Name: patterning cells on optically transparent indium tin oxide electrodes
Uploaded: 2007-08-20T00:00:00
Description: Watch this Scientific Journal Video about Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes at JoVE.com

<p class="jove_title">Part I: Patterning of the electrodes</p>
<ol>
<li>Clean and dehydrated ITO coated glass slides were coated with positive photoresist</li>
<li>The surface was baked at 100&deg;C to remove solvents from the resist</li>
<li>After baking, the surface was exposed to ultraviolet light through a photomask using the Canon Mask Aligner</li>
<li>The exposed regions were removed in a developer solution</li>
<li>The surface was hard baked to remove any remaining developer solution</li>
<li>The ITO regions not protected by photoresist ...

<ol>
	<li>Lee, J. Y., Jones, C., Zern, M. A., Revzin, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Local+Tissue-Specific+Gene+Expression+in+Cellular+Micropatterns.">Analysis of Local Tissue-Specific Gene Expression in Cellular Micropatterns.</a> <em style='font-style: italic'> Analytical Chemistry</em>. <b>78</b>, 8305-8312 (2006).</li></ol>

<p class="jove_content">In this video, we have demonstrated the cell patterning on optically transparent indium tin oxide electrodes.  After fabricating the ITO electrodes using photolithography, they were modified with a cell-resistive monolayer of PEG silane.  This monolayer was desorbed using electrochemistry switching the surface from cell-resistive to cell-adhesive.  Patterning of 3T3 murine fibroblasts has been shown in his video. We have also patterning hepatocytes, primary cells, and stellate cells using the same technique.  In addition...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>Indium Tin Oxide</td>
<td>Other</td>
<td>Delta Technologies</td>
<td>CB 40IN</td>
<td>&nbsp;</td>
<tr>
<td>PEG silane</td>
<td>Reagent</td>
<td>Gelest Inc.</td>
<td>SIM6492.66</td>
<td>&nbsp;</td>
<tr>
<td>Hydrochloric Acid</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>320331</td>
<td>&nbsp;</td>
<tr>
<td>Nitric Acid</td>
<td>Reagent</td>
<td>Sigma Aldrich</td>
<td>258121</td>
<td>&nbsp;</td>
<tr>
<td>Ethanol</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>459836</td>
<td>&nbsp;</td>
<tr>
<td>Acetone</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>650501</td>
<td>&nbsp;</td>
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<td>Toluene</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>34866</td>
<td>&nbsp;</td>
<tr>
<td>Media</td>
<td>Reagent</td>
<td>Invitrogen Technologies</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Collagen</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>C3867</td>
<td>&nbsp;</td>
<tr>
<td>3T3 murine Fibroblasts</td>
<td>Cell Line</td>
<td>ATCC</td>
<td>&nbsp;</td>
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patterning cells on optically transparent indium tin oxide electrodes

The ability to exercise precise spatial and temporal control over cell-surface interactions is an important prerequisite to the assembly of multi-cellular constructs serving as in vitro mimics of native tissues.  In this study, photolithography and wet etching techniques were used to fabricate individually addressable indium tin oxide (ITO) electrodes on glass substrates.  The glass substrates containing ITO microelectrodes were modified with poly(ethylene glycol) (PEG) silane to make them protein and cell resistive.  Presence of insulating PEG molecules on the electrode surface was verified by cyclic voltammetry employing potassium ferricyanide as a redox reporter molecule.  Importantly, the application of reductive potential caused desorption of the PEG layer, resulting in regeneration of the conductive electrode surface and appearance of typical ferricyanide redox peaks.    Application of reductive potential also corresponded to switching of ITO electrode properties from cell non-adhesive to cell-adhesive. Electrochemical stripping of PEG-silane layer from ITO microelectrodes allowed for cell adhesion to take place in a spatially defined fashion, with cellular patterns corresponding closely to electrode patterns.  Micropatterning of several cell types was demonstrated on these substrates.   In the future, the control of the biointerfacial properties afforded by this method will allow to engineer cellular microenvironments through the assembly of three or more cell types into a precise geometric configuration on an optically transparent substrate.

Watch this Scientific Journal Video about Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes at JoVE.com

Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes

<p class="jove_content">Non-fouling PEG silane monolayer was desorbed from individually addressable ITO electrodes on glass by application of a reductive potential.   Electrochemical stripping of PEG-silane layer from ITO microelectrodes allowed for cell adhesion to take place in a spatially defined fashion, with cellular patterns corresponding closely to electrode patterns.</p>

The ability to exercise precise spatial and temporal control over cell-surface interactions is an important prerequisite to the assembly of multi-cellular ...

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