Acquisition of Leica confocal/multiphoton microscope

 描述(由申请人提供)：我们申请资助一台共焦/多光子显微镜，该显微镜将满足15名教职员工的关键成像需求，他们中的许多人对依赖于再生医学、组织工程和各种疾病诊断和治疗应用的荧光和散射信号的组织细胞和基质成分的动态监测感兴趣。在过去的十年里，我们已经展示了丰富的组织结构和功能信息，可以通过非侵入性的高分辨率成像方法获得，例如共聚焦、双光子激发荧光和二次谐波产生显微镜。这些研究正在引领新的方向，需要：a)对同一样本进行长期(数小时到数天)的动态监测，b)监测非常快速的相互作用，c)检查大视野，d)增强对微弱内源信号的敏感性，和/或e)改善成像穿透深度。这些方向不能用塔夫茨梅德福德校区现在已经有十多年历史的一个共焦/多光子显微镜来追踪。2014年，即使对使用时间有严格的限制，目前的系统每周使用约100个小时。这阻止了所需研究的设计，这些研究需要几天的监测。此外，该系统已经过时，不允许对本提案中描述的许多研究所依赖的信号进行更敏感的检测的更新。出于这些原因，建议的仪器有望满足关键的显微成像需求。具体地说，这种新型显微镜将为众多项目提供长期或快速的高分辨率光学成像，这些项目旨在解决跨越微生物学、癌症生物学、代谢组学和组织工程等领域的重要生物医学问题。虽然长期研究将仅仅通过系统的可用性来实现，但它的许多功能是 这对于提升采集图像的功能内容至关重要，包括：a)提高了荧光检测的效率、灵敏度和灵活性，用于监测较弱和多变的发色团；b)能够在成像过程中自动控制入射强度来执行测量，以优化样品的活力和成像穿透深度；c)更自动化地检查厚和宽的样品；以及d)更快的图像采集。多学科调查团队代表了梅德福德和波士顿塔夫茨校区内的四所不同学校，展示了对广泛研究领域以及对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，她拥有确保创新和适当使用设备的专业知识和技术支持。有新装修的空间来存放仪器，重要的机构支持，以及维护和运行显微镜的长期可持续计划。最后，我们设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试验问题。

 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 In addition, the system is outdated and doesn't allow updates that enable more sensitive detection of the signals that many of the studies described in this proposal rely on.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.