The authors acknowledge the support of the Australian Research Council through the Centre of Excellence for Electromaterials Science.

The protocol has been approved by the La Trobe University (09-28P) and RMIT University Animal Ethics committees (1315).
1. Electrode Preparation and Preliminary in vitro Testing
<ol>
	<li>Prepare electrode coating deposition solutions; for instance 10 mM 3,4-ethylenedioxythiophene (EDOT) and 0.1 M sodium para-toluene sulfonate (Na2pTS) to form poly-3,4-ethylenedioxythiophene-pTS (PEDOT-pTS).</li>
	<li>Connect the electrode array to a potentiostat.</li>
	<li>Caref...

<ol>
	<li>Oluigbo, C. O., Rezai, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Addressing+Neurological+Disorders+With+Neuromodulation.">Addressing Neurological Disorders With Neuromodulation.</a> IEEE Trans. Biomed. Eng. 58, 1907-1917 (2011).</li><li>Shivdasani, M. N., Mauger, S. J., Rathbone, G. D., Paolini, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inferior+Colliculus+Responses+to+Multichannel+Microstimulation+of+the+Ventral+Cochlear+Nucleus:+Implications+for+Auditory+Brain+Stem+Implants.">Inferior Colliculus Responses to Multichannel Microstimulation of the Ventral Cochlear Nucleus: Implications for Auditory Brain Stem Implants.</a> J. Neurophysiol. 99, 1-13 (2008).</li><li>Perlmutter, J. S., Mink, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+Brain+Stimulation.">Deep Brain Stimulation.</a> Ann. Rev. Neurosci. 29, 229 (2006).</li><li>Weaver, F. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+Deep+Brain+Stimulation+vs+Best+Medical+Therapy+for+Patients+With+Advanced+Parkinson+Disease.">Bilateral Deep Brain Stimulation vs Best Medical Therapy for Patients With Advanced Parkinson Disease.</a> J. Am. Med. Assoc. 301, 63-73 (2009).</li><li>Biran, R., Martin, D. C., Tresco, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+cell+loss+accompanies+the+brain+tissue+response+to+chronically+implanted+silicon+microelectrode+arrays.">Neuronal cell loss accompanies the brain tissue response to chronically implanted silicon microelectrode arrays.</a> Exp. Neurol. 195, 115-126 (2005).</li><li>McConnell, G. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implanted+neural+electrodes+cause+chronic,+local+inflammation+that+is+correlated+with+local+neurodegeneration.">Implanted neural electrodes cause chronic, local inflammation that is correlated with local neurodegeneration.</a> J. Neural Eng. 6, (2009).</li><li>Liu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stability+of+the+interface+between+neural+tissue+and+chronically+implanted+intracortical+microelectrodes.">Stability of the interface between neural tissue and chronically implanted intracortical microelectrodes.</a> IEEE Trans. Rehab. Eng. 7, 315-326 (1999).</li><li>Rousche, P. J., Normann, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+recording+capability+of+the+Utah+Intracortical+Electrode+Array+in+cat+sensory+cortex.">Chronic recording capability of the Utah Intracortical Electrode Array in cat sensory cortex.</a> J. Neurosci. Methods. 82, 1-15 (1998).</li><li>Williams, J. C., Rennaker, R. L., Kipke, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+neural+recording+characteristics+of+wire+microelectrode+arrays+implanted+in+cerebral+cortex.">Long-term neural recording characteristics of wire microelectrode arrays implanted in cerebral cortex.</a> Brain Res. Protoc. 4, 303-313 (1999).</li><li>Harris, A. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conducting+polymer+coated+neural+recording+electrodes.">Conducting polymer coated neural recording electrodes.</a> J. Neural Eng. 10, (2013).</li><li>Bard, A. J., Faulkner, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electrochemical Methods. , (2001).</li><li>Ludwig, K. A., Uram, J. D., Yang, J., Martin, D. C., Kipke, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+neural+recordings+using+silicon+microelectrode+arrays+electrochemically+deposited+with+a+poly(3,4-ethylenedioxythiophene)+(PEDOT)+film.">Chronic neural recordings using silicon microelectrode arrays electrochemically deposited with a poly(3,4-ethylenedioxythiophene) (PEDOT) film.</a> J. Neural Eng. 3, 59 (2006).</li></ol>

This protocol provides a method for comparing neural recording electrode coatings within one animal.&#160;The electrode design used is ideal for implantation into a rat inferior colliculus (IC), with dimensions of a similar scale.&#160;Variations of this electrode such as more space between shanks would prevent all shanks being in the rat IC at the same time, while longer shanks and a larger pitch between electrodes increase the risk that the shank tips will come in contact with the base of the skull during insertion.&#1...

Neural implants are being used increasingly for research, controlling prosthetics and treatment of disorders such as Parkinson&#39;s disease, epilepsy, and sensory loss1,2.&#160;Measuring and/or controlling both the chemical and electrical composition of the brain is the basis for all neural implants.&#160;However, it is important to administer a treatment only when the neural tissue is in the aberrant state to reduce side effects3.&#160;For instance, deep brain stimulators for epilepsy treatment should only apply an electrical pulse to the brain during a seizure.&#160;Some side effects may be dystonia, loss of memory, disorientation, impaired cognitive function, induced hallucinations, depression or anti-depression3,4.&#160;In many devices, a closed loop system is therefore necessary to record electrical activity and to trigger stimulation when an abnormal state is detected.&#160;Recording electrodes are also used to control prosthetic devices.&#160;It is critical to record the target neural activity with the highest possible signal-to-noise ratio to achieve the most accurate triggering and device control.&#160;A large signal-to-noise ratio is also highly desirable for research applications, as more reliable data can be obtained, resulting in fewer required test subjects.&#160;This will also allow a greater understanding of the mechanisms and pathways involved in neural stimulation and recording.
After a neural implant has been placed into the brain, an immune response is triggered5,6.&#160;The time course of the response is generally divided into acute and chronic phases, each consisting of different biological processes7.&#160;The immune response can have dramatic effects on the performance of the implant, such as isolation of the electrodes from the target neurons by encapsulation in a glial scar or chemical degradation of the implant materials8.&#160;This can reduce the signal-to-noise ratio of a recording electrode and the power output of a stimulating electrode, and lead to electrode failure9.&#160;Careful choice of implant design and materials are necessary to prevent failure over the implant lifetime.
Many different materials and implant designs have been developed recently to improve the signal-to-noise ratio and implant stability for neural recording.&#160;Electrode materials have included platinum, iridium, tungsten, iridium oxide, tantalum oxide, graphene, carbon nanotubes, doped conducting polymers, and more recently hydrogels.&#160;Substrate materials tested also includes silicon, silicon oxide, silicon nitride, silk, Teflon, polyimide, and silicone.&#160; Various electrode modifications have also been investigated, using coatings such as laminin, neurotrophins, or self-assembled monolayers and treatments using electrochemical, plasma and optical techniques.&#160;Implant designs could be 1-, 2- or 3-dimensional with the electrodes generally at the tip of an insulating probe or along the edge of a shank for penetrating electrodes or in a 2-dimensional array for cortex surface implants.&#160;Regardless of electrode design or material, previous literature has typically demonstrated the performance of the new implant without reference to other implant constructs.&#160;This prevents a systematic evaluation of their properties.
This protocol provides a method for comparing different electrode materials via a range of analytical and electrophysiological techniques.&#160;It is based on a recently published article which compared 4 different doped conducting polymer coatings (polypyrrole (Ppy) and poly-3,4-ethylenedioxythiophene (PEDOT) doped with sulfate (SO4) or para-toluene sulfonate (pTS)) and 4 different coating thicknesses10.&#160;This article found one material, PEDOT-pTS with a 45 sec deposition time, had the highest signal-to-noise ratio and spike count with the smallest background noise and that these parameters were dependent on electrode impedance.&#160;PEDOT-pTS also displayed superior acute biostability compared to the other doped conducting polymers and bare iridium electrodes.&#160;The protocol allows the critical parameters controlling the signal-to-noise ratio and stability to be determined and used to further improve the performance of neural recording electrodes.

<table border="0" cellpadding="0" cellspacing="0" width="1217">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Programmable Attenuator</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">PA5</td>
			<td style="width:600px;">Controls the amplitude of the acoustic signal across frequencies</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Electrostatic speaker driver</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">ED1</td>
			<td>Drives the electrostatic speakers (EC1)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Coupled electrostatic speaker</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">EC1</td>
			<td>Delivers sound to the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Processing base station</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">RZ2</td>
			<td>Records neural activity from electrode array (using PZ2 preamplifier)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Preamplifier</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">PZ2-256</td>
			<td>256-channel high impedance preamplifier</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Multifunction Processor</td>
			<td style="width:195px;">TDT</td>
			<td style="width:179px;">RX6</td>
			<td>Used to generate acoustic stimuli</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:244px;">Multichannel electrode</td>
			<td style="width:195px;">NeuroNexus Technologies</td>
			<td style="width:179px;">A4 &#215; 8&#8211;5mm-200-200-413</td>
			<td>4-shank 32-channel electrode array</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Potentiostat</td>
			<td style="width:195px;">CH Instruments</td>
			<td style="width:179px;">CHI660B</td>
			<td>Deposits electrode coatings and performs cyclic voltammetry and EIS (used with CHI684)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Multiplexer</td>
			<td style="width:195px;">CH Instruments</td>
			<td style="width:179px;">CHI684</td>
			<td>Switches between electrodes on the potentiostat</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">di-sodium phosphate</td>
			<td style="width:195px;">Fluka</td>
			<td style="width:179px;">71644</td>
			<td>Used in the test solution</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:244px;">3,4-ethylenedioxythiophene (EDOT)</td>
			<td style="width:195px;">Sigma Aldrich</td>
			<td style="width:179px;">483028</td>
			<td>An electrode coating material</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:244px;">para-toluene sulfonate (Na2pTS)</td>
			<td style="width:195px;">Sigma Aldrich</td>
			<td style="width:179px;">152536</td>
			<td>An electrode coating material</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Urethane</td>
			<td style="width:195px;">Sigma Aldrich</td>
			<td style="width:179px;">U2500</td>
			<td>Used to anaesthetise the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Silver/Silver chloride electrode</td>
			<td style="width:195px;">CH Instruments</td>
			<td style="width:179px;">CHI111</td>
			<td>Used for testing the electrode in vitro</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Platinum electrode</td>
			<td style="width:195px;">CH Instruments</td>
			<td style="width:179px;">MW4130</td>
			<td>Used for testing the electrode in vitro</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Motorized microdrive</td>
			<td style="width:195px;">Sutter Instruments</td>
			<td style="width:179px;">DR1000</td>
			<td>To control the electrode array position during surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Enzymatic cleaner</td>
			<td style="width:195px;">Advanced Medical Optics</td>
			<td style="width:179px;">Ultrazyme</td>
			<td>Cleans the protein off the electrode array after implantation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Acoustic enclosure</td>
			<td style="width:195px;">TMC Ametek</td>
			<td style="width:179px;">83-501</td>
			<td>Isolates the animal from acoustic and electrical noise</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Stereotaxic frame</td>
			<td style="width:195px;">David Kopf Instruments</td>
			<td style="width:179px;">1430</td>
			<td>Secures and positions the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Temperature controller</td>
			<td style="width:195px;">World Precision Instruments</td>
			<td style="width:179px;">ATC1000</td>
			<td>Controls the animal temperature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Bone drill</td>
			<td style="width:195px;">KaVo Dental</td>
			<td style="width:179px;">K5Plus</td>
			<td>Used to perform the craniectomy</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Aspirator</td>
			<td style="width:195px;">Flaem</td>
			<td style="width:179px;">Suction pro</td>
			<td>Used to perform the craniectomy</td>
		</tr>
	</tbody>
</table>

a method for systematic electrochemical and electrophysiological evaluation of neural recording electrodes

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant characteristics.&#160;This precludes a rational selection of a particular implant as optimal for a particular application and the development of new materials based on the most critical performance parameters.&#160;This article develops a protocol for in vitro and in vivo testing of neural recording electrodes.&#160;Recommended parameters for electrochemical and electrophysiological testing are documented with the key steps and potential issues discussed.&#160;This method eliminates or reduces the impact of many systematic errors present in simpler in vivo testing paradigms, especially variations in electrode/neuron distance and between animal models.&#160;The result is a strong correlation between the critical in vitro and in vivo responses, such as impedance and signal-to-noise ratio.&#160;This protocol can easily be adapted to test other electrode materials and designs.&#160;The in vitro techniques can be expanded to any other nondestructive method to determine further important performance indicators.&#160;The principles used for the surgical approach in the auditory pathway can also be modified to other neural regions or tissue.

A typical electrode array used for this experimental protocol is shown in Figure 1.&#160;There are 32 iridium electrodes on 4 shanks with 413 &#956;m2 nominal geometric area and a 200 &#956;m pitch.&#160;Every second electrode on the array has been coated with one of four different electrode coatings, labeled 1-4.&#160;The coating materials have been carefully chosen for their chemical, mechanical and electrochemical properties.&#160;As mentioned previously10, incr...

Watch this Scientific Journal Video about A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes at JoVE.com

A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes

Different electrode coatings affect neural recording performance through changes to electrochemical, chemical and mechanical properties. Comparison of electrodes in vitro is relatively simple, however comparison of in vivo response is typically complicated by variations in electrode/neuron distance and between animals. This article provides a robust method to compare neural recording electrodes.

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant ...

a-method-for-systematic-electrochemical-electrophysiological

Research

JoVE Journal

Neuroscience

13.3K vistas.  La Trobe University.  Diferentes recubrimientos de electrodos afectan al rendimiento de grabación neural a través de cambios en las propiedades electroquímicas, químicas y mecánicas. Comparación de los electrodos in vitro es relativamente simple, sin embargo la comparación de la respuesta in vivo típicamente se complica por las variaciones en la distancia de los electrodos / neurona y entre animales. Este artículo proporciona un método robusto para comparar electro...