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The small Raman scattering cross section of hemoglobin (Hb) molecules limits its application using a Raman spectroscopy based optical biosensor. Label-free surface enhanced Raman scattering (SERS) detection and degradation of Hb have been achieved using 3D reusable superhydrophobic SERS substrates based on a Ag/ZnO/Ag hybrid structure. The fabrication process follows the decoration of thermally evaporated non-spherical like Ag nanoparticles on hydrothermally grown ZnO nanorods on a catalytic ultra-thin Ag film. From SERS point of view, these 3D SERS substrates exhibit four important characteristics such as a higher surface to volume ratio, surface plasmon resonance in the broad wavelength region of the visible spectrum, a strong electric field at the Ag–ZnO interface due to the formation of a Schottky barrier, and the superhydrophobic surface. The SERS substrates not only performed an outstanding Raman enhancement effect due to the above factors but also displayed multiple recyclabilities owing to their excellent self-cleaning property via a UV light assisted photocatalytic degradation process. The quantitative SERS analysis has been performed by a linear regression method and resulted in 10−13.42M and 10−7.24M limit of detection for Rhodamine 6G (Rh6G) and Hb molecules, respectively, with an enhancement factor of 6 × 1011. The effect of the 3D hybrid structure toward higher SERS activity has been compared with that of 2D SERS substrates, and the SERS mapping of Rh6G molecules proves good homogeneity of the 3D SERS substrates. These ultra-sensitive 3D SERS substrates with reusable capability open the possibility of their use toward biosensors for the early detection of diseases.

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