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: 10693270
• 负责人: Craig Prater
• 金额: $ 86.15万
• 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/10693270
• 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; High temperature of physical object; Image; In Situ; Individual; Infrared Rays; 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; 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; meter; microbial; 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.

本二期项目旨在开发和商业化荧光增强光热红外（FE-PTIR）成像技术

项目成果

Optical Photothermal Infrared Microspectroscopy with Simultaneous Raman - A New Non-Contact Failure Analysis Technique for Identification of <10 μm Organic Contamination in the Hard Drive and other Electronics Industries.

• DOI: 10.1017/s1551929520000917
• 发表时间: 2020-05
• 期刊: Microscopy today
• 作者: Kansiz M;Prater C;Dillon E;Lo M;Anderson J;Marcott C;Demissie A;Chen Y;Kunkel G
• 通讯作者: Kunkel G

Analysis of Fixed and Live Single Cells Using Optical Photothermal Infrared with Concomitant Raman Spectroscopy.

• DOI: 10.1021/acs.analchem.0c04846
• 发表时间: 2021-03-02
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Spadea A;Denbigh J;Lawrence MJ;Kansiz M;Gardner P
• 通讯作者: Gardner P