An autofocusing method for two-wavelength digital holographic microscopy (TWDHM) based on the wavelength dependence of the diffraction process is proposed. Red and green lights are employed for the illumination of the TWDHM, and the generated holograms are recorded simultaneously by a color CCD camera. Due to the wavelength dependency of the diffraction process, the farther the reconstruction plane is from the image plane, the larger the difference is between the red and green light distributions. Thus, the image plane can be determined by finding the minimum of the variation between the red and green lights on their amplitude distributions. The feasibility of the proposed method is demonstrated by simulation and experiment.
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