Very monodisperse water-soluble silica core-surfactant shell nanoparticles (SCSS NPs) doped with a rhodamine B derivative were prepared using micelles of F127 as nanoreactors for the hydrolysis and condensation of the silica precursor tetraethoxysilane (TEOS) The functionalization of the rhodamines with a triethoxysilane group allowed the covalent binding of the fluorophores to the silica core no leaking of the dye was observed when the NPs were purified either by ultrafiltration (UF) or dialysis The diameter of the core (d(c) = 10 +/- 1 nm) was determined by TEM and subtracted from the hydrodynamic diameter, measured by DLS, (d(H) = 24 nm PdI = 0 1) to calculate the shell thickness (similar to 7 nm) The presence of a single population of NPs with a radius compatible with the one measured by DLS after UF was confirmed by AF4-MALS-RI measurements The concentration of the NPs was measured by MALS-RI This allowed us to determine the average number of rhodamine molecules per NP (10) The ability of the NPs to host hydrophobic species as cyanines in the SS was confirmed by fluorescence anisotropy measurements Steady-state and time-resolved fluorescence measurements allowed us to observe the occurrence of a very efficient Forster resonance energy transfer process from the covalently linked rhodamines to the hosted cyanines In particular, the analysis of the TCSPC data and steady-state measurements revealed that the adsorption of a single cyanine molecule causes an almost complete quenching of the fluorescence of the NP Thanks to these observations, it was possible to easily determine the concentration of the NPs by fluorescence titration experiments Results are in good agreement with the concentration values obtained by MALS-RI Finally, the hosted cyanine molecule could be extracted with (+/-)-2-octanol, demonstrating the reversibility of the adsorption process