Electrophysiological measurement system in a planar patch-clamp chip integrating microfluidic intra- and extra-cellular solution exchange with optical detection
Date Issued
2009
Date
2009
Author(s)
Tu, Ting-Yuan
Abstract
Ion-channel plays an important role in signaling and functioning throughout human body. The most pervasive technique to interrogate ion-channel is patch-clamp owing to its potent and reliable information extracted from cell, but traditional patch-clamp technique requires high skill-dependent and laborious manipulation that impedes its throughput. Our lab have previously proposed a simple laser drilling technique on a plain cover glass in fabrication of planar patch-clamp chip in order to lessen traditional exquisite operation. This paper presents planar patch-clamp chip integrated with microfluidic system and optical detection for ion-channel recording. esults show that whole-cell recording is feasible for various cell lines, such as pancreatic beta cell of HIT-T15 and RIN-m5F, suspension lymphoma Jurkat cell, and HEK 293T cells that are generally used as transfection model for channel expression,. Intracellular solution exchange was performed via internal perfusion of perforated patch, and whole-cell current is consistent with typical patch recording via voltage zap. Extracellular solution was successfully exchanged from standard extracellular saline to symmetrical K+-containing solution. Fluorescence was measured via optical detection while extracellular solution exchanged with calcein AM stain. Nine cells were concurrently captured and aligned in 3x3 array configuration. It is believed to be the first proposition of economical fabrication, microfluidics integration, and optical detection, which is compatible with existing commercial products, and has great potential for drug-related applications.
Subjects
Ion-channel
Planar patch-clamp
Microfluidics
Fluorescence detection
Single-cell analysis
Chip
Type
thesis
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