拉曼显微成像技术无需样本制备，具有无损、无创、对水溶液不敏感的优点，可在微米或纳米尺度下表征样本的生化组分及分布，成为生命科学领域重要的研究工具。随着对复杂生物样本研究的不断深入，拉曼显微成像也被期待能够实现对生物样本中的分子组成与分布的动态立体观测。首先，系统性地梳理近年来三维拉曼显微成像技术的研究进展，包括基于自发拉曼散射、相干拉曼散射、表面增强拉曼散射以及拉曼标签的不同三维成像方法的技术手段、改进策略与实验结果。然后，总结了不同成像技术在细胞生物学、发育生物学等方面的应用进展。最后展望了不同三维拉曼显微成像技术在生物医学光学显微成像技术应用中所面临的挑战和发展前景。

Raman microscopic imaging has emerged as an important research tool in life sciences because it does not require sample preparation and is non-destructive, non-invasive, and insensitive to aqueous solutions. It enables the characterization of biochemical components and sample distributions at a micrometer or nanometer scale. As the research on complex biological samples increases, Raman microscopic imaging holds potential for the dynamic stereoscopic observation of molecular composition and distribution in biological samples. This paper systematically examined recent advancements in 3D Raman microscopic imaging, including technical approaches, improvement strategies, and experimental results of various 3D imaging methods based on spontaneous Raman scattering, coherent Raman scattering, surface-enhanced Raman scattering, and Raman tags. Further, the progress of the applications of different imaging techniques in cell biology and developmental biology was summarized. Finally, the challenges and development prospects of different 3D Raman microscopic imaging techniques in the application of biomedical optical microscopic imaging technology were outlined.