Prion diseases are a group of infectious neurodegenerative diseases that affect both animals and humans. A characteristic of prion diseases is the aggregation and accumulation of a disease-associated isoform of the prion protein in the brains of infected individuals. The amyloid imaging probe (trans,trans)-1-bromo-2,5-bis-(3-hydroxycarbonyl-4-hydroxy)styrylbenzene (BSB) has shown potential in the diagnosis of other amyloid disorders and we hypothesized that this compound would be effective in labeling prion protein plaques in vitro and in vivo. To test this, we compared BSB fluorescence to prion protein immunostaining on infected and uninfected brain tissue sections from scrapie-infected hamsters. We found that both methods labeled the same plaques in infected tissues while not substantially staining uninfected tissues. To test the potential of BSB as an in vivo label for prion aggregates, we perfused scrapie-infected animals with BSB and observed BSB labeled plaques co-stained with an anti-prion protein antibody. These results suggest that BSB may have use as a diagnostic tool for prion diseases. We were unable to detect BSB staining in preclinical scrapie-infected hamsters suggesting that the diagnostic potential of BSB could be limited in cases of prion disease that do not have plaques either due to a preclinical lack of pathology or disease agents like sporadic Creutzfeldt-Jakob disease (CJD), which generally lack prion plaques. However, BSB may be a useful for prion diseases where plaques are present, such as clinical variant CJD.