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，这在生物科学中尤为明显，因为活细胞成像正在探索亚细胞结构和功能的奥秘。成像技术正在给生物学带来革命性的变化，因为它允许实时观察发生在生物体内的事件。不幸的是，对活体的成像几乎保证了单线态氧的形成，这是一种高度活性和毒性的分子氧形式，对被观察的生物是危险的，并使从标本获得的数据恶化。 在我们对单线态氧的化学及其在细胞机制中的作用的兴趣的推动下，我们建议在成像细胞系统的同时利用单线态氧的产生。具体地说，我们将努力创造新的活细胞成像工具。这些新工具中的第一个将使用基因编码的荧光蛋白，这些蛋白在单线态氧存在下不起反应。这将减少在成像过程中观察到的人工制品的数量，并有助于保护活的有机体。本文提出的第二种工具是按需提供单线态氧的亚细胞来源，用于控制这种剧毒物种的时空释放，这将对细胞系统中单线态氧的形成提供更大的调节。 在我们寻求创造新的活细胞成像工具的过程中，我们将努力扩大目前与有机敏化剂和荧光蛋白质的化学和光化学/光物理有关的知识体系。我们将利用这些知识来开发新的工具，这些工具将对成像技术产生重大影响，进而加深我们对单线态氧在细胞机制中作用的理解。此外，这项研究将在加拿大生物医学研究和加拿大生物技术产业中培育新的机会。

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.