Novel tools for controlling singlet oxygen in cellular systems.

图像是人们感知宇宙的主要手段。目前的技术已经改变了这种看法，这在生物科学中尤其明显，因为活细胞成像深入研究了亚细胞结构和功能的奥秘。 成像技术通过允许实时观察生物体内部发生的事件来彻底改变生物学。 不幸的是，活生物体的成像几乎保证了单线态氧的形成，单线态氧是分子氧的高度反应性和毒性形式，其对被观察的活生物体是危险的，并且其使从样本获得的数据劣化。 由于我们对单线态氧的化学性质及其在细胞机制中的作用感兴趣，我们建议在对细胞系统成像的同时利用单线态氧的产生。具体而言，我们将努力创造用于活细胞成像的新工具。 这些新工具中的第一个将使用基因编码的荧光蛋白，这种荧光蛋白在单线态氧的存在下不起反应。 这将减少成像过程中观察到的伪影数量，并有助于保护生物体。本文提出的第二种工具是单线态氧的“按需”亚细胞来源，用于这种高毒性物质的受控空间和时间释放，这将提供对细胞系统中单线态氧形成的更大调节。 在我们寻求创造活细胞成像的新工具时，我们将奋进扩展与有机敏化剂和荧光蛋白的化学和光化学/光物理学有关的现有知识体系。 我们将利用这些知识开发新的工具，这些工具将强烈影响成像技术，进而进一步了解单线态氧在细胞机制中的作用。 此外，这项研究将促进加拿大生物医学研究和加拿大生物技术产业的新机会。

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.