Name: a microfluidic device with groove patterns for studying cellular behavior
Uploaded: 2007-08-30T00:00:00
Description: Watch this Scientific Journal Video about A Microfluidic Device with Groove Patterns for Studying Cellular Behavior at JoVE.com

<p class="jove_step">A. Microfabrication of the microfluidic device</p>
<ol>
<li>4-inch Si wafer is treated with reactive oxygen plasma (5 min at 30W, Harrick Scientific, NY).</li>
<li>Negative photoresist (SU-8 2015, Microchem, MA) is spin-coated at 900 rpm for 1 min on a Si wafer.</li>
<li>The wafer is soft baked at 95&deg;C for 6 min on a hotplate and is exposed to UV light (200W) for 4 min through a mask film containing microchannels.</li>
<li>The wafer is post baked at 95&deg;C for 6 min and is developed using SU-8 photoresist developer.</...

<ol>
	<li>Chung, B. G., Park, J. W., Hu, J. S., Huang, C., Monuki, E. S., Jeon, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+hybrid+microfluidic-vacuum+device+for+interfacing+with+conventional+cell+culture+platform.">A hybrid microfluidic-vacuum device for interfacing with conventional cell culture platform.</a> <em style='font-style: italic'>BMC Biotechnol</em>. <b>7</b>, (2007).</li><li>Khademhosseini, A., Yeh, J., Eng, G., Karp, J., Kaji, H., Borenstein, J., Farokhzad, O. C., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+docking+inside+microwells+within+reversibly+sealed+microfluidic+channels+for+fabricating+multiphenotype+cell+arrays.">Cell docking inside microwells within reversibly sealed microfluidic channels for fabricating multiphenotype cell arrays.</a> <em style='font-style: italic'>Lab Chip</em>. <b>5</b>, 1380-1386 (2005).</li><li>Whittemore, E. R., Loo, D. T., Watt, J. A., Cotman, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+detailed+analysis+of+hydrogen+peroxide-induced+cell+death+in+primary+neuronal+culture.">A detailed analysis of hydrogen peroxide-induced cell death in primary neuronal culture.</a> <em style='font-style: italic'>Neuroscience</em>. <b>67</b>, 921-932 (1995).</li></ol>

<p class="jove_content">Cells were immobilized and patterned within microgrooved substrates in a microfluidic device.  The apoptosis process of cells exposed to hydrogen peroxide was observed in real-time and analyzed by using Annexin V and propidium iodide.  Thus, this microfluidic device containing microgroove channels could be useful for high-throughput drug screening.</p>

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>PDMS</td>
<td>Reagent</td>
<td>K.R. Anderson Co. Inc. </td>
<td>2065622 </td>
<td>Poly(dimethylsiloxane), Dow Corning Sylgard 184 (8.6 lb)</td>
</tr>
<tr>
<td>DMEM</td>
<td>medium</td>
<td>Invitrogen</td>
<td>11965 </td>
<td>Dulbecco&#x2019;s Modified Eagle&#x2019;s Media </td>
</tr>
<tr>
<td>FBS</td>
<td>serum</td>
<td>Invitrogen</td>
<td>10082-147 </td>
<td>Fetal Bovine Serum
</td>
</tr>
<tr>
<td>Hydrogen peroxide</td>
<td>Reagent</td>
<td>Sigma Aldrich</td>
<td> H1009 </td>
<td>&nbsp;</td>
<tr>
<td>Apoptosis assay</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>V13242 </td>
<td>Annexin A, propidium iodide</td>
</tr>
<tr>
<td>Negative photoresist </td>
<td>&nbsp;</td>
<td>Microchem</td>
<td>SU-8 2015</td>
<td>&nbsp;</td>
<tr>
<td>Si wafer </td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>4 inch silicone wafer</td>
</tr>
<tr>
<td>Reactive oxygen plasma </td>
<td>Reagent</td>
<td>Harrick Scientific</td>
<td>&nbsp;</td>
<td>treat wafer 5 min at 30W</td>
</tr>
<tr>
<td>inverted microscope </td>
<td>Tool</td>
<td>Nikon </td>
<td>TE 2000</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
			<div>
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a microfluidic device with groove patterns for studying cellular behavior

<p class="jove_content">We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate.  To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate.  Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 &mu;m wide).  To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours.  We found that cells exposed to the oxidative stress became apoptotic.  These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process.  Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37&deg;C, 5% CO<sub>2</sub>).  Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.</p>

Watch this Scientific Journal Video about A Microfluidic Device with Groove Patterns for Studying Cellular Behavior at JoVE.com

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior

<p class="jove_content">We describe a protocol for the fabrication of microfluidic devices that can enable cell capture and culture.  In this approach patterned microstructures such as grooves within microfluidic channels are used to create low shear stress regions within which cell can dock.</p>

We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel ...

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