Ultrahigh-resolution and single-molecule stimulated Raman scattering (SRS) microscopy

超高分辨率单分子受激拉曼散射 (SRS) 显微镜

基本信息

批准号：
10377375
负责人：
Wei Min
金额：
$ 31.78万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10377375
关键词：
Address Adopted Algorithms Alkynes Amino Acids Amplifiers Area Basic Science Beds Biological Biophotonics Blinking Cancer Biology Carbon Cells Chemical Structure Chemicals Color Computational algorithm Coupling Crystallization Detection Development Dyes Fluorescence Fluorescence Microscopy Generations Glucose Goals Home Image Immunology Label Lasers Lipids Medical Research Methods Microscope Microscopy Molecular Target Neurosciences Optics Pharmaceutical Preparations Property Pump Radial Reporting Resolution Sampling Scheme Specificity Speed System Techniques Technology Testing Tubulin base bioimaging biological research biomaterial compatibility biomedical imaging complex biological systems design detector diffraction of light disease diagnostic imaging platform improved instrument instrumentation interdisciplinary approach invention molecular scale multiplexed imaging next generation novel precision medicine second harmonic single molecule vibration

项目摘要

Project summary Super-resolution optical microscopy promises to revolutionize biological imaging, as it enables non- invasive interrogation at molecular scale. Indeed, the emergence of super-resolution fluorescence microscopy has quickly impacted the way biologists study cells and subcellular phenomenon. However, super-resolution fluorescence microscopy has fundamental limitations due to the use the fluorescence as contrast mechanism. In particular, it has three major limitations: (1) it cannot reveal chemical composition of the sample; (2) it cannot interrogate small biomolecules due to the relatively bulky fluorescent tags; (3) it cannot image a large number of targets due to the color barrier (only 2~5 fluorescent colors can be practically resolved). The goal of this project is to develop a novel super-resolution imaging platform by exploiting stimulated Raman scattering (SRS) as the contrast mechanism. During the past 10 years since its invention in 2008, SRS microscopy has made widespread impact in biomedical imaging. Being a chemically sensitive method, SRS is well known for its label-free chemical analysis in a quantitative manner. With the recent development of tiny bio-orthogonal tags such as alkynes, SRS has been proven successful in interrogating a wide spectrum of small biomolecules such as lipids, glucose, amino acids, and drugs. Very recently, novel vibrational dye palettes with fine spectral resolution have been reported to achieve super-multiplex electronic pre-resonance (epr) SRS imaging of more than 20 targets simultaneously. Importantly, all these utilizes of SRS microscopy is limited by light diffraction. With SRS being a perfectly complementary contrast mechanism to the prevalent fluorescence, the current proposal aims to develop the necessary methods to bring SRS microscopy to the realm of super resolution. (1) How to improve the resolution for general chemical imaging and small biomolecule imaging; (2) how to break the diffraction limit of the super-multiplex epr-SRS imaging; (3) how to develop the matching vibrational dyes for single molecule SRS. Towards these goals, we had laid out a systematic plan as to how to crystallize this concept into a powerful technology platform. An inter-disciplinary approach has been planned out including instrumentation development, computational imaging, and novel probes synthesis. In Aim 1, we will develop and build new instrumentation. In Aim 2, we will explore new computational algorithm. In Aim 3, we will design next- generation vibrational probes. If successfully implemented, we will establish a transformative platform. The resulting super-resolution chemical imaging would find wide applications in systematically unraveling complex biological systems such as neuroscience, immunology and cancer biology for basic research, disease diagnostics and precision medicine. 1

项目摘要超分辨率光学显微镜有望彻底改变生物成像，因为它使得非 - 分子尺度上的侵入性询问。确实，超分辨率荧光显微镜的出现迅速影响了生物学家研究细胞和亚细胞现象的方式。然而，由于使用，超分辨率荧光显微镜具有基本的局限性荧光作为对比机制。特别是它具有三个主要局限性：（1）它无法透露化学样品的组成；（2）由于相对较大的荧光，它无法询问小的生物分子标签；（3）由于颜色屏障，它无法成像大量目标（只有2〜5荧光颜色可以是实际上解决了）。该项目的目的是通过利用刺激来开发一个新颖的超分辨率成像平台拉曼散射（SRS）作为对比机制。自2008年发明以来的过去10年中，SRS 显微镜已在生物医学成像中广泛影响。作为化学敏感的方法，SRS是以定量方式以其无标记化学分析而闻名。随着微小的最新发展 SRS等生物正交标签（如炔烃）已被证明在审问广泛的方面已被证明是成功的小的生物分子，例如脂质，葡萄糖，氨基酸和药物。最近，新颖的振动染料据报道，具有良好光谱分辨率的调色板可以实现超单型电子呼声（EPR）同时进行20多个目标的SRS成像。重要的是，所有这些利用SRS显微镜是受光衍射的限制。 SRS是与普遍的荧光的完全互补的对比机制，则当前的建议旨在开发必要的方法将SRS显微镜带入超级领域解决。（1）如何改善一般化学成像和小生物分子成像的分辨率；（2）如何打破超级型EPR-SRS成像的衍射极限；（3）如何发展匹配单分子SR的振动染料。朝向这些目标，我们制定了一个系统的计划，即如何将此概念结晶成一个强大的技术平台。已经计划了一种跨学科的方法开发，计算成像和新型探针合成。在AIM 1中，我们将开发并建立新的仪器。在AIM 2中，我们将探索新的计算算法。在AIM 3中，我们将设计接下来 - 一代振动探针。如果成功实施，我们将建立一个变革性平台。这由此产生的超分辨率化学成像将在系统揭开复合物中找到广泛的应用基础研究，疾病的神经科学，免疫学和癌症生物学等生物系统诊断和精度医学。 1