To clarify the function of caspase-1-like proteases in neuronal cell death, it is important to be able to detect the activity in living organs by microscopic visualization. In the present study, we synthesized a novel fluorescent substrate sensitive to the caspase-1-like activity, which is easily introduced into cells constituting living organs by extracellular application. As a result, the substrate was shown to be useful in imaging the caspase-1-like activity in rat hippocampal slice cultures. After induction of cell death with glutamate, a significant increase in the activity was observed, especially in the pyramidal cells, suggesting the association of the activity with promotion of cell death.