Research paper
Whole cell recording from neurons in slices of reptilian and mammalian cerebral cortex

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Abstract

We describe methods for obtaining stable, whole-cell recordings from neurons in brain hemispheres from turtles and in brain slices from rats and turtles. Synaptic currents and membrane properties of central neurons can be studied in voltage and current clamp in cells maintained within their endogenous synaptic circuits. The methods described here are compatible with unmodified dissecting microscopes and recording chambers, and with brain slices of standard thickness (400–500 μm).

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    However, since proteolytic enzymes may damage the proteins on the cell membrane of interest, different approaches were sought and developed. To enable direct contact between the tip of a patch pipette and the tissue-covered cell membrane, these approaches implemented either a two-step process in which a separate, “cleaning” pipette was first used to remove the part of the tissue covering the cell body of interest (Edwards et al., 1989) or a one-step process in which positive pressure was applied to a patch pipette as it was penetrating the tissue and approaching the cell membrane (Blanton et al., 1989). The integration of differential interference contrast (DIC) optics was another major advancement for enabling patch clamping in brain slices, as the improved imaging quality offered by DIC-based microscopy enabled visually-guided patching of soma as well as dendrites of targeted neurons in mammalian brain slices (Stuart et al., 1993).

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