In photosynthetic eukaryotes, photosynthesis occurs in the chloroplast, and more specifically in the thylakoid membranes. The thylakoids consist of two main domains: the grana, which are piles of membranes gathered together by stacking forces, and the stroma-lamellae, which are unstacked thylakoids connecting the grana. The major photosynthetic complexes are unevenly distributed, within these compartments, due to steric and electrostatic constraints and to their role in the photosynthetic process. To gain information on the sub-thylakoid distribution of these proteins (i.e. in the grana and in the stroma-lamellae) we combined experimental approaches to perform a complete survey of the protein composition of these thylakoid sub-compartments using i) thylakoid membrane fractionations to improve the dynamic range detection of minor thylakoid proteins and ii) a quantitative proteomic approach coupled to a dedicated data analysis pipeline and manual annotation to differentiate genuine grana and stroma-lamellae proteins from possible contaminants. Besides confirming previous information on the localization of photosynthetic proteins, these experiments provide some unexpected results, and allow revisiting the composition of the thylakoid micro-domains.