Development of planar patch clamp with potentiometric calcium ion-selective electrode

Kul Inn, Purdue University

Abstract

Ion channels are proteins in cell lipid bilayer membranes and act as pores which can adopt closed and open states, thus gating the flow of ions in and out of the cell. Patch clamp technology has been the proven standard for fundamental studies of ion channel activities. However, the technique has some basic limitations: low throughput, time consuming nature of its process, need of highly skilled personnel and inability to identify ionic composition of electrophysiological events. Many different materials and fabrication methods have been introduced to replace traditional patch clamp setup to overcome limitations. In this dissertation, a planar patch clamp device with calcium ion-selective electrode is developed in miniaturized form for high throughput cell electrophysiology, and screening of ion channel modulators as potential drug targets in an in vitro format. Femtosecond laser-drilling technique is newly introduced to fabricate the patch-pore and new design of planar ion-selective electrode is suggested for calcium ion measurement. By integrating a standard patch clamp electrophysiological interface with calcium ion-selective electrode on a single platform, it is possible to directly identify the ionic component of a whole-cell potential recording. This system is innovative because the focus is not entirely on increasing experimental throughput, but instead offers information on user specified target ion activities through ion channels.

Degree

Ph.D.

Advisors

Porterfield, Purdue University.

Subject Area

Biomedical engineering

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