MRI: Development of a High-Speed Light-Sheet Light-Field Microscope for Imaging in Materials Sciences, Physics, and Biology

非技术描述：光学显微镜为了解从简单的胶体悬浮液和生物细胞到组织和整个发育中的生物体的各种系统的微米级结构和动力学提供了一个重要的窗口。大多数光学显微镜研究仅限于感兴趣的二维区域。表征三维结构的典型方法依赖于共聚焦扫描技术，这是非常慢的。由于缺乏合适的技术，对大尺寸样品的快速动态的高空间分辨率成像仍然是未知领域。该项目为克服这一障碍铺平了道路。由此产生的一种名为光场光片显微镜的新型成像技术被用不同的样本进行测试和优化，从量化三维活性液晶的动力学和拓扑结构到成像果蝇在觅食过程中的全脑。显微镜的设计原则和相关的软件分析代码与科学界共享。这种成像技术还通过将其纳入每年一度的圣巴巴拉定量生物学暑期学校而得到普及，参与者在那里接受使用该仪器的培训。技术描述：通过无缝融合现有的多视角选择性平面照明显微镜和光场显微镜技术，科学家们正在开发一种新的光学显微镜，能够成像具有各向同性点扩散函数的毫米大小的样品。与最先进的选择性平面照明显微镜所能达到的最快成像时间相比，正在开发的技术将时间分辨率提高了近两个数量级。这为在高达5赫兹的频率下对宏观样品进行三维、高空间分辨率成像提供了可能。这种显微镜被用来研究从材料科学、细胞生物学到神经科学的各种样本。特别是，科学家使用显微镜来(1)阐明三维活动向列相液晶中运动向列环的拓扑结构和动力学之间的相互作用，(2)量化由单分子马达产生压力的活动凝胶中应变变形的时间传播，(3)在黑腹夜蛾幼虫寻找食物的过程中执行全脑成像，以及(4)描述整个神经系统在黑腹夜蛾胚胎发生过程中形成和激活的特征。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Nontechnical Description:Optical microscopy provides an essential window into the micrometer scale structure and dynamics of diverse systems ranging from simple colloidal suspension and biological cells to tissues and entire developing organisms. Most optical microscopy studies are limited to two-dimensional regions of interest. A typical method of characterizing three-dimensional structure relies on confocal point scanning techniques, which are very slow. Due to lack of appropriate techniques, high-spatial resolution imaging of fast dynamics of macro-sized samples remains uncharted territory. This project paves the way for overcoming this barrier. The resulting novel imaging technique named light-field light-sheet microscopy is tested and optimized with diverse samples ranging from quantifying dynamics and topological structure of three-dimensional active liquid crystals to imaging the whole-brain of a fruit fly during food-search. The design principles of the microscopy and the associated software analysis code are shared with scientific community. The imaging technique is also popularized through its inclusion in the Annual Santa Barbara Summer School in Quantitative Biology, where the participants receive the training in using the instrument. Technical Description:By seamlessly merging the existing techniques of the multi-view selective plane illumination microscopy and the light-field microscopy, scientists are developing a new optical microscope capable of imaging millimeter-sized samples with isotropic point spread function. When compared to the fastest imaging times accessible to state-of-the-art selective plane illumination microscopes, the technique being developed increases the temporal resolution by almost two orders of magnitude. This opens up the possibility of three-dimensional, high-spatial-resolution imaging of macroscopic samples at frequencies as high as 5 Hz. The microscope is used to study a variety of samples that range from materials science, cell biology and neuroscience. In particular, scientist use the microscope to (1) elucidate the interplay between the topological structure and dynamics of motile disclination loops in three-dimensional active nematic liquid crystals, (2) quantify temporal propagation of strain deformations in active gels where stresses are generated by single molecular motors, (3) perform whole-brain imaging during food-search in D. melanogaster larvae, and (4) characterize a whole nervous system as it forms and activates in the embryogenesis of D. melanogaster.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.