Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures

We report the use of a gold coating on microelectrode arrays (MEAs) to enable the use of the relatively reliable surface modification chemistry afforded by alkanethiol self-assembled monolayers (SAMs). The concept is simple and begins with planar MEAs, which are commercially available for neuronal cell culture and for brain slice studies. A gold film, with an intermediate adhesive layer of titanium, is deposited over the insulation of an existing MEA in a manner so as to be thin enough for transmission light microscopy as well as to avoid electrical contact to the electrodes. The alkanethiol-based linking chemistry is then applied for the desired experimental purpose. Here we show that polylysine linked to alkanethiol SAM can control the geometry of an in vitro hippocampal neuronal network grown on the MEA. Furthermore, recordings of neuronal action potentials from random and patterned networks suggest that the gold coating does not significantly alter the electrode properties. This design scheme may be useful for increasing the number of neurons located in close proximity to the electrodes. Realization of in vitro neuronal circuits on MEAs may significantly benefit basic neuroscience studies, as well as provide the insight relevant to applications such as neural prostheses or cell-based biosensors. The gold coating technique makes it possible to use the rich set of thiol-based surface modification techniques in combination with MEA recording.

[1]  B. Wheeler,et al.  Micrometer resolution silane-based patterning of hippocampal neurons: critical variables in photoresist and laser ablation processes for substrate fabrication , 1996, IEEE Transactions on Biomedical Engineering.

[2]  Y Shapira,et al.  Observations and modeling of synchronized bursting in two-dimensional neural networks. , 2001, Physical review. E, Statistical, nonlinear, and soft matter physics.

[3]  D Kleinfeld,et al.  Controlled outgrowth of dissociated neurons on patterned substrates , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[4]  D. Bertrand,et al.  A three-dimensional multi-electrode array for multi-site stimulation and recording in acute brain slices , 2002, Journal of Neuroscience Methods.

[5]  G J Brewer,et al.  Modulation of neural network activity by patterning. , 2001, Biosensors & bioelectronics.

[6]  David W. Tank,et al.  Sealing cultured invertebrate neurons to embedded dish electrodes facilitates long-term stimulation and recording , 1989, Journal of Neuroscience Methods.

[7]  Andreas Offenhäusser,et al.  Extended Gate Electrode Arrays for Extracellular Signal Recordings , 2000 .

[8]  Bruce C. Wheeler,et al.  Microelectrode Array Recordings of Patterned Hippocampal Neurons for Four Weeks , 2000 .

[9]  Daniel I. C. Wang,et al.  Engineering cell shape and function. , 1994, Science.

[10]  Bruce C. Wheeler,et al.  Current Source Density Estimation Using Microelectrode Array Data from the Hippocampal Slice Preparation , 1986, IEEE Transactions on Biomedical Engineering.

[11]  Y. Tai,et al.  The neurochip: a new multielectrode device for stimulating and recording from cultured neurons , 1999, Journal of Neuroscience Methods.

[12]  David A. Russell,et al.  Self-assembled monolayers: a versatile tool for the formulation of bio-surfaces , 2000 .

[13]  Gregory T. A. Kovacs,et al.  Cell-based sensor microelectrode array characterized by imaging x-ray photoelectron spectroscopy, scanning electron microscopy, impedance measurements, and extracellular recordings , 1998 .

[14]  D. Branch,et al.  Long-term stability of grafted polyethylene glycol surfaces for use with microstamped substrates in neuronal cell culture. , 2001, Biomaterials.

[15]  H. Robinson,et al.  Strengthening of synchronized activity by tetanic stimulation in cortical cultures: application of planar electrode arrays , 1998, IEEE Transactions on Biomedical Engineering.

[16]  T. Stieglitz,et al.  A biohybrid system to interface peripheral nerves after traumatic lesions: design of a high channel sieve electrode. , 2002, Biosensors & bioelectronics.

[17]  Milan Mrksich,et al.  A surface chemistry approach to studying cell adhesion , 2000 .

[18]  G. Brewer,et al.  Optimized survival of hippocampal neurons in B27‐supplemented neurobasal™, a new serum‐free medium combination , 1993, Journal of neuroscience research.

[19]  Christopher S. Chen,et al.  Microcontact Printing of Proteins on Mixed Self-Assembled Monolayers , 2002 .

[20]  U. Egert,et al.  A novel organotypic long-term culture of the rat hippocampus on substrate-integrated multielectrode arrays. , 1998, Brain research. Brain research protocols.

[21]  M. Grattarola,et al.  Modeling the neuron-microtransducer junction: from extracellular to patch recording , 1993, IEEE Transactions on Biomedical Engineering.

[22]  George M. Whitesides,et al.  Convenient methods for patterning the adhesion of mammalian cells to surfaces using self-assembled monolayers of alkanethiolates on gold , 1993 .

[23]  G. Gross,et al.  Drug evaluations using neuronal networks cultured on microelectrode arrays. , 2000, Biosensors & bioelectronics.

[24]  John C. Chang,et al.  An enhanced microstamping technique for controlled deposition of proteins , 2002, 2nd Annual International IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology. Proceedings (Cat. No.02EX578).

[25]  G. Gross,et al.  Stimulation of monolayer networks in culture through thin-film indium-tin oxide recording electrodes , 1993, Journal of Neuroscience Methods.

[26]  A. Aertsen,et al.  Two-dimensional monitoring of spiking networks in acute brain slices , 2001, Experimental Brain Research.

[27]  Bruce C. Wheeler,et al.  Electrical recordings with gold coated microelectrode arrays that permit the control of neuronal attachment , 2002, Proceedings of the Second Joint 24th Annual Conference and the Annual Fall Meeting of the Biomedical Engineering Society] [Engineering in Medicine and Biology.

[28]  Robert M. Corn,et al.  A Multistep Chemical Modification Procedure To Create DNA Arrays on Gold Surfaces for the Study of Protein−DNA Interactions with Surface Plasmon Resonance Imaging , 1999 .

[29]  Bruce C. Wheeler,et al.  Long-term maintenance of patterns of hippocampal pyramidal cells on substrates of polyethylene glycol and microstamped polylysine , 2000, IEEE Transactions on Biomedical Engineering.

[30]  Stefano Vassanelli,et al.  Transistor records of excitable neurons from rat brain , 1998 .

[31]  P. Bohn,et al.  Chemisorption and chemical reaction effects on the resistivity of ultrathin gold films at the liquid-solid interface. , 1999, Analytical chemistry.

[32]  M. Segal,et al.  GABA Withdrawal Modifies Network Activity in Cultured Hippocampal Neurons , 2000, Neural plasticity.

[33]  Robert M. Corn,et al.  Covalent Attachment and Derivatization of Poly(l-lysine) Monolayers on Gold Surfaces As Characterized by Polarization−Modulation FT-IR Spectroscopy , 1996 .

[34]  A. Offenhäusser,et al.  Electrical recordings from rat cardiac muscle cells using field-effect transistors. , 1999, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[35]  M Krause,et al.  Ordered networks of rat hippocampal neurons attached to silicon oxide surfaces , 2000, Journal of Neuroscience Methods.

[36]  John Chi-Hung Chang Technologies for and Electrophysiological Studies of Structured, Living, Neuronal Networks on Microelectrode Arrays , 2002 .