Advanced Imaging Core

先进的成像核心

• 批准号: 10491055
• 负责人: Gary CH Mo
• 金额: $ 34.97万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491055
• 关键词: Address; Algorithms; Animals; Anti-Inflammatory Agents; Biosensing Techniques; Biosensor; CREB1 gene; Cell Size; Cell surface; Cells; Color; Concentration Camps; Cues; Cyclic AMP; Development; Dissection; Environment; Family; Flow Cytometry; Functional Imaging; Genetic Transcription; Goals; Grain; Heterogeneity; Image; Immune; In Situ; In Vitro; Inflammation; Inflammatory; Injury; Janus kinase; Kinetics; Lung; Macrophage Activation; Metabolic; Metabolism; Methodology; Methods; Microscopy; Molecular; Monitor; Morphologic artifacts; Motion; Mus; Myelogenous; Myeloid Cells; Outcome; Paracrine Communication; Pathway interactions; Pattern; Phase; Phosphotransferases; Process; Property; Proteins; Reagent; Regulation; Resolution; STAT protein; Signal Pathway; Signal Transduction; Source; Substrate Interaction; Surface; Testing; Time; Tissues; Variant; Visual; base; beta catenin; cellular imaging; imaging approach; imaging study; in vivo; injury and repair; injury recovery; insight; interest; intravital imaging; intravital microscopy; lung imaging; lung injury; macrophage; novel; novel strategies; programs; protein kinase A kinase; respiratory; response; spatiotemporal; success; tool; transcription factor; two-photon

Abstract Advanced Imaging Core C is essential to accurately define spatial and temporal organization of processes governing macrophage plasticity. Surface markers determined by flow cytometry have been primarily used to identify and sort diverse macrophage subpopulations. However, each Project makes clear that macrophage diversity is fundamentally a consequence of specific spatiotemporal intracellular signaling and coordinated transcriptional programming dictated by the niche environment. These processes control macrophage plasticity during both inflammatory lung injury and its resolution. All Projects will address how inter-cellular and paracrine signals interact with intracellular cues to alter macrophage fate in response to lung injury and resolution. Thus, the function of Core C is to provide advanced reagents and imaging methodologies required for both in vitro and in vivo studies and enable testing of hypotheses in each Project. We will provide the tools to address the fundamental question that a given signaling pathway or transcriptional program is essential for the active regulation of macrophage differentiation, specialization, and function. To gain the mechanistic signaling insights needed for the success of each Project, Core C will provide methods at both conventional and super-resolution levels to interrogate the signaling pathways in live macrophages. Spatiotemporal metabolic, transcriptional, and signaling landscape changes in response to macrophage activation by ATP, β-catenin, CREB, STAT family transcription factors and other key signals will be studied by multi-color super-resolution microscopy and super- resolution activity imaging. These analyses of macrophages will permit a fine-grained assessment of the time- sequence and spatial organization of the programs employed by macrophage variants. Although intravital imaging is the most direct way to interrogate lung injury and recovery outcomes, respiratory motion in the live animal represents a major obstacle for obtaining meaningful results by in vivo lung imaging. Thus, a major Core C function will also be to provide advanced two-photon intravital lung imaging using a novel approach to reliably identify myeloid cells in living mouse lung (in vessels and tissue) and track single cell motion, allowing each Project to directly examine the functional effects of their pathway of interest during lung injury and repair phases. By providing a seamless visual examination of signaling activity in macrophages as they differentiate, migrate, and resolve immune challenges, Core C is essential for the Program’s overall success.

抽象的 高级成像核心C对于准确定义流程的空间和临时组织至关重要 管理巨噬细胞塑性。由流式细胞仪确定的表面标记已主要用于 识别和分类多种巨噬细胞亚群。但是，每个项目都清楚地表明巨噬细胞 多样性从根本上是特定空间颞内信号传导和协调的结果 转录编程由利基环境决定。这些过程控制巨噬细胞的塑性 在炎症性肺损伤及其分辨率期间。所有项目都将解决细胞间和旁分泌 信号与细胞内提示相互作用，以响应肺损伤和分辨率改变巨噬细胞命运。那， Core C的功能是提供体外和体外所需的高级试剂和成像方法 体内研究并启用每个项目中的假设。我们将提供解决问题的工具 给定的信号通路或转录程序对于活动的基本问题是必不可少的 调节巨噬细胞分化，专业化和功能。获得机械信号洞察 每个项目的成功所需 询问活巨噬细胞中信号通路的水平。时空代谢，转录和 信号传导景观响应于ATP，β-catenin，creb，stat家族激活巨噬细胞的变化 转录因子和其他关键信号将通过多色超分辨率显微镜和超级 - 分辨率活动成像。这些巨噬细胞的分析将允许对时间进行细粒度评估。 巨噬细胞变体携带的程序的序列和空间组织。虽然是插入的 成像是询问肺损伤和恢复结果的最直接方法，现场呼吸运动 动物代表了通过体内肺成像获得有意义结果的主要障碍。那是主要核心 C功能还将是使用一种新颖的方法来提供高级的两光弹药弹式肺成像 鉴定活小鼠肺（在血管和组织中）中的髓样细胞并跟踪单细胞运动，允许每个细胞 项目以直接检查其在肺损伤和修复阶段期间感兴趣途径的功能效应。 通过对巨噬细胞中的信号传导活性进行无缝的视觉检查，它们在区分，迁移时， 并解决免疫挑战，核心C对于该计划的整体成功至关重要。