RECONSTRUCTION OF PARTIALLY CONFOCAL IMAGES

共焦扫描显微镜(CSM)被广泛应用于三维成像 固定标本的可视化。然而，活的标本通常是 被所需的高强度激发光损坏和/或漂白 由CSM提供。此外，活标本中的时间变化通常也是如此 迅速被CSM成像。这严重限制了 CSM在活体标本成像中的应用。海流 对活体标本成像的首选技术是计算光学 分段(COSM)，它具有更大的集光能力 并允许以较少的光漂白进行快速图像收集，但受到 沿光轴的分辨率严重降低。这 光轴分辨率的下降可以部分地 通过计算密集的恢复方法进行补偿。 部分共焦扫描显微镜(PCSM)可以获得更快的图像 以降低光轴分辨率为代价进行记录。长的- 这里提出的研究的术语目标是使用计算方法 以找回丢失的光轴分辨率。通过组合使用 PCSM和计算方法，将有可能想象生活 样品的分辨率可与或优于 严格的共焦显微镜，计算时间短得多 比非共焦图像的COSM所要求的要高。 这一目标将实现如下：第一，现有的两个PCSM， 一种转盘PCSM和单缝PCSM，其特征在于 推导并验证每一个的理论点扩展函数。 这些显微镜的设计中的潜在限制将是 决心用金牌来提高自己的表现。第二, 为COSM开发的恢复方法将应用于PCSM和 他们负担得起的业绩增长将是其特征。第三，在 与华盛顿大学的同事合作，设计了一种 转盘PCSM将利用第一次试验中获得的信息进行改进 明确目标。最后，为了实现这些目标，轮换 必须对磁盘PCSM进行修改，以精确控制这三个组件的位置 轴(x、y和z)。

Confocal scanning microscopy (CSM) is widely used for three-dimensional visualization of fixed specimens. However, living specimens are often damaged and/or bleached by the high intensity excitation light required by CSM. Moreover, temporal changes in a living specimen are often too rapid to be imaged by a CSM. This has severely limited the applications of CSM to the imaging of living specimens. The current technique of choice for imaging living specimens is computational optical sectioning (COSM), which has a much greater light-collecting capability and permits rapid image collection with less photobleaching, but suffers from severely degraded resolution along the optical-axis. This degradation in optical axis resolution can be partially compensated for via computationally intensive restoration methods. Partially confocal scanning microscopes (PCSMs) allow faster image recording at the expense of degraded optical-axis resolution. The long- term goal of the research proposed here is to use computational methods to recover this lost optical-axis resolution. With the combination of PCSM and computational methods, it will be possible to image living specimens at resolutions comparable to or better than those of strictly confocal microscopes and with computation times much shorter than those required by COSM of non-confocal images. This goal will be achieved as follows: First, the two available PCSMs, a rotating disk PCSM and a single-slit PCSM, will be characterized by deriving and validating a theoretical point spread function for each. Potential limitations in the design of these microscopes will be determined with a gold to improving their performance. Second, restoration methods developed for COSM will be applied to PCSMs and the gain in performance that they afford will be characterized. Third, in collaboration with colleagues at Washington University, the design of a rotating-disk PCSM will be improved using information gained in the first specific aim. Finally, in order to achieve these goals, the rotating disk PCSM must be modified for precise position control over the three axes (x, y, and z).