Leica Stellaris 8 Confocal Microscope

项目概要/摘要 该提案要求为 Leica Stellaris 8 共焦显微镜系统提供资金，以便在 辛辛那提大学 (UC) 实时显微镜核心 (LMC)。加州大学医学院 (CoM) 的科学家们 极大地受益于 LMC 提供的成像服务，推动了 CoM 的生物医学研究。 在过去两年中，LMC 为 56 个实验室提供了服务，这些实验室由 35 项 NIH 拨款资助，导致超过 25 出版物。然而，澜湄合作的生产力和效率在过去逐渐下降 三年的原因是：1) 我们的共焦显微镜老化以及 2) 缺乏成像技术进步 能力。 目前，LMC 拥有两台共焦显微镜：Zeiss LSM510 NLO 和 Zeiss LSM710 （分别于 2004 年和 2010 年购买），两者都经历了越来越多的维护问题，并且 停机，给许多研究项目带来极大的不便和延误。此外，这些 旧仪器在速度、灵敏度、分辨率和多功能性方面存在技术限制。 NIH 资助的 加州大学的研究人员对先进的实时成像功能有强烈的需求，并且需要仪器来：1) 在活细胞、组织和 3D 类器官培养物中进行长时间延时荧光成像实验，2) 收集具有足够分辨率的 3D 荧光图像，以定位染色中感兴趣的蛋白质和结构 细胞、组织和类器官，3) 利用高效光谱同时检测大量荧光标记 分离。 为了满足这一迫切需求，该应用程序需要资金来购买先进的共焦显微镜 Leica Stellaris 8 系统是 LMC 现有显微镜的补充。这种新仪器将服务于 两个主要目的：增加我们目前过度使用的共焦系统的容量，并实现新的成像 现有技术无法完成的实验。具体来说，这种新型显微镜将极大地 提高样品长时间延时成像的可行性，增强检测微小颗粒的分辨率和灵敏度 亚细胞细节，并提高成像速度，以高效收集大型 3D 数据集并快速捕获 活样品中的动态过程。 Leica Stellaris 8 系统将安装在当前的中央 成像设施，将由在共聚焦各方面拥有 15 年经验的设施技术人员操作 显微镜实验和分析。该申请得到了实质性兴趣、需求和支持 大学的财政支持，并将为美国国立卫生研究院资助的大型科学界提供服务。我们已确保 来自 CoM 和 UC 研究办公室的资金保证，以确保仪器在服务范围内 始终签订合同，并制定财务计划，其中包含用于持续运营和人员的备用部门资金 日常操作显微镜的成本。所需的仪器非常适合解决 NIH 资助的 8 个主要用户的技术规范和容量问题。

PROJECT SUMMARY/ABSTRACT This proposal requests funding for a Leica Stellaris 8 confocal microscope system to be housed and used in the University of Cincinnati (UC) Live Microscopy Core (LMC). The scientists at UC College of Medicine (CoM) have greatly benefited from the LMC providing imaging services that have propelled biomedical research in the CoM. In the past two years, the LMC served 56 laboratories funded by 35 NIH grants, which resulted in more than 25 publications. The productivity and efficiency of the LMC, however, have been gradually diminished over the past three years due to: 1) the aging of our confocal microscopes and 2) lack of technical advancements of imaging capabilities. Currently, the LMC operates two confocal microscopes, Zeiss LSM510 NLO and Zeiss LSM710 (purchased in 2004 and 2010, respectively), both of which have experienced increasing maintenance issues and downtime, resulting in substantial inconveniences and delays in numerous research projects. Additionally, these older instruments have technical limitations in speed, sensitivity, resolution, and versatility. The NIH-funded investigators at UC have a strong need for advanced live imaging capabilities, and require instrumentation to: 1) perform long time lapse fluorescence imaging experiments in live cells, tissues, and 3D organoid cultures, 2) collect 3D fluorescence images with sufficient resolution to localize proteins and structures of interest in stained cells, tissues, and organoids, and 3) simultaneously detect numerous fluorescent markers with efficient spectral separation. To mitigate this critical need, this application requests funds to acquire an advanced confocal microscope system, the Leica Stellaris 8, to supplement the existing microscopes in the LMC. This new instrument will serve two major purposes: add capacity to our currently over-utilized confocal systems, and enable new imaging experiments that cannot be achieved with the existing technology. Specifically, this new microscope will greatly improve the viability of samples for long time lapse imaging, enhance resolution and sensitivity to detect small subcellular details, and improve imaging speed for efficient collection of large 3D data sets and capturing fast dynamic processes in live samples. The Leica Stellaris 8 system will be installed within the current central imaging facility and will be operated by the facility technician with 15 years of experience in all aspects of confocal microscopy experimentation and analysis. This application is supported by substantial interest, need, and financial support at the University, and will serve a large NIH-funded scientific community. We have secured funding guarantees from the CoM and UC Office of Research to ensure the instrument is covered under service contract at all times, and a financial plan with backup departmental funds for ongoing operational and personnel costs to operate the microscope on a day-to-day basis. The requested instrument is ideally suited to address both the technical specifications and capacity issues for the 8 NIH-funded Major Users on this grant.