Acquisition of Leica confocal/multiphoton microscope

 描述（由申请人提供）：我们请求资助共聚焦/多光子显微镜，将服务于15名教职员工的关键成像需求，其中许多人对组织细胞和基质成分的动态监测感兴趣，依赖于荧光和散射信号，用于再生医学，组织工程和各种疾病诊断和治疗应用。在过去的十年中，我们已经证明了丰富的组织结构和功能信息，可以从非侵入性，高分辨率成像方法，如共聚焦，双光子激发荧光和二次谐波发生显微镜。这些研究正在引导新的方向，需要：a）对相同样品的长期（多小时至数天）动态监测，B）对非常快速的相互作用的监测，c）大视场的检查，d）对弱内源性信号的增强的灵敏度，和/或e）改进的成像穿透深度。这些方向不能用一个共焦/多光子显微镜，可在塔夫茨梅德福校园，现在是十多岁。2014年，现有系统的使用率约为每周100小时，尽管对使用时间有严格限制。这妨碍了设计需要几天监测的必要研究。此外，该系统已经过时，不允许更新，使更灵敏的检测信号，在本提案中描述的许多研究依赖于这些原因，拟议的仪器预计将满足关键的显微成像需求。具体来说，新的显微镜将使长期或快速，高分辨率的光学成像的许多项目的性能，旨在解决重要的生物医学问题，跨越微生物学，癌症生物学，代谢组学和组织工程等领域。虽然系统的可用性将使长期研究成为可能，但其许多功能 在推进所采集图像的功能内容方面至关重要，包括：a）增强荧光检测的效率、灵敏度和灵活性，用于监测较弱和更可变的发色团，B）能够在成像过程中自动控制入射强度以优化样品活力和成像穿透深度的情况下进行测量，c）更自动化地检查厚和宽的样本，以及d）更快的图像获取。多学科的研究团队代表了梅德福和波士顿塔夫茨校区内的四所不同的学校，展示了对广泛的研究范围和学生培训和教育的重大影响。PI的主要研究兴趣是高分辨率光学成像领域，她拥有专业知识和技术支持，以确保设备的创新和适当使用。有新装修的空间来容纳仪器，重要的机构支持，以及维护和运行显微镜的长期可持续性计划。最后，我们建立了一个内部和外部咨询委员会，以协助解决可能出现的行政和实验问题。

 DESCRIPTION (provided by applicant): We request funding for a confocal/multi-photon microscope that will serve critical imaging needs of fifteen faculty members, many of whom are interested in the dynamic monitoring of tissue cell and matrix components relying on fluorescence and scattering signals for regenerative medicine, tissue engineering and various disease diagnostic and therapeutic applications. During the last ten years, we have demonstrated the wealth of tissue structural and functional information that can be acquired from non-invasive, high resolution imaging approaches, such as confocal, two-photon excited fluorescence and second harmonic generation microscopy. These studies are leading to new directions that require: a) long-term (multiple hours to days) dynamic monitoring of the same sample, b) monitoring of very fast interactions, c) examination of large fields of view, d) enhanced sensitivity to weak endogenous signals, and/or e) improved imaging penetration depths. These directions cannot be pursued with the one confocal/multi-photon microscope that is available at the Tufts Medford campus that is now over ten years old. The current system was utilized approximately 100 hours/week in 2014, even with strict limits imposed on usage time. This prevents the design of needed studies that require monitoring over several days. In addition, the system is outdated and doesn't allow updates that enable more sensitive detection of the signals that many of the studies described in this proposal rely on. For these reasons, the proposed instrument is expected to fulfill critical microscopic imaging needs. Specifically, the new microscope will enable the performance of long-term or fast, high-resolution optical imaging for numerous projects aiming to solve important biomedical problems spanning the fields of microbiology, cancer biology, metabolomics, and tissue engineering among others. While long-term studies will be enabled simply by the availability of the system, a number of its features are essential in advancing the functional content of the acquired images, including: a) enhanced efficiency, sensitivity and flexibility of fluorescence detection for monitoring weaker and more variable chromophores, b) ability to perform measurements with automated control over the incident intensity during imaging to optimize sample viability and imaging penetration depth, c) more automated examination of thick and wide specimens, and d) faster image acquisition. The multidisciplinary team of investigators represents four different Schools within the Medford and Boston Tufts campuses, demonstrating the significant impact on a broad research spectrum and on student training and education. The PI's main research interests are in the area of high-resolution optical imaging and she has the expertise and technical support to ensure innovative and appropriate use of the equipment. There is newly renovated space to house the instrument, significant institutional support, and a long-term sustainability plan for maintaining and running the microscope. Finally, we have established both an internal and an external advisory board to assist with potential administrative and experimental issues that arise.