OMX Blaze Structured-Illumination Microscope for a Core Facility

描述(由申请人提供)：作为已建立的核心设施的一部分，我们向应用精密公司申请一台OMX火焰结构照明显微镜(SIM)。关键的细胞过程发生在比共聚焦显微镜可分辨的结构和组件更小的结构和组件中，但仍然需要荧光标记的简单性和特异性。对于多个实验系统，我们的超分辨率需求有五个限制。(1)我们必须探索的体积往往在细胞和组织的深处，与盖板相距甚远。(2) 所有的研究都需要轴向超分辨率，以及横向超分辨率。(3)相关的结构，从大的蛋白质组装到细胞器到细胞核，通常是100 nm或更大的大分子组装。(4)这些组件经常与许多其他分子相互作用，需要强大的调色板才能同时可视化。(5)对于某些活细胞动力学，成像速度是重要的。经过多项技术的广泛测试，只有SIM适合我们的需求。此外，我们拥有所有用户的初步SIM卡数据，深入了解每个供应商的SIM卡实施情况。与约翰·霍普金斯大学的现有设备相比，拟议的仪器将提供数量级的优越性能，将使NIH资助的关键项目成为可能。

DESCRIPTION (provided by applicant): As part of an established core facility, we request an OMX Blaze structured-illumination microscope (SIM) from Applied Precision. Key cellular processes occur in structures and assemblies smaller than resolvable by confocal microscopy, but still requiring the ease and specificity of fluorescence labeling. Addressing multiple experimental systems, our super-resolution needs have five constraints. (1) The volumes we must explore are often deep within cells and tissues, significantly distant from the coverslip. (2) All studies require axial super-resolution, as well as lateral super-resolution. (3) The relevant structures, spanning large protein assemblies to organelles to the nucleus, are typically macromolecular assemblies 100 nm or larger. (4) The assemblies often interact with many other molecules, requiring a robust palette of colors for simultaneous visualization. (5) For some live-cell dynamics, speed of imaging is important. After extensive testing of multiple technologies, only SIM is appropriate for our needs. Furthermore, we have preliminary SIM data for all Users, thoroughly exploring each vendor's implementation of SIM. Compared to existing equipment anywhere at Johns Hopkins, the proposed instrument will provide an order-of-magnitude superior performance that will enable key NIH-funded projects.