Confocal multiphoton laser-scanning-microscope

本申请的目的是采购一台共焦多光子激光扫描显微镜(MPLSM)，用于活体和原位研究小鼠和人的胰腺。与MPLSM相关的工作重点将是研究胰岛在健康、糖尿病发病机制和治疗方面的生物学作用。在其具体配置中，MPLSM将作为我们团队开创和建立的两个创新技术平台的关键仪器，通过这两个平台，我们的实验室已经达到了全球独一无二的实验水平。这些平台包括眼前房内移植的朗格汉斯胰岛的无创纵向活体成像(Speier等人，2008a)，以及急性和培养组织切片的胰腺细胞生理原位成像(Cohars等人，2020b)。因此，眼台的前房首次允许在体内的系统环境中，在从几天到几个月的较长时间段内研究人类和啮齿动物的胰岛细胞生物学。此外，新的组织切片平台能够对啮齿动物胰腺进行原位研究，但更重要的是，它启动了从器官和组织捐赠者那里获得可存活的人类胰腺组织的第一次研究。除了内分泌胰腺生理学和病理生理学的研究外，这些平台还为研究潜在的细胞和药物疗法以及糖尿病的诊断工具提供了新的手段。因此，我们的项目将涉及三个主要领域：1)1型糖尿病(T1D)，2)2型糖尿病(T2D)和3)细胞治疗。目前的MPLSM已经成功地用于在小鼠模型中开发这些平台，并使该技术适用于人类组织的使用。这些研究已经发表了许多文章，其中几篇发表在影响力很大的研究期刊上。此外，它还帮助建立了许多成功的地方、国家和国际合作，并获得了大量的第三方资金。更换是必要的，因为大量使用和老化导致技术问题的数量和比率不断增加，同时制造商正在终止对这类机器的维护和维修服务选择。用技术上显著先进的MPLSM取代这一设备，不仅将确保继续使用我们的平台，而且还将扩大我们平台的成功使用。根据我们的经验，我们预计正在进行和计划中的项目，包括大量的地方、国家和国际合作，每年将使用大约4000个小时的所要求的MPLSM。

The present application aims at the procurement of a confocal and multiphoton laser scanning microscope (MPLSM) for the in vivo and in situ study of mouse and human pancreas. Main focus of the work related to the MPLSM will be the investigation of pancreatic islet of Langerhans biology in health, diabetes pathogenesis and therapy. In its specific configuration the MPLSM will serve as the key instrumentation of two innovative technical platforms our group has pioneered and established, and through which our laboratory has reached a worldwide unique experimental profile. These platforms include the non-invasive longitudinal in vivo imaging of transplanted islets of Langerhans within the anterior chamber of the eye (Speier et al., 2008a), and the in situ imaging of pancreas cell physiology in acute and cultured tissue slices (Cohrs et al., 2020b). Hereby, the anterior chamber of the eye platform allows for the first time to study human and rodent islet cell biology over prolonged time periods, ranging from several days to multiple months, in a systemic setting in vivo. Moreover, the novel tissue slice platform enables in situ investigations of the rodent pancreas, but more importantly, has initiated the first studies of viable human pancreas tissue from organ and tissue donors. In addition to the study of endocrine pancreas physiology and pathophysiology these platforms provide new means to study potential cellular and medicinal therapies as well as diagnostic tools for diabetes. Thereby, our projects will address aspects in three main areas: 1) type 1 diabetes (T1D), 2) Type 2 diabetes (T2D) and 3) Cell Therapy.The here requested MPLSM is essential as replacement for the currently utilized imaging system. The current MPLSM has been successfully employed for more than a decade to develop these platforms in mouse models and adapt the technology to the use of human tissue. These studies have led to numerous publications, several in high-impact research journals. Furthermore, it has helped to establish numerous successful local, national and international collaborations and acquire significant third-party funding. The replacement is necessary as heavy use and age are leading to an increasing number and rate of technical issues while at the same time maintenance and repair service options for this type of machine are being ended by the manufacturer. Replacement of this device with a technically significantly advanced MPLSM will not only ensure continuation, but also expansion of the successful use of our platforms. Based on our experience, we anticipate the ongoing and planned projects, including a significant number of local, national and international collaborations, to amount to an annual use of approximately 4000 hours of the requested MPLSM.