Laser microsurgery permits fungal plasma membrane single-ion-channel resolution at the hyphal tip

Abstract

A method for formation of high-electrical-resistance seals on the Neurospora crassa plasma membrane, allowing resolution of single-ion-channel activity by patch clamp electrophysiology, is reported. Laser microsurgery permits access to the hyphal apex without enzymatic cell wall digestion and loss of morphological polarity. Cell wall reformation is delayed by brefeldin. The method can allow full characterization of apical plasma membrane channels, which are implicated in tip growth.