High-resolution electrophysiology on a chip: Transient dynamics of alamethicin channel formation.
Microstructured planar substrates have been shown to be suitable for patch clamp recording from both whole cells and isolated patches of membrane, as well as for measurements from planar lipid bilayers. Here, we further explore this technology with respect to high-resolution, low noise single-channel recording. Using solvent-free lipid bilayers from giant unilamellar vesicles obtained by electro-swelling, we recorded channels formed by the peptaibol alamethicin, a well-studied model system for voltage-dependent channels, focusing on the transient dynamics of single-channel formation upon application of a voltage step. With our setup, we were able to distinctly resolve dwell times well below 100 mus and to perform a thorough statistical analysis of alamethicin gating. Our results show good agreement with models that do not rely on the existence of non-conducting preaggregate states. Microstructured apertures in glass substrates appear promising with respect to future experiments on cellular ion channels reconstituted in suspended lipid membranes.
Sondermann
Markus
M
Department of Physiology, Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Str. 7, 79104 Freiburg, Germany.
George
Michael
M
Fertig
Niels
N
Behrends
Jan C
JC
D000408:Alamethicin / Q000737:chemistry*; D004594:Electrophysiology / Q000379:methods; D007473:Ion Channels / Q000502:physiology; D008051:Lipid Bilayers; D008081:Liposomes / Q000737:chemistry; D018408:Patch-Clamp Techniques; D040081:Protein Array Analysis; D012680:Sensitivity and Specificity
D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D000408:Alamethicin