A single-ended perfusion method for large, cylindrical cells.

Abstract

This article describes a method for the intracellular perfusion of large, cylindrically shaped cells with access to the intracellular compartment at only one point. A single cut is made in the cell membrane at some distance from the site to be studied. A glass cannula is then inserted into the cut and the tip is advanced to the study site. Intracellular perfusion solution flows out of the tip and fills the cell as it washes back along the outside of the cannula, to finally exit through the original cut. The technique is applicable to long cells with blind ends, such as segmented axons. It is also useful for the study of synapses which occur between pairs of cells because the minimal dissection required by the technique tends to leave the synaptic structure undisrupted. The method of fabrication of the glass cannula is discussed, along with applicability to electrophysiological methods such as voltage clamp. Examples are presented of perfused axons from the crayfish and earthworm ventral nerve cords.