Use of confocal microscopy for three-dimensional imaging of neurons in the spinal cord

Confocal microscopy provides a powerful and efficient tool for studying the morphology of cells. Here we describe its use to study the morphology of neurons in the dorsal horn of the spinal cord either following electrophysiological studies in live tissue slices or in neurons filled with dye in fixed tissue sections following identification using retrograde tracing. The methods are broadly applicable to other cell types and can be combined with multiple label immunohistochemistry to study cellular constituents or with subsequent DAB staining to produce a permanent mount.