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Intracellular filling and reconstruction of identified neurons in fixed rat brain slices
1Department of Psychology and Neuroscience, University of Lethbridge, Alberta, Canada. prusky@uleth.ca
Brain Research. Brain Research Protocols
|February 12, 1999
Abstract:
A simple technique for the morphological characterization of pre-labelled neurons in fixed brain slices is described. Neurons are retrogradely-labeled with a carbocyanine dye and the tissue is fixed and sliced. Individual labeled cells from the interior of a slice are then visualized on an upright fluorescence microscope and impaled with a micropipet containing rhodamine or fluorescein-peroxidase conjugates. The cells are filled by iontophoresis, postfixed, and the peroxidase is oxidized into a permanent, opaque reaction product. The dendritic morphology of the neurons is then reconstructed under camera lucida and quantified.