Development of confocal microscope using random mask and active filter

本研究的目的是在作者以前发展的莫尔荧光显微镜的基础上，利用常规光源构建“明亮”的共焦成像系统。对三维光学传递函数进行了理论分析，相对于Petran型真实的时间荧光共聚焦显微镜，其在Nipkow盘上具有随机图案掩模而不是常规针孔阵列。分析表明，在使用随机掩模图案的情况下，“亮”共焦图像中的常规图像分量的混合是不可避免的。因此，使用以前开发的真实的时间有源3D滤波器（真实的时间3D有源卷积器）是非常可取的，以消除传统的图像分量。共聚焦荧光显微镜系统（基于商业Petran型共聚焦荧光显微镜，K2-BIO）装备的真实的时间3D有源卷积器和相关的控制电路的开发已在下一阶段（1993年）完成。从目前的评估实验中已经证实了在使用3D主动卷积器时3D成像特性的改善，尽管所应用的3D加权函数不适合于此时的真实的时间处理。因此，我们提出了另一种方法，用于消除传统的图像分量，组合的空间分裂的替代掩模模式（随机和均匀）的Nipkow盘和真实的时间图像相减。这种方法的可能性也已在目前的评价实验中得到证实。

The present study has aimed to construct 'bright' confocal imaging system using conventional light source, under the basis of the moire fluorescent microscopy previously developed by authors.At the first stage (1992), theoretical analysis has been carried out for the 3-dimensional optical transfer function (3D OTF) with respect to the Petran type real time fluorescent confocal microscope with a random pattern mask instead of the conventional pin hole array on the Nipkow disk. The analysis shows that mixing of the conventional image components in the 'bright' confocal image is unavoidable under the usage of the random mask pattern. Therefore, the use of the previously developed real time active 3D filter (real time 3D active convolver) is highly desirable to eliminate the conventional image components.The development of the confocal fluorescent microscope system (based on the commercial Petran type confocal fluorescent microscope, K2-BIO) quipped the real time 3D active convolver and related control circuits has been done at the next stage (1993). Improvement of the 3D imaging characteristics have been confirmed at the usage of the 3D active convolver from present evaluation experiments, although the applied 3D weighting function is not suitable for the real time processing in this time. We have, therefore, proposed another approach for the elimination of the conventional image components, combination of a spatially split alternative mask pattern (random and uniform) for the Nipkow disk and the real time image subtraction. The possibility of this approach has been also confirmed at the present evaluation experiments.