Authors have no one to acknowledge.

1. Device Fabrication
<ol>
	<li>Design two photomasks, the first for patterning the surface acoustic wave (SAW) layer, and the second for the polydimethylsiloxane (PDMS) microchannel mold.
	<ol>
		<li>The first photomask has a pair of opposing interdigital transducers (IDTs)&#8212;also known as a SAW delay line&#8212;and markers for channel alignment and spatial reference during microscopy. In our standard device we have single-electrode IDTs with a finger width p = 10 &#956;m, aperture of 750 &#956;m...

<ol>
	<li>Masini, L., Cecchini, M., Girardo, S., Cingolani, R., Pisignano, D., Beltram, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface-acoustic-wave+counterflow+micropumps+for+on-chip+liquid+motion+control+in+two-dimensional+microchannel+arrays.">Surface-acoustic-wave counterflow micropumps for on-chip liquid motion control in two-dimensional microchannel arrays.</a> Lab on a Chip. 10 (15), 1997-2000 (2010).</li><li>Travagliati, M., De Simoni, G., Lazzarini, C. M., Piazza, V., Beltram, F., Cecchini, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction-free,+automatic,+on-chip+fluid+routing+by+surface+acoustic+waves.">Interaction-free, automatic, on-chip fluid routing by surface acoustic waves.</a> Lab on a Chip. 12 (15), 2621-2624 (2012).</li><li>Wixforth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustically+driven+planar+microfluidics.">Acoustically driven planar microfluidics.</a> Superlattices and Microstructures. 33 (5), 389-396 (2003).</li><li>Friend, J., Yeo, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscale+acoustofluidics:+Microfluidics+driven+via+acoustics+and+ultrasonics.">Microscale acoustofluidics: Microfluidics driven via acoustics and ultrasonics.</a> Reviews of Modern Physics. 83 (2), 647-64 (2011).</li><li>Tan, M. K., Yeo, L. Y., Friend, J. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LabCD:+a+centrifuge-based+microfluidic+platform+for+diagnostics.">LabCD: a centrifuge-based microfluidic platform for diagnostics.</a> Proceedings of SPIE. 3259, 80 (1998).</li><li>Cecchini, M., Girardo, S., Pisignano, D., Cingolani, R., Beltram, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustic-counterflow+microfluidics+by+surface+acoustic+waves.">Acoustic-counterflow microfluidics by surface acoustic waves.</a> Applied Physics Letters. 92 (10), 104103 (2008).</li><li>Campbell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Surface acoustic wave devices for mobile and wireless communications. 1, (1998).</li><li>Hashimoto, K. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Surface acoustic wave devices in telecommunications: modelling and simulation. , (2000).</li><li>Royer, D., Dieulesaint, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elastic+Waves+in+Solids+II.">Elastic Waves in Solids II.</a> Generation, Acousto-Optic Interaction, Applications. 2, (2000).</li><li>Renaudin, A., Sozanski, J. P., Verbeke, B., Zhang, V., Tabourier, P., Druon, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+SAW-actuated+microdroplets+in+view+of+biological+applications.">Monitoring SAW-actuated microdroplets in view of biological applications.</a> Sensors and Actuators B: Chemical. 138 (1), 374-382 (2009).</li><li>Glass, N. R., Tjeung, R., Chan, P., Yeo, L. Y., Friend, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organosilane+deposition+for+microfluidic+applications.">Organosilane deposition for microfluidic applications.</a> Biomicrofluidics. 5 (3), 036501 (2011).</li><li>Wereley, S. T., Meinhart, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+micro-particle+image+velocimetry.">Recent advances in micro-particle image velocimetry.</a> Annual Review of Fluid Mechanics. 42, 557-576 (2010).</li><li>Augustsson, P., Barnkob, R., Wereley, S. T., Bruus, H., Laurell, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+and+temperature-controlled+micro-PIV+measurements+enabling+long-term-stable+microchannel+acoustophoresis+characterization.">Automated and temperature-controlled micro-PIV measurements enabling long-term-stable microchannel acoustophoresis characterization.</a> Lab on a Chip. 11 (24), 4152-4164 (2011).</li><li>Hebert, B., Costantino, S., Wiseman, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+image+correlation+spectroscopy+(STICS)+theory,+verification,+and+application+to+protein+velocity+mapping+in+living+CHO+cells.">Spatiotemporal image correlation spectroscopy (STICS) theory, verification, and application to protein velocity mapping in living CHO cells.</a> Biophysical Journal. 88 (5), 3601 (2005).</li><li>Rossow, M., Mantulin, W. W., Gratton, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+image+correlation+spectroscopy+measurements+of+flow+demonstrated+in+microfluidic+channels.">Spatiotemporal image correlation spectroscopy measurements of flow demonstrated in microfluidic channels.</a> Journal of Biomedical Optics. 14 (2), 024014 (2009).</li><li>Schindelin, J., Arganda-Carreras, I., Frise, E., Kaynig, V., Longair, M., Pietzsch, T., Preibisch, S., Rueden, C., Saalfeld, S., Schmid, B., 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=Fiji:+an+open-source+platform+for+biological-image+analysis.">Fiji: an open-source platform for biological-image analysis.</a> Nature Methods. 9 (7), 676-682 (2012).</li><li>Rogers, P. R., Friend, J. R., Yeo, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploitation+of+surface+acoustic+waves+to+drive+size-dependent+microparticle+concentration+within+a+droplet.">Exploitation of surface acoustic waves to drive size-dependent microparticle concentration within a droplet.</a> Lab on a Chip. 10 (21), 2979-2985 (2010).</li><li>Muller, P. B., Barnkob, R., Jensen, M. J. H., Bruus, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+numerical+study+of+microparticle+acoustophoresis+driven+by+acoustic+radiation+forces+and+streaming-induced+drag+forces.">A numerical study of microparticle acoustophoresis driven by acoustic radiation forces and streaming-induced drag forces.</a> Lab on a Chip. 12 (22), 4617-4627 (2012).</li><li>Luo, J. K., Fu, Y. Q., Li, Y., Du, X. Y., Flewitt, A. J., Walton, A. J., Milne, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moving-part-free+microfluidic+systems+for+lab-on-a-chip.">Moving-part-free microfluidic systems for lab-on-a-chip.</a> Journal of Micromechanics and Microengineering. 19 (5), 054001-05 (2009).</li><li>Lindken, R., Rossi, M., Gro&#223;e, S., Westerweel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micro-particle+image+velocimetry+(&#956;PIV):+recent+developments,+applications,+and+guidelines.">Micro-particle image velocimetry (&#956;PIV): recent developments, applications, and guidelines.</a> Lab on a Chip. 9 (17), 2551-2567 (2009).</li><li>Gedge, M., Hill, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustofluidics+17:+Theory+and+applications+of+surface+acoustic+wave+devices+for+particle+manipulation.">Acoustofluidics 17: Theory and applications of surface acoustic wave devices for particle manipulation.</a> Lab on a Chip. 12 (17), 2998-3007 (2012).</li></ol>

One of the greatest challenges faced by the microfluidic community is the realization of an actuation platform for truly portable point-of-care devices. Among the proposed integrated micropumps23,those based on surface acoustic waves (SAWs) are particularly attractive due to their associated capabilities in fluid mixing, atomization and particle concentration and separation4. In this paper we have demonstrated how to fabricate and operate a lab-on-chip device in which fluid is steered in a closed PD...

One of the continued challenges facing the microfluidic community is the need to have an efficient pumping mechanism that can be miniaturized for integration into truly portable micro-total-analysis systems (&#956;TAS&#39;s). Standard macroscopic pumping systems simply fail to provide the portability required for &#956;TAS&#39;s, owing to the unfavorable scaling of the volumetric flow rates as the channel size decreases down to the micron range or below. On the contrary, SAWs have gained increasing interest as fluid actuation mechanisms and appear as a promising avenue for the solution of some of these problems1,2.
SAWs were shown to provide a very efficient mechanism of energy transport into fluids3. When a SAW propagates onto a piezoelectric substrate, e.g. lithium niobate (LN), the wave will be radiated into any fluid in its path at an angle known as the Rayleigh angle &#952;R = sin&#8722;1 (cf /cs), owing to the mismatch of sound velocities in the substrate, cs, and the fluid cf. This leakage of radiation into the fluid gives rise to a pressure wave which drives acoustic streaming in the fluid. Depending on the device geometry and power applied to the device, this mechanism was shown to actuate a wide variety of on-chip processes, such as fluid mixing, particle sorting, atomization, and pumping1,4. Despite the simplicity and effectiveness of actuating microfluids with SAW, there are only a small number of SAW driven microfluidic pumping mechanisms that have been demonstrated to date. The first demonstration was the simple translation of free droplets placed in the SAW propagation path on a piezoelectric substrate3. This novel method generated much interest in using SAWs as a microfluidic actuation method, however there was still a need for fluids to be driven through enclosed channels&#8212;a more difficult task. Tan et al. demonstrated pumping within a microchannel that was laser ablated directly into the piezoelectric substrate. By geometric modification with respect to the channel and IDT dimensions, they were able to demonstrate both uniform and mixing flows5. Glass et al. recently demonstrated a method of moving fluids through microchannels and microfluidic components by combining SAW actuated rotations with centrifugal microfluidics, as a demonstration of true miniaturization of the popular Lab-on-a-CD concept6,7. However, the only fully enclosed SAW driven pumping mechanism that has been demonstrated remains to be Cecchini et al.&#39;s SAW-driven acoustic counterflow8&#8212;the focus of this video. It exploits the atomization and coalescence of a fluid to pump it through a closed channel in the direction opposing the propagation direction of the acoustic wave. This system can give rise to surprisingly complex flows within a microchannel. Moreover, depending on the device geometry, it can provide a range of flow schemes, from laminar flows to more complex regimes characterized by vortices and particle-accumulation domains. The ability to easily influence the flow characteristics within the device shows opportunities for advanced on-chip particle manipulation.
In this protocol we wish to clarify the main aspects of practical SAW-based microfluidics: device fabrication, experimental operation, and flow visualization. While we are explicitly describing these procedures for the fabrication and operation of SAW-driven acoustic counterflow devices, these sections can easily be modified for their application to a range of SAW-driven microfluidic regimes.

<table border="1">
	<tbody>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Double side polished 128&#176; YX lithium niobate wafer</td>
			<td>Crystal Technology, LLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon wafer</td>
			<td>Siegert Wafers</td>
			<td></td>
			<td>We use &#60;100&#62;</td>
		</tr>
		<tr>
			<td>IDT Optical lithography mask with alignment marks (positive)</td>
			<td>Any vendor</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Channel Optical lithography mask (negative)</td>
			<td>Any vendor</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Positive photoresist</td>
			<td>Shipley</td>
			<td>S1818</td>
			<td></td>
		</tr>
		<tr>
			<td>Positive photoresist developer</td>
			<td>Microposit</td>
			<td>MF319</td>
			<td></td>
		</tr>
		<tr>
			<td>Negative tone photoresist</td>
			<td>Allresist</td>
			<td>AR-N-4340</td>
			<td></td>
		</tr>
		<tr>
			<td>Negative tone photoresist developer</td>
			<td>Allresist</td>
			<td>AR 300-475</td>
			<td></td>
		</tr>
		<tr>
			<td>SU8 thick negative tone photoresist</td>
			<td>Microchem</td>
			<td>SU-8 2000 Series</td>
			<td></td>
		</tr>
		<tr>
			<td>SU8 thick negative tone photoresist developer</td>
			<td>Microchem</td>
			<td>SU-8 developer</td>
			<td></td>
		</tr>
		<tr>
			<td>Hexadecane</td>
			<td>Sigma-Aldrich</td>
			<td>H6703</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon tetrachloride (CCl4)</td>
			<td>Sigma-Aldrich</td>
			<td>107344</td>
			<td></td>
		</tr>
		<tr>
			<td>Octadecyltrichlorosilane (OTS)</td>
			<td>Sigma-Aldrich</td>
			<td>104817</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone CMOS grade</td>
			<td>Sigma-Aldrich</td>
			<td>40289</td>
			<td></td>
		</tr>
		<tr>
			<td>2-propanol CMOS grade</td>
			<td>Sigma-Aldrich</td>
			<td>40301</td>
			<td></td>
		</tr>
		<tr>
			<td>Titanium</td>
			<td>Any vendor</td>
			<td>99.9% purity</td>
			<td></td>
		</tr>
		<tr>
			<td>Gold</td>
			<td>Any vendor</td>
			<td>99.9% purity</td>
			<td></td>
		</tr>
		<tr>
			<td>PDMS</td>
			<td>Dow Corning</td>
			<td>Sylgard 184 silicone elastomer kit with curing agent</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Any vendor</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>5 mm ID Harris Uni-Core multi-purpose coring tool</td>
			<td>Sigma-Aldrich</td>
			<td>Z708895</td>
			<td>Any diameter greater than 2 mm is suitable</td>
		</tr>
		<tr>
			<td>Acoustic absorber</td>
			<td>Photonic Cleaning Technologies</td>
			<td>First Contact regular kit</td>
			<td></td>
		</tr>
		<tr>
			<td>RF-PCB</td>
			<td>Any vendor</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spinner</td>
			<td>Laurell technologies corporation</td>
			<td>WS-400-6NPP</td>
			<td>Any spinner can be used</td>
		</tr>
		<tr>
			<td>UV Mask aligner</td>
			<td>Karl Suss</td>
			<td>MJB 4</td>
			<td>Any aligner can be used</td>
		</tr>
		<tr>
			<td>Thermal evaporator</td>
			<td>Kurt J. Lesker</td>
			<td>Nano 38</td>
			<td>Any thermal, e-beam evaporator or sputtering system can be used</td>
		</tr>
		<tr>
			<td>Oxygen plasma asher</td>
			<td>Gambetti Kenologia Srl</td>
			<td>Colibr&#236;</td>
			<td>Any plasma asher or RIE machine can be used</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Eppendorf</td>
			<td>5810 R</td>
			<td>Any centrifuge can be used</td>
		</tr>
		<tr>
			<td>Wire bonder</td>
			<td>Kulicke &#38; Soffa</td>
			<td>4523AD</td>
			<td>Any wire bonder can be used if the PCB is used without pogo connectors</td>
		</tr>
		<tr>
			<td>Contact Angle Meter</td>
			<td>KSV</td>
			<td>CAM 101</td>
			<td>Any contact angle meter can be used</td>
		</tr>
		<tr>
			<td>Spectrum analyzer</td>
			<td>Anristu</td>
			<td>56100A</td>
			<td>Any spectrum or network analyzer can be used</td>
		</tr>
		<tr>
			<td>RF signal generator</td>
			<td>Anristu</td>
			<td>MG3694A</td>
			<td>Any RF signal generator can be used</td>
		</tr>
		<tr>
			<td>RF high power amplifier</td>
			<td>Mini Circuits</td>
			<td>ZHL-5W-1</td>
			<td>Any RF high power amplifier can be used</td>
		</tr>
		<tr>
			<td>Microbeads suspension</td>
			<td>Sigma-Aldrich</td>
			<td>L3280</td>
			<td>Depending on the experimental purpose different suspension of different diameter and different material properties can be used</td>
		</tr>
		<tr>
			<td>Optical microscope</td>
			<td>Nikon</td>
			<td>Ti-Eclipse</td>
			<td>Any optical microscope with spatial resolution satisfying experimental purposes can be used</td>
		</tr>
		<tr>
			<td>Video camera</td>
			<td>Basler</td>
			<td>A602-f</td>
			<td>Any video camera that has enough frame rate and sensitivity satisfying experimental purposes can be used</td>
		</tr>
		<tr>
			<td>Camera acquisition software</td>
			<td>Advanced technologies</td>
			<td>Motion Box</td>
			<td>Any software enabling high and controlled frame rate acquisition can be used</td>
		</tr>
	</tbody>
</table>

fabrication, operation and flow visualization in surface-acoustic-wave-driven acoustic-counterflow microfluidics

Surface acoustic waves (SAWs) can be used to drive liquids in portable microfluidic chips via the acoustic counterflow phenomenon. In this video we present the fabrication protocol for a multilayered SAW acoustic counterflow device. The device is fabricated starting from a lithium niobate (LN) substrate onto which two interdigital transducers (IDTs) and appropriate markers are patterned. A polydimethylsiloxane (PDMS) channel cast on an SU8 master mold is finally bonded on the patterned substrate. Following the fabrication procedure, we show the techniques that allow the characterization and operation of the acoustic counterflow device in order to pump fluids through the PDMS channel grid. We finally present the procedure to visualize liquid flow in the channels. The protocol is used to show on-chip fluid pumping under different flow regimes such as laminar flow and more complicated dynamics characterized by vortices and particle accumulation domains.

Figure 2 shows representative results of device RF testing which were taken prior to bonding the LN layer to the microchannel layer: typical S11 and S12 spectra are reported in panel a) and b) respectively. The depth of the valley at central frequency in S11 spectrum is related to the efficiency of conversion of RF power in SAW mechanical power. Hence, for a fixed number of IDT finger pairs, a reduction in the valley minimum will result i...

Watch this Scientific Journal Video about Fabrication, Operation and Flow Visualization in Surface-acoustic-wave-driven Acoustic-counterflow Microfluidics at JoVE.com

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

In this video we first describe fabrication and operation procedures of a surface acoustic wave (SAW) acoustic counterflow device. We then demonstrate an experimental setup that allows for both qualitative flow visualization and quantitative analysis of complex flows within the SAW pumping device.

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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16.9K ビュー.  Istituto Italiano di Tecnologia. このビデオでは、第1の弾性表面波（SAW）音響向装置の製作および操作手順を説明する。我々はその後、質的流れの可視化とSAWポンプ装置内の複雑な流れの定量分析の両方を可能にした実験のセットアップを示しています。