A Confocal Multiphoton/Atomic Force Microscopy System

描述(由申请人提供)：本申请请求支持购买将与原子力显微镜(AFM)系统相结合的共焦/多光子显微镜系统。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，无论有没有AFM技术，都可以使用满足用户需求的AFM组件进行操作。这项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，单一焦点接触水平上的焦点接触和细胞骨架蛋白对机械负荷的响应，血管和淋巴管中的细胞信号反应，AFM对机械刺激的内皮细胞的响应，分离的和功能的小动脉和淋巴管壁上三维平滑肌肉组织的可塑性，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管网络，以及交叉上核(SCN)单个细胞中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快的速度和更少的细胞和组织损伤随着时间的推移进行重复测量的能力。这种混合成像系统使我们能够进行这些研究的独特能力包括：1)对活细胞和组织的生物损伤有限；2)聚焦，厚样品的光学切片；3)深入切片和组织的能力；4)对各种荧光分子成像的能力；5)提高从细胞和组织获取三维数据集的能力，并随着时间的推移做到这一点。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.