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为解决光片显微镜成像视场范围小的问题,通过光束波前相位调制与图像拼接技术实现了光片显微镜对样本的宽视场成像。数值模拟了照明光束在波前相位调制后物镜聚焦面处的光强分布。搭建了光片显微镜光路系统,并对荧光微球、菊花花粉进行成像实验。采用片状照明光束对不同聚焦位置处的样本进行成像,再将图像进行剪裁、拼接得到宽视场成像结果。实验结果表明,488nm激发光通过数值孔径为0.3的物镜照明样本成像,可在保持光片厚度为0.81μm的情况下达到31.93μm的成像视场,视场扩展到原视场的3倍以上。仿真和实验表明,采用光束波前相位调制与图像拼接技术可在不损失层切能力的前提下扩展光片显微镜的成像视场。
In order to solve the problem of small field of view of light microscopy imaging, a wide field imaging of the specimen by light microscope is realized by the wavefront phase modulation and image splicing technique. The light intensity distribution of the illumination beam at the objective focal plane after wavefront phase modulation is numerically simulated. The optical system of optical microscope was built, and the imaging experiments of fluorescent microspheres and chrysanthemum pollen were carried out. The sheet illumination beam is used to image the samples at different focus positions, and then the images are cut and spliced to obtain a wide field imaging result. The experimental results show that 488 nm excitation light passes through an objective lens with a numerical aperture of 0.3 and can achieve an imaging field of 31.93 μm while maintaining the thickness of the light plate at 0.81 μm. The field of view extends to more than three times of the original field of view. The simulation and experiments show that using the wavefront phase modulation and image stitching technology can expand the imaging field of light microscope without losing the layer cutting ability.