Elyra 7 Microscope with Lattice SIM2

项目摘要/摘要 加州大学圣迭戈分校医学院显微镜核心是校园内最大的光学显微镜核心设施，拥有 为加州大学圣地亚哥分校的研究人员服务了18年。加州大学圣迭戈分校的所有研究人员都可以访问它，无论 部门隶属关系，目前在整个校园拥有200多个注册用户实验室。核心享受 来自教职员工和我们机构领导层的大力支持。目前由12台仪器组成的核心阵列 包括多共焦系统、多光子、光片、Gatan 3查看串行块人脸扫描EM、幻灯片 扫描，以及其他。尖端的超分辨率活细胞领域存在着迫切的需求 成像，这是目前当地缺乏的。目前，我们唯一的超分辨率系统是POINT 扫描速度较慢且大量漂白的共焦平台。在这里，我们寻求资金以 从蔡司购买一台带有晶格SIM2的Elyra 7系统作为我们主要的超分辨率活细胞成像 系统。该系统还可用于正在发育的模式生物和厚固定标本的成像。这个 开创性地将采样效率提高2倍于经典SIM(结构照明显微镜) 实现快速、温和的3D超分辨率成像，穿透深度可与点扫描共聚焦媲美 显微镜。我们的现场仪器演示验证了最初的Elyra 7系统与Lattice SIM(版本 1)作为超分辨率活细胞成像的实用解决方案，它也带有一些创新的 进一步提高速度和通用性的处理模式。随着最近推出的DUAL 迭代重建算法，新升级的Elyra7晶格SIM2大幅提高了 该系统的分辨率从120 nm XY和300 nm Z到显著的60 nm XY和200 nm Z。 升级后的SIM2系统也更加灵活，需要更少的相移来进行图像重建。我们的 拟议的用户实验室说明了当地调查人员在推进拟议系统方面的迫切需要 他们由美国国立卫生研究院资助的研究。他们来自校园的各个角落，他们的研究项目都集中在一起 涵盖一系列生物医学研究领域，包括许多疾病领域，包括癌症、糖尿病、 心血管、肺部、代谢、感染性、炎症性、神经和精神疾病 其他。如果获得资金，Elyra 7 Lattice SIM2技术平台将被提议的用户广泛使用 此外，还将覆盖潜在的更大用户基础，因为我们的核心已被证明能够传播 将尖端显微技术推广到广泛的本地研究社区。有足够的空间和 基础设施、有能力的员工、强有力的教师监督和强大的机构承诺以及轨道 计划领导的核心赠款引用记录将确保有效和高效地使用拟议的 未来几年的乐器。Elyra 7晶格SIM2系统将填补我们仪器中的一个当前空白 并对加州大学圣地亚哥分校的生物医学研究组合产生持续、强大的影响。

PROJECT SUMMARY / ABSTRACT The UCSD School of Medicine Microscopy Core is the largest light microscopy core facility on campus and has been serving UCSD researchers for 18 years. It is accessible by all UCSD researchers regardless of department affiliation and currently has over 200 registered user labs across the campus. The Core enjoys strong support from the faculty and our institutional leadership. The current array of 12 instruments at the Core includes multiple confocal systems, multiphoton, light sheet, Gatan 3View serial block face scanning EM, slide scanning, as well as others. A critical need exists in the realm of cutting-edge super-resolution live cell imaging, which is currently lacking locally. At the moment, our only super-resolution systems are point scanning confocal platforms which are slow and heavily photo-bleach samples. Here we seek funding to acquire an Elyra 7 system with Lattice SIM2 from Zeiss as our primary super-resolution live cell imaging system. This system can also be used to image developing model organisms and thick fixed specimens. The groundbreaking 2-fold increase in sampling efficiency over classic SIM (Structured Illumination Microscopy) enables fast, gentle 3D super-resolution imaging at penetrating depths on par with point scanning confocal microscopy. Our on-site instrument demonstration validated the initial Elyra 7 system with Lattice SIM (Version 1) as a practical solution for super-resolution live cell imaging, which also comes with some innovative processing modalities that further enhance speed and versatility. With the recent introduction of the dual iterative reconstruction algorithm, the newly upgraded Elyra 7 Lattice SIM2 has substantially improved the resolution of the system from 120 nm XY and 300 nm Z to a remarkable 60 nm XY and 200 nm Z. The upgraded SIM2 system is also more flexible requiring fewer phase shifts for image reconstruction. Our proposed user labs illustrate a critical need among the local investigators for the proposed system in advancing their NIH-funded research. They come from all corners of the campus, and their research programs collectively cover a range of biomedical research fields with many disease areas represented, including cancer, diabetes, cardiovascular, pulmonary, metabolic, infectious, inflammatory, neurological and psychiatric diseases among others. If funded, the Elyra 7 Lattice SIM2 technology platform will be extensively utilized by proposed users and additionally will reach a potentially much larger user base as our core has proven capable of disseminating cutting-edge microscopy technologies to the wide local research community. Adequate space and infrastructure, capable staff, robust faculty oversight, and strong institutional commitment along with a track record of program-leading core grant citations will ensure the effective and efficient use of the proposed instrument for years to come. The Elyra 7 Lattice SIM2 system will fill a current void in our instrumentation portfolio and generate a sustained, powerful influence on the biomedical research portfolio at UC San Diego.