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.

项目摘要 在原位观察代谢活动对于理解许多生物过程是至关重要的。它 是执行基因蓝图和维持生命的新陈代谢。糖，如 葡萄糖是人类和动物必不可少的营养物质，不仅提供能量，而且 也是合成新生物质的基石，如蛋白质、脂肪、RNA/DNA和 糖原。我们提出的研究是开发和优化新一代氢化物 葡萄糖([D]-葡萄糖)三维体积复合受激拉曼散射(SRS)研究 成像平台。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，以可视化代谢动力学和异质性 动物。新合成的分子中的碳-氢化学键 由于C-D标记的蛋白质、脂类、DNA/RNA、碳水化合物会在“细胞中沉默” 拉曼光谱的“区域”。此外，根据代谢物的不同光谱模式 来自不同的同位素，代谢物的合成将在多个时间点 在一张图像中可视化。在AIM II中，新一代增强组织清除配方 将针对大规模3D体积成像进行筛选和设计(穿透深度增加20倍) [D]-葡萄糖在动物模型中的代谢活性和异质性。在《目标3》中， 将建立定制设计的光谱分解算法和协议，以最大限度地 检测20多个新合成的C-D标记分子的潜力和最小化 背景信号。我们的计算资源和算法将优化众多 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 动态化和异质性。利用资源和算法，我们将优化一个平台 解开多个新老合成分子并原位定量解剖 有害的东西。拟议研究的完成将可能揭示机械学的基本原理 在不同器官中与葡萄糖代谢相关的多个分子通路。