The luciferin of the bioluminescent decapod shrimp, Oplophorus gracilorostris, was purified and studied with respect to u.v. spectrum, fluorescence spectrum, mass spectrum and luminescent cross-reaction with the enzyme luciferase of the bioluminescent ostracod, Cypridina hilgendorfii. On the basis of these results, an empirical formula C10H13N3O3 and an imidazo [1,2-a]pyrazin-3-one structure are proposed for luciferin. Of three model luciferin compounds, 3-hydroxy-2-methylimidazo[1,2-a]pyridine is biologically active with both Oplophorus and Cypridina luciferase, indicating that a pyrazine structure is not essential for biological activity with Cypridina luciferase.