Two-photon microscopy (2P), coupled to fluorescence lifetime imaging microscopy (FLIM) is a stateof-the-art method in cancer, stem cell and developmental biology fields. It enables for deep tissue imaging (down to 100-200 um) and, coupled with genetically encoded, staining probes and autofluorescence, enables for non-destructive analysis of the living tissue morphology, extracellular matrix, physiological and metabolic activities. In particular, label-free optical metabolic imaging, based on 2P-FLIM, is currently the method of choice for studying immunometabolism, quality of the pre-implantation embryos, long-term imaging of adult stem cell-, iPSC-derived organoid cultures, and intravital high-resolution cancer imaging. This upgrade is essential for UGent-based core facilities and the applying consortium, as it will positively impact and boost capabilities of the Ghent Light Microscopy core, MREB3 and 4Brain core facilities and research activities at the several UGent faculties and UZ Gent, including CRIG, GATE Health and GGIG. The upgrade will enable to deepen and expand collaborations with other Flemish institutions, including VIB, VUB, UA, UHasselt and KU Leuven (Flanders Bio-Imaging) as well as international research and training networks, including the Euro-Bioimaging and Horizon-funded FLIMagin3D MSCA PhD network.