Hematopoietic stem cells(HSCs)in the bone marrow(BM)are maintained in distinct microenvironments called niches. Technological advances in in vivo imaging have enabled dynamic analyses of BM cells. This in vivo imaging can be a key tool to elucidate the mechanisms underlying HSC maintenance in the BM through analysis of HSC motility and evaluation of the relationship between HSCs and their niche factors over time. Furthermore, application of this imaging technology to leukemia research can lead to new discoveries in leukemic stem cell maintenance and the development of novel antileukemic drugs. In this review, we summarize recent advances in dynamic analyses of HSCs and leukemic cells in the BM by intravital imaging.