Tunable White Light Laser Confocal/Spectral Imaging Microscopy

描述（由申请人提供）：W.M. Keck细胞成像中心（KCCI）是国际公认的先进光学显微镜成像中心，尤其以蛋白质-蛋白质相互作用成像而闻名，该中心正在申请Leica TCS SP 5X“可调谐白色激光共聚焦/光谱成像显微镜”系统。KCCI是一个大学范围内的成像设施，所要求的仪器将有利于该大学的基础和临床科学研究人员，特别是8名参与联邦资助的研究人员。仪器将放置在KCCI中，位于所有参与研究者的步行距离内。在所要求的Leica TCS SP 5X上进行的研究的中心主题是研究活细胞中的蛋白质-蛋白质相互作用和其他细胞成像，包括用于监测垂体GHFT 1细胞中动态蛋白质相互作用的光活化GFP（PA_GFP）、IQGAP和阿尔茨海默病的细胞生物学、新FRET对的表征、谷氨酰胺FRET传感器、肌动蛋白在膜上组装的机制研究、以及在胚胎发育期间监测细胞生物系统的多个分子组分的动态行为。所要求的仪器Leica SP 5X系统是独一无二的，代表了共聚焦显微镜的一次飞跃，为用户提供了调整激发和发射波长的能力，使其与实验样品中的荧光团完全匹配。用户不再需要使实验适应共焦系统上可用的激光线;可调谐白色光激光系统适应特定实验。目前在弗吉尼亚大学还没有其他系统具有可调谐（470-670 nm）白色激光共焦/光谱成像能力。这对我们的核心研究小组至关重要，对培训其他对活细胞和组织中蛋白质相关性研究感兴趣的研究人员也至关重要。该KCCI是公认的年度实践培训课程，在过去七年中在弗吉尼亚大学举行。迄今为止，来自美国的304名科学家以及来自23个国家的科学家参加了KCCI研讨会。总之，所要求的仪器将为弗吉尼亚大学的研究界提供目前尚不具备的成像能力，这将大大加强正在进行的和未来的研究。公共卫生相关性：许多不同颜色的荧光蛋白使活细胞内生物事件的非侵入性检测成为可能，但这种方法需要高度适应性的显微镜系统。Leica TCS SP 5X可调谐白色激光系统是一个独特的系统，允许用户调整激发和发射波长，以精确匹配实验样品中的荧光团。如果我们必须了解疾病过程并设计治疗策略，那么继续开发这些活细胞成像技术就非常重要，而不同颜色荧光蛋白和Leica SP 5X的组合代表了该领域的重大进步。

DESCRIPTION (provided by applicant): The W.M. Keck Center for Cellular Imaging (KCCI), an internationally recognized center for advanced light microscopy imaging that is particularly known for protein-protein interaction imaging, is requesting a Leica TCS SP5X "Tunable White Light Laser Confocal/Spectral Imaging Microscopy" system. KCCI is a University wide imaging facility and the requested instrument will benefit the basic and clinical science investigators of this university, and particularly the 8 participating federally funded investigators. The instrument will be housed in the KCCI, located within walking distance of all the participating investigators. The central theme of the research to be performed on the requested Leica TCS SP5X is the study of protein-protein interactions in live cells and other cellular imaging including Photoactivated GFP (PA_GFP) to monitor the dynamic protein interactions in pituitary GHFT1 cells, IQGAPs and the Cell Biology of Alzheimer's Disease, characterization of new FRET pairs, glutamine FRET sensors, study of the mechanism of actin assembly at membranes, and monitoring dynamic behavior of multiple molecular components of the cell biological system during embryonic development. The requested instrument, the Leica SP5X system is unique and represents a quantum leap in confocal microscopy, providing users the ability to tune the excitation and emission wavelengths to match exactly the fluorophores in their experimental samples. The user no longer needs to adapt the experiment to the laser lines available on the confocal system; the tunable white light laser system adapts to the particular experiment. There is currently no other system available at the University of Virginia that has a tunable (470-670 nm) white light laser confocal/spectral imaging capability. This is essential for our core investigator group, and will also be critical for training other investigators interested in the study of protein associations in living cells and tissues. The KCCI is recognized for its annual hands-on-training course, held for the past seven years at the University of Virginia. To date, 304 scientists, from the United States as well as scientists from 23 countries have participated in the KCCI workshop. In summary, the requested instrument will provide the research community at the University of Virginia with a level of imaging capability that is not currently available, and this will enhance significantly ongoing and future research. PUBLIC HEALTH RELEVANCE: The non-invasive detection of biological events inside the living cell was made possible by the many different color fluorescent proteins, but this approach requires a highly adaptable microscope system. The requested Leica TCS SP5X tunable white light laser system is a unique system that permits the user to tune the excitation and emission wavelengths to match exactly the fluorophores in their experimental samples. If we have to understand disease processes and design therapeutic strategies, it is important to continue to develop these live-cell imaging technologies, and the combination of different color fluorescent proteins and the requested Leica SP5X represents a substantial advance in this area.