Leica Stellaris 8 Confocal Microscope System

7.项目摘要/摘要 我们理解健康和疾病中细胞功能的能力，从宏观到微观，再到纳米，甚至到 原子尺度取决于最先进的仪器设备的能力。美国国立卫生研究院资助的项目 此S10应用程序中描述的应用程序需要增强的成像分辨率和多光谱功能 荧光显微镜的应用将更上一层楼。徕卡Stellaris 8激光扫描共聚焦 需要显微镜并将其放置在UMKC共焦和多光子显微镜核心设施中。这 仪器将取代和升级我们目前的徕卡TCS II SP5共焦显微镜，它作为一个共享的 用于支持NIH资助的主要用户以及次要用户和其他用户核心群体的研究计划的资源 UMKC牙科、药学、医学、生物科学、计算机和工程学院的用户，以及 附近的儿童慈悲医院。用户群体在肌肉骨骼研究方面有不同的研究重点， 神经/视觉研究，发育生物学，颅面和牙科研究，分子生物学，酵母遗传学， 神经退行性变、衰老、癌症和药物输送、材料科学，甚至石化研究。至 为了适应广泛的项目应用，该仪器将配置为在 固定细胞和组织标本，并用于活细胞成像、多光谱成像、荧光恢复后 光漂白(FRAP)、光学切片/3D成像和反射成像。采用这种技术的项目有三个 技术正在研究肌肉和骨骼之间信号串扰的机制，这可能调节与年龄有关的 骨质疏松和骨质疏松症。这些项目正在使用活细胞成像来监测线粒体功能和 细胞外小泡介导的肌肉-骨骼通讯，并使用3D共聚焦成像来评估细胞 运动前后增龄小鼠和功能丧失小鼠的骨组织结构。一 该项目正在使用活细胞成像来检查控制钙信号以防止神经退化的机制 在早期青光眼和两个项目正在开发药物输送和组织工程方法来治疗 老年性黄斑变性。其他项目主要集中在腭裂的分子和细胞机制上。 形成，心脏收缩功能和钙信号，发育的颌骨形态发生，单细胞和空间 儿童骨组织基因组学，骨细胞机械转导和骨组织的有限元和流体动力学模型 用于癌症和纤维化药物输送的纳米颗粒系统。Stellaris 8共焦系统将加速 通过提供3D单元所需的卓越分辨率和灵敏度来推动这些项目的进展 矿化组织和其他组织中的组织成像和活细胞成像。的多光谱成像能力 Stellaris 8，其激发和检测能力的多功能性，其增强的灵敏度和TauSense/ 荧光寿命技术是将这项研究推进到下一个水平的关键，并将允许扩展 我们的多路复用、3D空间映射和透明组织成像能力。收购这项技术将 加速在肌肉骨骼健康、发育、衰老、视力、牙齿修复和药物输送方面的发现。

7. Project Summary/Abstract Our ability to understand cellular function in health and disease from the macro to micro to nano and even to the atomic scale depend on the capabilities of state-of-the-art advanced instrumentation. The NIH funded projects described in this S10 application require enhanced imaging resolution and multi-spectral capabilities for their fluorescence microscopy applications to advance to the next level. A Leica Stellaris 8 laser scanning confocal microscope is requested and will be located in the UMKC Confocal and Multiphoton Microscopy Core Facility. This instrument will replace and upgrade our current Leica TCS II Sp5 confocal microscope that operates as a shared resource to support the research programs of a core group of NIH funded major users as well as minor and other users in the UMKC Schools of Dentistry, Pharmacy, Medicine, Biological Sciences, Computing and Engineering, and the nearby Children’s Mercy Hospital. The user group has a diverse research emphasis on musculoskeletal research, neural/vision research, developmental biology, craniofacial and dental research, molecular biology, yeast genetics, neurodegeneration, aging, cancer and drug delivery, materials science, and even petrochemical research. To accommodate the wide-ranging project applications, the instrument will be configured for confocal microscopy on fixed cell and tissue specimens, and for live cell imaging, multispectral imaging, fluorescence recovery after photobleaching (FRAP), optical sectioning/3D imaging, and reflection imaging. Three of the projects employing this technology are examining mechanisms of signaling crosstalk between muscle and bone that may regulate age related osteoporosis and sarcopenia. The projects are using live cell imaging to monitor mitochondrial function and extracellular vesicle-mediated muscle-bone communication and are using 3D confocal imaging to assess the cellular and structural organization of bone in aged gain and loss of function mouse models with and without exercise. One project is using live cell imaging to examine mechanisms controlling calcium signaling to prevent neurodegeneration in early stage glaucoma and two projects are developing drug delivery and tissue engineering approaches to treat age related macular degeneration. Other projects are focused on molecular and cellular mechanisms of cleft palate formation, heart contractile function and calcium signaling, developmental jaw morphogenesis, single cell and spatial genomics in pediatric bone tissues, finite element and fluid dynamic modeling of osteocyte mechanotransduction and nanoparticle systems for drug delivery in cancer and fibrosis. The Stellaris 8 confocal system will accelerate the progress of these projects by providing an exceptional level of resolution and sensitivity that is needed for 3D cell and tissue imaging and live cell imaging in mineralized and other tissues. The multi-spectral imaging capabilities of the Stellaris 8, the versatility of its excitation and detection capabilities, its enhanced sensitivity and its TauSense/ fluorescence lifetime technology are critical to advance this research to the next level, and will allow for expansion of our capabilities for multiplexing, 3D spatial mapping and cleared tissue imaging. Acquisition of this technology will accelerate discoveries in musculoskeletal health, development, aging, vision, dental restorations, and drug delivery.