3D fluorescence microscope for virus-host interaction studies

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Bats are ecologically important as pollinators and insect controllers. They are natural reservoirs for several emerging and re-emerging viruses, including coronaviruses such as SARS-CoV-2, that are causative agents of important human and agricultural infections. These viruses are speculated to have spilled over or "jumped" from bats to humans and other animals, causing severe and often fatal disease. Despite evidence of bats harboring these viruses, they do not appear to show overt symptoms or clinical signs of infection. Little is understood why bats are not susceptible to these viral infections and the physiological characteristics that allow them to be carriers or reservoirs of these viruses. The Mossman lab has over 20 years of experience studying virus-host interactions and along with clinical colleagues in Toronto, were the first to isolate and characterize SARS-CoV-2 from two patients, which has led to over 20 publications. Our Discovery Grant has generated a unique and innovative program that leverages our expertise (virus-host interactions and innate immunity) and that of Dr. Faure who runs the world's only bat colony containing wild caught and bred animals (McMaster Bat Lab). We have developed cell lines and tools with which to study mechanisms of virus-bat interactions, including primary cultures and 3D organoids, the latter composed of multiple cell types from intestinal stem cells. We routinely use fluorescence microscopy to monitor how viruses invade cells and how cells respond to viral invasion. As our current inverted fluorescence microscope is obsolete due to lack of availability of components for repair (it was purchased 19 years ago), we are requesting a new fluorescence microscope. The requested infrastructure, a THUNDER 3D Imaging System, uses new computational clearing technologies to generate high resolution and high contrast images from thick samples, such as organoid cultures. It allows for acquisition of images taken deep within tissues such as organoids. The THUNDER system is a Leica technology, as is our current microscope, allowing for recycling of some components. This technology will allow us to monitor the kinetics and complexity of virus-host interactions in real time, while maintaining the integrity of sensitive samples. Understanding how bats harbor and spread viruses with an apparent lack of symptoms has implications for disease control in animals and humans. As big brown bats naturally occur in Canada, our findings will directly impact Canadian agricultural and health sectors. Moreover, findings will inform global strategies for disease control and development of novel antiviral therapies, particularly for new emerging viruses that bats appear to carry. Finally, this program will lay the foundation for training of highly qualified personnel in a multidisciplinary environment, working at the forefront of science and impacting on critical areas such as pandemic preparedness and understanding.