Study of gigaseal formation in patch clamping using nanotechnology

Abstract

This PhD project is set out to study the mechanism of gigaseal’ formation and to provide techniques which enable the frequent formation of high resistance seals. Such techniques are highly desirable at both research and industrial levels. The research can be divided into three main sections. In the first section the most important factors in seal formation were identified. Then a group of novel approaches have been developed to alter these factors and achieve better sealing conditions. In the second section the effect of roughness, hydrophilicity and tip size on gigaseal’ formation has been studied. Patch clamp experiments were carried out using conventional and treated pipettes. In the third section of this research, glass micropipettes were characterized using various measurement techniques. The proposed techniques in this thesis not only explain the gigaseal formation in more detail, but also provide techniques to increase both the seal resistance and probability of seal formation. FIB polished pipettes improved the seal resistance significantly and chemically treated pipettes resulted in the formation of seals for more than 80% of trials. Therefore the research aims have been successfully met.