Current progress in our understanding of neurological diseases is tremendously facilitated by the development of new transgenic mouse models, as well as by advanced electrophysiological and imaging techniques. Here, we describe such a combinatorial approach to study the mouse spinal cord. Adult transgenic mice with cell-type specific fluorescent protein (FP) expression were anaesthetized; vital functions were monitored and could be maintained for up to 11 h. During anaesthesia, a laminectomy was performed at the main input region of the hind leg, either at the dorsal root ganglion L4 or at the spinal cord segment L4. After surgery the mouse was placed underneath the objective of the laser-scanning microscope (one-and two-photon excitation). To minimize movements in the region of interest, the adjacent spines were rigidly fixed. Due to the FP expression, neurons, astrocytes or microglia could be imaged at high spatial resolution. It was also possible to record from fluorescent Ca²⁺ indicator-labelled cells in spinal ganglia in which extracellular field potentials could be evoked by stimulation of the hind foot. We are now in a position to investigate the signalling mechanisms of the excitation spread from the periphery to the spinal cord. This might be particularly interesting in the context of pain perception.