Biomedical subjects

Yoonkey Nam

Publications and source records attributed to Yoonkey Nam.

4 recordsLinked to original sources

PubMed2006

Epoxy-silane linking of biomolecules is simple and effective for patterning neuronal cultures.

Surface chemistry is one of the main factors that contributes to the longevity and compliance of cell patterning. Two to three weeks are required for dissociated, embryonic rat neuronal cultures to mature to the point that they regularly produce spontaneous and evoked responses. Though proper surface chemistry can be achieved through the use of covalent protein attachment, often it is not maintainable for the time periods necessary to study neuronal growth. Here we report a new and effective covalent linking approach using (3-glycidoxypropyl) trimethoxysilane (3-GPS) for creating long term neuronal patterns. Micrometer scale patterns of cell adhesive proteins were formed using microstamping; hippocampal neurons, cultured up to 1 month, followed those patterns. Cells did not grow on unmodified 3-GPS surfaces, producing non-permissive regions for the long-term cell patterning. Patterned neuronal networks were formed on two different types of MEA (polyimide or silicon nitride insulation) and maintained for 3 weeks. Even though the 3-GPS layer increased the impedance of metal electrodes by a factor of 2-3, final impedance levels were low enough that low noise extracellular recordings were achievable. Spontaneous neural activity was recorded as early as 10 days in vitro. Neural recording and stimulation were readily achieved from these networks. Our results showed that 3-GPS could be used on surfaces to immobilize biomolecules for a variety of neural engineering applications.

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PubMed2006

Neural recording and stimulation of dissociated hippocampal cultures using microfabricated three-dimensional tip electrode array.

There is increasing interest in interfacing dissociated neuronal cultures with planar multielectrode arrays (MEAs) for the study of the dynamics of neuronal networks. Here we report on the successful use of three-dimensional tip electrode arrays (3D MEAs), originally developed for use with brain slices, for recording and stimulation of cultured neurons. We observed that many neurons grew directly on protruding electrode surface, appearing to make excellent contact. A larger than usual portion of extracellular spikes had large positive peaks, while most of the spikes from conventional two-dimensional electrode arrays had large negative spikes. This may be due to the direct capacitive coupling situation provided by relatively large electrode surface area.

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PubMed2006

Application of a PDMS microstencil as a replaceable insulator toward a single-use planar microelectrode array.

Here we present a novel idea for a replaceable insulator, and thus advance toward the goal of a single-use planar microelectrode array (MEA) for the study of electrogenic tissues. The concept of a replaceable insulator is motivated by insulator degradation after repeated usage of an MEA. Instead of fabricating a more durable insulator for repeated MEA usage, we propose replacing the insulator and effectively producing a fresh MEA for each experiment. We chose a polydimethylsiloxane (PDMS) microstencil as a candidate for the replaceable insulator as it is biocompatible, shows reversible adhesion to surfaces, and can be easily and controllably fabricated. As a proof-of-concept, we demonstrate two applications using microstencils: the rejuvenation of an old MEA and the fabrication of a single-use MEA. These MEAs were tested with dissociated neural cell cultures and neural recordings were performed at 14 days in vitro. Inexpensive and quick supply of insulators with micrometer-sized holes provides a way of constructing an MEA that can be treated as a disposable component in high throughput cell-based biosensor applications.

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PubMed2004

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.

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