Recent work with B. anthracis Delta Sterne spores demonstrated that Raman spectroscopy could be used to discriminate between viable and gamma deactivated spores and provided initial insight into the probable source of discrimination found in the spores. From this previous work, we believe through Raman spectral analyses of viable and deactivated spore samples, significant changes in spectral response can be resolved and ascribed to classes of biomolecules affected by the deactivation processes. We expanded upon this study to include four different Bacillus spores (B. anthracis, B. megaterium, B. thuringiensis, and B. atrophaeus) and probe de-activation techniques to include gamma radiation UV radiation, chemical, and thermal methodologies. We used sequential Raman imaging scanning electron (RISE) microscopy to determine chemical (Raman spectral information) and physical (SEM imaging) variance between viable and deactivated spore samples. Additional use of machine learning algorithms to
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