Signaling Core

总结 单细胞团细胞术最近已经导致大大增强的高维、单细胞、定量的流式细胞术。 分析细胞群上和细胞群内的生物活性分子。抗体识别表面标记， 描述细胞类型或细胞内信号分子或转录因子， 功能如磷酸信号传导、细胞凋亡、DNA损伤、代谢和细胞周期。通过测量所有 这些参数的同时，可以在“网络”处测量单个小区的信令状态。 水平”。虽然在这个深度测量单细胞是高度信息化的，但基因组和蛋白质组分析表明， 原位组织化仍然是一个重要的目标。核心已经开发了多路离子束成像 （MIBI）是一种目前能够同时分析多达45个目标的技术， 标准福尔马林固定、石蜡包埋（FFPE）组织标本，以及最常见的样本类型 在世界各地的临床储存库中。MIBI平台具有独特的能力， 单分子检测以及实体组织的单细胞3D数据可视化。核心已经延伸 这项工作的创建CODEX-一个基于荧光的高通量系统能够渲染50-100 参数在3-6小时（可扩展到数百个参数）。CODEX平台几乎可以将任何 一个荧光范围为~ 10，000美元的高维成像设备-一个关键的实用程序，这个U19计划。 鉴于MIBI和CODEX的能力，并认识到国际生物医学和 在免疫学、诊断学和药物开发的成像应用方面的制药兴趣--这一核心 将扩展组织和细胞的CODEX和MIBI深度表型分析的当前特征。新 将开发用于特征提取和与临床状态相关的算法。核心将配置文件， 在大多数情况下多达50个参数（在其他情况下更多），免疫和组织细胞结构的变化， 由遗传和病原体引起的扰动引起。从这个前所未有的深度提取的特征 将提供数据以理解发生的大规模和微小的组织改变 在单细胞水平绘制“组织组学”图谱。对这些数据的分析将在 生物信息学核心。这项工作所支持的数据集将提供定量的鲁棒性， 这个计划的基础。更一般地说，这项工作将建立新的平台， 这是对免疫功能和发病机制的前所未有的看法，可以在未来的工作中很容易地适应， 在全球范围内了解免疫功能。

Summary Single cell mass cytometry has recently led to greatly enhanced high-dimensional, single-cell, quantitative analysis of bioactive molecules on and within cell populations. The antibodies recognize surface markers that delineate cell types or intracellular signaling molecules or transcription factors that demarcate multiple cell functions such as phospho-signaling, apoptosis, DNA damage, metabolism, and cell cycle. By measuring all these parameters simultaneously, the signaling state of an individual cell can be measured at the “network level”. Though measuring single cells at this depth is highly informative, genomic and proteomic profiling of tissue organization in situ remains an important goal. The Core has developed Multiplexed-ion Beam Imaging (MIBI) a technology currently capable of analyzing up to 45 targets simultaneously and which is compatible with standard formalin-fixed, paraffin-embedded (FFPE) tissue specimens, and the most common sample type in clinical repositories worldwide. The MIBI platform has the unique ability to provide extraordinarily sensitive single molecule detection as well as single cell 3D data visualization for solid tissues. The Core has extended this work to the creation of CODEX—a fluorescence based high throughput system capable rendering 50-100 parameters in 3-6 hours (scalable to hundreds of parameters). The CODEX platform converts nearly any fluorescence scope for ~ $10,000 into a high dimensional imaging device – a key utility for this U19 program. Given the capabilities of MIBI and CODEX--and recognizing a growing international biomedical and pharmaceutical interest in imaging applications to immunology, diagnostics, and drug development-- this Core will extend the current features of CODEX and MIBI deep phenotypic profiling of tissues and cells. New algorithms will be developed for feature extraction and correlation to clinical status. The Core will profile, with up to 50 parameters in most cases (and more in others), the immune and tissue cell architectural changes that arise from genetic and pathogen-induced perturbations. Features extracted from this unprecedentedly deep data will be provided for understanding of wholesale and minor tissue alterations that occur—enabling a first ever map of “tissue-omics” at the single cell level. Analysis of these data will be expanded upon in the Bioinformatics Core. The datasets enabled by this work will provide the quantitative robustness that is a foundation of this program. More generally, this work will establish novel platforms for gaining an unprecedented view into immune function and pathogenesis that could be easily adapted in future work to understand immune function at a global level.