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Development of a new method for brief compound exposures to ion channels proteins using an automated patch clamp platform (NanionLigand)
Date du début: 15 juin 2009, Date de fin: 14 juin 2012 PROJET  TERMINÉ 

The gold standard for obtaining data on ion channel function is the patch clamp method. The patch clamp technique is very labor intensive and has ultra low data throughput which has been highly motivating for the development of automated patch clamp systems.Nanion has developed a technology for performing automated patch clamp recordings on planar glass substrates, with comparable recording quality as with conventional patch clamping, however, lacking the possibility to very briefly expose the cells to different solutions. If exposure times are too long, this can have adverse effects when investigating ion channels that activate on the addition of agonists (ligand gated ion channels). Ligand gated ion channels are gaining more attention in research and drug development because of their involvement in disease. These ion channels often display desensitization, resulting in a diminishing compound response, during constant stimulation. Recovery kinetics often depends on exposure time, thus are brief exposure times desirable. We aim at developing a method for very short compound exposure using automated patch clamp by combining microfluidic structures for liquid administration to the patch clamp substrate. A robotic pipettor arm applies all solutions to the microchannels and by subsequently aspirating two differnt solutions of buffer into the pipette, and immediately adding this “stack“ of solutions to the cell, the cell experiences a very brief exposure of agonist, directly followed by wash buffer. This way, exposure times are anticipated to be shorter than 1 second, with switching times in the order of 50 ms.This would allow for accurate experiments on ligand gated ion channels that exhibit rapid desensitization , which in turn would be useful in ion channel research on channelopathies, and to facilitate the drug development of compounds modulating ligand gated ion channels.

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