Fluorescence Enhanced Photothermal Infrared Spectroscopy (FE-PTIR) - breakthrough for simultaneous fluorescence microscopy and sub-micron IR spectroscopy

荧光增强光热红外光谱 (FE-PTIR) - 同步荧光显微镜和亚微米红外光谱的突破

• 批准号: 10543927
• 负责人: Craig Prater
• 金额: $ 86.54万
• 依托单位: PHOTOTHERMAL SPECTROSCOPY CORP.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-02 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10543927
• 关键词: Alzheimer' s Disease; Amyloid; Antimicrobial Resistance; Antimicrobial susceptibility; Area; Biochemical; Biocompatible Materials; Biological; Biological Sciences; Biology; Biomedical Engineering; Biomedical Research; Cells; Chemical Structure; Chemicals; Classification; Computer software; Confocal Microscopy; Data; Data Analyses; Dependence; Detection; Development; Disease Resistance; Fluorescence; Fluorescence Microscopy; Glucose; Goals; Heating; Image; In Situ; Individual; Infrared Rays; Institutes; Isotopes; Label; Lasers; Light; Lipids; Malignant Neoplasms; Maps; Measurement; Measures; Metabolism; Microscope; Molecular; Molecular Analysis; Molecular Structure; Multimodal Imaging; Neurodegenerative Disorders; Neurons; Optical Instrument; Pharmaceutical Preparations; Pharmacotherapy; Phase; Photobleaching; Physiologic pulse; Plant Roots; Positioning Attribute; Protein Analysis; Proteins; Publications; Research; Resolution; Sampling; Small Business Innovation Research Grant; Source; Specimen; Spectrum Analysis; Structure; Techniques; Technology; Temperature; Testing; Tissues; United States National Institutes of Health; Visible Radiation; absorption; antimicrobial; biological research; biomedical imaging; brain tissue; cancer cell; commercialization; data acquisition; design; engineering design; fluorescence imaging; fluorophore; glucose metabolism; improved; infrared microscopy; infrared spectroscopy; innovation; insight; interest; microbial; microscopic imaging; multimodality; novel; optical imaging; protein folding; protein misfolding; prototype; response; spectroscopic imaging; submicron; tau Proteins; vibration

1 This Phase II project aims to develop and commercialize Fluorescence Enhanced Photothermal Infrared (FE-PTIR) imaging 2 and spectroscopy. The proposed FE-PTIR will use fluorescence microscopy to map the distribution of fluorescently labeled 3 regions of cells and tissue and then provide chemical structural analysis of the labeled regions using photothermal infrared 4 spectroscopy. Fluorescence microscopy is a cornerstone technique in biological research, allowing sensitive mapping of 5 specifically targeted biomolecules within cells and tissue, but it does not provide information about their molecular 6 structure. Infrared (IR) spectroscopy can provide rich analysis of molecular structure and has been used in life sciences 7 research to study tissue classification, drug/tissue interaction, neurodegenerative diseases, cancer and other areas. 8 Conventional IR spectroscopy, however, has a fundamental spatial resolution limit (i.e. roughly how small an object it can 9 analyze) of around 10 micrometers, similar to the size of an average biological cell. Thus conventional IR spectroscopy has 10 been extremely limited for many biomedical applications where the structures of interest are smaller than a cell. 11 The FE-PTIR technique illuminates a fluorescently labeled sample with UV/visible light which results in fluorescent 12 emission from fluorescently tagged molecules in the sample. A tunable infrared laser source also illuminates the sample, 13 causing localized heating in the sample if the IR laser is tuned to a wavelength that excites molecular vibrations in the 14 sample. Using a camera or other sensitive photodetector is used to record the fluorescent emission from different regions 15 of the sample generates a map of the distribution of fluorescently labeled biomolecules. A key innovation of this proposal 16 was the recognition that common fluorophores have an emission efficiency that is highly temperature dependent. Thus 17 when the sample is also irradiated with infrared light at wavelengths corresponding to molecular vibrations, localized 18 heating from IR absorption by the sample causes a significant change in the fluorescent emission. Recording the emission 19 change as a function of sample position or IR wavelength produces IR absorption images and IR absorption spectra, 20 respectively. Phase I research demonstrated the following key advances: (1) ability of FE-PTIR to map of target 21 biomolecules with fluorescence and analyze the molecular structural of the target molecules; (2) achieve submicron 22 spatial resolution for both fluorescence imaging and infrared spectroscopy; (3) demonstrated a 100X improvement in 23 measurement sensitivity; (4) application of FE-PTIR to study of protein misfolding relevant to neurodegenerative disease 24 research; (5) demonstrated FE-PTIR on individual bacterial and live cancer cells with subcellular resolution. This project is 25 well aligned with NIH goals as it incorporates several key thrusts of the National Institute of Biomedical Imaging and 26 Bioengineering, including optical imaging and spectroscopy, IR imaging, confocal microscopy, and multimodal imaging. FE- 27 PTIR will be extremely useful for example in analyzing the molecular structure/folding/aggregation of fluorescence- 28 localized proteins. Protein misfolding/aggregation is a root cause of many neurodegenerative diseases (e.g. Alzheimer’s). 29 Completion of this Phase II project will lead to the commercialization of a new multimodal microscope that will offer 30 profound benefits for biomedical research including neurodegenerative diseases and antimicrobial resistance research.

1该阶段II项目旨在开发和商业化荧光增强光热红外（FE-PTIR）成像 2和光谱法。拟议的Fe-Ptir将使用荧光显微镜来绘制荧光标记的分布 3个细胞和组织区域，然后使用光热红外进行化学结构分析 4光谱法。荧光显微镜是生物学研究中的基石技术，允许对 5特异性靶向细胞和组织中的生物分子，但不提供有关其分子的信息 6结构。红外（IR）光谱可以提供对分子结构的丰富分析，并已用于生命科学 7研究组织分类，药物/组织相互作用，神经退行性疾病，癌症和其他领域的研究。 8但是，常规的红外光谱法具有基本的空间分辨率极限（即，它可以大约有多小的物体 9分析）约10微米，类似于平均生物细胞的大小。那个常规的红外光谱具有 在许多生物医学应用中，有10个非常有限的，在许多生物医学应用中，感兴趣的结构小于细胞。 11 Fe-Ptir技术用紫外线/可见光照亮了荧光标记的样品，从而导致荧光 样品中荧光标记的分子的12个排放。可调的红外激光源也照亮了样本， 13如果将IR激光调谐到波长，则在样品中引起局部加热。 14个样本。使用摄像头或其他灵敏的光电探测器来记录不同区域的荧光发射 15个样品生成了荧光标记的生物分子的分布图。该提议的关键创新 16是认识到普通荧光团具有高度依赖性的排放效率。那 17当样品还以对应于分子振动对应的波长的红外光线照射时， 18通过样品滥用IR滥用加热会导致荧光发射的显着变化。记录排放 19随着样品位置或IR波长的变化产生IR抽象图像和IR抽象光谱， 分别为20。第一阶段研究证明了以下关键进展：（1）Fe-Ptir能够映射目标的能力 21荧光生物分子并分析靶分子的分子结构； （2）实现亚微米 22荧光成像和红外光谱的空间分辨率； （3）证明了100倍的改善 23测量灵敏度； （4）Fe-PTIR应用于与神经退行性疾病有关的蛋白质错误折叠的研究 24研究； （5）在单个细菌和具有亚细胞分辨率的活细胞上证明了Fe-PTIR。这个项目是 25与NIH目标保持一致，因为它结合了美国国家生物医学成像研究所的几个关键推力和 26生物工程，包括光学成像和光谱，IR成像，共聚焦显微镜和多模式成像。 fe 27 PTIR将非常有用，例如分析荧光 - 分子结构/折叠/聚集 - 28个局部蛋白质。蛋白质不形成/聚集是许多神经退行性疾病（例如阿尔茨海默氏症）的根本原因。 29该阶段项目的完成将导致新的多模式显微镜的商业化，该显微镜将提供 30生物医学研究的深刻益处，包括神经退行性疾病和抗菌耐药性研究。