Silver film over nanospheres (AgFONs) were successfully employed as surface-enhanced Raman spectroscopy (SERS) substrates to characterize several artists' red dyes including: alizarin, purpurin, carminic acid, cochineal, and lac dye. Spectra were collected on sample volumes (1 × 10<sup>–6</sup> M or 15 ng/μL) similar to those that would be found in a museum setting and were found to be higher in resolution and consistency than those collected on silver island films (AgIFs). In fact, to the best of the authors' knowledge, this work presents the highest resolution spectrum of the artists' material cochineal to date. In order to determine an optimized SERS system for dye identification, experiments were conducted in which laser excitation wavelengths were matched with correlating AgFON localized surface plasmon resonance (LSPR) maxima. Enhancements of approximately two orders of magnitude were seen when resonance SERS conditions were met in comparison to non-resonance SERS conditions. Finally, because most samples collected in a museum contain multiple dyestuffs, AgFONs were employed to simultaneously identify individual dyes within several dye mixtures. These results indicate that AgFONs have great potential to be used to identify not only real artwork samples containing a single dye but also samples containing dyes mixtures.

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