Focal depth extension and stereo viewing apparatus for high resolution microscope based on active image processing method

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A new technique of the focal depth extension for the high resolution optical microscope, with the aid of the active image processing method, has been proposed in the present study ( 1987 - 1988, Grant-in-Aid for Developmental Scientific Research 2 ).This focal depth extension technique is essentially based on Hausler's method ( 1972 )where moving defocus averaging and high pass filtering are applied to the observed microscopic image. The present technique, however, uses real time active deconvolution technique ( proposed by one of the authors ( 1985 ) as the active image processing method ) instead of the conventional high pass filter used in the Hausler's method. Main aims of this study are to construct a prototype focal depth extension system and to confirm its practicability.At the first stage of the study ( 1987 ), a fast 3D movable sample stage using piezo driving elements has been assembled and attached to the conventional optical microscope ( Olympus BHS ). Next, the previously constructed active image processing system ( include a frame integrator with bipolar weighting function and a personal computer ) has been connected to this sample stage via interface circuits. A stereo viewing subsystem using liquid crystal glasses has been also included in the active image processing system. By using this system, focal depth extension and stereo viewing experiments have been successfully achieved to transmission and reflection mode images of the microscope. Focal depth extension about 20 um defocus range has been obtained for the objective lens of N.A.= 0.7. Focal depth extension has been also applied to the phase microscope at the next stage ( 1988 ). Although the image formation of the phase microscope is some more complicated, successful focal depth extension and stereo viewing have been achieved with phase object by using a similar filtering function.