The electrospinning of rhodamine B (RhB)-doped poly(methyl methacrylate) (PMMA) is presented as a promising technique for processing photoluminescent nanofibers. Nanofibers with concentrations of RhB in PMMA between 0.05 and 2.0 wt. % were studied. The FTIR spectrum, fiber morphology, thermal properties and optical properties of the nanofibers were investigated. Smooth, bead-free and nonporous nanofibers were obtained. Differential scanning calorimetry (DSC) results revealed that the glass transition temperature of the PMMA (T-g) increased with the increasing of RhB content up to 0.5 wt. % and thereafter decreased. The laser induced fluorescence (LIF) technique was used to study the fluorescence properties of the nanofibers. The maximum fluorescence emission was observed at a concentration of RhB of 0.2 wt. %.