Localized electrical stimulation of in vitro neurons using an array of sub-cellular sized electrodes

• Published in: Biosensors & bioelectronics Vol. 26; no. 4; pp. 1474 - 1477
• Main Authors: Braeken, Dries, Huys, Roeland, Loo, Josine, Bartic, Carmen, Borghs, Gustaaf, Callewaert, Geert, Eberle, Wolfgang
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Kidlington Elsevier B.V 15.12.2010; Elsevier
• Subjects: Aniline Compounds; Animals; Biological and medical sciences; Biosensing Techniques - instrumentation; Biotechnology; Calcium imaging; Cell Line; Electric Stimulation - instrumentation; Electroporation; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; Hippocampal neurons; Hippocampus - cytology; Hippocampus - physiology; In Vitro Techniques; Membrane Potentials; Mice; Microelectrode array; Microelectrodes; Microscopy, Electron, Scanning; Nerve Net - cytology; Nerve Net - physiology; Neurons - physiology; Neurons - ultrastructure; NG108-15 cell line; Patch-Clamp Techniques; Semiconductors; Single-Cell Analysis - instrumentation; Xanthenes; Calcium imaging; NG108-15 cell line; Electroporation; Hippocampal neurons; Microelectrode array; Calcium; Electrical stimulus; Imagery; In vitro; Electrodes; Array; Cell line; Neuron; Imaging; Microelectrode
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2010.07.086

The investigation of single-neuron parameters is of great interest because many aspects in the behavior and communication of neuronal networks still remain unidentified. However, the present available techniques for single-cell measurements are slow and do not allow for a high-throughput approach. We present here a CMOS compatible microelectrode array with 84 electrodes (with diameters ranging from 1.2 to 4.2 μm) that are smaller than the size of cell, thereby supporting single-cell addressability. We show controllable electroporation of a single cell by an underlying electrode while monitoring changes in the intracellular membrane potential. Further, by applying a localized electrical field between two electrodes close to a neuron while recording changes in the intracellular calcium concentration, we demonstrate activation of a single cell (∼270%, DF/F 0), followed by a network response of the neighboring cells. The technology can be easily scaled up to larger electrode arrays (theoretically up to 137,000 electrodes/mm 2) with active CMOS electronics integration able to perform high-throughput measurements on single cells.