A technique for recovering functional synaptoneurosomes containing vesicularized elements of the presynapse and postsynapse into an enriched fraction has been modified to allow for small amounts of starting brain tissue. Single 400 microm rat hippocampal slices were homogenized and sequentially filtered in a 1 cc tuberculin syringe to produce an enriched synaptoneurosome fraction. Data from Western immunoblots for specific synaptic proteins suggest that these fractions are neurochemically similar to synaptosome fractions generated by sucrose gradients. Electron micrographs show that the 'small scale' preparations contain an abundant population of fused presynaptic and postsynaptic vesicularized bodies as previously published for synaptoneurosome fractions prepared from relatively large amounts of starting tissue. The single slice synaptoneurosome preparation is a quick, easy and reliable method for use in the study of synaptic function.