Tunable White Light Laser Confocal/Spectral Imaging Microscopy

描述（由申请人提供）：wm 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，以蛋白质-蛋白质相互作用成像而闻名，现申请徕卡TCS SP5X“可调谐白光激光共聚焦/光谱成像显微镜”系统。KCCI是一个大学范围内的成像设备，所要求的仪器将使该大学的基础和临床科学研究人员受益，特别是8名参与联邦资助的研究人员。该仪器将被安置在KCCI，位于所有参与调查人员的步行距离内。在Leica TCS SP5X上进行的研究的中心主题是活细胞中蛋白质-蛋白质相互作用的研究和其他细胞成像，包括光激活GFP (PA_GFP)，以监测垂体GHFT1细胞中的动态蛋白质相互作用，IQGAPs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机制的研究。并监测胚胎发育过程中细胞生物系统中多个分子组分的动态行为。所要求的仪器，徕卡SP5X系统是独一无二的，代表了共聚焦显微镜的一次飞跃，为用户提供了调节激发和发射波长的能力，以准确匹配实验样品中的荧光团。用户不再需要调整实验以适应共聚焦系统上可用的激光线；该可调谐白光激光系统能适应特殊的实验要求。目前，弗吉尼亚大学还没有其他系统具有可调谐（470-670 nm）白光激光共聚焦/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关联研究感兴趣的研究人员也至关重要。KCCI因其每年一次的实践培训课程而获得认可，该课程在弗吉尼亚大学举办了七年。迄今为止，有来自美国和23个国家的304名科学家参加了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无法获得的成像能力水平，这将大大加强正在进行和未来的研究。公共卫生相关性：通过许多不同颜色的荧光蛋白，可以对活细胞内的生物事件进行非侵入性检测，但这种方法需要高度适应性的显微镜系统。所要求的徕卡TCS SP5X可调谐白光激光系统是一种独特的系统，允许用户调整激发和发射波长，以准确匹配实验样品中的荧光团。如果我们必须了解疾病过程和设计治疗策略，继续发展这些活细胞成像技术是很重要的，不同颜色荧光蛋白和要求的徕卡SP5X的组合代表了这一领域的重大进步。

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.