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Zeiss LSM 710 Confocal Microscopy System for Imaging of Mineralized Tissues

用于矿化组织成像的蔡司 LSM 710 共焦显微镜系统

基本信息

批准号：
8050238
负责人：
SARAH L DALLAS
金额：
$ 44.6万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2012-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8050238
关键词：
Bone Matrix Brightfield Microscopy Calcium Cell physiology Cherubism Computers and Advanced Instrumentation Confocal Microscopy Connective Tissue Diseases Dendrites Development Differentiation Antigens Disease Environment Extracellular Matrix Fluorescence Microscopy Fluorescence Recovery After Photobleaching Fluorescent Probes Funding Health Homeostasis Image Inflammatory Inherited Kansas Lasers Lesion Life Mechanics Medical center Microscope Molecular Molecular Probes Muscle Musculoskeletal Nature Optics Osteoblasts Osteocytes Osteomalacia Osteoporosis Pathogenesis Patients Process Property Proteins Reporter Research Research Support Resolution Resource Sharing Role Scanning School Dentistry School Nursing Signal Transduction Specimen System Technology Three-Dimensional Imaging Time Tissues Transgenic Mice Translating United States National Institutes of Health Universities bone cell fixing cell motility cell type cellular imaging information gathering inorganic phosphate insight instrument nano novel programs research study response skeletal time use

项目摘要

DESCRIPTION (provided by applicant): Our capacity 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 advanced instrumentation. The NIH funded projects described in this application require increased imaging resolution and multi-spectral capabilities for their brightfield and fluorescence microscopy applications to advance to the next level. A Zeiss LSM 710 34 channel laser scanning confocal microscope is requested. The instrument will be operated as a shared resource to support the research programs of a core group of NIH funded major and secondary users in the UMKC School of Dentistry, UMKC School of Nursing and University of Kansas Medical Center. The user group has a research emphasis on mineralized tissues and musculoskeletal research. To accommodate the range of project applications, the instrument will be configured for basic confocal microscopy on fixed cell and tissue specimens, and for live cell imaging, multispectral imaging, fluorescence recovery after photobleaching (FRAP) and optical sectioning/3D imaging. One of the projects that will employ this technology is investigating the mechanisms of assembly of bone extracellular matrix (ECM), using time lapse imaging with novel fluorescent probes for various bone ECM components. These studies are providing novel insights into the dynamic nature of bone matrix assembly and the role of cell motility in the assembly process. The role of the osteocyte in regulating skeletal responses to mechanical loading is the focus of three of the participating projects. Until recently, little was known about the function of this elusive cell type, due to its inaccessibility, embedded within bone. With the recent identification of osteocyte differentiation markers and development of transgenic mice with fluorescent osteocyte lineage reporters, it is possible to image these cells within their mineralized environment to an unprecedented level. One project examines the dynamic properties of osteoblasts and osteocytes using lineage reporters combined with time lapse imaging to gain insight into the process by which osteoblasts differentiate into osteocytes. Another project examines the role of the osteocyte protein, E11/gp38, in dendrite formation and in osteocyte responses to mechanical loading. A third project examines the molecular mechanisms by which osteocytes perceive mechanical loading signals and translate them into bone formative responses. Other projects involve calcium imaging in muscle, determining the molecular mechanisms regulating phosphate homeostasis and determining the molecular pathogenesis of bone inflammatory lesions in Cherubism patients. The Zeiss LSM 710 system will advance the progress of these projects by providing an unsurpassed level of resolution that is needed for 3D imaging of osteocyte networks in mineralized tissues. The multi-spectral imaging capabilities of the LSM 710 and its enhanced sensitivity for live imaging, are essential to advance this research to the next level, and will allow for simultaneous imaging of multiple molecular probes, maximizing the information gathered from each experiment. Acquisition of this technology will advance discoveries regarding musculoskeletal health and will have implications for diseases, such as osteoporosis, osteomalacia and inherited connective tissue disorders. PUBLIC HEALTH RELEVANCE: This application is for a Zeiss LSM 710 34 channel laser scanning confocal microscope, to be operated as a shared resource that will support the research programs of a core group of NIH funded major users and secondary users, located in the UMKC School of Dentistry, UMKC School of Nursing and University of Kansas Medical Center. The research emphasis of the core users is on mineralized and musculoskeletal tissues and this state of the art instrumentation will enhance their ability to generate high resolution images of muscle cells and bone cells in culture and in situ within the mineralized tissue to advance the research goals of the participating projects. Acquisition of this technology will result in new discoveries regarding bone and muscle health and will have major implications for diseases of mineralized tissues, such as osteoporosis, osteomalacia and inherited connective tissue disorders.

描述（由申请人提供）：我们从宏观到微观到纳米甚至到原子尺度了解健康和疾病中细胞功能的能力取决于先进仪器的能力。本申请中描述的NIH资助的项目需要提高成像分辨率和多光谱能力，以使其明场和荧光显微镜应用更上一层楼。需要Zeiss LSM 710 34通道激光扫描共聚焦显微镜。该仪器将作为共享资源运行，以支持UMKC牙科学院、UMKC护理学院和堪萨斯大学医学中心的NIH资助的主要和次要用户核心小组的研究计划。用户组的研究重点是矿化组织和肌肉骨骼研究。为了适应项目应用的范围，该仪器将被配置为用于固定细胞和组织标本的基本共聚焦显微镜，以及活细胞成像，多光谱成像，光漂白后荧光恢复（FRAP）和光学切片/3D成像。将采用该技术的项目之一是研究骨细胞外基质（ECM）的组装机制，使用新型荧光探针的时间推移成像用于各种骨ECM成分。这些研究为骨基质组装的动态性质和组装过程中细胞运动的作用提供了新的见解。骨细胞在调节骨骼对机械负荷的反应中的作用是三个参与项目的重点。直到最近，人们对这种难以捉摸的细胞类型的功能知之甚少，因为它嵌入骨骼中。随着最近骨细胞分化标记的鉴定和具有荧光骨细胞谱系报告基因的转基因小鼠的开发，有可能将这些细胞在其矿化环境中成像到前所未有的水平。其中一个项目使用谱系报告结合延时成像检查成骨细胞和骨细胞的动态特性，以深入了解成骨细胞分化为骨细胞的过程。另一个项目研究骨细胞蛋白E11/gp 38在树突形成和骨细胞对机械负荷的反应中的作用。第三个项目研究骨细胞感知机械载荷信号并将其转化为骨形成反应的分子机制。其他项目包括肌肉中的钙成像，确定调节磷酸盐稳态的分子机制，以及确定巨颌症患者骨炎症病变的分子发病机制。蔡司LSM 710系统将通过提供矿化组织中骨细胞网络3D成像所需的无与伦比的分辨率水平来推进这些项目的进展。LSM 710的多光谱成像能力及其增强的实时成像灵敏度对于将这项研究推进到下一个水平至关重要，并将允许同时对多个分子探针进行成像，最大限度地提高从每个实验中收集的信息。获得这项技术将推动有关肌肉骨骼健康的发现，并将对骨质疏松症、骨软化症和遗传性结缔组织疾病等疾病产生影响。公共卫生相关性：此应用程序是一个蔡司LSM 710 34通道激光扫描共聚焦显微镜，将作为一个共享资源，将支持一个核心组的NIH资助的主要用户和二级用户的研究计划，位于牙科，护理和堪萨斯大学医学中心的UMKC学校。核心用户的研究重点是矿化和肌肉骨骼组织，这种最先进的仪器将提高他们在培养和矿化组织内原位生成肌细胞和骨细胞的高分辨率图像的能力，以推进参与项目的研究目标。获得这项技术将导致有关骨骼和肌肉健康的新发现，并将对矿化组织疾病，如骨质疏松症，骨软化症和遗传性结缔组织疾病产生重大影响。