In this paper, a depth-sensing method employing active irradiation of a semi-annular beam is proposed for observing the multi-layered reflective surfaces of transparent samples with higher resolutions and lower interference. To obtain the focusing resolution of the semi-annular aperture diaphragm system, a model for computing the diffracted optical energy distribution of an asymmetric aperture diaphragm is constructed, and mathematical formulas are deduced for determining the system resolution based on the position of the first dark ring of the amplitude distribution. Optical simulations were performed under specific conditions; the lateral resolution δr of the depth-sensing system was determined to be 0.68 μm, and the focusing accuracy δz was determined to be 0.60 μm. An experimental platform was established under the same conditions, and the results were in accord with those of the simulation results, which validated the correctness of the formula for calculating the amplitude distribution of the diffracted light from the asymmetric aperture diaphragm.
Keywords: depth-sensing system; microscopy; semi-annular diaphragm; transparent biological samples.