Monitoring the Kinetics ofBacillus subtilisEndospore Germination via Surface-Enhanced Raman Scattering Spectroscopy

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Abstract
Mirror sandwich SERS substrates (M3S) were used to monitor the kinetics of Bacillus subtilis endospore germination. The sandwich configuration of the substrates allows real-time observation of germination in samples that contained only several hundred endospores. The enhancement provided by the substrates is attributed to the enhanced local electromagnetic field that originates from coupling between the Ag nanoparticles and the underlying metal film as well as from coupling between the Ag nanoparticles themselves. The germination kinetics at varying concentrations of l-alanine and different temperatures were studied by monitoring the intensity and growth of the Raman peak at 1010 cm-1, which is characteristic of dipicolinic acid. A total of four concentrations (50, 75, 100, and 150 mM) of l-alanine and three different temperatures (30, 37, and 55 °C) were investigated.