Super-resolution Workstation for Imaging Live Biological Nanostructure

描述（由申请人提供）：用于活体生物纳米结构成像的超分辨率工作站。将开发一种新的研究仪器，用于可视化和测量低于传统光学显微镜分辨率限制的活体生物结构。这种超分辨率显微镜在图像采集过程中不需要任何机械光学调整，因此可以快速成像无固有机械工件的亚分辨率结构。该仪器将结合两种已建立的技术-驻波显微镜的空间分辨率增强和声光激光扫描的时间分辨率增强。该仪器的性能将是独一无二的，在研究亚分辨率活生物结构动力学的应用中将具有特别的优势。PI先前构思并构建了一系列先进的成像仪器，这是他长期生物学研究目标所必需的，以了解单个神经元和小神经元群体的信息处理。所有开发的仪器都利用了PI在衍射光学元件方面的专业知识，特别是声光器件。这些元件的光学特性可以快速调节，即在无线电频率范围内使用电子产生的声波，使声光设备成为先进成像仪器的独特组成部分。拟议的成像工作站将分两步开发，从而提高三维空间分辨率。通过声光器件对必要的照明模式进行无惯性控制，将产生具有优越机械稳定性和成像速度的高度通用性仪器。所提出的快速超分辨率成像工作站在生物医学研究中具有重要意义。这将极大地改善重要细胞内结构的可视化和功能监测，包括线粒体、内质网和微管。特别是在实验神经科学中，这样的仪器将支持突触传递的各个方面的研究，包括突触前囊泡簇和突触后树突棘颈。例如，脊柱颈部脆弱的亚分辨率结构在发育和可塑性过程中容易发生变化，也容易受到许多神经系统疾病的影响。总的来说，提议的超分辨率成像能力的可用性将是变革性的，并使生物医学领域的大型社区受益。

DESCRIPTION (provided by applicant): Super-resolution Workstation for Imaging Live Biological Nanostructures. A novel research instrument will be developed for visualizing and measuring living biological structures that are below the resolution limit of conventional light microscopy. This super-resolution microscope does not require any mechano-optical adjustments during image acquisition and will thus allow for fast imaging of sub-resolution structures free of inherent mechanical artifacts. The instrument will combine two established techniques -- the spatial resolution enhancement of Standing Wave Microscopy with the temporal resolution enhancement of Acousto-Optic Laser Scanning. The proposed instrument will be unique in its performance and will be of particular advantage in applications where the dynamics of sub-resolution living biological structures are to be studied. The PI has previously conceived and constructed a series of advanced imaging instruments necessary for his long-term biological research goal to understand information processing in single neurons and small neuronal populations. All developed instruments utilized the PI's expertise with Diffractive Optical Elements, specifically Acousto-Optic Devices. The optical properties of these elements are rapidly adjustable, i.e. with electronically produced sound waves in the radio frequency range, making acousto-optic devices unique building blocks for advanced imaging instrumentation. The proposed imaging workstation will be developed in a two-step approach, resulting in improved spatial resolution in three dimensions. The inertia-free control of the necessary illumination patterns by acousto-optic devices will result in a highly versatile instrument with superior mechanical stability and imaging speed. The proposed workstation for fast super-resolution imaging would be of high importance in biomedical research. It would vastly improve the way important intracellular structures can be visualized and their function monitored, including mitochondria, endoplasmic reticulum, and microtubules. Specifically in experimental Neuroscience such an instrument would support the study of various aspect of synaptic transmission, including presynaptic vesicle clusters and postsynaptic dendritic spine necks. For example, the fragile sub-resolution structure of spine necks is susceptible to changes during development and plasticity, but also to a number of neurological diseases. In general, the availability of the proposed super-resolution imaging capability would be transformational and benefit large communities in the biomedical field. PUBLIC HEALTH RELEVANCE (provided by the applicant): Although the proposed imaging workstation was conceived for Biomedical Research, it has also great potential as a diagnostic tool. Changes in subcellular structure and function often coincide with various states of numerous diseases. The proposed instrument will allow microscopic inspection and functional testing of subcellular structures in live, non-fixed cellular specimen at unparalleled spatio- temporal resolution.