Hyperspectral microscopy imaging (HMI) is an efficient and non-destructive method to detect microbial contaminants in food, as it can provide both spatial morphology and spectral signature. Aims at reducing thermal effect, low cost, and improving spectral resolution in testing, a pipeline-operated LEDs monochromatic illumination mode is proposed, which integrates the design concepts of both grating-based and LED-based HMI systems. By design of the LED set, shared grating monochromatic optical path, and coordinated control system, an HMI system has been developed that could obtain the hyperspectral data cube with 101 bands in 400–700 nm. Hyperspectral datasets of three species of Aspergillus are prepared using the prototype, and efficient results have been achieved in the training and testing of classical classification algorithms (1D-CNN (97.33 %), k-NN (96.33 %), SVM (97.67 %) and ResNet-18 (95.67 %)). The results demonstrate that the proposed monochromatic illumination mode and associated system are potential detection solutions for foodborne microbial contaminants with low-cost and high-accurate.
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