DELTAVISION MICROSCOPE SYSTEM

这项提案要求支持华盛顿大学现有的共享成像资源Keck成像设备购买一台名为Deltavision的高分辨率光学显微镜。Deltavision是指一种集成的显微镜系统，它通过数学去卷积来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获得的最高分辨率(到75 nm点源检测)，用户详细说明的应用表明对该分辨率的明显需求。量化是非常好的，因为Deltavision提供了线性范围的数据采集和显示，比共焦显微镜大10倍。此外，使用低水平紫外光照射来激发样品，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/或使用活细胞记录技术监测细胞死亡。由于使用的是高能激光，后一种效应是共焦显微镜的显著缺点。由于一系列滤光片与UV光源结合使用，因此采样是多波长的，不受激光线路等限制。最后，配备精密工作台微操作器和Deltavision软件的显微镜提供了重建三维生物样本的最佳机会。NIH资助的使用者和其他资助团体建议使用显微镜观察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、神经递质的运输和分泌、神经肌肉连接的组织以及毛细胞结构。这种广泛的用途表明了该仪器的许多应用，其他研究人员将通过凯克中心获得这些应用。描述了设备的操作和管理、内部咨询委员会以及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.