Perforated patch recording with β-escin

Perforated patch recording with nystatin, amphotericin B and gramicidin can be more difficult in the hands of some investigators than others. In addition, it is difficult to introduce low molecular weight substances such as dyes into the cytoplasm in such experiments. We have determined that beta-escin represents a convenient, easy-to-use alternative to less water-soluble ionophores. ports describing long waiting times and inconvenience of working with ionophores that first need solubilization in nonaqueous solvents prior to dissolution in the pipette solution. Other methods have been used to permeabilize cells, including digitonin and saponin, which spare intracellular membranes. beta-escin, a saponin derivative used initially to permeabilize smooth muscle [4], has been reported to form holes quite permeable to high molecular weight substances [5]. We report here our very positive experiences with beta-escin as an alternative for perforated patch recording, particularly when there is a desire to introduce indicators or other substances into cells via the patch pipette for long-duration experiments.