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。上皮是动物身体的主要组成部分，也是皮肤、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展，重点是控制上皮细胞极化结构的机制，以及它们之间的粘连。极性和粘附性是理解形态发生、组织生长和基于干细胞的上皮细胞更新的关键。进行这一研究计划的一项绝对必要的技术是共焦显微镜，它需要使用激光扫描共焦显微镜(LSCM)。目前的提案寻求获得资金，为这些显微镜更换磨损的镜片。此外，我们需要为立体显微镜的新荧光光源提供资金，该光源用于样品选择和共聚焦显微镜的准备。需要新镜头的两台LSCM(徕卡TCS SP8)是在2013年收购的，资金来自加拿大创新基金会授予Tepass博士的资金。这两种仪器对我实验室中10-12名HQP进行的研究都是必不可少的。一种仪器非常适合于胚胎和组织的活体成像，而第二种仪器主要用于固定样本的成像。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部门的成像设施中。其他部门和外部研究人员可通过在线预订系统获取这些仪器。自2013年以来，近300名研究人员接受了使用LSCM的培训。我们预计，在可预见的未来，这种使用模式将继续下去。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。经过制造商的彻底评估后，建议进行几次维修以使LSCM恢复规范。其中一些修复工作已经完成，公务员事务局和文理学院投资了10万美元。在这里，我正在寻求资金，以更换两个磨损的40x和两个63x的油浸式镜头。此外，我请求支持购买LED荧光光源，以更换现有立体显微镜上损坏的单元。这台仪器是在2005年购买的，支持NSERC-RTI，除此之外，它的光学和机械状况良好。将激光扫描共聚焦显微镜和荧光立体显微镜恢复到最先进的光学质量和工作条件，对于我的实验室的研究计划是必不可少的。这些仪器帮助大量不同的受训人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.