To address the negative impact of moving samples on image quality and sample integrity in traditional light-sheet microscopy, a three-dimensional light-sheet imaging system incorporating a liquid zoom lens and galvanometer mirror is designed and constructed. Imaging experiments are performed on opaquestandard matte ceramic spheres, 3D printed translucent polylactic acid (PLA) samples, fluorescent microspheres, and apple pollen. The light-sheet microscopy three-dimensional imaging system is used to obtain images of different sections of the samples. By adjusting the galvanometer mirror and liquid zoom lens, a series of changing images of the samples were obtained. The feasibility of using scanning galvanometer mirror and liquid zoom lens to achieve 3D imaging of immovable samples is verified. The images of the sections are reconstructed in three dimensions to achieve the three-dimensional imaging of the samples. Extract, normalize, and fit the brightness information of fluorescent microsphere images.The experimental results of fluorescent microspheres and apple pollen show that the system has a lateral resolution of 10 μm and an axial resolution of 19 μm.