Technical Abstract: Pathogenic fungi, including Aspergillus flavus, are mycotoxin-producing and can contaminate grain products, such as corn. The contaminated food poses a threat to humans and animals. Traditionally, corn kernels have been examined for evidence of bright greenish-yellow fluorescence (BGYF), indicative of the presence of A. flavus, when illuminated with a high-intensity ultra-violet light. The BGYF approach is time and labor intensive and somewhat inaccurate. Several previous studies have examined spectral-based, non-destructive methods for the detection of fungi and toxins. The present report focused on using spectral image data for fungal detection. A tabletop hyperspectral imaging system, VNIR-100E, was used for high spectral and high spatial resolution data acquisition. A total of five toxin producing fungal species were used in a two-part experiment: Penicillium chrysogenum, Fusarium moniliforme, Aspergillus parasiticus, Trichoderma viride and Aspergillus flavus. All fungal isolates were cultured on agar in Petri-dishes under lab conditions and were imaged on day 5 of growth. The objective of the study was to use hyperspectral imagery for classification of different fungal species. Results indicate that the five fungi are separable to a high degree of accuracy. In addition, all five toxigenic fungi could be classified by using only three narrow bands (bandwidth = 2.43 nm) centered at 743 nm, 458 nm and 541 nm.