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

Ringraziamo la Fondazione Lindback per il sostegno finanziario, e il dottor Alexander Sidorenko e Elza Chu per le discussioni fruttuose e assistenza tecnica, e il professor Robert Smith per l&#39;assistenza con la C. saggi elegans.

NOTA: Nelle procedure descritte di seguito, le linee guida di sicurezza di laboratorio devono essere sempre seguite. Di particolare importanza è la sicurezza quando si tratta di alta tensione (&gt; 500 V) e prodotti chimici di trattamento. 1. Rivestimento di un substrato conduttivo con la candela fuliggine <ol><li> Taglio di metallo in rame in rettangoli (75 x 43 mm, spessore di 0,5 mm). Pulire ogni substrato di rame per immersione in mordenzante di rame per circa 30 secondi, lavare con a...

<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>

La fase più critica del protocollo è la protezione dello strato di fuliggine, direttamente associata con il successo in goccioline movimento. Metallizzazione lo strato di fuliggine (precedenti metodi 1) permette di quasi il 100% di successo fabbricazione. Tuttavia, il tempo massimo di funzionamento è di circa 10 minuti; eventualmente, le frazioni delle gocce sono bagnare la fuliggine attraverso buchi nello strato di metallo. Rivestimento lo strato di fuliggine con il liquido fluorurato è l&#39;alternativa più sempl...

La miniaturizzazione dei dispositivi che funzionano con i liquidi è di fondamentale importanza per lo sviluppo di piattaforme &quot;lab-on-a-chip&quot;. In questo senso, gli ultimi due decenni hanno visto un significativo progresso nel campo della microfluidica, con una varietà di applicazioni. 1-5 contrasto con il trasporto di liquido nei canali chiusi (microfluidica canale), DMF manipola goccioline su array di elettrodi. Uno dei meriti più interessanti di questa tecnica è l&#39;assenza di parti mobili per il trasporto di fluidi, e il movimento è immediatamente fermato spegnendo segnali elettrici. Tuttavia, il movimento delle gocce dipende contenuti gocciolina, certamente una caratteristica indesiderabile per una piattaforma universale &quot;lab-on-a-chip&quot;. Goccioline contenenti proteine ​​e altri analiti aderire alle superfici dei dispositivi, diventando inamovibile. Probabilmente, questo è stato il limite maggiore per ampliare la gamma di applicazioni DMF; 6-8alternative per ridurre al minimo la formazione di incrostazioni di superficie indesiderato comportano l&#39;aggiunta di specie chimiche in più per la goccia o dei suoi dintorni, che potrebbe potenzialmente influenzare il contenuto delle gocce. In precedenza, il nostro gruppo ha sviluppato un dispositivo per consentire il trasporto di cellule e proteine ​​in DMF, senza additivi aggiunti (dispositivi di campo-DW). 9 Ciò è stato ottenuto combinando una superficie basata sulla candela fuliggine, 10 con una geometria dispositivo che favorisce goccia di rotolamento e conduce ad una forza verso l&#39;alto sul droplet, diminuendo ulteriormente l&#39;interazione gocciolina superficie. In questo approccio, il movimento gocciolina non è associato con bagnatura superficiale. 11 L&#39;obiettivo del metodo dettagliato di seguito descritto è quello di realizzare un dispositivo DMF in grado di trasportare goccioline contenenti proteine, cellule e interi organismi, senza additivi supplementari. I dispositivi di campo-DW aprono la strada per le piattaforme interamente controllate di lavoro in gran parte indipendentemente goccia chimicory. Qui, anche simulazioni presenti mostrando che, nonostante l&#39;alta tensione necessaria per il funzionamento del dispositivo, la caduta di tensione attraverso la goccia è una piccola frazione della tensione applicata, indicando effetti trascurabili sulla bioanalytes all&#39;interno della gocciolina. Infatti, prove preliminari con Caenorhabditis elegans (C. elegans), un nematode utilizzato per una varietà di studi in biologia, mostrano che i vermi nuotano indisturbati come vengono applicate tensioni.

<table border="0" cellpadding="0" cellspacing="0" width="1230">
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			<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>
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			<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>
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		<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>
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		<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>
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			<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>
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		<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>
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		<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>
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		<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>
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		<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>
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			<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>
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			<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>
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		<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>
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		<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>
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		<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>
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		<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>
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		<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.

In precedenza, abbiamo utilizzato i dispositivi di campo-DW per consentire il movimento delle proteine ​​in DMF. In particolare, goccioline con albumina di siero bovino (BSA) possono essere spostati ad una concentrazione di 2000 volte superiore a quanto precedentemente riportato da altri autori (senza additivi). Ciò è dovuto alla ridotta interazione tra la goccia e la superficie; la figura 4 mostra una gocciolina contenente fluorescente-tag BSA (vedi Freire et al 9 per ulterior...

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.

Approfittando di ridotta interazione Droplet-superficie per ottimizzare il trasporto di Bioanalytes in microfluidica digitale

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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Fabrication, Operation and Flow Visualization in Surface-acoustic-wave-driven Acoustic-counterflow Microfluidics

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Surface acoustic waves (SAWs) can be used to drive liquids in portable microfluidic chips via the acoustic counterflow phenomenon. In this video we ...

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