Raman discrimination of bacterial strains using multilayered microcavity substrates

Surface-enhanced Raman scattering (SERS) utilizing colloidal silver and gold has been demonstrated to provide a rapid means of measuring the Raman spectra of microorganisms in the fingerprint region. In this study, we have introduced microcavity substrates coated with alternating layers of silver and gold thin films for measuring the Raman spectra of four strains of E. coli. These microcavitiy substrates have been prepared by placing glass microspheres between two polished aluminum substrates and pressing them together using a standard lab press. After removing the glass microspheres from the substrates, the substrates have been coated with 15 to 70 nm thick films of chromium, silver and gold in a precise order. The cavities were evaluated for SERS enhancement by measuring Raman spectra of dilute rhodamine 6G (R6G) down to 10-8 M. With these microcavities, we have investigated the SERS spectra of four chemically competent strains of E. coli (One Shot OmniMAX 2-T1, Mach1-T1, Stbl3, and TOP10). Replicate SERS spectra of all the four e-coli strains show excellent reproducibility. Visual examination of the spectra, however, reveals differences in the spectra of these strains. To confirm this observation, we have used multivariate analysis for positive identification and discrimination between the strains.