Cytometry & Cell Sorting Core

细胞计数法

• 批准号: 9918400
• 负责人: Remi J Creusot
• 金额: $ 14.9万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918400
• 关键词: Address; Adipose tissue; Alpha Cell; Beta Cell; Budgets; Cell Separation; Cell surface; Cells; Chemicals; Chimeric Proteins; Complex; Cytometry; Data; Development; Diabetes Mellitus; Disease model; Dyes; Equipment; Fees; Flow Cytometry; Fluorescence-Activated Cell Sorting; Funding; Gene Expression Profiling; Genetic Transcription; Grant; Hepatocyte; Hour; Housing; Human; Immune; Immunologics; Immunology; In Vitro; Individual; Insulin-Dependent Diabetes Mellitus; Investments; Lipids; Lysosomes; Measures; Medicine; Metabolic; Mitochondria; Molecular; Monitor; Names; National Institute of Diabetes and Digestive and Kidney Diseases; Neurons; Pancreas; Pathway interactions; Phenotype; Phosphorylation; Ploidies; Population; Procedures; Proteins; Publications; Quality Control; Reproducibility; Research; Research Personnel; Resolution; Resources; Role; Services; Signal Transduction; Signaling Molecule; Stains; T-Cell Activation; Techniques; Technology; Therapeutic; Tissues; United States National Institutes of Health; Universities; animal tissue; antibody conjugate; base; cell type; cost; cytokine; design; experience; experimental study; fluorescence imaging; genetic profiling; human tissue; humanized mouse; implantation; in vivo; instrumentation; islet; member; migration; mouse model; new technology; programs; protein purification; recruit; release of sequestered calcium ion into cytoplasm; stem cells; tool; transcription factor

Flow cytometry is an important tool in quantifying and phenotyping the cells that define pathways relevant to diabetes. Previously limited to the analysis of circulating immune cells, advances in cell extraction from tissues has allowed flow cytometry to expand its capabilities to the study of developmental, metabolic and signaling programs in a wide-variety of cell types. The Cytometry & Cell Sorting Core (CCSC) has established itself as a vital and growing component of the Columbia University Diabetes Research Center (DRC), as it addressed the research needs of a growing number of DRC investigators who study immunological, developmental, and metabolic aspects of diabetes. Created in 2010 in partnership with the Columbia Center for Translational Immunology (CCTI) and the Department of Medicine (DOM) with a substantial University investment as well as NIH support, in the past 5 years CCSC has assisted 22 DRC users currently funded by 55 NIH grants, including 30 NIDDK grants. The data generated with the CCSC contributed to 63 publications and 17 new grants. The implementation of CCSC has been essential to support many aspects of diabetes research at Columbia. Since its establishment, the demand for CCSC services has steadily increased, from 60 hours per week with two analyzers to over 130 hours per week with three analyzers, and from 25 hours per week with one sorter to over 60 hours per week with two sorters. We expect usage to continue to grow as new recruits join the DRC, and established investigators take advantage of resources and technologies offered by the Core. Overall, CCSC will assist investigators with the following two aims: Aim 1 – To quantify and phenotypically characterize cell populations that contribute to the metabolic, immunologic and developmental programs of diabetes and its complications. CCSC leverages advanced technologies in fluorescent imaging at the single cell resolution to support the analysis of many different cell types in both human and animal tissues, as well as humanized mouse models of Type 1 diabetes. Aim 2 – To purify populations of cells of relevance to diabetes and its complications. Using fluorescence-activated cell sorting, CCSC supports DRC investigators to purify individual populations of cells for in vitro culture, in vivo implantation or molecular characterization including genetic and transcriptional profiling, protein purification and signaling studies, and for functional analysis including T cell activation, proliferation and migration studies. To achieve these aims, the CCSC has established standard operating procedures to: (i) Assure quality control and reproducibility; (ii) Prioritize investigator use; (iii) Monitor Core use; and (iv) Adapt to new technologies and to the needs of the Columbia Research Base.

流式细胞仪是量化和表型的重要工具，该细胞定义了与 糖尿病。以前仅限于分析循环免疫细胞，从组织中提取细胞的进展 允许流式细胞仪扩展其能力，以研究发育，代谢和信号传导 各种细胞类型的程序。细胞仪和细胞分类核心（CCSC）已将自己确立为 哥伦比亚大学糖尿病研究中心（DRC）的重要组成部分，因为它解决了 研究免疫，发育和 糖尿病的代谢方面。与哥伦比亚转化中心合作创建于2010年 免疫学（CCTI）和医学系（DOM）具有大量的大学投资以及 NIH的支持，在过去的5年中 包括30个NIDDK赠款。 CCSC生成的数据贡献了63个出版物和17个 新赠款。 CCSC的实施对于支持糖尿病研究的许多方面至关重要 哥伦比亚。自成立以来，对CCSC服务的需求已悄悄地增加，从60小时 一周，两名分析仪每周超过130小时，三个分析仪，每周25小时 每周有两个分类器每周一次超过60个小时。我们预计随着新兵的使用率将继续增长 加入刚果民主共和国，成熟的调查人员利用核心提供的资源和技术。 总体而言，CCSC将协助调查人员以下两个目标：目标1 - 量化和表型 表征有助于代谢，免疫和发育的细胞群体 糖尿病计划及其并发症。 CCSC利用荧光的高级技术 在单细胞分辨率下成像，以支持人类和人类中许多不同细胞类型的分析 动物组织以及1型糖尿病的人源化小鼠模型。目标2 - 净化人口 与糖尿病及其并发症相关的细胞。使用荧光激活的细胞分选，CCSC 支持DRC研究人员净化细胞的个体种群，以进行体外培养，体内植入或 分子表征，包括遗传和转录分析，蛋白质纯化和信号传导 研究，以及用于功能分析，包括T细胞激活，增殖和迁移研究。实现 这些目的，CCSC已建立了标准操作程序：（i）确保质量控制和 可重复性； （ii）优先使用研究者使用； （iii）监视核心使用； （iv）适应新技术和 哥伦比亚研究基础的需求。