Single cell and neural process experimentation using laterally applied electrical fields between pairs of closely apposed microelectrodes with vertical sidewalls.
As biomedical research has moved increasingly towards experimentation on single cells and subcellular structures, there has been a need for microscale devices that can perform manipulation and stimulation at a correspondingly small scale. We propose a microelectrode array (MEA) featuring thickened microelectrodes with vertical sidewalls (VSW) to focus electrical fields horizontally on targets positioned in between paired electrodes. These microelectrodes were fabricated using gold electroplating that was molded by photolithographically patterned SU-8 photoresist. Finite element modeling showed that paired VSW electrodes produce more uniform electrical fields compared to conventional planar microelectrodes. Using paired microelectrodes, 3 microm thick and spaced 10 microm apart, we were able to perform local electroporation of individual axonal processes, as demonstrated by entry of EGTA to locally chelate intra-axonal calcium, quenching the fluorescence of a pre-loaded calcium indicator dye. The same electrode configuration was used to electroporate individual cells, resulting in the targeted transfection of a transgene expressing a cytoplasmically soluble green fluorescent protein (GFP). In addition to electroporation, our electrode configuration was also capable of precisely targeted field stimulation on individual neurons, resulting in action potentials that could be tracked by optical means. With its ability to deliver well-characterized electrical fields and its versatility, our configuration of paired VSW electrodes may provide the basis for a new tool for high-throughput and high-content experimentation in broad areas of neuroscience and biomedical research.
Chang
Wesley C
WC
Department of Ophthalmology, University of California, K110, Box 0730, UC San Francisco, 10 Koret Way, K-110, San Francisco, CA 94143, USA. changw@vision.ucsf.edu
Sretavan
David W
DW
D000818:Animals; D002469:Cell Separation / Q000295:instrumentation*; D002478:Cells, Cultured; D004558:Electric Stimulation / Q000295:instrumentation*; D004574:Electromagnetic Fields; D018274:Electroporation / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D051379:Mice; D008839:Microelectrodes; D008846:Micromanipulation / Q000295:instrumentation*; D009474:Neurons / Q000502:physiology* / Q000528:radiation effects*