Rapid and high-contrast photothermal microscopy with a novel tunable ZGP source

具有新型可调谐 ZGP 光源的快速高对比度光热显微镜

• 批准号: 10600781
• 负责人: Adam M Hanninen
• 金额: $ 30.56万
• 依托单位: TRESTLE OPTICS LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-05 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10600781
• 关键词: Address; Algae; Back; Bacteria; Biological Assay; Biology; Biopsy; Cell Line; Cell Separation; Cells; Cellular biology; Chemicals; Comparative Study; Complement; Dark-Field Microscope; Data; Development; Diagnostic; Engineering; Fingerprint; Fourier Transform; Germanium; Goals; Health; Health Promotion; Heating; Histopathology; Image; Imaging Techniques; Industrialization; Label; Lasers; Light; Light Microscope; Lipids; Malignant Neoplasms; Mammary Gland Parenchyma; Maps; Medicine; Methods; Microbe; Microfluidic Microchips; Microfluidics; Microscopic; Microscopy; Molecular Probes; Molecular Profiling; Morphology; Natural Products; Noise; Nutraceutical; Optics; Pain; Performance; Phenotype; Physiologic pulse; Physiological; Play; Polyunsaturated Fatty Acids; Population; Predictive Value; Process; Production; Productivity; Protocols documentation; Public Health; Pump; Qualifying; Rapid screening; Refractive Indices; Research; Research Personnel; Resolution; Role; Sampling; Science; Signal Transduction; Source; Specialist; Specimen; Spectrum Analysis; Speed; System; Techniques; Technology; Tissue Microarray; Tissue imaging; Tissues; Visible Radiation; Visualization; Yeasts; Zinc; absorption; biomedical imaging; commercialization; contrast imaging; design; detection limit; detector; experimental study; fundamental research; high standard; imaging approach; imaging modality; improved; infrared microscopy; innovation; lens; light microscopy; long chain fatty acid; novel; prevent; prototype; quantum; screening; submicron; tool

Abstract. Natural products offer excellent sources of health-promoting medicines yet challenges in efficiently and economically sourcing these value-add bio-compounds have prevented their greater availability and promotion to public health. We address this problem by developing a novel image-based screening technology that offers highly sensitive, phenotypic bioassays of metabolite concentrations on live single cells. This allows a greatly improved method for identification of better natural product producers, as well as quantitative information about the production per cell and production distribution across the population. This high-resolution, non- destructive, label-free cell screening and cell sorting system is based on vibrational photothermal microscopy which uses mid-infrared light to probe molecular vibrational absorptions at resolutions comparable to visible light microscopes. To demonstrate the utility of the approach we focus on the production of essential polyunsaturated fatty acids (PUFAs) in lipid droplets sourced from microalgae (thraustochytrids) – a natural producer of PUFAs. These long-chain fatty acids play a vital role in the physiological health of cells and tissues throughout the body are commercialized as a prominent nutraceutical. To develop enriched cell lines, we will engineer and prototype a multichannel microfluidic chip with an active piezoelectric actuator to selectively save cells identified through photothermal as being highly productive. Our goal is to provide an attractive method to non-destructively evaluate metabolite content in engineered microbes including bacteria and yeast, not just algae, and for a variety of high-value metabolites. To accomplish these goals, we have established a team of qualified experts including specialists in advanced microscopy, microfluidics design, and algae cultivation for industrial applications. Ultimately, the methods developed here will showcase a roadmap toward rare-cell discovery and active cell sorting based on chemical mapping allowed through the photothermal imaging contrast.

抽象的。 天然产品为促进健康的药物提供了极好的来源，但在 有效和经济地采购这些增值生物化合物， 更大的可用性和促进公共卫生。我们通过开发一种新的 基于图像的筛选技术，提供高灵敏度的表型生物测定， 活单细胞上的代谢物浓度。这允许一种大大改进的方法， 确定更好的天然产品生产者，以及关于 每个细胞的产量和整个群体的产量分布。这种高分辨率，非 破坏性的，无标记的细胞筛选和细胞分选系统是基于振动光热 使用中红外光探测分子振动的分辨率的显微镜 与可见光显微镜相当。为了证明我们所关注的方法的实用性， 在脂滴中产生必需多不饱和脂肪酸（PUFA）， 微藻（破囊壶菌）-PUFA的天然生产者。这些长链脂肪酸发挥着 在整个身体的细胞和组织的生理健康中的重要作用被商业化 作为一种著名的营养品。为了开发富集的细胞系，我们将设计和制作一种 具有主动压电致动器以选择性地保存细胞的多通道微流体芯片 通过光热技术鉴定为高产。我们的目标是提供一个有吸引力的 一种非破坏性评估工程化微生物中代谢物含量的方法， 细菌和酵母，而不仅仅是藻类，以及各种高价值的代谢产物。完成 为了实现这些目标，我们建立了一支由包括高级专家在内的合格专家团队 显微镜，微流体设计和藻类培养的工业应用。最终 这里开发的方法将展示稀有细胞发现和活性细胞的路线图 通过光热成像对比度允许的基于化学映射的分选。