Abstract: With the advantages of high resolution, high sensitivity, high molecule specificity and non-invasive, fluorescence microscopy can characterize the morphological and molecular functional information of samples at the micron or even nanometer scale, making it an important tool for life science research. As microbiology research continues to advance, fluorescence microscopy is expected to provide dynamic and 3D observation of microscopic biological structures and molecular events. This paper systematically reviews the research progress of fast 3D fluorescence microscopy in recent years, including the main technical means, improvement strategies and representative research results of point-scan imaging, wide-field imaging, and projection tomography to improve the imaging speed, expand the imaging dimension and enhance the imaging quality. In the end, we look forward to the future challenges and prospects of fast 3D fluorescence microscopic imaging techonology.