INCUBATION TYPE PLANAR PATCH CLAMP AS A NEW POTENTIAL TECHNOLOGY FOR DEVELOPING NEURONAL NETWORK HIGH THROUGHPUT SCREENING DEVICES

Abstract

Ion-channel current recordings based on an incubation type planar patch clamp were first reported in 2008, using HEK293 cells expressed with TRPV1 channels and capsaicin as ligand molecules. At first the success probability (number of devices which normally worked/total number of devices fabricated) was extremely low (several %). Several years later, we have succeeded in significantly decreasing the base line noise by using a salt-bridge-type Ag/AgCl electrode and successfully demonstrated the application of an incubation type planar patch clamp to ligand gated ion-channel biosensors and light gated ion-channel biosensors using HEK293 cells expressed with ChRWR. Furthermore, a spontaneous ion-channel current from a neuronal network was successfully observed by using a planar patch clamp chip, on which the neuronal network was occasionally formed with a soma of a neuron on a micro through-hole. Although the neuronal network was not controlled, this success shows the high potential of realizing a high throughput screening device on the basis of channel current measurements, which contain the most important information on network conditions.