Optical imaging of small bio-molecules in living cells and tissues by nonlinear Raman microscopy coupled with vibrational tags

通过非线性拉曼显微镜结合振动标签对活细胞和组织中的小生物分子进行光学成像

• 批准号: 9298651
• 负责人: Wei Min
• 金额: $ 32.14万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9298651
• 关键词: Adopted; Affect; Alkynes; Alpha Cell; Alzheimer' s Disease; Amino Acids; Animals; Biological; Brain Diseases; Carbon; Cells; Characteristics; Chemicals; Color; Coupled; Crystallization; Data; Deoxyribonucleosides; Deuterium; Development; Diffuse; Dyes; Exhibits; Fatty Acids; Glucose; Goals; Health; Hippocampus (Brain); Huntington Disease; Huntington gene; Image; Imaging technology; Inclusion Bodies; Isotopes; Label; Lasers; Libraries; Lighting; Maps; Metabolic; Metabolism; Microscopic; Microscopy; Modernization; Molecular; Monitor; Mus; Nervous system structure; Neurodegenerative Disorders; Neurons; Neurotransmitters; Nucleic Acids; Organism; Parkinson Disease; Pharmaceutical Preparations; Phenotype; Phenylalanine; Physiologic pulse; Protein Biosynthesis; Proteins; Proteome; Protocols documentation; Publications; Sampling; Sensitivity and Specificity; Slice; Source; Study models; Techniques; Tissues; Width; base; biomaterial compatibility; brain tissue; chemical bond; fluorescence imaging; imaging platform; improved; in vivo; insight; instrumentation; light microscopy; mutant; neural stimulation; neurogenesis; novel; optical imaging; protein aggregation; protein degradation; protein metabolism; public health relevance; small molecule libraries; tumor; tumor metabolism; uptake; vibration

 DESCRIPTION (provided by applicant): The goal of the project is to develop a general optical imaging technology for studying vital small bio-molecules (e.g., amino acids, nucleic acids, fatty acids, glucose, neurotransmitters and drugs) inside live cells and tissues, an important but otherwise intractable goal. To do so, we propose to couple the emerging stimulated Raman scattering (SRS) microscopy, which is capable of producing concentration maps of chemical bonds in biological samples, with three distinct classes of small vibrational tags with characteristic Raman transitions, including alkyne moieties (i.e., C¿C triple bond), deuterium isotope and 13C isotope. While alkyne tags are generally applicable to any small biomolecules, deuterium isotope and 13C isotope will be particularly useful for labeling amino acids. When spectrally targeting these vibrational tags labeled to small bio-molecules, SRS microscopy is ideally suited for probing functional metabolism of living systems at microscopic level, as proved in our recent publications. We have laid out systematic plans as to how to crystallize this concept into a mature and general technical platform. Accompanied by the technical development, several biomedical applications are being proposed (some with exciting preliminary data) including imaging neurogenesis in living brain tissues by monitoring the metabolic incorporation of alkyne-tagged deoxyribonucleoside into newly born neurons, multicolor chemical imaging by developing a color palette of metabolite library, monitoring protein synthesis in nervous systems with deuterium-labeled amino acids, and probing intracellular protein degradation during Huntingtin aggregation with 13C-labeled phenylalanine. If successfully implemented, we will establish a new imaging platform of bioorthogonal nonlinear Raman microscopy that could allow us to interrogate a broad spectrum of small bio-molecules with superb sensitivity, specificity, biocompatibility and multiplex ability. The resulting bioorthogonal nonlinear Raman microscopy might do for small bio-molecules what fluorescence imaging of organic dyes and fluorescent proteins has done for larger molecular species, bringing small bio-molecules under the illumination of modern light microscopy.

 描述（由适用提供）：该项目的目的是开发一种通用的光学成像技术，用于研究重要的生物分子（例如，氨基酸，核酸，脂肪酸，葡萄糖，神经递质和药物）在活细胞和组织内部，是一个重要的目标。为此，我们提议将紧急刺激的拉曼散射（SRS）显微镜结合在一起，该显微镜能够在生物学样品中产生化学键的浓度图，并具有三种不同类别的小振动标签，具有特征性的拉曼过渡，包括碱基部分，包括碱基部分（即，C⿣CCTriple Bond），Deuterium Ispope和13c ISOPOPE。虽然炔烃标签通常适用于任何小型生物分子，但同位素和13C同位素对于标记氨基酸特别有用。当将这些标记为小生物分子标记的这些振动标签瞄准这些振动标签时，SRS显微镜非常适合在我们最近的出版物中证明，SRS显微镜在显微镜水平上探测生存系统的功能代谢。我们已经制定了有关如何将这一概念结晶成成熟和一般技术平台的系统计划。伴随着技术开发，提出了几种生物医学应用（一些具有令人兴奋的初步数据），包括通过监测新生出生的神经元的代谢掺入中的代谢性脱氧神经元的代谢成像，通过开发具有多微型化学成像的多体化学成像，以监测Amminio palete of Sermaborab syntripers syntrys syntrys syntrys syntrys syntrys syntrys syntrys protthers syntrys syntry protthers syntrys syners syntry protther，该神经元将其成像，该神经元成像，该神经元成像。并用13C标记的苯丙氨酸探测细胞内蛋白质降解。如果成功实施，我们将建立一个新的成像平台的生物正交非线性拉曼显微镜，该平台可以使我们能够以出色的灵敏度，特异性，生物相容性和多重能力询问广泛的小型生物分子。产生的生物正交非线性拉曼显微镜可能对小生物分子有机染料和荧光蛋白的荧光成像对大分子物种做出了什么作用，从而使小生物分子在现代光学显微镜的照明下。