Advanced Imaging Core

先进的成像核心

• 批准号: 10701926
• 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/10701926
• 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; Diameter; 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; Macrophage Activation; Metabolic; Metabolism; Methodology; Methods; Molecular; Monitor; Morphologic artifacts; Motion; Mus; Myelogenous; Myeloid Cells; Outcome; Paracrine Communication; Pathway interactions; Pattern; Phase; Phosphorylation; Phosphotransferases; Process; Property; Proteins; Reagent; Recovery; Regulation; Resolution; STAT protein; Signal Pathway; Signal Transduction; Sorting; Source; Substrate Interaction; Surface; Testing; Time; Tissues; Variant; Visual; beta catenin; cellular imaging; imaging approach; imaging study; in vivo; insight; interest; intravital imaging; intravital microscopy; lung imaging; lung injury; lung repair; migration; novel; novel strategies; programs; protein kinase A kinase; respiratory; response; spatiotemporal; success; superresolution microscopy; tool; transcription factor; two-photon; ultra high resolution

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.

摘要 高级成像核心C对于准确定义过程的空间和时间组织至关重要 控制巨噬细胞的可塑性通过流式细胞术测定的表面标志物主要用于 鉴定和分类不同巨噬细胞亚群。然而，每个项目都明确指出，巨噬细胞 多样性从根本上说是特定时空细胞内信号传导的结果， 转录编程由生态位环境决定。这些过程控制巨噬细胞的可塑性 在炎症性肺损伤及其消退期间。所有项目将解决如何细胞间和旁分泌 信号与细胞内信号相互作用以改变巨噬细胞对肺损伤和消退的反应。因此，在本发明中， 核心C的功能是提供先进的试剂和成像方法， 体内研究，并使每个项目的假设测试。我们将提供工具来解决 一个基本的问题是，一个给定的信号通路或转录程序是必不可少的活性 调节巨噬细胞分化、特化和功能。为了获得机械信号的见解， 为了保证每个项目的成功，核心C将提供常规和超分辨率的方法 水平来询问活巨噬细胞中的信号通路。时空代谢，转录，和 ATP、β-catenin、CREB、STAT家族对巨噬细胞活化的信号通路改变 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分辨率显微镜进行研究。 分辨率活动成像。对巨噬细胞的这些分析将允许对时间进行精细的评估- 巨噬细胞变体所采用的程序的序列和空间组织。尽管活体内 成像是询问肺损伤和恢复结果、活体中的呼吸运动 动物是通过体内肺成像获得有意义结果的主要障碍。一个主要的核心 C功能还将提供先进的双光子活体肺成像，使用一种新的方法， 识别活小鼠肺（血管和组织）中的髓样细胞，并跟踪单个细胞的运动， 在肺损伤和修复阶段直接检查其感兴趣途径的功能影响的项目。 通过提供巨噬细胞在分化、迁移时信号传导活性的无缝视觉检查， 和解决免疫挑战，核心C是该计划的整体成功至关重要。