Advanced Imaging Core

先进的成像核心

• 批准号: 10170861
• 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/10170861
• 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 Advanced Imaging Core C 对于准确定义流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记物主要用于 识别和分类不同的巨噬细胞亚群。然而，每个项目都明确表明巨噬细胞 多样性从根本上来说是特定时空细胞内信号传导和协调的结果 由利基环境决定的转录编程。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过程中。所有项目都将讨论细胞间和旁分泌如何 信号与细胞内信号相互作用，改变巨噬细胞的命运，以响应肺损伤和消退。因此， Core C的功能是提供体外和体外实验所需的先进试剂和成像方法。 体内研究并能够测试每个项目中的假设。我们将提供工具来解决 一个基本问题是，给定的信号通路或转录程序对于活性物质至关重要。 巨噬细胞分化、特化和功能的调节。获得机制信号洞察 为了每个项目的成功，Core C 将提供传统和超分辨率的方法 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-连环蛋白、CREB、STAT 家族激活巨噬细胞后信号格局发生变化 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 分辨率活动成像。这些对巨噬细胞的分析将允许对时间进行细粒度的评估。 巨噬细胞变体所使用的程序的序列和空间组织。虽然在活体内 成像是询问肺损伤和恢复结果、实时呼吸运动的最直接方法 动物是通过体内肺部成像获得有意义的结果的主要障碍。因此，一个主要的核心 C 的功能还将是使用一种可靠的新方法提供先进的双光子活体肺部成像 识别活体小鼠肺（血管和组织中）中的骨髓细胞并跟踪单细胞运动，使每个 项目旨在直接检查其感兴趣的通路在肺损伤和修复阶段的功能影响。 通过对巨噬细胞分化、迁移时的信号活动进行无缝视觉检查， 并解决免疫挑战，核心 C 对于该计划的整体成功至关重要。