Foodborne pathogens have become ongoing threats in the food industry, whereas their rapid detection and classification at an early stage are still challenging. To address early and rapid detection, hyperspectral microscope imaging (HMI) technology combined with convolutional neural networks (CNN) was proposed to classify foodborne bacterial species at the cellular level. HMI technology can simultaneously obtain both spatial and spectral information of different live bacterial cells, while two CNN frameworks, U-Net and one-dimensional CNN (1D-CNN), were employed to accelerate the data analysis process. U-Net was used for automating cellular regions of interest (ROI) segmentation, which generated accurate cell-ROI masks in a shorter timeframe than the conventional Otsu or Watershed methods. The 1D-CNN was employed for classifying the spectral profiles extracted from cell-ROI and resulted in a higher accuracy (90%) than k-nearest neighbor (81%) and support vector machine (81%). Overall, the CNN-assisted HMI technology showed potential for foodborne bacteria detection.