Functions of immune cells are linked to metabolic profiles, which are affected by changes in the tissue microenvironment. Focusing on bone regeneration after injury, we aim to understand how the microenvironment affects the metabolism of myeloid cells in the bone marrow over time, and how metabolic profiles impact on the function of those cells. We developed a lens implant for longitudinal intravital imaging of the mouse femur, to enable longitudinal imaging at subcellular resolution, allowing us to analyze cellular dynamics in the fracture gap over the whole time course of bone healing. Recently, we combined this with NAD(P)H-fluorescence lifetime imaging, a label-free technique which enables metabolic profiling of single myeloid cells in vivo. Our studies demonstrate a high degree of spatiotemporal metabolic flexibility of myeloid cells over the course of regeneration.