The in vitro slice preparation for combined morphological and electrophysiological studies of rat visual cortex

Abstract

The morphological condition of slices of rat visual cortex, maintained in vitro in an interface-type recording chamber, was assessed. In addition, neurones in these slices were impaled with glass micropipettes for intracellular recording and horseradish peroxidase (HRP) injection. After fixation and embedding, slices were examined by light and electron microscopy. Slices sectioned orthogonally to the original plane of cutting showed a vertical zonation of tissue preservation. The upper zone contained dense and flattened neuronal somata, although the neuropil appeared normal. The central zone was well preserved, with the appearance of most somata, dendrites, axons and synapses comparing favourably with perfusion-fixed material. The lower zone contained many abnormal, vacuolated somata. The morphology of HRP-injected neurones was assessed by light microscopy. Dendrites could be visualised in great detail and spines were clearly visible. Local axon arbors were well represented. There was good correlation between electrophysiological and morphological criteria for the assessment of the condition of the slice. We conclude that, provided the extent of degeneration within the slice is monitored and appreciated, slices of visual cortex can provide both electrophysiological and morphological data of high quality.