Name: taking advantage of reduced droplet-surface interaction to optimize transport of bioanalytes in digital microfluidics
Uploaded: 2014-11-10T15:00:00
Description: Watch this Scientific Journal Video about Taking Advantage of Reduced Droplet-surface Interaction to Optimize Transport of Bioanalytes in Digital Microfluidics at JoVE.com

We thank the Lindback Foundation for financial support, and Dr. Alexander Sidorenko and Elza Chu for fruitful discussions and technical assistance, and Professor Robert Smith for assistance with the C. elegans assays.

NOTE: In the procedures described below, laboratory safety guidelines must always be followed. Of particular importance is the safety when dealing with high voltage (&#62; 500 V) and handling chemicals.
1. Coating of a Conductive Substrate with Candle Soot
<ol>
	<li>Cut copper metal into rectangles (75 x 43 mm, 0.5 mm thick). Clean each copper substrate by immersion in copper etchant for about 30 sec, wash with tap water for about 20 sec, and dry with paper. 
	NOTE: If using Method 1 be...

<ol>
	<li>Fair, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digital+microfluidics:+is+a+true+lab-on-a-chip+possible.">Digital microfluidics: is a true lab-on-a-chip possible.</a> Microfluid Nanofluid. 3 (3), 245-281 (2007).</li><li>Gupta, S., Alargova, R. G., Kilpatrick, P. K., Velev, O. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On-Chip+Dielectrophoretic+Coassembly+of+Live+Cells+and+Particles+into+Responsive+Biomaterials.">On-Chip Dielectrophoretic Coassembly of Live Cells and Particles into Responsive Biomaterials.</a> Langmuir. 26 (5), 3441-3452 (2009).</li><li>Shih, S. 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=Dried+blood+spot+analysis+by+digital+microfluidics+coupled+to+nanoelectrospray+ionization+mass+spectrometry.">Dried blood spot analysis by digital microfluidics coupled to nanoelectrospray ionization mass spectrometry.</a> Anal Chem. 84 (8), 3731-3738 (2012).</li><li>Gorbatsova, J., Borissova, M., Kaljurand, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrowetting-on-dielectric+actuation+of+droplets+with+capillary+electrophoretic+zones+for+off-line+mass+spectrometric+analysis.">Electrowetting-on-dielectric actuation of droplets with capillary electrophoretic zones for off-line mass spectrometric analysis.</a> J Chromatogr. 1234, 9-15 (2012).</li><li>Qin, J., Wheeler, 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=Maze+exploration+and+learning+in+C.+elegans.">Maze exploration and learning in C. elegans.</a> Lab Chip. 7 (2), 186-192 (2007).</li><li>Koc, Y., de Mello, A. J., McHale, G., Newton, M. I., Roach, P., Shirtcliffe, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nano-scale+superhydrophobicity:+suppression+of+protein+adsorption+and+promotion+of+flow-induced+detachment.">Nano-scale superhydrophobicity: suppression of protein adsorption and promotion of flow-induced detachment.</a> Lab Chip. 8 (4), 582-586 (2008).</li><li>Perry, G., Thomy, V., Das, M. R., Coffinier, Y., Boukherroub, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibiting+protein+biofouling+using+graphene+oxide+in+droplet-based+microfluidic+microsystems.">Inhibiting protein biofouling using graphene oxide in droplet-based microfluidic microsystems.</a> Lab Chip. 12 (9), 1601-1604 (2012).</li><li>Kumari, N., Garimella, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrowetting-Induced+Dewetting+Transitions+on+Superhydrophobic+Surfaces.">Electrowetting-Induced Dewetting Transitions on Superhydrophobic Surfaces.</a> Langmuir. 27 (17), 10342-10346 (2011).</li><li>Freire, S. L. S., Tanner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Additive-Free+Digital+Microfluidics.">Additive-Free Digital Microfluidics.</a> Langmuir. 29 (28), 9024-9030 (2013).</li><li>Deng, X., Mammen, L., Butt, H. -. J., Vollmer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candle+Soot+as+a+Template+for+a+Transparent+Robust+Superamphiphobic+Coating.">Candle Soot as a Template for a Transparent Robust Superamphiphobic Coating.</a> Science. 335, 67-70 (2011).</li><li>Kang, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+Electrostatic+Fields+Change+Contact+Angle+in+Electrowetting.">How Electrostatic Fields Change Contact Angle in Electrowetting.</a> Langmuir. 18 (26), 10318-10322 (2002).</li><li>Abdelgawad, M., Watson, M. W. L., Young, E. W. K., Mudrik, J. M., Ungrin, M. D., Wheeler, 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=Soft+lithography:+masters+on+demand.">Soft lithography: masters on demand.</a> Lab Chip. 8 (8), 1379-1385 (2008).</li><li>Barbulovic-Nad, I., Yang, H., Park, P. S., Wheeler, 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=Digital+microfluidics+for+cell-based+assays.">Digital microfluidics for cell-based assays.</a> Lab Chip. 8 (4), 519-526 (2008).</li><li>Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+Caenorhabditis+elegans.">The genetics of Caenorhabditis elegans.</a> Genetics. 77 (1), 71-94 (1974).</li></ol>

The most critical step of the protocol is the protection of the soot layer, directly associated with the success in moving droplets. Metallizing the soot layer (method 1 above) allows close to 100% of fabrication success. However, the maximum operation time is about 10 min; possibly, droplet fractions are wetting the soot through holes in the metal layer. Coating the soot layer with the fluorinated liquid is the easiest and fastest alternative, and requires minimum resources, but only 40&#8211;50% of the fabricated subst...

The miniaturization of devices that work with liquids is of paramount importance for the development of &#8220;lab-on-a-chip&#8221; platforms. In this direction, the last two decades have witnessed a significant progress in the field of microfluidics, with a variety of applications.1-5 Contrasting with the transport of fluid in enclosed channels (channel microfluidics), DMF manipulates droplets on arrays of electrodes. One of the most attractive merits of this technique is the absence of movable parts to transport fluids, and motion is instantly stopped by turning off electric signals.
However, droplet motion is dependent on droplet contents, certainly an undesirable characteristic for a universal &#8220;lab-on-a-chip&#8221; platform. Droplets containing proteins and other analytes stick to device surfaces, becoming unmovable. Arguably, this has been the major limitation for broadening the scope of DMF applications;6-8 alternatives to minimize the unwanted surface fouling involve the addition of extra chemical species to the droplet or its surroundings, which could potentially affect droplet content.
Previously, our group developed a device to allow the transport of cells and proteins in DMF, without extra additives (Field-DW devices).9 This was achieved by combining a surface based on candle soot,10 with a device geometry that favors droplet rolling and leads to an upward force on the droplet, further decreasing droplet-surface interaction. In this approach, droplet motion is not associated with surface wetting.11
The goal of the detailed method described below is to produce a DMF device capable of transporting droplets containing proteins, cells, and entire organisms, without extra additives. The Field-DW devices pave the way for fully controlled platforms working largely independently of droplet chemistry.
Here, we also present simulations showing that, despite the high voltage required for device operation, the voltage drop across the droplet is a small fraction of the applied voltage, indicating negligible effects on bioanalytes inside the droplet. In fact, preliminary tests with Caenorhabditis elegans (C. elegans), a nematode used for a variety of studies in biology, show that worms swim undisturbed as voltages are applied.

<table border="0" cellpadding="0" cellspacing="0" width="1230">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Name of Material/ Equipment</td>
			<td style="width:119px;">Company</td>
			<td style="width:161px;">Catalog Number</td>
			<td style="width:720px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Paraffin candle</td>
			<td style="width:119px;"></td>
			<td style="width:161px;"></td>
			<td>Any paraffin candle</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Sputtering system</td>
			<td style="width:119px;">Denton Vacuum, Moorestown, NJ</td>
			<td style="width:161px;"></td>
			<td>Sputter coater Desk V HP equipped with an Au target.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">1-dodecanethiol</td>
			<td style="width:119px;">Sigma-Aldrich</td>
			<td align="right" style="width:161px;">471364</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Teflon</td>
			<td style="width:119px;">Dupont</td>
			<td style="width:161px;">AF-1600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Fluorinert FC-40</td>
			<td style="width:119px;">Sigma-Aldrich</td>
			<td style="width:161px;">F9755</td>
			<td>Fluorinated liquid: Prepare Teflon-AF resin in Fluorinert FC-40, 1:100 (w/w), to create the hydrophobic coating.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Graphic design software -Adobe Illustrator</td>
			<td style="width:119px;">Adobe Systems</td>
			<td style="width:161px;"></td>
			<td>Other softwares might be used as well.</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:231px;">Copper laminate</td>
			<td style="width:119px;">Dupont</td>
			<td style="width:161px;">LF9110</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:231px;">Laser Printer</td>
			<td style="width:119px;">Xerox</td>
			<td style="width:161px;">Phaser 6360 or similar</td>
			<td>Check for the compatibility with &#34;rich black&#34; or &#34;registration black&#34; (see text).</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Copper Etchant</td>
			<td style="width:119px;">Transene</td>
			<td style="width:161px;">CE-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Perfluoroalkoxy (PFA) film</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td style="width:161px;">84955K22</td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:231px;">Breadboard</td>
			<td style="width:119px;">Allied Electronics</td>
			<td style="width:161px;">70012450 or similar</td>
			<td>Large enough to allow the assemble of 10 drivers.</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:231px;">Universal circuit board</td>
			<td style="width:119px;">Allied Electronics</td>
			<td style="width:161px;">70219535 or similar</td>
			<td></td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:231px;">Connector</td>
			<td style="width:119px;">Allied Electronics</td>
			<td style="width:161px;">5145154-8 or similar</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Control board and control program (LabView software)</td>
			<td style="width:119px;">National Instruments</td>
			<td style="width:161px;">NI-6229 or similar</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">High-voltage amplifier</td>
			<td style="width:119px;">Trek</td>
			<td style="width:161px;">PZD700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Resistor R 27 k&#8486;, 1/4 W</td>
			<td style="width:119px;">Allied&#160;</td>
			<td align="right" style="width:161px;">2964762</td>
			<td></td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:231px;">Capacitors C and C1, 100 nF, 60 V</td>
			<td style="width:119px;">Allied&#160;</td>
			<td align="right" style="width:161px;">8817183</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Transistor T, NPN</td>
			<td style="width:119px;">Allied&#160;</td>
			<td align="right" style="width:161px;">9350289</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Diode D, 1N4007</td>
			<td style="width:119px;">Allied&#160;</td>
			<td align="right" style="width:161px;">2660007</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Relay&#160;</td>
			<td style="width:119px;">Allied&#160;</td>
			<td align="right" style="width:161px;">8862527</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Visualization system</td>
			<td style="width:119px;">Edmund Optics</td>
			<td style="width:161px;">VZM 200i or similar</td>
			<td>System magnification 24X- 96X. It is combined with a Hitachi KP-D20B 1/2 in CCD Color Camera.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Recorder</td>
			<td style="width:119px;">Sony</td>
			<td style="width:161px;">GV-D1000 NTSC or similar</td>
			<td>It is connected to the camera by an S-video cable.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:231px;">Simulations</td>
			<td style="width:119px;">COMSOL Multiphysics</td>
			<td style="width:161px;">V. 4.4</td>
			<td></td>
		</tr>
	</tbody>
</table>

taking advantage of reduced droplet-surface interaction to optimize transport of bioanalytes in digital microfluidics

Digital microfluidics (DMF), a technique for manipulation of droplets, is a promising alternative for the development of &#8220;lab-on-a-chip&#8221; platforms. Often, droplet motion relies on the wetting of a surface, directly associated with the application of an electric field; surface interactions, however, make motion dependent on droplet contents, limiting the breadth of applications of the technique.
Some alternatives have been presented to minimize this dependence. However, they rely on the addition of extra chemical species to the droplet or its surroundings, which could potentially interact with droplet moieties. Addressing this challenge, our group recently developed Field-DW devices to allow the transport of cells and proteins in DMF, without extra additives.
Here, the protocol for device fabrication and operation is provided, including the electronic interface for motion control. We also continue the studies with the devices, showing that multicellular, relatively large, model organisms can also be transported, arguably unaffected by the electric fields required for device operation.

Previously, we have used Field-DW devices to allow the motion of proteins in DMF. In particular, droplets with bovine serum albumin (BSA) could be moved at a concentration 2,000 times higher than previously reported by other authors (without additives). This was due to the reduced interaction between droplet and surface; Figure 4 shows a droplet containing fluorescently-tagged BSA (see Freire et al.9 for more information on the experiments). The first picture on the left shows the dro...

Watch this Scientific Journal Video about Taking Advantage of Reduced Droplet-surface Interaction to Optimize Transport of Bioanalytes in Digital Microfluidics at JoVE.com

Taking Advantage of Reduced Droplet-surface Interaction to Optimize Transport of Bioanalytes in Digital Microfluidics

The protocol for fabrication and operation of field dewetting devices (Field-DW) is described, as well as the preliminary studies of the effects of electric fields on droplet contents.

Digital microfluidics (DMF), a technique for manipulation of droplets, is a promising alternative for the development of &#8220;lab-on-a-chip&#8221; ...

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