Nonlinear optical microscopy for ultrastructural bioimaging

用于超微结构生物成像的非线性光学显微镜

• 批准号: RGPIN-2017-06923
• 负责人: Barzda, Virginijus
• 金额: $ 4.08万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=712503
• 关键词: Nonlinear; optical; microscopy; ultrastructural; bioimaging

Advancement of nonlinear optical microscopy technology and application for biological imaging is proposed in this Discovery program. Novel optical microscopy modalities will be developed for ultrastructural investigations of ordered molecular structures within each focal volume of the imaged structure, beyond the diffraction limited spatial resolution. Nonlinear quantum microscopy modalities based on the interactions of entangled-photons with the sample will be developed. The microscopic imaging will also be employed using classical ultrashort pulsed lasers for obtaining vibrational sum-frequency generation and stimulated Raman scattering image contrasts that provide chemical specificity for imaging based on the selected molecular vibrations. Polarimetric microscopy measurements will be employed using nonlinear Stokes-Mueller formalism recently developed by our group, to investigate ultrastructural organization of biological samples. The ultrastructural microscopy will be employed to study collagen organization in the extracellular matrix of biological tissue, myosin nanomotor organization in muscle cells and synchronization of nanomotor activity during muscle contraction. The ultrastructure of starch granules, crystalline cellulose and photosynthetic pigment aggregates will also be investigated. Ordered molecular assemblies of proteins, polysaccharides and pigments play important structural and functional role in the living organisms. For example, the organization of collagen fibers in the extracellular matrix determines optical and mechanical properties of the tissue, myosin nanomotors assembled into myofilaments in the striated muscles cyclically change their conformation to produce contractions for movement of the organisms, starch granules are organized in intricate crystalline structures for efficient storage and release of carbohydrates, and ordered pigment aggregates are used for efficient solar energy collection. Therefore, it is important to develop non-invasive, chemically-specific and label-free imaging tools to visualize in vivo biological structures and reveal the nanoscopic ultrastructural organization of the molecular assemblies. The structural information is important for biophysics, biomedical and plant biology researchers, pathologists, and bio-nanotechnology investigators. The ultrastructural information about the organization of specific molecular groups in biological tissue will validate the structural models drawn from previous nonlinear microscopy studies. The novel techniques will stimulate the development of new in vivo tissue imaging tools, provide new methods to study muscular disorders, and facilitate investigations in solar energy conversion and storage in plants, with applications to artificial photosynthesis, solar cell development and biomimetic nanotechnology.

在此发现程序中提出了非线性光学显微镜技术的进步和生物成像的应用。将开发出新的光学显微镜模式，以对成像结构的每个焦点体积内有序分子结构进行超微结构研究，超出衍射有限的空间分辨率。将开发基于纠缠量与样品的相互作用的非线性量子显微镜模态。微观成像还将使用经典的超短脉冲激光器来获得振动总和频率产生和刺激拉曼散射图像对比度，这些对比度基于所选分子振动为成像提供化学特异性。我们小组最近开发的非线性Stokes-Mueller形式主义将采用偏光显微镜测量，以研究生物样品的超微结构组织。超微结构显微镜将用于研究生物组织外基质，肌肉细胞中肌球蛋白纳米运动组织的胶原蛋白组织，并在肌肉收缩过程中纳米运动活性同步。还将研究淀粉颗粒，结晶纤维素和光合色素骨料的超微结构。蛋白质，多糖和色素的有序分子组件在生物体中起重要的结构和功能作用。例如，细胞外基质中胶原蛋白纤维的组织决定组织的光学和机械性能，肌球蛋白纳米运动体在横纹肌肉中组装成肌膜中的肌球蛋白纳米运动，循环地改变其构象以产生宫缩，以产生缩水，以使生物体运动，淀粉颗粒在杂质的结晶中组织起来有效储存和释放碳水化合物和有序的色素骨料的结构用于有效的太阳能收集。因此，重要的是开发非侵入性，化学特异性和无标签的成像工具来可视化体内生物学结构并揭示分子组件的纳米镜头超微结构组织。该结构信息对于生物物理学，生物医学和植物生物学研究人员，病理学家和生物 - 纳米技术研究者很重要。有关生物组织中特定分子基团的组织的超微结构信息将验证从以前的非线性显微镜研究中得出的结构模型。这种新型技术将刺激新的体内组织成像工具的发展，提供研究肌肉疾病的新方法，并促进植物中太阳能转化和储存的研究，并适用于人工光合作用，太阳能细胞发育和生物模拟纳米技术。