IMAGING FACILITY FOR NEUROBIOLOGY

在目前的申请中，该部门的五名教员 亚利桑那大学的神经生物学试图建立一种 最先进的生物成像设备，以加强他们对 实验有利因素的发展及其功能组织 昆虫的神经系统。该司目前的成像设施，配备了工作人员 由一名全职助理科学家，拥有一台Bio-Rad MRC-600激光器 扫描共焦显微镜，这是这五个主要用户 提案大量用于美国国立卫生研究院资助的研究。MRC-600效率低下 与较新的共焦系统相比，它是如此流行，以至于现在人们不得不 提前3-4周报名，制作共焦显微镜 这是我们大多数实验中的限速步骤。此外，还有一些 共聚焦显微镜项目已经发展到了另一种情况 分辨率，可视化荧光标记的结构的能力 在广泛的亮度范围内，以及成像生活的能力 在非常弱的光照下进行长时间的准备工作将 加大力度。因此，PI建议对MRC-600进行升级 共焦显微镜到MRC-1024，并购买去卷积 显微镜系统。共焦升级将提供更大的像素 用于更大视场、更多滤镜集和三个滤镜的阵列 用于改进多个荧光团检测的光电倍增管， 以及更高效的计算机和软件包。反卷积，而 表面上类似于共聚焦显微镜的产生能力 明显高于标准分辨率的荧光图像 荧光显微镜，与共聚焦显微镜的区别在于 重要方法：不要用针孔来拒绝失焦的光线 在探测器中，它收集尽可能多的来自 物镜，并用它来数学重建明亮的 对象；而不是使用光电倍增管，因为光电倍增管噪音大， 其输出与输入不是线性相关的，它使用ccd摄像机来 敏感、线性的报道。因此，反卷积系统将满足 我们许多新出现的需求。

With the current application, five faculty in the Division of Neurobiology at the University of Arizona seek to establish a state-of-the-art biological imaging facility to enhance their studies of the development and functional organization of experimentally favorable insect nervous systems. The Division's current Imaging Facility, staffed by a full-time Assistant Staff Scientist, has a Bio-Rad MRC-600 laser scanning confocal microscope, which the five Major Users of this proposal use heavily for NIH-funded research. The MRC-600 is inefficient compared to newer confocal systems and is so popular that one now has to sign up for time 3 - 4 weeks in advance, making the confocal microscope the rate-limiting step in most of our experiments. Moreover, some confocal-microscopy projects have advanced to the point where additional resolution, the capability to visualize fluorescently labeled structures across a broad range of brightness, and the capability to image living preparations over long time periods under very low light levels would enhance our efforts. Therefore, the PI proposes to upgrade the MRC-600 confocal microscope to a MRC-1024, and to purchase a deconvolution microscopy system. The confocal upgrade will provide a larger pixel array for larger fields of view, more filter sets and three photomultiplier tubes for improved detection of multiple fluorophores, and a more efficient computer and software package. Deconvolution, while superficially similar to confocal microscopy in its ability to produce fluorescence images of significantly higher resolution than a standard fluorescence microscope, differs from the confocal microscope in two important ways: rather than rejecting out-of-focus light with a pinhole in the detector, it collects as much light as possible from the objective lenses and uses it to mathematically reconstruct the bright object; and rather than using a photomultiplier tube, which is noisy and whose output is not linearly related to input, it uses a CCD camera for sensitive, linear reporting. Thus, the deconvolution system will meet many of our emerging needs.