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

基本信息

批准号：
DGDND-2017-00099
负责人：
Barzda, Virginijus
金额：
$ 2.91万
依托单位：
University of Toronto
依托单位国家：
加拿大
项目类别：
DND/NSERC Discovery Grant Supplement
财政年份：
2017
资助国家：
加拿大
起止时间：
2017-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=617773
关键词：
Nonlinear optical microscopy ultrastructural bioimaging

项目摘要

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

该发现计划提出了非线性光学显微镜技术及其在生物成像中的应用的进展。将开发新型光学显微镜模式，用于成像结构的每个焦点体积内有序分子结构的超微结构研究，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。显微成像还将使用经典的超短脉冲激光器来获得振动和频生成和受激拉曼散射图像对比度，从而为基于所选分子振动的成像提供化学特异性。我们小组最近开发的非线性 Stokes-Mueller 形式主义将采用偏振显微镜测量来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细胞外基质中的胶原蛋白组织、肌肉细胞中的肌球蛋白纳米运动组织以及肌肉收缩期间纳米运动活动的同步。淀粉颗粒、结晶纤维素和光合色素聚集体的超微结构也将被研究。蛋白质、多糖和色素的有序分子组装在生物体中发挥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特性，肌球蛋白纳米马达组装成横纹肌中的肌丝，周期性地改变其构象以产生生物体运动的收缩，淀粉颗粒被组织成复杂的晶体用于有效储存和释放碳水化合物的结构，以及有序颜料聚集体用于有效收集太阳能。因此，开发非侵入性、化学特异性和无标记成像工具来可视化非常重要