Current procedures for microbial testing typically involve a homogenizing step. These methods give valuable information on the presence/absence of a bacterial contaminant, but not where the contaminant was in the original sample. Spatial information could be useful in troubleshooting sources of bacterial contamination in a plant. For example, if the contaminant was localized on the top of a cheese, this might indicate dripping condensate along a specific processing line as its source. The objective of this proof-of-concept study was to evaluate the use of a T7 bacteriophage engineered to overexpress the luciferase NanoLuc to reveal the spatial location of Escherichia coli on Luria-Bertani agar (LBA) and queso fresco (QF). Four scenarios were tested to explore how phage may be applied, with a blue bioluminescent signal revealing the spatial location of contaminants: 1) Phage applied topically via molten soft agar to E. coli-inoculated a) LBA or b) QF and 2) Phage incorporated within a) LBA or b) QF, then inoculated with E. coli. Each was tested in triplicate. Cultures of BL21 E. coli grown for 18hr were serially diluted in phosphate-buffered saline and inoculated onto 8 ± 0.5g of LBA or QF in 6-well plates. Plates were incubated at 37C for 8hr for condition 1a and 24 h for 1b, 2a, and 2b. For 1a and 1b, stock phage was added to molten soft agar, applied topically, and incubated for 2 additional hours to allow for E. coli infection. After incubation, 100µL of the substrate NanoGlo was added to cover the surface of the agar or cheese, and imaged immediately in a dark box using a Canon EOS Rebel T6 camera and long exposure to capture the bioluminescent signal. Photos capture small blue spots where the incubated cfu are located. The lowest inoculum level of E. coli detected for each scenario was 0.64 ± 0.11, 2.73 ± 0.16, 0.67 ± 0.8, and 1.16 ± 0.54 logCFU/well, for 1a, 1b, 2a, and 2b, respectively. These data demonstrate the reporter phage proof-of-concept can be used as a forensic tool to visualize the spatial location of bacteria in a cheese matrix. Future work will translate this concept to dairy relevant phage-pathogen systems.