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 资助项目需要提高成像分辨率和多光谱功能，以将其明场和荧光显微镜应用提升到一个新的水平。需要 Zeiss LSM 710 34 通道激光扫描共焦显微镜。该仪器将作为共享资源运行，以支持 NIH 资助的 UMKC 牙科学院、UMKC 护理学院和堪萨斯大学医学中心的主要和次要用户核心小组的研究项目。用户组的研究重点是矿化组织和肌肉骨骼研究。为了适应各种项目应用，该仪器将配置用于固定细胞和组织样本的基本共焦显微镜检查，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，使用延时成像和新型荧光探针来检测各种骨 ECM 成分。这些研究为骨基质组装的动态性质以及细胞运动在组装过程中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，因为它难以接近，嵌入骨骼中。随着最近对骨细胞分化标记物的鉴定以及具有荧光骨细胞谱系报告基因的转基因小鼠的开发，可以将这些细胞在其矿化环境中的成像达到前所未有的水平。一个项目利用谱系报告基因结合延时成像来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。另一个项目研究了骨细胞蛋白 E11/gp38 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负荷信号并将其转化为骨形成反应的分子机制。其他项目涉及肌肉钙成像、确定调节磷酸盐稳态的分子机制以及确定 Cherubism 患者骨炎症病变的分子发病机制。蔡司 LSM 710 系统将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，从而推动这些项目的进展。 LSM 710 的多光谱成像功能及其增强的实时成像灵敏度对于将这项研究提升到一个新的水平至关重要，并且将允许同时对多个分子探针进行成像，从而最大限度地利用每个实验收集的信息。获得这项技术将推进有关肌肉骨骼健康的发现，并对骨质疏松症、骨软化症和遗传性结缔组织疾病等疾病产生影响。公共健康相关性：此应用程序适用于 Zeiss LSM 710 34 通道激光扫描共焦显微镜，作为共享资源运行，支持位于 UMKC 学校的 NIH 资助的主要用户和二级用户核心小组的研究项目牙科、UMKC 护理学院和堪萨斯大学医学中心。核心用户的研究重点是矿化组织和肌肉骨骼组织，这种最先进的仪器将增强他们在培养物和矿化组织内原位生成肌肉细胞和骨细胞高分辨率图像的能力，以推进以下研究目标：参与的项目。获得这项技术将带来有关骨骼和肌肉健康的新发现，并对矿化组织疾病（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。