Many studies have revealed that Macrophages play an important role in the pathogenesis of many diseases，and different phenotypes and functions can be obtained on exposure to different tissue environ⁃ ments，especially in tumor tissues. Previously known surface markers cannot distinguish macrophage subsets in tumors. The recent rapid development of mass cytometry and single⁃cell RNA sequencing，has made it possible to comprehensively map and study various cells in tumors and their phenotypes and functions. In this review，we summarized recent advances applying these technologies in defining macrophage subsets，further discussed the distinctive characteristic phenotypes and function of tumor ⁃associated macrophages in tumors and searched for new biological markers with therapeutic or prognostic potential to provide a theoretical basis for clinical immunotherapy of tumors in the future.