The hippocampal slice is a popular model system in which to study the cellular properties of long-term potentiation (LTP). Synaptogenesis induced by exposure to ice-cold artificial cerebrospinal fluid (ACSF), however, raises the concern that morphological correlates of LTP might be obscured, especially in mature slices. Here we demonstrate that preparation of mature hippocampal slices at room temperature (approximately 25 degrees C) maintains excellent ultrastructure and a synapse density comparable to perfusion-fixed hippocampus. These results suggest that slices prepared at room temperature might provide a better basis from which to detect LTP-related changes in synapse number and morphology.