As the basic structural and functional unit of life, cells have very important research significance in biology, medicine and other fields. With the development of modern science and technology, scientists have a very clear understanding of the spatial structure of cells and organelles with the help of electron microscopes. However, very little is known about their functions and the interactions between cells, and this is precisely the information that disease treatment and drug development urgently need to know. Therefore, the study of subcellular organelles in vitro living cells and in vivo living cells have become very important. However, the structure of many organelles in living cells are at the nanoscale. Traditional optical imaging techniques cannot observe nanoscale biological structures due to the limitation of the optical diffraction limit. Therefore, optical super-resolution imaging technology is an effective tool to study the structure and function of subcellular organelles. Among all optical super-resolution microscopy techniques, stimulated emission depletion (STED) super-resolution imaging technology has the capabilities of real-time, three-dimensional super-resolution and tomographic imaging. Therefore, the STED is very suitable for nanoscale live cell and vivo imaging studies. Moreover, STED super-resolution imaging technology has been widely used for super-resolution dynamic observation of living cells and even living mouse cells after decades of development. This paper summarized the research progress of STED super-resolution imaging in the fields of in vitro living cells and in vivo mouse neurons in recent years, and introduces the development status of fluorescent dyes and fluorescent proteins for STED super-resolution imaging of in vitro and in vivo living cells. In vivo cells super-resolution imaging is very meaningful for understanding the nature of cells, but it has been more than 20 years since the STED super-resolution imaging technology was proposed to the present, and there are still very few literatures on in vivo cells super-resolution imaging. The main problem is that very few probes are available for in vivo cells super-resolution imaging. At the same time, the depth of in vivo cells super-resolution imaging is also very limited, mainly due to the lack of fluorescent probes in the infrared band. Finally, the future application prospects of in vivo cells super-resolution imaging are prospected.