Name: time-dependent increase in the network response to the stimulation of neuronal cell cultures on micro-electrode arrays
Uploaded: 2017-05-29T15:00:00
Duration: 10 min 45 s
Description: Watch this Scientific Journal Video about Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays at JoVE.com

This work was funded by the National Science Foundation grant CMMI-1300007. We would like to acknowledge previous lab members, who have helped with the design of these protocols and with the maintenance of the cultures for over five years at George Mason University: Dr. Joseph J. Pancrazio, Dr. Hamid Charkhkar, Dr. Gretchen Knaack, Dr. Franz Hamilton, Michael Maquera, and Robert Graham.

<ol>
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Guidelines for Euthanasia of Rodents Using Carbon Dioxide Available from: <a target="_blank" href="https://oacu.oir.nih.gov/sites/default/files/uploads/arac-guidelines/rodent_euthanasia_adult.pdf">https://oacu.oir.nih.gov/sites/default/files/uploads/arac-guidelines/rodent_euthanasia_adult.pdf</a> (2016)</li><li>Banker, G., Goslin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Culturing Nerve Cells: Business Source. , (1998).</li><li>Gullo, F., Maffezzoli, A., Dossi, E., Wanke, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-latency+cross-and+autocorrelation+identify+clusters+of+interacting+cortical+neurons+recorded+from+multi-electrode+array.">Short-latency cross-and autocorrelation identify clusters of interacting cortical neurons recorded from multi-electrode array.</a> J. 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The authors have nothing to disclose.

The steps outlined in this protocol provide sufficient detail for the beginner to plate his/her own neuronal cultures on MEAs and to record network activity. This protocol will help to ensure that the cultures adhere properly, forming a carpet layer of cells over the electrode arrays, and remain healthy and contaminant-free for months.
Although it is best to adhere to all parts of the protocol, there are steps throughout the process that are critical to the successful outcome. The use of aseptic technique throughout the entire process is imperative to prevent the cultures from becoming contaminated. New MEAs must be made hydrophilic, as described in the protocol, or else poor cell adhesion will result. Avoiding harsh pipetting and the formation of air bubbles during dissociation will reduce the number of damaged cells plated and will lead to a higher and healthier yield. Switching from DMEM 5/5 to DMEM+ after the first feeding is also important. DMEM 5/5 contains horse serum, which will cause glial cells to dominate the culture if used continuously and will result in poor neuronal activity, although the cultures will appear healthy17. Feeding the cultures as scheduled and keeping them in proper incubating conditions is also crucial.
Plating cell cultures on MEAs involves many variables that can lead to less-than-optimal results. Although the goal is a perfect &#34;carpet&#34; of cells, failure to address the critical steps mentioned above will result in poor cell maturation or in contamination. Poor cell adhesion, which is different from poor cell maturation, is also a concern. This can be caused by several factors, including poor MEA preparation prior to plating or the use of old medium. If old medium containing a stabilized form of L-glutamine and serum-free supplement for neural cell culture (see Materials Table) is used, the cells initially adhere but then float away after about two weeks. If bacterial contamination is a persistent problem, an antibiotic, such as ampicillin or pen-strep, can be added to the medium. There are also fungicides available to treat fungal contamination. These are some of the more common variables that can affect the outcome of the cultures. There are many others that will only be encountered after time and experience.
In comparison to the use of glass microelectrodes, this technique is excellent for studying network dynamics and pharmacological responses. It enables the use of many different spatio-temporal stimulation patterns and allows for the recording of neuronal responses from multiple areas at once. Previous groups have demonstrated interesting results using protocols similar to the ones described here18. Since the cultures last for weeks or months and the same cultures can be reused, this technique also allows for multiple experiments over time on the same network.
However, there are limitations to this technique. MEAs are non-invasive. Therefore, they can only record extracellular activity, as opposed to patch-clamping or intracellular recording with pipettes. Moreover, since each electrode in an array is covered by several cells, it is not possible to resolve the activity of a single neuron. Conversely, because these are in vitro cultures, they cannot fully reproduce the structural properties of networks in the brain. Also, activity can only be recorded for less than 30 min at a time without some mechanism providing a CO2 atmosphere for the cells to maintain their pH balance.
Once this technique is mastered,pharmacological manipulations with or without electrical stimulation can be explored. New protocols to probe learning and memory formation in neuronal networks can also be designed and tested, along with protocols for hippocampal or spinal cord networks. Protocols for the stimulation and training of networks have been previously published, and some of these were further developed into in vivo protocols, such as the &#34;selective adaptation&#34; proposed by Eytan et al.19. Several protocols were tested. However, only results from a modification to the tetanus procedure proposed by Ruaro in 2005 are presented here14,20.

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures1. In contrast to more traditional methods-such as patch-clamping, which can only record activity from a single cell, or field recording with a glass pipette, which can record extracellular responses from the neurons surrounding the electrode at a single site-MEAs can simultaneously record from multiple sites in a cell culture without requiring the arduous task of placing each electrode individually. This allows for the study of the dynamic interactions between groups of cells that form a network within that culture. Moreover, the effects of electrical stimulation on network firing patterns2,3,4,5 and network control6 in neuronal cultures have been well documented, and numerous configurations of electrical stimulation and controls can be easily applied within the same experimental setup, allowing for a broad range of spatio-temporal dynamics to be explored.
One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning7,8,9,10,11,12,13. The Peixoto Lab previously examined the effects of high-frequency training signals, as described in Ruaro et al.14, on networks of mouse neurons plated on microelectrode arrays15. In these experiments, networks displayed an increase in response following training induced by electrical stimulation. The increased response was considered a form of learning via stimulus recognition, whereby the networks responded in a consistent manner to a change in stimulus after the application of a specific stimulation (i.e., training) protocol.
This protocol demonstrates how to culture neuronal cells on MEAs, successfully record from over 95% of the plated dishes, establish a protocol to train the networks to respond to patterns of stimulation, sort single-unit activity, plot histograms, and interpret the results from such experiments. The use of a proprietary system (see the Table of Materials) for the stimulation and recording of neuronal cultures is demonstrated, as well as the application of software packages (see the Table of Materials) to sort neuronal units. A custom-designed graphical user interface (see the Table of Materials) is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol.

<table border="0" cellpadding="0" cellspacing="0" width="1135">
	<colgroup>
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		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Ascorbic Acid</td>
			<td style="width:204px;">Sigma Aldrich</td>
			<td style="width:155px;">A4403</td>
			<td style="width:356px;">Make 4mg/mL aliquots</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">B-27</td>
			<td>Invitrogen</td>
			<td>17504-044</td>
			<td>Serum-free supplement for neural cell culture. Make 1 mL aliquots and freeze</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Beakers - glass - 100 mL</td>
			<td>VWR</td>
			<td>13912-160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Beakers - glass - 500 mL</td>
			<td>VWR</td>
			<td>10754-956</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Blunt-tipped thumb forceps</td>
			<td>World Precision Instruments</td>
			<td>500365-G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Cryogenic vial - sterile, 2 mL</td>
			<td>Fisher Scientific</td>
			<td>10-500-26</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Cryogenic vial - sterile, 5 mL</td>
			<td>Fisher Scientific</td>
			<td>10-500-27</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">DMEM with Glutamax</td>
			<td>Gibco Life Technology</td>
			<td>10569-010</td>
			<td>Cell culture media that contains a stabilized form of L-glutamine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">DNase</td>
			<td>Worthington Biochemical</td>
			<td>LK003172</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Ethanol</td>
			<td>Fisher Scientific</td>
			<td>04-355-451</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Fetal bovine serum (FBS)</td>
			<td>ATCC</td>
			<td>30-2030</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Fine-tipped thumb forceps</td>
			<td>World Precision Instruments</td>
			<td style="width:155px;">501324-G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Glass Pipets</td>
			<td>VWR</td>
			<td>14673-010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">GlutaMAX -- I (100X)</td>
			<td>Gibco Life Technology</td>
			<td>35050-061</td>
			<td>A stabilized form of L-glutamine used as a suplement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Hemocytometer chip</td>
			<td>Fisher Scientific</td>
			<td>22-600-101</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:420px;">Hibernate EB complete</td>
			<td>BrainBits</td>
			<td>D00118</td>
			<td>Ambient CO2 cell storage media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Horse Serum</td>
			<td>Atlanta Biologicals</td>
			<td>S12195H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Laminin</td>
			<td>Sigma Aldrich</td>
			<td>L2020-1MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS filter amplifier</td>
			<td>MultiChannel System</td>
			<td>FA60SBC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS headstage/pre-amplifier</td>
			<td>MultiChannel System</td>
			<td>MEA1060-INV</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:420px;">MCS microelectrode array</td>
			<td>MultiChannel System</td>
			<td>60MEA200/10iR-ITO</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS power supply</td>
			<td>MultiChannel System</td>
			<td>PS20W</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS signal divider</td>
			<td>MultiChannel System</td>
			<td>SDMEA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS stimulus generator</td>
			<td>MultiChannel System</td>
			<td>STG4002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">MCS temperature controller</td>
			<td>MultiChannel System</td>
			<td>TC02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Media storage bottle - glass 500 mL</td>
			<td>VWR</td>
			<td>10754-818</td>
			<td>Autoclave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Microcentrifuge tubes - 0.4 mL</td>
			<td>Thermo Fisher</td>
			<td>3485</td>
			<td>Autoclave</td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:420px;">Microcentrifuge tubes - 2 mL</td>
			<td>Thermo Fisher</td>
			<td>3434ECONO</td>
			<td>Autoclave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipette tips - 10 uL&#160;</td>
			<td>VWR</td>
			<td>37001-166</td>
			<td>Autoclave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipette tips - 1000 uL&#160;</td>
			<td>VWR</td>
			<td>13503-464</td>
			<td>Autoclave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipette tips - 200 uL&#160;</td>
			<td>VWR</td>
			<td>37001-596</td>
			<td>Autoclave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipetter - 10 uL&#160;</td>
			<td>Thermo Fisher</td>
			<td>4641170N</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipetter - 1000 uL&#160;</td>
			<td>Thermo Fisher</td>
			<td>4641210N</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Micropipetter - 200 uL&#160;</td>
			<td>Thermo Fisher</td>
			<td>4641230N</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Papain - 0.22 Filtered</td>
			<td>Worthington Biochemical</td>
			<td>LK003178</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Penicillin-Streptomycin (Pen Strep)</td>
			<td>Thermo Fisher</td>
			<td>15070063</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Petri dishes -100mm disposable</td>
			<td>Fisher Scientific</td>
			<td>08-757-100D</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Petri dishes -100mm glass</td>
			<td>VWR</td>
			<td>10754-788</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Petri dishes -35 mm&#160;</td>
			<td>Fisher Scientific</td>
			<td>08-757-100A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Phosphate buffer saline (PBS) (1X)</td>
			<td>Corning</td>
			<td>21-040-CV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Plasma Cleaning System</td>
			<td>Plasma Etch, Inc.</td>
			<td>PE-50</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Poly-D-lysine hydrobromide (PDL)</td>
			<td>Sigma Aldrich</td>
			<td>P6407-5MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Polyethylene conical (centrifuge) tube -15 mL</td>
			<td>Fisher Scientific</td>
			<td>05-527-90</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Polypropylene conical (centrifuge) tube -50 mL</td>
			<td>Falcon</td>
			<td>352070</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Procedure masks</td>
			<td>Imco</td>
			<td>1530-imc</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Pyruvate</td>
			<td>Sigma Aldrich</td>
			<td>P4562-5G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Scalpel blades</td>
			<td>World Precision Instruments</td>
			<td>&#160;500237-G</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:420px;">Scissors - iris</td>
			<td>World Precision Instruments</td>
			<td>14111-G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Scissors - large</td>
			<td>World Precision Instruments</td>
			<td>14214</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Scissors - small surgical</td>
			<td>World Precision Instruments</td>
			<td>&#160;501733-G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Serological pipette - sterile, 1 mL&#160;</td>
			<td>VWR</td>
			<td>89130-892</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Serological pipette - sterile, 2 mL&#160;</td>
			<td>VWR</td>
			<td>89130-894</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Serological pipette - sterile, 25 mL&#160;</td>
			<td>VWR</td>
			<td>89130-900</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Serological pipette - sterile, 50 mL&#160;</td>
			<td>VWR</td>
			<td>89130-902</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software: Matlab GUI</td>
			<td>Peixoto Lab</td>
			<td>npeixoto@gmu.edu</td>
			<td>Used to analyze .mat files.&#160; Available for free upon request.&#160; Contact npeixoto@gmu.edu to request a copy.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software: MCS MC_Rack</td>
			<td>MultiChannel System</td>
			<td>N/A</td>
			<td>Used for data acquisition</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software: NeuroExplorer</td>
			<td>Plexon&#160;</td>
			<td>http://www.plexon.com/products/neuroexplorer</td>
			<td>Used to convert .nex files to .mat</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software: Offline Sorter</td>
			<td>Plexon&#160;</td>
			<td>http://www.plexon.com/products/offline-sorter</td>
			<td>Used to sort neural spikes and convert data files to .plx and then to .nex</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software Manual: MCS MC_Rack</td>
			<td>MultiChannel System</td>
			<td></td>
			<td>https://www.multichannelsystems.com/sites/multichannelsystems.com/files/documents/manuals/MC_Rack_Manual.pdf</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software Manual: NeuroExplorer</td>
			<td>Plexon&#160;</td>
			<td></td>
			<td>https://www.neuroexplorer.com/downloads/Nex3Manual.pdf</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Software Manual: Offline Sorter</td>
			<td>Plexon&#160;</td>
			<td></td>
			<td>www.plexon.com/system/files/downloads/Offline%20Sorter%20v2.8%20Manual.pdf</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Spatula - small double-ended</td>
			<td>World Precision Instruments</td>
			<td>&#160;503440</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Stericup 0.22 &#181;m pore filter - 250 mL</td>
			<td>Millipore</td>
			<td>SCGVU02RE</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Transfer pipette - large bore&#160;</td>
			<td>Thermo Fisher</td>
			<td>335-1S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Transfer pipette - small bore&#160;</td>
			<td>Thermo Fisher</td>
			<td>242-1S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Trypan blue</td>
			<td>Sigma Aldrich</td>
			<td>T8154-20ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:420px;">Whatman filter paper</td>
			<td>Whatman</td>
			<td>1442 150</td>
			<td>Cut into 8 pie wedges and autoclave in a glass Petri dish</td>
		</tr>
	</tbody>
</table>

time-dependent increase in the network response to the stimulation of neuronal cell cultures on micro-electrode arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Watch this Scientific Journal Video about Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays at JoVE.com

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

Research

JoVE Journal

Neuroscience

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Sex Stratified Neuronal Cultures to Study Ischemic Cell Death Pathways

Sex differences in neuronal susceptibility to ischemic injury and neurodegenerative disease have long been observed, but the signaling mechanisms ...

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

Preparation of Neuronal Co-cultures with Single Cell Precision

Microfluidic embodiments of the Campenot chamber have attracted great interest from the neuroscience community. These interconnected co-culture platforms ...

Electroretinogram Analysis of the Visual Response in Zebrafish Larvae

The electroretinogram (ERG) is a noninvasive electrophysiological method for determining retinal function. Through the placement of an electrode on the ...

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model

Currently, large-scale networks derived from dissociated neurons growing and developing in vitro on extracellular micro-transducer devices are the ...

Investigating Functional Regeneration in Organotypic Spinal Cord Co-cultures Grown on Multi-electrode Arrays

Adult higher vertebrates have a limited potential to recover from spinal cord injury. Recently, evidence emerged that propriospinal connections are a ...

Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis

During neural development, growing axons extend to multiple synaptic partners by elaborating axonal branches. Axon branching is promoted by extracellular ...