Stretch and poke stimulation for characterizing mechanically activated ion channels

Quantitative functional characterization of mechanically activated ion channels is most commonly achieved by a combination of patch-clamp electrophysiology and stimulation by stretch (or pressure-clamp) and poke (or cell-indentation). A variety of stretch and poke protocols can be used to make measurements of many ion channel properties, including channel number, unitary conductance, ion selectivity, stimulus threshold and sensitivity, stimulus adaptation, and gating kinetics (activation, deactivation, inactivation, recovery from inactivation). Here, we review the general methods of stretch and poke stimulation and discuss the advantages and disadvantages of each. We use the vertebrate mechanically activated ion channel Piezo1 to explain equipment components and calibration, demonstrate experimental protocols and data analyses, and discuss underlying concepts and mechanistic interpretations.