AOTF hyperspectral microscopic imaging for foodborne pathogenic bacteria detection

Hyperspectral microscope imaging (HMI) method which provides both spatial and spectral information can be effective for foodborne pathogen detection. The AOTF-based hyperspectral microscope imaging method can be used to characterize spectral properties of biofilm formed by Salmonella enteritidis as well as Escherichia coli. The intensity of spectral imagery and the pattern of spectral distribution varied with system parameters (integration time and gain) of HMI system. The preliminary results demonstrated determination of optimum parameter values of HMI system and the integration time must be no more than 250 ms for quality image acquisition from biofilm formed by S. enteritidis. Among the contiguous spectral imagery between 450 and 800 nm, the intensity of spectral images at 498, 522, 550 and 594 nm were distinctive for biofilm; whereas, the intensity of spectral images at 546 nm was distinctive for E. coli. For more accurate comparison of intensity from spectral images, a calibration protocol, using neutral density filters and multiple exposures, need to be developed to standardize image acquisition. For the identification or classification of unknown food pathogen samples, ground truth regions-of-interest pixels need to be selected for "spectrally pure fingerprints" for the Salmonella and E. coli species.