Molecular Imaging of Biomaterials - Single Cells

生物材料的分子成像 - 单细胞

• 批准号: 8761423
• 负责人: NICHOLAS WINOGRAD
• 金额: $ 46.13万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8761423
• 关键词: 3-Dimensional; Alzheimer' s Disease; Alzheimer' s disease model; Architecture; Biocompatible Materials; Biological; Biological Models; Biological Process; Brain; Cell Communication; Cell Membrane Structures; Cell model; Cell physiology; Cells; Chemicals; Chemistry; Collection; Communication; Communities; Data Collection; Development; Device or Instrument Development; Dimensions; Effectiveness; Event; Exocytosis; Fluorescence Microscopy; Freezing; Gases; Image; Ions; Kinetics; Knowledge; Label; Lasers; Lateral; Left; Lipid Biochemistry; Lipids; Liposomes; Mass Spectrum Analysis; Membrane; Metabolic; Methodology; Methods; Mission; Modeling; Molecular; Molecular Weight; Neurons; Neurotransmitters; PC12 Cells; Pathway interactions; Pharmacology; Play; Property; Proteins; Protocols documentation; Protons; Publishing; Regulation; Research; Research Design; Resolution; Role; Sampling; Shapes; Site; Source; Spatial Distribution; Specificity; Spectrometry, Mass, Secondary Ion; Spottings; Staging; Structure; Surface; System; Testing; Three-Dimensional Imaging; Tissue membrane; Variant; Vesicle; Work; base; biological research; brain tissue; cellular imaging; cryogenics; design; direct application; improved; indexing; instrumentation; ion source; ionization; mass analyzer; mass spectrometer; mast cell; molecular imaging; nanoscale; neurotoxic; neurotransmitter release; new technology; novel; particle; peer; protein aggregation; public health relevance; public health research; research study; skills; small molecule; stable isotope; theories; tool

DESCRIPTION (provided by applicant): This research is aimed to push the boundaries of imaging mass spectrometry with unique biological applications directed toward the chemical characterization of single biological cells. A special emphasis is placed upon probing the lipid distributions and the role of small molecule neurotransmitters embedded in neurotransmitter vesicles primed for exocytosis and brain lipids in Alzheimer's disease. Model systems include liposome cell mimics, mast cells, PC12 cells and brain tissue where membrane structure is changing. The central approach is to utilize an advanced bio-analytical mass spectrometry-based protocol to acquire two and three dimensional molecule-specific image information at the nano-scale level. Transformational mass spectrometry instrumentation for secondary ion mass spectrometry (SIMS) is proposed to complete this mission. Special emphasis is placed upon implementing a novel gas cluster ion beam (GCIB), consisting of an Ar4000+ projectile, which has been shown to desorb molecules without breaking bonds and without chemical damage buildup on the sample. There are four specific aims for this proposal. First, we propose to combine 3- dimensional imaging with a tightly focused C60+ ion source and a GCIB source especially tuned by chemistry to dramatically enhance spatial resolution and ionization efficiency in imaging. This will increase sensitivity and therefore spatial resolution as we push t the limits obtained by dynamic SIMS. Second, the effectiveness of these protocols will be verified using well-characterized liposome particles as stand-ins for biological cells. Third, developments in the first two aims will be implemented to image the substructure of vesicles in single cells in order to gain a better understanding of what regulates chemical communication via exocytosis. These studies will provide fundamentally important information about how neurotransmitter release is regulated for plasticity and pharmacology. Fourth, metabolic labeling with stable isotope-encoded lipid and protein species will be used to elucidate spatial and temporal changes in the molecular architecture of single nerve cells. We will then use this imaging methodology to study protein aggregation in single nerve cells and a model for Alzheimer's disease. The overarching objective is to establish the unique imaging strategy proposed here as a valuable new tool for use by the larger biological community.

描述（由申请人提供）：这项研究旨在通过针对单个生物细胞的化学表征的独特生物学应用来推动成像质谱的边界。特别强调了探测脂质分布和嵌入在阿尔茨海默氏病中胞吐作用和脑脂质的神经递质囊泡中的小分子神经递质的作用。模型系统包括脂质体细胞模拟物，肥大细胞，PC12细胞和脑组织正在发生变化的脑组织。中心方法是利用基于先进的生物分析质谱法方案在纳米级级别获取两个和三维分子特异性图像信息。提出了用于完成此任务的次级离子质谱法（SIMS）的转换质谱仪器。特别重点是实施新的气体簇离子束（GCIB），该束由AR4000+弹丸组成，该弹丸已显示出无需断裂键并且在样品上没有化学损害的情况下解除分子。 该提案有四个具体的目标。首先，我们建议将3维成像与紧密焦点的C60+离子源和GCIB源相结合，尤其是化学调节的GCIB源，以显着提高空间分辨率和成像中的电离效率。这将增加灵敏度，因此随着我们推动动态模拟人生获得的限制的空间分辨率。其次，这些方案的有效性将使用特征良好的脂质体颗粒作为生物细胞的固定剂进行验证。第三，将实施前两个目标的发展，以对单个细胞中囊泡的亚结构进行图像，以便更好地了解通过胞吐作用调节化学通信的方法。这些研究将提供有关如何调节可塑性和药理学的神经递质释放的根本重要信息。第四，具有稳定的同位素编码脂质的代谢标记和蛋白质物种将用于阐明单神经细胞分子结构的空间和时间变化。然后，我们将使用这种成像方法研究单神经细胞中的蛋白质聚集，并研究了阿尔茨海默氏病的模型。总体目标是建立此处提出的独特成像策略，作为较大生物群落使用的宝贵新工具。