Multiplexing with SERS labels using mixed SAMs of Raman reporter molecules

Surface-enhanced Raman scattering (SERS) offers a tremendous multiplexing capacity for the selective detection of biomolecules in targeted research. SERS labels comprising self-assembled monolayers (SAMs) of Raman reporter molecules on the surface of metal nanoparticles are sensitive and robust probes. Advantages of a SAM include maximum sensitivity, minimal unwanted co-adsorption of molecules from the surroundings, and reproducible SERS spectra with only few dominant Raman bands--all of these independent of a particular SERS substrate. We demonstrate experimentally how to increase the multiplexing capacity of SERS labels by using mixed SAMs with up to three different Raman reporter molecules on the surface of the metal colloid. Type and stoichiometry of a particular Raman label in a multi-component SAM are additional parameters compared with one-component SAMs. All one-, two-, and three-component SAMs on gold nanospheres can be easily discriminated, either by their original SERS spectra or the corresponding bar codes.