Moire super resolution fluorescent microscopy based on active image processing

本研究旨在建立新提出的莫尔超分辨荧光显微镜的理论背景。原型系统的构建和成像特性的评价也是本研究的重要内容。在第一阶段(1988)，我们开展了将莫尔超分辨技术应用于使用一维和二维投影型掩模的荧光显微镜的理论分析。分析表明，莫尔荧光显微镜具有面内超分辨率和光轴方向真3D分辨率的特点，这与共焦荧光显微镜的特性相似。接下来，通过在常规荧光显微镜上安装可移动平台和莫尔/重建掩模，构建了一个原型系统。基于活动图像Proc…的一维蒙版重建实验，基于活动图像处理的蒙版双动法更多的技术，已经尝试到生物样本。通过本实验，很好地证实了光轴方向的真三维分辨率。另一方面，人们还发现，目前的莫尔荧光显微镜非常灵敏，导致莫尔/重建掩模之间的对比度反转。对这种影响进行了分析和修正，并进行了二维掩模重建实验，验证了下一阶段()的真实三维分辨率。通过计算观测到的低谷散焦序列的三维光谱，对三维光学传递函数进行了评价。为了提高光学系统的亮度，引入了位相掩模来代替原来的幅度掩模，虽然超分辨率的确认还有待于未来的研究，但目前的莫尔荧光显微镜有望在生物医学领域中成为有用的手段。较少

The present study has aimed to establish the theoretical background of the newly proposed moire super resolution fluorescent microscopy. Construction of the prototype system and the evaluation of the imaging characteristics have been also important subject of this study.At the first stage ( 1988 ), theoretical analysis has been developed to apply the moire super resolution technique to the fluorescent microscopy, which uses both 1-dimensional and 2-dimensional projection type masks. The analysis shows that the moire fluorescent microscopy has the characteristics of the in-plane super resolution and the true 3D resolution along the optical axis, which are similar to the characteristics of the confocal fluorescent microscope. Next, a prototype system has been constructed by attaching movable stages and moire/reconstruction masks to a conventional fluorescent microscope. Reconstruction experiment using 1-dimensional masks, under the mask double moving method based on the active image proc … More essing technique, has been attempted to biotic samples. From this experiment, the true 3D resolution along the optical axis has been well confirmed. On the other hand, it has been also found that the present moire fluorescent microscope is very sensitive and results in the contrast inversion with the misalignment between moire/reconstruction masks. The analysis and the proposal of the correction with this effect have been presented.The reconstruction experiment using 2-dimensional masks has been achieved and also confirmed the true 3D resolution at the next stage ( 1989 ) of the study. The evaluation of 3D OTF has been attempted by calculating the 3D spectra from observed trough defocus series. To increase brightness of the optical system, a phase mask has been introduced instead of the original amplitude mask.Although the confirmation of the super resolution rests in the future study in this time, the present moire fluorescent microscopy is expected to be useful means in the field of the biomedical application. Less