Sugar Probed SRS Volumetric imaging of Metabolic Activities

代谢活动的糖探针 SRS 体积成像

• 批准号: 10639208
• 负责人: Lingyan Shi
• 金额: $ 39.41万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-21 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639208
• 关键词: 3-Dimensional; Adipose tissue; Aging; Algorithm Design; Algorithmic Analysis; Algorithms; Animal Model; Animals; Biological Process; Biomass; Brain; Carbohydrates; Carbon; Cells; Chemicals; Computational algorithm; Computing Methodologies; Custom; DNA; Data; Data Analyses; Data Set; Deuterium; Development; Fingerprint; Generations; Genetic; Glucose; Glycogen; Goals; Heterogeneity; Homeostasis; Human; Image; Imaging Techniques; In Situ; Label; Life; Lipids; Liver; Measures; Metabolic; Metabolic Pathway; Metabolism; Methods; Microscopy; Molecular; Mus; Nutrient; Optics; Organ; Pancreas; Pathway interactions; Pattern; Penetration; Play; Printing; Proteins; Protocols documentation; Publishing; RNA; Radiation; Recipe; Regression Analysis; Reporting; Resolution; Resources; Role; Signal Transduction; Source; Spatial Distribution; Stimulus; Techniques; Technology; Three-Dimensional Imaging; Time; Tissues; Urea; Visualization; algorithmic methodologies; chemical bond; computing resources; design; glucose metabolism; imaging platform; imaging probe; in vivo; innovation; macromolecule; metabolic abnormality assessment; metabolic imaging; multiplexed imaging; response; sugar; three-dimensional visualization; tool

Project Summary Visualizing metabolic activities in situ is critical to understanding many biological processes. It is the metabolism which carries out the genetic blue print and maintain life. Sugar such as glucose is an essential nutrient for human being and animals, not only provides energy, but also is building blocks for synthesizing new biomass such as protein, lipid, RNA/DNA, and glycogen. Our proposed study is to develop and optimize a new generation of deuterated glucose ([D]-glucose) probed 3D volumetric multiplex stimulated Raman scattering (SRS) imaging platform. In aim I, various types of [D]-glucose including [D1], [D2], [D5], [D7], and [D12]- glucose will be studied and optimized for visualizing metabolic dynamics and heterogeneity in animals. The carbon-deuterium (C-D) chemical bond in the newly synthesized molecules such as C-D labeled proteins, lipids, DNA/RNA, carbohydrates will be imaged in the “cell silent region” of Raman spectra. In addition, based on diverse spectral patterns of metabolites derived from different isotopologues, metabolite synthesis in multiple time points will be visualized in a single image. In Aim II, a new generation of enhanced tissue clearing recipe will be screened and designed for large scale 3D volumetric imaging (20x deeper penetration) of [D]-glucose probed metabolic activities and heterogeneity in animal models. In Aim III, custom designed spectral unmixing algorithm and protocol will be established to maximize the potential of detecting more than 20 newly synthesized C-D labeled molecules and minimize the background signal. Our computational resources and algorithms will optimize numerous variables in complicated hyperspectral imaging datasets for studying organ specific metabolic dynamics and heterogeneity. Using the resources and algorithms, we will optimize a platform to unmix multiple old and newly synthesized molecules and quantitatively dissect the in situ detritions. Completion of the proposed study will potentially reveal mechanistic fundamentals of multiple molecular pathways related to glucose metabolism in different organs.

项目概要 原位可视化代谢活动对于理解许多生物过程至关重要。 是执行遗传蓝图并维持生命的新陈代谢，例如糖。 葡萄糖是人类和动物必需的营养素，不仅提供能量，而且 也是合成新生物质的基础材料，如蛋白质、脂质、RNA/DNA 和 我们提出的研究是开发和优化新一代氘代糖原。 葡萄糖（[D]-葡萄糖）探测 3D 体积多重受激拉曼散射 (SRS) 在目标 I 中，各种类型的 [D]-葡萄糖，包括 [D1]、[D2]、[D5]、[D7] 和 [D12]- 将研究和优化葡萄糖，以可视化代谢动态和异质性 新合成的分子中的碳-氘（C-D）化学键，例如 因为 C-D 标记的蛋白质、脂质、DNA/RNA、碳水化合物将在“细胞沉默”中成像 此外，基于代谢物的不同光谱模式。 衍生自不同的同位素体，多个时间点的代谢物合成将 在 Aim II 中，新一代增强的组织清除配方被可视化。 将针对大规模 3D 体积成像进行筛选和设计（穿透深度 20 倍） [D]-葡萄糖在动物模型中探测代谢活动和异质性。 将建立定制设计的光谱分解算法和协议，以最大化 检测 20 多种新合成的 C-D 标记分子的潜力，并最大限度地减少 我们的计算资源和算法将优化许多背景信号。 用于研究器官特异性代谢的复杂高光谱成像数据集中的变量 利用资源和算法，我们将优化平台。 分解多个旧的和新合成的分子并定量剖析原位 完成拟议的研究将有可能揭示机械原理。 不同器官中与葡萄糖代谢相关的多个分子途径。