Fluorescence microscopy is a vital tool in biomedical research, enabling high-resolution imaging by using fluorescent dyes or proteins to label specific cells or molecules, which then emit fluorescence under a microscope. Light-sheet fluorescence microscopy (LSFM) is an emerging three-dimensional imaging technology that achieves high-throughput, high-resolution 3D imaging by rapidly scanning thin samples. This technique offers advantages such as low phototoxicity and photobleaching, high photon efficiency, fast imaging speed, and high resolution. It is widely used in fields such as neuroscience, cell biology, and pathology. LSFM shows great potential in three-dimensional pathological analysis. Unlike traditional two-dimensional pathology slides, 3D pathological analysis provides complete spatial information on tissue structures, aiding in a more comprehensive understanding of disease mechanisms. The development of 3D pathological analysis significantly advances pathological research and clinical diagnostics, offering strong support for early disease detection, precise treatment, and personalized medicine. We first introduce the development of light-sheet microscopy and its applications in the pathological field, then discuss the main current approaches and methodologies for 3D pathological analysis. We focus on the potential applications of emerging multimodal large language models in pathological analysis.