This work was supported by the Ministry of Science and Technology of Taiwan under Grant Number 101-2221-E-002-064-MY3.

1. Fabrication of Microfluidic Device
<ol>
	<li>Use a graphic layout software (e.g., AutoCAD) to draw the outline of a T-microchannel. For the T-microchannel, the two feed channels are 90 &#181;m wide and 2,500 &#181;m long, and the confluence channel is 180 &#181;m wide and 3,000 &#181;m long. Connect the end of each channel to an individual circle with a diameter of 1,100 &#181;m.</li>
	<li>Mark &#8216;clear&#8217; and &#8216;dark&#8217; for the exposure and covered areas, respectively. For a negative photor...

<ol>
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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+mixing+in+a+microfluidic+oscillator+using+an+impinging+jet+on+a+concave+surface.">Effective mixing in a microfluidic oscillator using an impinging jet on a concave surface.</a> Microsystem Technologies. , (2011).</li><li>Strook, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chaotic+mixer+for+microchannels.">Chaotic mixer for microchannels.</a> Science. 295 (5555), 647-651 (2002).</li><li>Wheat, P. M., Posner, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+mixing+using+equilibrium+reactions.">Quantifying mixing using equilibrium reactions.</a> Physics of Fluids. 21 (3), 037101 (2009).</li><li>Liu, R. 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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=Out-of-focus+effects+on+particle+image+visibility+and+correlation+in+microscopic+particle+image+velocimetry.">Out-of-focus effects on particle image visibility and correlation in microscopic particle image velocimetry.</a> Experiments in Fluids. 29 (1), S166-S174 (2000).</li><li>Friend, J., Yeo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+microfluidic+devices+using+polydimethylsiloxane.">Fabrication of microfluidic devices using polydimethylsiloxane.</a> Biomicrofluidics. 4 (2), 026502 (2010).</li><li>Wohlfarth, C., Lechner, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Landolt-B&#246;rnstein:+Numerical+Data+and+Functional+Relationships+in+Science+and+Technology.">Landolt-B&#246;rnstein: Numerical Data and Functional Relationships in Science and Technology.</a> Optical Constants: Refractive Indices of Pure Liquids and Binary Liquid Mixtures (Supplement to III/38). 47, (2008).</li><li>Heller, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remarks+on+refractive+index+mixture+roles.">Remarks on refractive index mixture roles.</a> Journal of Physical Chemistry. 69 (4), 1123-1129 (1965).</li><li>Korczyk, P. M., Cybulski, O., Makulska, S., Garstecki, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+unsteadiness+of+the+rates+of+flow+on+the+dynamics+of+formation+of+droplets+in+microfluidic+systems.">Effects of unsteadiness of the rates of flow on the dynamics of formation of droplets in microfluidic systems.</a> Lab on a Chip. 11 (1), 173-175 (2011).</li><li>Dreher, S., Engler, M., Kockmann, N., Woias, P., Bockhorn, H., Mewes, D., Peukert, W., Warnecke, H. .. -. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theoretical+and+experimental+investigations+of+convective+micromixers+and+microreactors+for+chemical+reactions.">Theoretical and experimental investigations of convective micromixers and microreactors for chemical reactions.</a> Micro and Macro Mixing: Analysis, Simulation and Numerical Calculation. , (2010).</li><li>Huang, C. Y., Gregory, J. W., Sullivan, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Modified+schlieren+technique+for+micro+flow+visualization.">A Modified schlieren technique for micro flow visualization.</a> Measurement Science &amp; Technology. 18 (5), N32-N34 (2007).</li><li>Tyn, M. T., Calus, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature+and+concentration+dependence+of+mutual+diffusion+coefficients+of+some+binary+liquid+systems.">Temperature and concentration dependence of mutual diffusion coefficients of some binary liquid systems.</a> Journal of Chemicaland Engineering Data. 20 (3), 310-316 (1975).</li><li>Celik, I. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Procedure+for+estimation+and+reporting+of+uncertainty+due+to+discretization+in+CFD+applications.">Procedure for estimation and reporting of uncertainty due to discretization in CFD applications.</a> Journal of Fluids Engineering. 130 (7), 078001 (2008).</li><li>Tasi&#263;, A. &. #. 3. 8. 1. ;., Djordjevi&#263;, B. D., Grozdani&#263;, D. K., Radojkovi&#263;, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+mixing+rules+in+predicting+refractive+indices+and+specific+refractivities+for+some+binary+liquid+mixtures.">Use of mixing rules in predicting refractive indices and specific refractivities for some binary liquid mixtures.</a> Journal of Chemical and Engineering Data. 37 (3), 310-313 (1992).</li><li>Hsiao, P. -. J., Chen, S. -. T., Hsiao, T. -. H., Sun, C. -. 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+comparison+of+coarse-resolution+numerical+simulation+with+experimental+measurements+of+wind+turbine+aerodynamic+performance.">A comparison of coarse-resolution numerical simulation with experimental measurements of wind turbine aerodynamic performance.</a> , (2013).</li><li>Sun, C. -. l., Huang, C. -. 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For fluidic mixing in a microfluidic device, the microscale schlieren technique is able to measure the magnitude of concentration gradient through quantifying change in light intensity. Because the principle of this technique relies on detecting the alternation of light propagation, the working fluids and the microfluidic device have to be transparent to the incident light. In addition, the protocol requires a linear relationship between the refractive index of the solution and its composition so that preliminary assessm...

Fluid mixing is an important issue that is found in many industrial processes and biological systems. With the emergence of microfluidics, mixing in microscale has brought much attention due to its challenge in diffusion domination among the mass transport mechanisms. Since designing an effective micromixer required quantitative validation, several measuring methods were developed5-7. Nevertheless, the three-dimensional structure, commonly found in efficient micromixers5, demands a more accurate representation of the concentration field that the common measuring techniques fail to deliver. Due to the limit of viewing angle8 or reaction kinetics6, the aforementioned methods may produce misleading results that do not correctly account for the homogeneity of the mixture.
For optically transparent fluids mixing in optically transparent microstructures, microscale schlieren technique3,9-14 provides an attractive alternative to analyze mixing inhomogeneity. In the past, microscale schlieren technique has been mostly used to visualize compressible flow9-13,15 or phase gradient16. Microscale schlieren technique benefits from both a simple optical layout and high sensitivity and enables not only the non-invasive investigation of specific flow feature that causes optical disturbance but is well suited for use in assessing mixing. In this paper, we construct the microscale schlieren system by inserting a knife-edge in the back focal plane of the objective of a microscope, describe a calibration procedure to realize quantitative analysis, and report a validation measurement in a microfluidic oscillator4. To implement measurements, the working fluids are properly selected so that the refractive index of the mixed fluids varies linearly with the composition, and the thickness of the target microfluidic device is identical to the one used in calibration. Besides species concentration, microscale schlieren technique can be extended to measure the gradient of other scalar quantity that is linearly correlated to the index of refraction, such as temperature or salinity.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Permanent Epoxy Negative Photoresist</td>
			<td>MicroChem</td>
			<td>SU-8 2150</td>
			<td></td>
		</tr>
		<tr>
			<td>single side polished silicon wafer</td>
			<td>Light Technology</td>
			<td>S4W1PP5SABUP1</td>
			<td>p-type, diameter: 100&#177;0.5 mm, thickness: 525 &#177;25 mm, orientation: (1 0 0)</td>
		</tr>
		<tr>
			<td>syringe pump</td>
			<td>kdScientific</td>
			<td>kds210</td>
			<td></td>
		</tr>
		<tr>
			<td>syringe, i.e. 10 ml</td>
			<td>Terumo</td>
			<td>SS-10L2138</td>
			<td></td>
		</tr>
		<tr>
			<td>Hoffman modulation contrast microscope</td>
			<td>Leica Microsystems</td>
			<td>DM IL LED</td>
			<td></td>
		</tr>
		<tr>
			<td>5X objective lens</td>
			<td>Leica Microsystems</td>
			<td>N PLAN</td>
			<td>NA = 0.12</td>
		</tr>
		<tr>
			<td>knife-edge</td>
			<td></td>
			<td></td>
			<td>custom made part</td>
		</tr>
		<tr>
			<td>camera, i.e. high speed</td>
			<td>Integrated Design Tools</td>
			<td>NX7-S1</td>
			<td></td>
		</tr>
		<tr>
			<td>C-mount adapter HC 0.63x</td>
			<td>Leica Microsystems</td>
			<td>541537</td>
			<td></td>
		</tr>
		<tr>
			<td>camera operating software</td>
			<td>Integrated Design Tools</td>
			<td>MotionPro X Studio 2.02.01</td>
			<td></td>
		</tr>
		<tr>
			<td>polydimethylsiloxane (PDMS)</td>
			<td>Dow Corning</td>
			<td>Sylgard-184 silicone elastomer kit</td>
			<td></td>
		</tr>
		<tr>
			<td>Teflon tubing, i.e. O.D. x I.D. 1/16 in. x 0.031 in.</td>
			<td>Supelco</td>
			<td>58700-U</td>
			<td></td>
		</tr>
		<tr>
			<td>micro glass slide</td>
			<td>Matsunami Glass</td>
			<td>S2215</td>
			<td></td>
		</tr>
		<tr>
			<td>hot plate</td>
			<td>Yeong-Shin</td>
			<td>HP-303DN</td>
			<td></td>
		</tr>
		<tr>
			<td>distilled water, i.e. HPLC grade</td>
			<td>Alps Chemicals</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ethyl alcohol, i.e. reagent grade</td>
			<td>Nihon Shiyaku Reagent</td>
			<td>EA448652</td>
			<td></td>
		</tr>
		<tr>
			<td>image processing software</td>
			<td>Mathworks</td>
			<td>MATLAB R2009a</td>
			<td></td>
		</tr>
		<tr>
			<td>computational fluid dynamics package</td>
			<td>ESI group</td>
			<td>CFD-ACE+ 2008</td>
			<td></td>
		</tr>
	</tbody>
</table>

analyzing mixing inhomogeneity in a microfluidic device by microscale schlieren technique

In this paper, we introduce the use of microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. The microscale schlieren system is constructed from a Hoffman modulation contrast microscope, which provides easy access to the rear focal plane of the objective lens, by removing the slit plate and replacing the modulator with a knife-edge. The working principle of microscale schlieren technique relies on detecting light deflection caused by variation of refractive index1-3. The deflected light either escapes or is obstructed by the knife-edge to produce a bright or a dark band, respectively. If the refractive index of the mixture varies linearly with its composition, the local change in light intensity in the image plane is proportional to the concentration gradient normal to the optical axis. The micro-schlieren image gives a two-dimensional projection of the disturbed light produced by three-dimensional inhomogeneity.
To accomplish quantitative analysis, we describe a calibration procedure that mixes two fluids in a T-microchannel. We carry out a numerical simulation to obtain the concentration gradient in the T-microchannel that correlates closely with the corresponding micro-schlieren image. By comparison, a relationship between the grayscale readouts of the micro-schlieren image and the concentration gradients presented in a microfluidic device is established. Using this relationship, we are able to analyze the mixing inhomogeneity from associate micro-schlieren image and demonstrate the capability of microscale schlieren technique with measurements in a microfluidic oscillator4. For optically transparent fluids, microscale schlieren technique is an attractive diagnostic tool to provide instantaneous full-field information that retains the three-dimensional features of the mixing process.

The grayscale ratio I/I0 under different Reynolds number for both positive and negative gradients of mass fraction is shown (Figure 2) with a symmetrical band appearing in the middle of the T-microchannel. At low Reynolds number, the tail of the schlieren band is expanded and blurred due to the dispersion across the mixing interface. As the Reynolds number increases, the diffusion length shortens leading to a narrower band. At different downstream locations, the variations of...

Watch this Scientific Journal Video about Analyzing Mixing Inhomogeneity in a Microfluidic Device by Microscale Schlieren Technique at JoVE.com

Analyzing Mixing Inhomogeneity in a Microfluidic Device by Microscale Schlieren Technique

Herein, we describe a procedure that employs microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. Through calibration, distribution of concentration gradient can be derived from the micro-schlieren image.

In this paper, we introduce the use of microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. The microscale schlieren ...

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8.8K Views.  National Taiwan University. Herein, we describe a procedure that employs microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. Through calibration, distribution of concentration gradient can be derived from the micro-schlieren image. 