; Techniques; Tissues; Training; United States National Institutes of Health; Work; autoimmune uveitis; coronavirus disease; cytokine; data acquisition; equipment acquisition; exosome; genome-wide; human tissue; instrument; instrumentation; neuroinflammation; operation; recruit; retinal rods; 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 work of the cytometry core was tremendously impacted by the emergence of the COVID-19 pandemic. The laboratory was secured and closed in March 2020. In May 2020, the core partially restarted for a Covid related project. In August, the biologist in the core moved to NIAID. As of September, users of the core have not been able to restart their regular operations. SERVICES PROVIDED THE CORE: This year, fifty-fivindividuals from twentyfour different laboratories used the facility. These services and collaborative services were performed for 10 Principal Investigators (PIs) from 5 NEI labs (ERPD, LI, LRCMB, MSF, N-NRL, RN, OGVFB and UNGIRD), plus 15 other laboratories at NIH. This year the core performed 750 hours of sorting and over 9,000 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. The following are examples of the publications that used NEI Flow Cytometry Core resources: Corso-Daz X, Gentry J, Rebernick R, et al. Genome-wide Profiling Identifies DNA Methylation Signatures of Aging in Rod Photoreceptors Associated with Alterations in Energy Metabolism. Cell Rep. 2020;31(3):107525. doi:10.1016/j.celrep.2020.107525 Kang M, Choi JK, Jittayasothorn Y, Egwuagu CE. Interleukin 35-Producing Exosomes Suppress Neuroinflammation and Autoimmune Uveitis. Front Immunol. 2020;11:1051. Published 2020 May 29. doi:10.3389/fimmu.2020.01051 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。 COVID-19大流行的出现极大地影响了细胞仪核的工作。该实验室于2020年3月确保并关闭。2020年5月，核心部分重新启动了 相关项目。八月，核心的生物学家搬到了Niaid。截至9月，核心用户无法重新启动其常规操作。 服务提供了核心： 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来自5个NEI Labs（ERPD，LI，LRCMB，MSF，N-NRL，RN，RN，OGVFB和UNGIRD）的10位首席研究人员（PIS）进行的，以及NIH的其他15个实验室。今年，核心进行了750小时的分类，并分析了9,000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。以下是使用NEI流式细胞仪核心资源的出版物的示例： Corso-Daz X，Gentry J，Rebernick R等。全基因组的分析鉴定了与能量代谢改变有关的杆感光体衰老的DNA甲基化特征。 CellRep。2020; 31（3）：107525。 doi：10.1016/j.celrep.2020.107525 Kang M，Choi JK，Jittayasothorn Y，Egwuagu CE。白介素35产生的外泌体抑制神经炎症和自身免疫性葡萄膜炎。前免疫。 2020; 11：1051。出版于2020年5月29日。doi：10.3389/fimmu.2020.01051 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。