title: Microwell device for targeting single cells to electrochemical microelectrodes for high-throughput amperometric detection of quantal exocytosis.
abstract: Electrochemical microelectrodes are commonly used to detect spikes of amperometric current that correspond to exocytosis of oxidizable transmitter from individual vesicles, i.e., quantal exocytosis. We are developing transparent multielectrochemical electrode arrays on microchips in order to automate measurement of quantal exocytosis. Here, we report development of an improved device to target individual cells to each microelectrode in an array. Efficient targeting (~75%) is achieved using cell-sized microwell traps fabricated in SU-8 photoresist together with patterning of poly(l-lysine) in register with electrodes to promote cell adhesion. The surface between electrodes is made resistant to cell adhesion using poly(ethylene glycol) in order to facilitate movement of cells to electrode "docking sites". We demonstrate the activity of the electrodes using the test analyte ferricyanide and perform recordings of quantal exocytosis from bovine adrenal chromaffin cells on the device. Multiple cell recordings on a single device demonstrate the consistency of spike measurements, and multiple recordings from the same electrodes demonstrate that the device can be cleaned and reused without degradation of performance. The new device will enable high-throughput studies of quantal exocytosis and may also find application in rapidly screening drugs or toxins for effects on exocytosis.
authors.affiliation: Dalton Cardiovascular Research Center, University of Missouri, Columbia, Missouri 65211, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Syed
authors.initials: S
authors.lastname: Barizuddin
authors.forename: Wonchul
authors.initials: W
authors.lastname: Shin
authors.forename: Cherian J
authors.initials: CJ
authors.lastname: Mathai
authors.forename: Shubhra
authors.initials: S
authors.lastname: Gangopadhyay
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Gillis
mesh_terms: D000818:Animals; D002395:Catecholamines / Q000378:metabolism; D002417:Cattle; D019439:Chromaffin Cells / Q000378:metabolism; D004563:Electrochemistry / Q000295:instrumentation*; D004867:Equipment Design; D017743:Equipment Reuse; D005089:Exocytosis; D008839:Microelectrodes; D059010:Single-Cell Analysis / Q000295:instrumentation*; D013499:Surface Properties
chemical_list: D002395:Catecholamines