The new co-fluorescence enhancement system was found in polymer matrix and studied with a combinatorial approach based on Re(DBM)3phen (Re3+: Tb3+, La3+, Gd3+, Y3+; dbm: dibenzoylmethane; phen: phenanthroline) and Sm3+ complex co-doped poly(methyl methacrylate)(PMMA). We have presents a new methodology for the rapid optimization of the luminescence conditions of thin-film sample in arrays of microwells. Two libraries were generated in order to study the effect of the species and content of enhancing ions on the luminescence enhancing efficiency, respectively. At the optimal content of 20 wt.% Sm(DBM)3phen, the maximum sensitization efficiency of Tb(DBM)3phen is about nine times. The intramolecular and intermolecular energy transfer processes in rare-earth complex-doped PMMA are discussed. The energy transfer processes make the co-fluorescence effect come true. Although, the methodology described here was implemented for optimization of co-fluorescence conditions, it is can be further implemented for a variety of applications in which optimization of parameters can be studied in situ using spectroscopic tool.