Grant: Research Tools and Instruments

The ability to peer inside the human body has revolutionized our understanding of the spectrum of human health. Similarly, the ability to peer inside the cells that make up the body - some 30-40 trillion in total - and visualize their architecture and dynamics has transformed our understanding of cellular function. Advances in microscopy have allowed us to identify specific molecules, or biomarkers, that may reveal previously unknown cellular functions and further our understanding of human health. But merely identifying the presence or absence of a particular biomarker is not sufficient: we need to know how cellular function and behaviour changes over time, and to measure these parameters in living cells. These cellular events may occur at the cell surface or deep inside the cells, and occur on the order of microseconds. There are new technologies that extend the capabilities of microscopy to enable the fast, high-throughput imaging, thus capturing cellular events that characterize normal physiology. This proposal seeks to add functionalites to our existing microscope which will enable imaging of these cellular dynamics. The first is Total Internal Reflection Fluorescence (TIRF) for the visualization of fast events that occur at the cell surface, allowing for the real-time detection of hormone secretion or cell-cell communication. The second is a 20X long working distance (LWD) objective lens and high-content software for rapid screening of several biochemical processes. The third will be a near-infrared (NIR) laser that will enable the imaging of structures and cellular events deep within cells and tissues that are not accessible by our current complement of lasers. Finally, we will add artificial intelligence-based image processing software (NIS.ai) that will rapidly analyse large datasets at faster speeds. These upgrades will address significant gaps in our city-wide hospital-based research programs that are focussed on furthering our understanding of fundamental biochemical and cellular processes of human physiology. Examples are: visualization of the dynamics of hormone secretion from individual cells (Metabolism and Diabetes Program); detection of biochemical communication and cell-cell interactions between the microbiome and human tissue (Microbiome and Probiotics Program); generation of bioengineered contrast agents for cellular magnetic resonance imaging (MRI) (Imaging Research Program); and the study of tissue injury and self-repair in the musculoskeletal system (Critical Illness Research Program). Therefore, the requested upgrades to the confocal will benefit a number of research programs that address scientific innovations across the spectrum of human health. The upgrades will also allow us to train our students and research personnel on leading edge technologies in fluorescence microscopy so that they will be competitive for jobs in the health and medical sciences, educational and industrial sectors.