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FIGURE 5–5. Intracellular staining of neuron recorded intracellularly.

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FIGURE 5–5. Intracellular staining of neuron recorded intracellularly.During intracellular recordings, the recording pipette is filled with an electrolyte to enable the transmission of membrane voltage deflections to the preamplifier. The electrode may also be filled with substances, such as a morphological stain, for injection into the impaled neuron. In this example, the electrode was filled with the highly fluorescent dye Lucifer yellow. Because this dye has a negative charge at neutral pH, it may be ejected from the electrode by applying a negative current across the electrode, with the result that the Lucifer yellow carries the negative current flow from the electrode and into the neuron. Because this dye diffuses rapidly in water, it quickly fills the entire neuron impaled. The tissue is then fixed in a formaldehyde compound, the lipids clarified by dehydration-defatting or by using dimethylsulfoxide (Grace and Llinás 1985), and the tissue examined under a fluorescence microscope. In this case, a brightly fluorescing pyramidal neuron in layer 3 of the neocortex of a guinea pig is recovered.

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