Two-photon microscope，as the most important optical microscope technology，showed remarkable advantages of 3D imaging，in vivo imaging，photobleaching and low phototoxicity.It has been utilized in cell imaging，tissue imaging and in vivo imaging related to traditional Chinese medicine (TCM) to provide a more scientific and intuitive theoretical basis for the treatment of related diseases by TCM.This paper summarizes the application research of the existing two-photon microscope technology in the field of TCM，in the hope of providing new directions for the research of TCM.