Ultra-resolution optical microscopy, which breaks through the diffraction limit, is typically used to observe structural characteristics and interactions of subcells. This method has great significance for the study of genomes and tackling major diseases. This paper begins by introducing the working principles of four typical super-resolution microscopic imaging techniques. Subsequently, research progress in the areas of multi-color fluorescence imaging and three-dimensional imaging is emphasized. Finally, recent applications of super-resolution optical imaging for cell activity observation, bacterial cell research, and cytoskeleton observation are reviewed both domestically and abroad. The main factors reportedly affecting the imaging quality are poor light stability of the fluorescent protein, low light activation rate, and weak fluorescence intensity. Solution of the above problems will lead to the widespread use of super-resolution optical imaging for the three-dimensional imaging of thick samples, multi-color fluorescence imaging, and fast imaging of living cells, ultimately furthering the development of life science and materials science.