PERFORATED-PATCH RECORDING WITH BETA-ESCIN

Perforated patch recording with nystatin, amphotericin B and gramicidi n can be more difficult in the hands of some investigators than others . In addition, it is difficult to introduce low molecular weight subst ances such as dyes into the cytoplasm in such experiments. We have det ermined that beta-escin represents a convenient, easy-to-use alternati ve to less water-soluble ionophores. ports describing long waiting tim es and inconvenience of working with ionophores that first need solubi lization in nonaqueous solvents prior to dissolution in the pipette so lution. 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 w eight substances [5]. We report here our very positive experiences wit h beta-escin as an alternative for perforated patch recording, particu larly when there is a desire to introduce indicators or other substanc es into cells via the patch pipette for long-duration experiments.