Nikon Multiphoton (MP) Imaging System

项目摘要/摘要 请提供资金购买尼康A1R多光子(MP)成像系统，将其集成到 Vanderbilt细胞成像共享资源(CISR)。范德比尔特在生物医学成像方面有着卓越的历史， 成像科学研究所拥有出色的人类和活体动物成像设施， 质谱学研究中心内突破性的分子成像和尖端设备 先进的光学显微镜在国际科学研究中心。然而，将这些杰出的资源整合到 所有尺度上的图像生物过程--即从整个有机体到亚细胞结构--都有 由于缺乏将活体动物成像与组织和体外成像联系起来的成像解决方案而受到限制 细胞。所提出的MP系统能够可视化生命组织深处的细胞事件 通过活体成像，在体内和体外规模之间提供了关键的联系。12个人的小组 NIH资助的范德比尔特地区的用户群体最初将得到这一工具的支持。这些调查人员已经 未得到满足的成像需求，以支持NIH资助的横跨传染病、免疫学、神经生物学、 消化系统疾病、上皮生物学、糖尿病、肺损伤和癌症。 MP系统的拟议配置以用户群体的需求为基础，包括：1)双 用一次可调谐光束(800-1300 nm)和二次固定光束(1040 Nm)支持成像的输出激光器 深度高达1.4 mm，2)高达720fps动态成像的高速共振扫描仪，3)完整 自动化的大幅面舞台和直立显微镜支架，可适应麻醉和活体中 准备工作，以及4)为支持拟议的用户项目而特别选择的目标。 此外，在范德比尔特感染、免疫学和炎症研究所(VI4)的支持下，我们 已经建立了专门的生物安全二级成像套件，包括辅助麻醉、手术和 灌流设备，以容纳MP系统，并确保我们可以容纳对生物有害的标本。 范德比尔特正在提供财政支持，以促进该工具的长期运作，包括工资 用于技术人员和必要的服务合同。将促进对新用户的技术支持和培训 由CSR内的杰出工作人员提供，该中心已有20多年的历史，帮助300多个实验室 校园里。此外，位于中国国际贸易研究中心内的尼康卓越中心提供了一个专门的当地团队 专家支持校园内研究人员的显微镜需求。总体而言，拟议的MP制度， 再加上出色的机构和核心设施支持，将使建立 范德比尔特的活体成像。

PROJECT SUMMARY / ABSTRACT Funds are requested to purchase a Nikon A1R Multiphoton (MP) Imaging System, to be integrated into the Vanderbilt Cell Imaging Shared Resource (CISR). Vanderbilt has a history of excellence in biomedical imaging, with outstanding facilities for human and live animal imaging within the Institute of Imaging Sciences, groundbreaking molecular imaging within the Mass Spectrometry Research Center, and cutting-edge equipment for advanced light microscopy within the CISR. However, the ability to integrate these outstanding resources to image biologic processes across all scales – i.e. from the whole organism down to subcellular structure – has been limited by the lack of an imaging solution to bridge live animal imaging to ex vivo imaging of tissues and cells. The proposed MP System is capable of visualizing cellular events deep within the tissues of living organisms through intravital imaging, providing a critical link between in vivo and ex vivo scales. A group of 12 NIH-funded User Groups across Vanderbilt will initially be supported by this instrument. These investigators have unmet imaging needs to support NIH-funded projects spanning infectious diseases, immunology, neurobiology, digestive diseases, epithelial biology, diabetes, lung injury, and cancer. The proposed configuration of the MP System is based on the needs of the User Groups and includes: 1) a dual- output laser with primary tunable beam (800-1300 nm) and secondary fixed (1040 nm) beam to support imaging depth of up to 1.4mm, 2) a high-speed resonant scanner for dynamic imaging of up to 720fps, 3) a fully automated, large-format stage and upright microscope stand to accommodate anesthesia and intravital preparations, and 4) a selection of objectives specifically chosen to support the proposed User projects. Additionally, with support from the Vanderbilt Institute for Infection, Immunology, and Inflammation (VI4), we have already established a dedicated biosafety level 2 imaging suite with ancillary anesthesia, surgical, and perfusion equipment to house the MP System and ensure that we can accommodate biohazardous specimens. Vanderbilt is providing financial support to promote the long-term operation of the instrument, including salary for technical staff and necessary service contracts. Technical support and training of new users will be facilitated by the exceptional staff within the CISR, which has a greater than 20-year history of assisting over 300 labs on campus. Moreover, the Nikon Center of Excellence housed within the CISR provides a dedicated local team of specialists to support the microscopy needs of investigators on campus. Collectively, the proposed MP System, together with outstanding institutional and core facility support, will enable the long-term objective of establishing intravital imaging at Vanderbilt.