We describe a preparation of arterially-perfused spinal cord with attached hindquarters, taken from adult mouse. This is the first preparation of adult mammalian spinal cord tissue to have the advantages of an in vitro approach whilst retaining intact intraspinal circuitry, sensory inputs, and somatic and sympathetic segmental outputs. The functional integrity of the preparation has been demonstrated by the motor and sympathetic reflexes that can readily be evoked by peripheral noxious thermal, mechanical and electrical stimuli, and also by bladder distension. The mechanical stability of the preparation allows intracellular recordings to be made from spinal dorsal or ventral horn neurones. The intact connectivity permits synaptic responses to be evoked by stimulation of functionally-defined peripheral sensory receptors. The preparation is relatively quick to set up and remains viable for more than 6 h. This model offers the opportunity to perform complex electrophysiological and pharmacological studies on functionally characterised synaptic responses of mature spinal neurones. The choice of the mouse will furthermore permit studies to be performed on genetically mutant strains.