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

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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.