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Indium-tin-oxide (ITO) nanoparticles were annealed in vacuum or reducing atmosphere to obtain different surface structures and investigate their influence on the adsorptive character and conformation of cytochrome c (Cyt c) molecule. Annealing-induced morphometric or structural changes of ITO nanoparticles were characterized by instruments of transmission electron microscopy, x-ray diffraction, and Raman scattering. Semiconductor ITO nanoparticle-enhanced Raman scattering of Cyt c was observed and the enhanced efficiency was found to closely depend on the surface structures which control the adsorbance of buffer anions needed for Cyt c loading. Direct electron transfer between Cyt c and ITO surface at the moment of molecular elastic collision was found and a reverse electron transfer process for O-terminated surface and metal-terminated surface was observed, according to absorption spectroscopic measurement on the residual solution.

This work reports an efficient method to fabricate large-area flexible substrates for surface enhanced Raman scattering (SERS) application. Our technique is based on a single-step direct imprint process via porous anodic alumina stamps. Periodic hexagonal arrangements of porous anodic alumina stamps are transferred to the polyethylene terephthalate substrates by mechanically printing process. Printed nanocaps will turn into “hot spots” for electromagnetic enhancement with a deposited gold film by high vacuum evaporation. The gaps between the nanocaps are controllable with a tight correspondence to the thickness of the deposited gold, which dramatically influence the enhancement factor. After covered with a single-layer graphene sheet, the gold nanocap substrate can be further optimized with an extra enhancement of Raman signals, and it is available for the trace detection of probe molecules. This convenient, simple, and low-cost method of making flexible SERS-active substrates potentially opens a way towards biochemical analysis and disease detection.