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.

本发现计划提出了非线性光学显微技术和生物成像应用的进展。将开发新的光学显微镜模式，用于成像结构的每个焦点体积内的有序分子结构的超微结构调查，超出衍射限制的空间分辨率。将开发基于纠缠光子与样品相互作用的非线性量子显微镜模式。还将使用经典超短脉冲激光器来采用显微成像，以获得振动和频产生和受激拉曼散射图像对比度，所述图像对比度基于所选分子振动为成像提供化学特异性。偏振显微镜测量将采用非线性斯托克斯-米勒形式主义最近开发的我们的小组，调查超微结构组织的生物样品。超微结构显微镜将用于研究生物组织细胞外基质中的胶原组织，肌细胞中的肌球蛋白纳米组织以及肌肉收缩过程中纳米活性的同步化。此外，还将研究淀粉粒、结晶纤维素和光合色素聚集体的超微结构。由蛋白质、多糖和色素组成的有序分子组装体在生物体内起着重要的结构和功能作用。例如，细胞外基质中胶原纤维的组织决定了组织的光学和机械性质，在横纹肌中组装成肌丝的肌球蛋白纳米马达周期性地改变它们的构象以产生用于生物体运动的收缩，淀粉颗粒以复杂的晶体结构组织以有效储存和释放碳水化合物，并且有序的颜料聚集体用于有效的太阳能收集。因此，重要的是开发非侵入性的，化学特异性的和无标记的成像工具，以可视化在体内的生物结构，并揭示分子组装体的纳米超微结构组织。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人员是重要的。在生物组织中的特定分子组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得出的结构模型。这些新技术将刺激新的体内组织成像工具的开发，提供研究肌肉疾病的新方法，并促进太阳能转换和植物储存的研究，并应用于人工光合作用，太阳能电池开发和仿生纳米技术。