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（由申请人提供）：该项目的目标是开发一种通用光学成像技术，用于研究活细胞和组织内的重要生物小分子（例如氨基酸、核酸、脂肪酸、葡萄糖、神经递质和药物）为了实现这一目标，我们建议将新兴的受激拉曼散射（SRS）显微镜与三种不同类型的化学键结合起来，该显微镜能够生成生物样品中键的浓度图。具有特征拉曼跃迁的小振动标签，包括炔部分（即 C¿C 三键）、氘同位素和 13C 同位素虽然炔标签通常适用于任何小生物分子，但氘同位素和 13C 同位素对于标记氨基特别有用。当对这些标记有小生物分子的振动标签进行光谱定位时，SRS 显微镜可以发挥作用。非常适合在微观水平上探索生命系统的功能代谢，正如我们最近的出版物所证明的那样，我们已经制定了系统计划，以将这一概念具体化为成熟且通用的技术平台，伴随着技术的发展，一些生物医学应用。正在提出（一些具有令人兴奋的初步数据），包括通过监测炔标记的脱氧核糖核苷代谢掺入新生神经元来对活体脑组织中的神经发生进行成像，通过开发代谢物库的调色板进行多色化学成像，用氘标记的氨基酸监测神经系统中的蛋白质合成，并用 13C 标记的苯丙氨酸探测亨廷顿蛋白聚集过程中的细胞内蛋白质降解。如果成功实施，我们将建立一个新的生物正交非线性拉曼显微镜成像平台，使我们能够探究由此产生的生物正交非线性拉曼显微镜可能具有出色的灵敏度、特异性、生物相容性和多重能力的广谱生物小分子。对小生物分子的作用就像有机染料和荧光蛋白的荧光成像对较大分子种类的作用一样，将小生物分子置于现代光学显微镜的照明下。