A honeycomb-shaped gold substrate was developed for surface-enhanced Raman imaging (SERI). The honeycombs are composed of clusters of 50鈥?0 nm gold nanoparticles and exhibit high Raman enhancement efficiency. An average surface enhancement factor (ASEF) of 1.7 脳 106 was estimated for a monolayer of l-cysteine molecules adsorbed to gold via a thiol linkage. The presence of a linear relationship in the low concentration region was observed in SERI detection of malachite green isothiocyanate (MGITC). These results together with the high reproducibility and simple and cost-effective fabrication of this substrate suggest that it has utility for applications of surface-enhanced Raman scattering (SERS) in quantitative diagnoses and analyte detection.