Core B: Nanoscopy

项目总结/摘要：核心B纳米显微镜 超分辨率光学显微镜或纳米显微镜（即具有纳米级分辨率的显微镜）是 定义为成像的分辨率低于传统光学显微镜的衍射极限，其为~200 在赤道（xy）维度上约为500 nm，在轴向（z）维度上约为500 nm。最近开发的超分辨率 允许光学显微镜成像亚细胞结构和分子定位在10 nm的方法 规模已经彻底改变了神经生物学，使神经科学家能够研究神经胶质，轴突， 树突和突触在前所未有的细节，以前只能使用电子 显微镜（EM）。特别是，超分辨率荧光成像比EM具有许多优点 包括更容易的样品制备和染色程序，以及应用于非 不仅是固定的，还有活的细胞和组织。几种不同的超分辨荧光成像方法 已经开发了诸如STimulated-Emission Depletion（STED）显微镜和相关的测量方法。 随机光学重建显微镜（STORM）和光活化定位显微镜（PALM）。 这些纳米方法中的每一种都具有优点和局限性，这取决于纳米材料的性质。 生物样品和待成像的荧光团。因此，有利地并且甚至必要地采用 在研究任何给定的科学问题时，这些不同的方法不止一种。因此 RMNDC Nanoscopy Core B将提供最近获得的仪器和技术支持， 科罗拉多大学安舒茨医学院的NINDS资助和其他神经科学研究人员 （UC-AMC）进行最先进的STED和STORM/PALM超分辨率成像（Aim 1）， 基于互补福斯特共振能量转移（FRET）的成像（Aim 2）。

Project Summary/Abstract: Core B Nanoscopy Super-resolution light microscopy or nanoscopy (i.e. microscopy with nanometer-scale resolution) is defined as imaging with a resolution below the diffraction limit of conventional light microscopy, which is ~200 nm in the equatorial (xy) and ~500 nm in the axial (z) dimension. Recently developed super-resolution methods that allow light microscopy to image subcellular structure and molecular localization on the 10s of nm scale have revolutionized neurobiology by allowing neuroscientists to study the inner workings of glia, axons, dendrites, and synapses in unprecedented detail that had previously only been possible using electron microscopy (EM). In particular, super-resolution fluorescence imaging has many advantages over EM including much easier sample preparation and staining procedures as well as the ability to be applied to not only fixed, but also living cells and tissues. Several different super-resolution fluorescence imaging methods have been developed, such as STimulated-Emission Depletion (STED) microscopy and the related methods of Stochastic Optical Reconstruction Microscopy (STORM) and Photo-activation Localization Microscopy (PALM). Each of these nanoscopy methods has advantages and limitations depending on the properties of the biological sample and fluorophores to be imaged. Thus, it is advantageous and even necessary to employ more than one of these different methods when investigating any given scientific question. Accordingly, the RMNDC Nanoscopy Core B will provide access to recently acquired instrumentation and technical support for NINDS-funded and other neuroscience investigators at the University of Colorado-Anschutz Medical Campus (UC-AMC) to perform state-of-the-art STED and STORM/PALM super-resolution imaging (Aim 1) and complementary Forster-resonance energy transfer (FRET)-based imaging (Aim 2).