DELTAVISION MICROSCOPE SYSTEM

该提案要求支持为华盛顿大学现有的共享成像资源-凯克成像设施-购置一台名为Deltavision的高分辨率光学显微镜。Deltavision是一种集成的显微镜系统，通过数学解卷积实现荧光标记样品的高分辨率。获得的分辨率是光学显微镜可用的最高分辨率（75 nm点源检测），用户详细介绍的应用表明了对该分辨率的明确需求。定量是优秀的，因为Deltavision提供了一个线性范围的数据采集和显示10倍以上的共聚焦显微镜。此外，低水平UV光照明用于激发样品，最大限度地减少光漂白的影响，光漂白可能意味着信号完全丢失和/或用活细胞记录技术监测的细胞死亡。这些后者的影响是共焦显微镜的显着缺点，由于高能量的激光器。由于一系列滤光片与紫外光源结合使用，因此采样是多波长的，不受激光线等限制。最后，显微镜及其精确的载物台显微操作器和Deltavision软件为三维重建生物样品提供了最佳机会。NIH资助的用户和其他资助的团体建议使用显微镜来可视化活细胞中的着丝粒结构，活线粒体，胶原纤维结构，细胞核内的转录单位神经递质运输和分泌，神经肌肉接头的组织和毛细胞结构。这种广泛的用途表明，该仪器的许多应用将通过凯克中心提供给其他研究人员。描述了设备的操作和管理、内部咨询委员会和对Deltavision的机构支持。

This proposal requests support for the acquisition of a high resolution light microscope known as Deltavision to an existing shared imaging resource, the Keck Imaging Facility, at the University of Washington. Deltavision refers to an integrated microscopy system which achieves high resolution of fluorescently labeled samples by mathematical deconvolution. The resolution obtained is the highest available by light microscopy (to 75nm point source detection), and the applications detailed by the users demonstrates a clear need for this resolution. Quantitation is excellent, because Deltavision offers a linear range of data acquisition and display 10-fold greater than confocal microscopy. In addition, low level UV light illumination is used to excite the sample, minimizing the effects of photobleaching which can mean loss of the signal altogether and/or death of cells being monitored with live cell recording techniques. These latter effects are significant drawbacks of confocal microscopy, due to the high energy lasers employed. Because a range of filters are used in combination with the UV light source, sampling is multiwavelength and unlimited by constraints such as laser lines. Finally, the microscope with its precise stage micromanipulator and the Deltavision software offers the best opportunity to reconstruct biological samples in three dimensions. NIH-funded users and other funded groups propose to use the microscope to visualize centromere structure in living cells, living mitochondria, collagen fibril structure, transcription units within cell nuclei neurotransmitter transport and secretion, the organization of the neuromuscular junction, and hair cell structure. This broad range of uses indicates the many applications of the instrument which will be available to other investigators through the Keck Center. The operation and management of the equipment, the internal advisory committee, and institutional support for Deltavision are described.