Single Cell and Immunoanalysis Core

单细胞和免疫分析核心

• 批准号: 10455094
• 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/10455094
• 关键词: ATAC-seq; Antibodies; Biological Models; Biology; Biometry; Biopsy; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; 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; Ships; Skin; Stains; T cell receptor repertoire sequencing; T-Cell Receptor Genes; T-Lymphocyte; Techniques; Time; Tissues; Transgenic Mice; Transgenic Model; XCL1 gene; advanced analytics; antibody conjugate; base; cellular imaging; cost; image reconstruction; insight; mouse model; next generation; service utilization; 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 对人类皮肤生物学和炎症性皮肤病的研究由于缺乏转基因技术而进展缓慢。 模型系统和有限的样本，可以从活的病人。先进的分析 已经开发出从小样本中提供最大信息的技术， 来增进我们对人类皮肤病的了解。这个核心使关键的突破性技术 包括单细胞RNA测序（scRNA-seq），能够批量处理样品，发现TCR基因， 用抗体标记细胞（CITE-seq），以及单细胞ATAC-seq，通过流式细胞术的单细胞成像， 飞行时间流式细胞术（CyTOF）和高通量T细胞受体测序可供研究人员在 任何希望进行人类皮肤病研究的机构。该中心还提供援助， 实验计划，关键试剂和数据分析的每项技术的援助。比如说 中心提供协议，允许皮肤活检从遥远的地方运送和可行的冷冻保存 在scRNA-seq研究之前;这一进展使研究人员可以使用单细胞分析来研究样本 从临床试验和未充分研究的皮肤病中。该中心还提供CyTOF染色方案， CyTOF抗体的验证面板，CyTOF抗体的定制缀合，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， CyTOF研究。该中心在实验设计、DNA提取、 协议，DNA样品制备服务，运输和数据分析援助，将使 研究人员成功地利用高通量TCR测序来分析皮肤免疫反应， 在多个时间点和组织中追踪致病性T细胞克隆。我们已经包括了36个研究项目 希望利用本核心服务的调查人员。总而言之，该核心提供了对单个 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