Single Cell and Immunoanalysis Core

单细胞和免疫分析核心

• 批准号: 10615219
• 负责人: JAMES A. LEDERER
• 金额: $ 14.11万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-19 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615219
• 关键词: ATAC-seq; Acceleration; Antibodies; Biological Models; Biology; Biometry; Biopsy; Cells; Clinical Trials; Clone Cells; Cryopreservation; Custom; Cytometry; DNA; Data Analyses; Distant; Experimental Designs; Flow Cytometry; Geographic Locations; Human; Human Biology; Image; Immune; Immune response; Immunobiology; Inflammatory; Institution; Location; Pathogenicity; Patients; Preparation; Proteins; Protocols documentation; Rare Earth Metals; Reagent; Research; Research Design; Research Personnel; Research Project Grants; Resources; Sampling; Sampling Studies; Services; Shipping; Skin; Stains; T cell receptor repertoire sequencing; T-Cell Receptor Genes; T-Lymphocyte; Techniques; Time; Tissues; Transgenic Mice; Transgenic Model; advanced analytics; antibody conjugate; cellular imaging; cost; image reconstruction; insight; mouse model; next generation; service utilization; single cell sequencing; single-cell RNA sequencing; skin disorder

Project Summary/Abstract: The Single Cell and Immunoanalysis Core Research into human skin biology and inflammatory skin diseases has been slowed by the lack of transgenic model systems and the limited samples that can be obtained from living patients. However, advanced analytic techniques that provide maximal information from small samples have been developed that have the potential to advance our understanding of human skin disease. This Core makes critical groundbreaking techniques including single cell RNA sequencing (scRNA-seq) with the ability to batch samples, discover TCR genes and tag cells with antibodies (CITE-seq), as well as single cell ATAC-seq, single cell imaging by flow cytometry, cytometry by time of flight (CyTOF) and high throughput T cell receptor sequencing available to investigators at any institution who wish to carry out human skin disease research. The Center also provides assistance with experimental planning, key reagents and assistance with data analysis for each technique. For example, the Center provide protocols that allow skin biopsies to be shipped from distant locations and viably cryopreserved before scRNA-seq studies; this advance makes single cell profiling available to investigators to study samples from clinical trials and from understudied skin diseases. The Center also provides CyTOF staining protocols, validated panels of CyTOF antibodies, custom conjugation of CyTOF antibodies, advice on experimental design and data analysis, and all services needed for outside investigators to send their cells to the Center for CyTOF studies. The Center provides comprehensive support in the experimental design, DNA extraction protocols, DNA sample preparation services, shipping, and data analysis assistance that will enable researchers to successfully utilize high throughput TCR sequencing to profile skin immune responses and track pathogenic T cell clones across multiple timepoints and tissues. We have included 36 research projects from investigators who wish to utilize the services of this Core. In summary, this core provides access to single cell approaches that have the potential to provide unprecedented insights into the biology of human skin disease.

项目摘要/摘要：单细胞和免疫分析核心 缺乏转基因的人类皮肤生物学和炎症性皮肤疾病的研究已减慢 模型系统和可以从活着的患者获得的有限样品。但是，高级分析 已经开发了从小样本中提供最大信息的技术 促进我们对人皮肤病的理解。这个核心是批判性的开创性技术 包括具有分类样品的能力，发现TCR基因和 带有抗体（Cite-Seq）的标记细胞以及单细胞ATAC-SEQ，流式细胞仪的单细胞成像， 通过飞行时间（Cytof）和高吞吐量T细胞受体测序的细胞仪 任何希望进行人类皮肤病研究的机构。该中心还提供帮助 实验计划，关键试剂和每种技术的数据分析的帮助。例如， 中心提供可以从遥远的位置运送皮肤活检并进行冷冻保存的协议 在SCRNA-SEQ研究之前；此进步使研究人员可以使用单细胞分析来研究样品 来自临床试验和研究的皮肤疾病。该中心还提供染色方案， 经过验证的细胞抗体面板，定制抗体的定制结合，实验建议 设计和数据分析以及外部调查人员将其细胞发送到中心所需的所有服务 细胞研究。该中心在实验设计中提供了全面的支持，DNA提取 协议，DNA样本准备服务，运输和数据分析协助，将启用 研究人员成功利用高吞吐量TCR测序到剖面皮肤免疫反应和 在多个时间点和组织中跟踪致病性T细胞克隆。我们包括36个研究项目 来自希望利用此核心服务的调查人员。总而言之，此核心提供了对单个的访问 细胞方法有可能为人类皮肤生物学提供前所未有的见解 疾病。