A Confocal Multiphoton/Atomic Force Microscopy System

描述（由申请人提供）：本申请要求支持购买共聚焦/多光子显微镜系统，该系统将与原子力显微镜（AFM）系统相结合。这两种成像方式的结合是独特的和可逆的，只涉及显微镜阶段的转换。因此，它可以使用或不使用AFM组件，满足用户的需要，使用显微镜系统有或没有AFM技术。该技术的主要用途将包括研究细胞膜整合素及其与细胞外基质蛋白的相互作用、单点接触水平上对机械负荷的反应的局灶接触和细胞骨架蛋白、血管和淋巴平滑肌中的细胞信号反应、内皮细胞对机械刺激的反应。分离的功能性小动脉和淋巴管管壁中平滑肌三维组织的可塑性、完整血管壁中细胞外基质蛋白的三维组织特征、人工细胞外基质中的三维毛细血管网络以及体外视交叉上核（SCN）单个细胞实时昼夜节律基因表达的空间调控。所要求的系统将极大地促进我们以更高的速度和更少的细胞和组织损伤随时间重复测量的能力。这种混合成像系统的独特功能将使我们能够进行这些研究，包括：1)对活细胞和组织的生物损伤有限；2)在焦点上，厚样品的光学切片；3)能够深入观察切片和组织；4)对多种荧光分子成像的能力；5)增强从细胞和组织中获取三维数据集的能力，并随着时间的推移这样做。这项新技术将为教师、博士后研究员和学生提供独特的、最先进的成像设备，并将促进主要和次要用户正在进行的研究。

DESCRIPTION (provided by applicant): This application is requesting support for the purchase of a confocal / multiphoton microscopy system that will be combined with an atomic force microscope (AFM) system. The combination of these two imaging modalities is unique and reversible involving only conversion of the microscope stages. Consequently, it can be operated with or without the AFM components satisfying the needs of users to use the microscope system with or without the AFM technology. Major uses of this technology will include studies of cell membrane integrins and their interactions with extracellular matrix proteins, focal contact and cytoskeletal proteins in response to mechanical loading at the level of a single focal contact, cell signaling responses in vascular and lymphatic smooth muscle, endothelial ceils in response to mechanical stimulation with the AFM, plasticity of three-dimensional smooth muscle organization in the wall of isolated and functional arterioles and lymphatic vessels, three-dimensional organizational characteristics of extracellular matrix proteins in the intact vascular wall, three-dimensional capillary tube networks in artificial extracellular matrices and spatial regulation of real time circadian gene expression in individual cells from suprachiasmatic nuclei (SCN) in vitro. The requested system will greatly facilitate our ability to perform repeated measurements over time at higher speeds and with less cellular and tissue damage. The unique capabilities of this hybrid imaging system that will allow us to perform these studies include: 1) limited biological damage to living cells and tissues; 2) in focus, optical sections of thick samples; 3) ability to view deep into slices and tissues; 4) the ability to image a variety of fluorescent molecules; and 5) increased ability to obtain three-dimensional data sets from cells and tissues and do this over time. This new technology will provide faculty, postdoctoral fellows and students with access to a unique state-of-the-art imaging facility and will facilitate the ongoing research of the major and minor users.