Grant: Research Tools and Instruments

We request funds to replace the Keyence microscope, an integrated part of the PhenoCycler multiplex imaging system. The PhenoCycler system enables the imaging of >100 proteins simultaneously in various tissue samples - critical to better understand the mechanisms of cell interaction within the entire tissue landscape. Although specialized in different tissues, including blood vessels, hearts, and brains, our team has a shared research interest in understanding the mechanism of tissue homeostasis via cell-cell/microenvironment interaction. Our team of early, mid-, and established career investigators has published in top-tier journals detailing novel methods and analysis pipelines to investigate these interactions, and revealed several key mechanisms of cell degradation, memory retention, and energy homeostasis. Our recent projects featured multiplex imaging PhenoCycler technology brought to Canada in 2022 via a CFI-JELF awarded to the principal applicant. The PhenoCycler is the only system available to us to assess the activity of signaling cascades at the post-translational level. Due to inflation and the budget cap of the JELF award, we did not purchase the PhenoImager microscope, an integrated component of PhenoCycler, designed for best performance and compatibility. Instead, we assembled a functional system using an in-house Keyence microscope. In less than two years, we used this system to become the first and only researchers in Canada to develop an imaging pipeline to answer research questions (published in May 2024), and to launch a spatial biology research community to nurture expertise in this quickly evolving field. This community is now building a training hub for `Omics' in the structural context. The scope of our studies has, however, been limited by our lack of a compatible and user-friendly microscope to leverage the high-throughput capacity of PhenoCycler. System compatibility has become an obvious and immediate issue to resolve in 2025. The Keyence microscope has to be replaced because: 1) it has recently been phased out and no manufacturers will provide future service; 2) it triples our processing time, delaying critical experiments and making training of new users challenging; 3) sample transfer is not user-friendly, complicating access for off-site HQP to the instrument. Once the PhenoImager replaced the Keyence microscope, the PhenoCycler will be used extensively to increase our research impact and provide a deep and broad understanding of the signaling cascades triggered by cell-cell/microenvironment interactions. The more compatible, efficient, and user-friendly microscope, coupled with our team's robust experience in providing hands-on training, will provide HQP with a leading edge in their research. Our PhenoCycler training hub will help stimulate their scientific creativity, critical thinking, communication, and collaboration skills to prepare them to excel in the competitive job market.