Perforated Whole-Cell Patch-Clamp Technique: A User’s Guide
The patch-clamp technique has revolutionized the study of membrane physiology, enabling unprecedented resolution in recording cellular electrical responses and underlying mechanisms. The perforated-patch variant of whole-cell patch-clamp recording was developed to overcome the dialysis of cytoplasmic constituents that occurs with traditional whole-cell recording. With perforated-patch recordings, perforants, such as the antibiotics nystatin and gramicidin, are included in the pipette solution and form small pores in the membrane attached to the patch pipette. These pores allow certain monovalent ions to permeate, enabling electrical access to the cell interior, but prevent the dialysis of larger molecules and other ions. In this review we give a brief overview of the key features of some of the perforants, present some practical approaches to the use of the perforated patch-clamp mode of whole-cell (PPWC) recordings, and give some typical examples of neuronal responses obtained with the PPWC recording that highlight its utility as compared to the traditional whole-cell patch recording configuration.
KeywordsPipette Solution Patch Pipette Access Resistance Polyene Antibiotic Cellular Excitability