OMX BLAZE High Speed Super Resolution Imaging System

描述（由申请人提供）：我们对基本细胞生物学过程的理解既受到我们可视化细胞和组织内结构和分子组织的能力的驱动，也受到其限制。“超分辨率”荧光成像方法的发展，规避了传统光学显微镜的分辨率限制，实现了d100 nm的横向分辨率，正在改变细胞生物学研究。超分辨率荧光成像技术对于希望推动细胞生物学研究前沿的科学家来说至关重要。该提案要求提供资金购买OMX BLAZE 2D、3D-SIM快速超分辨率成像系统（Applied Precision，Inc.）。这种超分辨率系统可以实现横向和轴向分辨率的两倍，传统的光学显微镜的衍射极限，并能够远远超过盖玻片的多通道超分辨率。OMX BLAZE的快速成像能力也旨在克服3D活细胞成像的速度限制。这种先进的成像系统将是一种共享资源，位于斯坦福大学一个完善的多用户显微镜设施：细胞科学成像设施。所要求的OMX BLAZE 3D-SIM成像系统将支持NIH资助的14名研究人员的项目。这些项目调查了广泛的主题，包括：减数分裂染色体分离（维勒纳夫）;基因重组（维勒纳夫）;中心体结构、功能和复制（Stearns）;着丝粒和动粒组装初级纤毛的生物发生和功能（Nachury，Rohatgi，Stearns）;神经回路的结构（Smith）;原肠胚形成期间上皮细胞重排的分子机制（纳尔逊）;肌动蛋白丝组装和动力学（纳尔逊）;分泌囊泡对接和融合的分子机制（Pfeffer）;神经突触的发育（Shen）;触觉受体神经元中的机械-电转导（Goodman）;重组和基因工程（Porteus）;发育和癌症中的刺猬途径信号传导（Chen，Scott，Beachy，Rohatgi）。这些研究调查了有关基本细胞生物学过程的关键功能和结构问题，并涵盖了NIH研究领域，对人类健康和疾病的各个方面产生影响，包括癌症，出生缺陷，肥胖，肾囊肿（遗传性Bardet-Biedl综合征）和神经退行性疾病。所有这些项目都需要多通道超分辨率成像和同步多通道快速成像; OMX BLAZE 3D SIM系统能够最有效地提供这种功能组合。

DESCRIPTION (provided by applicant): Our understanding of fundamental cell biological processes is both driven by and limited by our ability to visualize the organization of structures and molecules within cells and tissues. The development of "super-resolution" fluorescence imaging methods that circumvent the resolution limits of conventional light microscopy, achieving lateral resolution of d100 nm, are transforming cell biological research. Access to super-resolution fluorescence imaging technology is now essential for scientists wishing to push the frontiers of cell biological research. This proposal requests funds to purchase the OMX BLAZE 2D, 3D-SIM fast super-resolution imaging system (Applied Precision, Inc.). This super-resolution system can achieve both lateral and axial resolution at twice the diffraction limit of conventional light microscopy and is capable of multi- channel super-resolution far beyond the cover glass. The fast imaging capability of OMX BLAZE is also designed to overcome speed limitations for 3D live-cell imaging. This advanced imaging system will be a shared resource, located in a well-established, multi-user microscopy facility at Stanford: the Cell Sciences Imaging Facility. The requested OMX BLAZE 3D-SIM imaging system will support NIH funded projects from 14 researchers. These projects investigate a wide range of topics, including: meiotic chromosome segregation (Villeneuve); genetic recombination (Villeneuve); centrosome structure, function and duplication (Stearns); centromere and kinetochore assembly (Straight); biogenesis and function of the primary cilium (Nachury, Rohatgi, Stearns); architecture of neural circuits (Smith); molecular mechanisms underlying epithelial cell rearrangements during gastrulation (Nelson); actin filament assembly and dynamics (Nelson); molecular mechanisms of secretory vesicle docking and fusion (Pfeffer); development of neural synapses (Shen); mechano-electrical transduction in touch receptor neurons (Goodman); recombination and genetic engineering (Porteus); Hedgehog pathway signaling in development and cancer (Chen, Scott, Beachy, Rohatgi). These studies investigate critical functional and structural questions regarding fundamental cell biological processes and cover NIH research areas with implications for diverse aspects of human health and disease, including cancer, birth defects, obesity, kidney cysts (in the inherited Bardet-Biedl Syndrome) and neurodegenerative disease. All these projects require multi-channel super-resolution imaging and simultaneous multi-channel fast imaging; this combination of capabilities is most effectively provided by the requested OMX BLAZE 3D SIM system.