Novel tools for controlling singlet oxygen in cellular systems.

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Images are the primary means by which people perceive the universe. Current technology has transformed this perception, as is particularly evident in the biological sciences, as live cell imaging delves into the mysteries of subcellular architecture and function. Imaging techniques are revolutionizing biology by permitting the observation of events in real-time, as they occur inside of living organisms. Unfortunately, the imaging of living organisms almost guarantees the formation of singlet oxygen, a highly reactive and toxic form of molecular oxygen, which is dangerous to the living organism being observed and which deteriorates the data obtained from the specimen. Driven by our interest in the chemistry of singlet oxygen and its role in cellular mechanisms, we propose to exploit the production of singlet oxygen while imaging cellular systems. Specifically, we will strive to create new tools for live cell imaging. The first of these new tools will use genetically encoded fluorescent proteins that are unreactive in the presence of singlet oxygen. This will reduce the number of artifacts observed during the imaging process, and help protect the living organism. The second tool proposed herein is an "on demand" subcellular source of singlet oxygen for controlled spatial and temporal release of this highly toxic species that will afford greater regulation of singlet oxygen formation in the cellular system. In our quest to create new tools for live cell imaging, we will endeavor to expand the current body of knowledge pertaining to the chemistry and photochemistry/photophysics of organic sensitizers and fluorescent proteins. We will capitalize on this knowledge in the development of novel tools that will strongly impact imaging techniques, and in turn further our understanding of the role of singlet oxygen in cellular mechanisms. Furthermore, this research will foster new opportunities in Canadian biomedical research and the Canadian biotechnology industry.