Upgrades for essential imaging platforms at a multi-user microscope facility

我的实验室进行细胞和发育生物学的研究，重点是形成和维持上皮组织的基本机制。上皮细胞是动物身体的关键组成部分，是皮肤、肠道和许多其他器官的重要组成部分。我们的研究探索上皮细胞的形成和发育，重点是控制上皮细胞极化结构的机制，以及它们之间的粘附。极性和粘附是理解上皮形态发生、组织生长和干细胞更新的关键。进行这一研究项目的一项绝对必要的技术是共聚焦显微镜，它需要使用激光扫描共聚焦显微镜（LSCMs）。目前的提案旨在为这些显微镜更换磨损的镜片争取资金。此外，我们需要为用于共聚焦显微镜的标本选择和制备的立体显微镜的新荧光光源提供资金。两台需要新镜头的LSCMs（徕卡TCS SP8）是在2013年获得的，资金来自加拿大创新基金会授予Tepass博士的资助。这两种仪器对于我实验室中10-12个HQPs进行的研究都是必不可少的。一种仪器非常适合胚胎和组织的实时成像，而另一种仪器主要用于固定样品的成像。我的实验室是这些显微镜的主要使用者，这些显微镜被安置在一个部门的成像设施里。其他部门和外部研究人员可以通过在线预订系统访问这些仪器。自2013年以来，近300名研究人员接受了LSCM使用培训。我们预计这种使用模式将在可预见的未来继续下去。由于大量使用，这两种显微镜都出现了一些性能问题，导致图像质量和可用性的不可接受的损失，严重干扰了我们的研究。经过制造商的全面评估后，建议进行几次维修，以使lscm恢复到规格。在CSB系和文理学院投资10万美元的情况下，其中一些维修工作已经实现。在这里，我寻求资金更换两个磨损的40倍和63倍油浸镜头。此外，我请求支持购买一个LED荧光光源，以替换现有立体显微镜上的一个损坏的单元。该仪器是在NSERC-RTI的支持下于2005年购买的，其他方面都处于良好的光学和机械状态。将LSCMs和荧光立体显微镜恢复到最先进的光学质量和工作状态对我实验室的研究项目至关重要。这些仪器帮助大量不同的学员开展他们的研究项目，并获得先进成像技术的经验。

My laboratory carries out research in cell and developmental biology with a focus on fundamental mechanisms that form and maintain epithelial tissues. Epithelia are key building blocks of animal bodies and are prominent components of the skin, gut, and many other organs. Our research explores the formation and development of epithelia with an emphasis on mechanisms that govern the polarized structure of epithelial cells, and adhesion between them. Polarity and adhesion are key to understanding morphogenesis, tissue growth, and stem cell-based renewal of epithelia. An absolutely essential technology for conducting this research program is confocal microscopy that requires the use of Laser Scanning Confocal Microscopes (LSCMs). The present proposal seeks to secure funding for replacing worn-out lenses for these microscopes. In addition, we require funding for a new fluorescent light source for a stereo microscope that is used in specimen selection and preparation for confocal microscopy. The two LSCMs (Leica TCS SP8) that require new lenses were acquired in 2013 with funds from a Canadian Foundation for Innovation grant awarded to Dr. Tepass. Both instruments are essential for the research carried out by 10-12 HQPs in my laboratory. One instrument is highly suitable for live-imaging of embryos and tissues, whereas the second is used predominantly for the imaging of fixed samples. My laboratory is the main user of these microscopes, which are housed in a departmental imaging facility. The instruments are accessible through an online booking system to other departmental and external researchers. Since 2013 nearly 300 researchers were trained in the use of a LSCM. We anticipate that this usage pattern will continue for the foreseeable future. As a result of heavy use, both microscopes have developed a number of performance issues that have led to an unacceptable loss of image quality and usability that severely interferes with our research. After a thorough assessment from the manufacturer several repairs were recommended to bring the LSCMs back to specification. Some of these repairs have already been realized with an investment of >$100,000 from the CSB Department and the Faculty of Arts & Science. Here, I am seeking funds for the replacements of two worn-out 40x and two 63x oil immersion lenses. In addition, I am requesting support for the purchase of an LED Fluorescent Light Source to replace a broken unit on an existing stereo microscope. This instrument, which was purchased in 2005 with NSERC-RTI support is otherwise in good optical and mechanical condition. Returning the LSCMs and the fluorescent stereo microscope to state-of-the art optical quality and working condition is essential for the research programs of my laboratory. These instruments help a large number of diverse trainees carry out their research project and acquire experience with advanced imaging technologies.