Development of confocal microscope using random mask and active filter

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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.