Localized electrical stimulation of in vitro neurons using an array of sub-cellular sized electrodes.
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.
Braeken
Dries
D
Bioelectronics Group, Imec, Kapeldreef 75, 3001 Heverlee, Belgium. dries.braeken@imec.be
Huys
Roeland
R
Loo
Josine
J
Bartic
Carmen
C
Borghs
Gustaaf
G
Callewaert
Geert
G
Eberle
Wolfgang
W
D000814:Aniline Compounds; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D004558:Electric Stimulation / Q000295:instrumentation*; D018274:Electroporation; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D066298:In Vitro Techniques; D008564:Membrane Potentials; D051379:Mice; D008839:Microelectrodes; D008855:Microscopy, Electron, Scanning; D009415:Nerve Net / Q000166:cytology / Q000502:physiology; D009474:Neurons / Q000502:physiology* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques; D012666:Semiconductors; D059010:Single-Cell Analysis / Q000295:instrumentation*; D014966:Xanthenes
D000814:Aniline Compounds; C409648:Fluo 4; D005456:Fluorescent Dyes; D014966:Xanthenes