Frequency-domain Fluorescence Lifetime Imaging Microscopy (FD-FLIM)

描述（由申请人提供）：W.M.凯克细胞成像中心（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，正在申请“频域荧光寿命成像显微镜（FD-FLIM）”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7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FD-FLIM上进行的研究的中心主题是研究活细胞和其他细胞成像中的蛋白质-蛋白质相互作用，包括IQGAP和阿尔茨海默病的细胞生物学，新FRET参考标准的表征，谷氨酰胺FRET传感器，RNA-蛋白质相互作用，以及监测聚合物中的氧气，pH传感器。所要求的仪器，共焦FD-FLIM系统是独一无二的，为用户提供了调谐激发波长（通过软件）的能力，以准确匹配他们的实验样品中的荧光团。用户不再需要根据FD-FLIM系统上可用的激光线调整实验;计算机化的单色可调谐超连续激光系统可以适应特定的实验。目前在弗吉尼亚大学还没有一个系统具有可调谐（400-2500 nm）激光FD-FLIM成像能力，并在几秒钟内收集FLIM图像。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FRET课程，在过去八年中在弗吉尼亚大学举行。迄今为止，来自美国的224名科学家（336名注册）以及来自25个国家的科学家参加了FRET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。

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 "Frequency-domain Fluorescence Lifetime Imaging Microscopy (FD-FLIM)" 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 7 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 FD-FLIM is the study of protein-protein interactions in live cells and other cellular imaging including IQGAPs and the Cell Biology of Alzheimer's Disease, characterization of new FRET reference standards, glutamine FRET sensors, RNA-protein interactions, and monitoring oxygen, pH sensors in polymers. The requested instrument, confocal FD-FLIM system is unique and, providing users the ability to tune the excitation wavelengths (through software) 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 FD-FLIM system; the computerized monochromator tunable supercontimuum laser system adapts to the particular experiment. There is currently no system available at the University of Virginia that has a tunable (400-2500 nm) laser FD-FLIM imaging capability and collects FLIM images in few seconds. 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 FRET course, held for the past eight years at the University of Virginia. To date, 224 scientists (336 registered), from the United States as well as scientists from 25 countries have participated in the FRET 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.