Name: 气动驱动的微流体平台，用于微颗粒浓缩 (视频) | JoVE Journal
Uploaded: 2022-02-01T14:15:00.000+00:00
Duration: 8 min 43 s
Description: 学习气动微流控平台技术，用于高效浓缩微颗粒并防止堵塞与细胞损伤。该方法采用PDMS结构和气动阀门控制，适用于生物医学工程中的生物样本处理与分析。

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This work was supported by the National Research Foundation of Korea(NRF) grant funded by the Korea government(Ministry of Science and ICT). (No. NRF-2021R1A2C1011380).

1. Designing the microfluidic platform for particle concentration
<ol>
	<li>Design the pneumatic microfluidic platform consisting of one pneumatic valve for fluid flow in the 3D flow network and three pneumatic valves for sieve (Vs), fluid (Vf), and particle (Vp) valve operation (Figure 1). 
	NOTE: Vs blocks concentrate particles from the liquid, and Vf and Vp allow fluid and particle release after concentration. Three pneumatic ports provide compressed air from the fluid/pneumatic supply layer (normally open) and the pneumatic valve light outlet to actuate the valve. The microfluidic channel network is designed with a CAD program18,19.</li>
	<li>Design the channel to be a pneumatic supply layer and a 3D channel network layer (Figure 2). 
	NOTE: The fluid network is interconnected with the curved channels in the anterior part and the rectangular chamber in the posterior region. Vs block the inlet, and particles accumulate in the collection area of the curved fluid channel. Particle-free fluids (particle-free liquids) are exited through the Qf outlet and the concentrated particles through the Qp outlet (Figure 3).</li>
	<li>According to the above conditions, prepare four types of SU-8 molds. 
	NOTE: The four molds include a mold that allows the valve to be controlled via pneumatics, two molds that create fluid channels, and a clean mold without shape (Figure 4 and Table 1). The four types of molds mentioned are fabricated using standard photolithography processes. This mold making consists of a SU-8 mold on a silicon wafer as per previously published reports18,19. Figure 5 depicts the device chip.</li>
</ol>
2. Fabrication of the microfluidic platform for particle concentration
NOTE: Figure 6 illustrates the fabrication of a microfluidic platform that concentrates particles.
<ol>
	<li>Replicate the PDMS layer using a prepared pneumatic valve channel SU-8 mold (step 1.3) for pneumatically controlling the valve.
	<ol>
		<li>Pour 10 mL of liquid PDMS and 1 mL of curing agent (see Table of Materials) into a prepared pneumatic valve channel mold (step 1.3) and heat-activate at 90 &#176;C for 30 min.</li>
		<li>After the PDMS structures are cured, separate the SU-8 mold of step 2.1.1.</li>
		<li>Punch three 1.5 mm pneumatic ports (Vs, Vf, and Vp) into the pneumatic valve channel manufactured according to step 2.1.2 using a 1.5 mm puncture (see Table of Materials).</li>
		<li>Pour 10 mL of liquid PDMS and 1 mL of curing agent into a prepared clean SU-8 mold prepared in step 1.3 and spin-coat at 1,500 rpm for 15 s using a spin coater (see Table of Materials). Then heat-activate at 90 &#176;C for 30 min.</li>
		<li>After the PDMS structures are cured, separate the SU-8 mold of step 2.1.4. 
		NOTE: The valve diaphragm layer controls fluid flow according to the pneumatic pressure.</li>
		<li>Treat atmospheric plasma (see Table of Materials) to the PDMS structures prepared in steps 2.1.3 and 2.1.5 for 20 s.</li>
		<li>Align directly plasma-treated PDMS structures from step 2.1.6 according to the channel structure by checking with a microscope.</li>
		<li>Bond the aligned PDMS structures prepared in step 2.1.7 by heating at 90 &#176;C for 30 min.</li>
		<li>Punch a 1.5 mm diameter hole in the fluid channel inlet (Qfp) and fluid channel outlets (Qf and Qp) within the pneumatic channel part to which the thin diaphragm layer is bonded, using a 1.5 mm puncture.</li>
	</ol>
	</li>
	<li>Replicate both sides of the PDMS layer using two SU-8 molds to make a microfluidic channel. Use a curved and rectangular microfluidic channel mold on the front and a microfluidic interconnection channel mold on the rear.
	<ol>
		<li>Pour 10 mL of liquid PDMS and 1 mL of curing agent into the curved and rectangular microfluidic channel mold and spin-coat at 1,200 rpm for 15 s. Then create molds for the curved fluid chamber and fluid channels by thermal activation at 90 &#176;C for 30 min (Figure 6A).</li>
		<li>Separate the PDMS layer on which the microfluidic channel is formed, then make a heat-activated mold covering the sealed vent wall by bonding to the glass wafer by treating atmospheric plasma for 20 s (Figure 6B).</li>
		<li>Pour 3 mL of liquid PDMS into the interconnection channel of the SU-8 mold (Figure 6C).</li>
		<li>Arrange the structure fabricated in step 2.2.2 with the interconnection channel mold in liquid PDMS on the microfluidic interconnect channel mold, and dry the superimposed structure at 130 &#176;C for 30 min (Figure 6D). 
		NOTE: While curing the rear structure, the PDMS mold fabricated in step 2.2.2 is inflated by the thermal pressure of the air layer, and the deformed PDMS layer is thermally activated (Figure 6E)16.</li>
		<li>After curing, remove the front SU-8 mold from the microfluidic channel network layer and carefully peel off the rear PDMS mold (Figure 6F). 
		NOTE: The 3D fluidic network layer allows the creation of an anterior curved fluid chamber and microfluidic channels.</li>
		<li>Pour 10 mL of liquid PDMS and 1 mL of curing agent into a clean SU-8 mold. Then heat-activate at 90 &#176;C for 30 min.</li>
		<li>After the PDMS structures are cured, separate the SU-8 mold. 
		NOTE: This step creates the additional sealing layer.</li>
		<li>Treat the atmospheric plasma to PDMS structures prepared in steps 2.2.3 and 2.2.7 for 20 s.</li>
		<li>Align directly plasma-treated PDMS structures according to the channel structure by checking with a microscope.</li>
		<li>Bond the aligned PDMS structures by heating at 90 &#176;C for 30 min.</li>
	</ol>
	</li>
	<li>Align the PDMS structures prepared in steps 2.1 and 2.2 according to the channel structure and bond them by treating atmospheric plasma for 20 s.</li>
</ol>
3. Setting up the device
NOTE: Figure 7 shows fabricating a microfluidic platform that concentrates particles.
<ol>
	<li>Manually fill the microfluidic channel with bubble-free demineralized water using a 10 mL syringe.</li>
	<li>To control the P_Qfp and the three pneumatic valves (P_Vs, P_Vf, and P_Vp) that control the microbead flow, insert a precision pressure controller with four or more output channels (see Table of Materials) for the working fluid (Qfp) into the microfluidic platform. 
	NOTE: A precision pressure controller with four output channels can be replaced with multiple precision pressure controllers. In this experiment, the operating pressure of P_Qfp was 10 kPa, P_Vs was 15 kPa, and P_Vf and P_Vp were both 18 kPa (Figure 8 and Table 2). Figure 8 shows the working fluid flow rate over time as particles are concentrated by the microfluidic platform with P_Vs of 15 kPa, and Table 2 shows the actuation results according to the pneumatic valves.</li>
	<li>Prepare carboxyl polystyrene test particles of various sizes in distilled water (see Table of Materials). 
	NOTE: The particle sizes used in this experiment were 24.9, 8.49, and 4.16 &#181;m; particles of various sizes can be used depending on the pressure of P_Vs.</li>
	<li>To control the flow rate of the working fluid, fill a glass bottle half full with water (working fluid) and connect the glass bottle cap to the controller output channel and microvalve. 
	NOTE: Connect one tube to the microvalve to receive compressed air from the controller and the other tube to inject water.</li>
	<li>Observe platform operation through an inverted microscope for all platform operations and measure the operating flow rate over time at the outlet by a liquid flow meter (see Table of Materials).</li>
</ol>
4. Operation of the device
<ol>
	<li>Inject the particle/fluid mixture under pressure at the inlet (Qfp) with Vp (Figure 9A). 
	NOTE: The flow of particles and clean fluid from the outlet through the interconnected channels are controlled via Vp and Vf, respectively (Table 2).</li>
	<li>Apply pressure to Vs at 15 kPa and Vp at 18 kPa to actuate the valve. 
	NOTE: At this time, the diaphragm is deformed, the particles of the fluid Qfp are blocked in the contact space between the curved fluid channel and the curved fluid cantilever, and the unwanted Qfp fluid is released through the open Qf (Figure 9B,C).</li>
	<li>When the particles are concentrated, apply pressure only to Vf. 
	NOTE: At this time, when pressure is applied only to Vf, the clogged particles are released through Qp (Figure 9D).</li>
</ol>

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<li>Irimia, D., Toner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cell+handling+using+microstructured+membranes%5BTitle%5D)+AND+%22Lab+on+a+Chip%22%5BJournal%5D)">Cell handling using microstructured membranes.</a> Lab on a Chip. 6 (3), 345-352 (2006).</li>
<li>Huang, S. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+tunable+micro+filter+modulated+by+pneumatic+pressure+for+cell+separation%5BTitle%5D)+AND+%22Sensors+and+Actuators+B%3A+Chemical%22%5BJournal%5D)">A tunable micro filter modulated by pneumatic pressure for cell separation.</a> Sensors and Actuators B: Chemical. 142 (1), 389-399 (2009).</li>
<li>Chang, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+tunable+microfluidic-based+filter+modulated+by+pneumatic+pressure+for+separation+of+blood+cells%5BTitle%5D)+AND+%22Microfluidics+and+Nanofluidics%22%5BJournal%5D)">A tunable microfluidic-based filter modulated by pneumatic pressure for separation of blood cells.</a> Microfluidics and Nanofluidics. 12 (1-4), 85-94 (2012).</li>
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<li>McDonald, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Poly%28dimethylsiloxane%29+as+a+material+for+fabricating+microfluidic+device%5BTitle%5D)+AND+%22Accounts+of+Chemical+Research%22%5BJournal%5D)">Poly(dimethylsiloxane) as a material for fabricating microfluidic device.</a> Accounts of Chemical Research. 35 (7), 491-499 (2002).</li>
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</ol>

The authors have no conflicts of interest to disclose.

This platform provides a simple way to purify and concentrate particles of various sizes. Particles are accumulated and released through pneumatic valve control, and no clogging is observed because there is no passive structure. Using this device, the concentration of particles of three sizes is presented. However, the operating pressure, the time required for concentration, and the rate may vary depending on the device dimensions, particle size magnification, and the pressure at Vs18,<...

Due to the importance of biological analysis, microfluidic and biomedical microelectromechanical systems (BioMEMS) technologies1,2 are used to develop and study devices for the purification and collection of micromaterials2,3,4.&#160;Particle capture is categorized as active or passive. Active traps have been used for external dielectric5, magnetophoretic6, auditory7, visual8, or thermal9 forces acting on independent particles, enabling precise control of their movements. However, an interaction between the particle and external force is required; thus, the throughput is low. In microfluidic systems, controlling the flow rate is very important because the external forces are transmitted to the target particles.
In general, passive microfluidic devices have micropillars in microchannels10,11. Particles are filtered through interaction with a flowing fluid, and these devices are easy to design and inexpensive to manufacture. However, they cause particle clogging in micro-pillars, so more complex devices have been developed to prevent particle clogging12. Microfluidic devices with complex structures are generally suitable for managing a limited number of particles13,14,15,16,17,18.
This article describes a method to fabricate and operate a pneumatically driven microfluidic platform for large particle concentrations that overcomes the shortcomings18 as mentioned above. This platform can block and concentrate particles by deformation and actuation of the thin diaphragm layer of the sieve valve (Vs) plate that accumulates in curved microfluidic channels. Particles accumulate in curved microfluidic channels, and the concentrated particles can separate by discharging the working fluid via the actuation of two PDMS seals on/off valves18. This method makes it possible to process a limited number of particles or concentrate a large number of small particles. Operating conditions such as the magnitude of flow rate and compressed air pressure can prevent unwanted cell damage and increase cell trapping efficiency.

<table><tbody><tr><td>1.5 mm puncture</td><td>Self procduction</td><td>Self procduction</td><td>This puncture was made by requesting a mold maker based on the Miltex&amp;reg; Biopsy Punch with Plunger (15110-15) product.</td></tr><tr><td>4 inch Silicon Wafer/SU-8 mold</td><td>4science</td><td>29-03573-01</td><td>4 inch (100) Ptype silicon wafer/SU-8 mold</td></tr><tr><td>Carboxyl Polystyrene Crosslinked Particle(24.9 &amp;mu;m)</td><td>Spherotech</td><td>CPX-200-10</td><td>Concentrated bead sample1</td></tr><tr><td>Flow meter</td><td>Sensirion</td><td>SLI-1000</td><td>Flow measurement</td></tr><tr><td>High-speed camera</td><td>Photron</td><td>FASTCAM Mini</td><td>Observation of concentration</td></tr><tr><td>Hot plate</td><td>As one</td><td>HI-1000</td><td>heating plate for curing of liquid PDMS</td></tr><tr><td>KOVAX-SYRINGE 10 mL/Syringe</td><td>Koreavaccine</td><td>22G-10ML</td><td>Fill the microfluidic channel with bubble-free demineralized water.</td></tr><tr><td>Laboratory Conona treater/Atmospheric plasma</td><td>Electro-Technic</td><td>BD-20AC</td><td>Chip bonding/atmospheric plasma</td></tr><tr><td>Liquid polydimethylsiloxane, PDMS</td><td>Dow Corning Inc.</td><td>Sylgard 184</td><td>Components of chip</td></tr><tr><td>Microscope</td><td>Olympus</td><td>IX-81</td><td>Observation of concentration</td></tr><tr><td>PEEK Tubes</td><td>SAINT-GOBAIN PPL CORP.</td><td>AAD04103</td><td>Inject or collect particles</td></tr><tr><td>Polystyrene Particle(4.16 &amp;mu;m)</td><td>Spherotech</td><td>PP-40-10</td><td>Concentrated bead sample3</td></tr><tr><td>Polystyrene Particle(8.49 &amp;mu;m)</td><td>Spherotech</td><td>PP-100-10</td><td>Concentrated bead sample2</td></tr><tr><td>Pressure controller/&amp;mu;flucon</td><td>AMED</td><td>&amp;mu;flucon</td><td>Control of air pressure</td></tr><tr><td>Spin coater</td><td>iNexus</td><td>ACE-200</td><td>spread the liquid PDMS on SU-8 mold</td></tr></tbody></table>

pneumatically driven microfluidic platform for micro-particle concentration

Pneumatically Driven Microfluidic Platform for Micro-Particle Concentration

The present article introduces a method for fabricating and operating a pneumatic valve to control particle concentration using a microfluidic platform. This platform has a three-dimensional (3D) network with curved fluid channels and three pneumatic valves, which create networks, channels, and spaces through duplex replication with polydimethylsiloxane (PDMS). The device operates based on the transient response of a fluid flow rate controlled by a pneumatic valve in the following order: (1) sample loading, (2) sample blocking, (3) sample concentration, and (4) sample release. The particles are blocked by thin diaphragm layer deformation of the sieve valve (Vs) plate and accumulate in the curved microfluidic channel. The working fluid is discharged by the actuation of two on/off valves. As a result of the operation, all particles of various magnifications were successfully intercepted and disengaged. When this technology is applied, the operating pressure, the time required for concentration, and the concentration rate may vary depending on the device dimensions and particle size magnification.

Figure 8 shows the flow rate of the fluid rates for a four-stage platform operation, as mentioned in Table 2. The first stage is the loading state (a state). The platform was supplied with fluid with all valves open, and the working fluid (Qf) and particles (Qp) are almost identical as the microfluidic channel network exhibits structural symmetry. In the second stage (b state), compressed air was transported to Vs to block the particles, and as the Vs diaphragm deformed, the...

The present protocol describes a pneumatic microfluidic platform that can be used for efficient microparticle concentration.

The present article introduces a method for fabricating and operating a pneumatic valve to control particle concentration using a microfluidic platform. ...

This protocol describe our method to fabricate and operate on pneumatically-driven microfluidic platform for multi-particle concentrations that overcomes shortcomings such as particle clogging and complex structures. This methods makes it possible to process an unlimited number of particles, concentrate on a large number of small particles, and prevent unwanted cell damage, and increases entropy efficiency. Due to the importance of biological analysis, microfluidic and biomedical microelectro mechanical systems technologies are used to develop and study devices for the purification and collection of micro-materials.To begin, use a pre-prepared pneumatic valve channel SU8 mold to replicate the PDMS layer for pneumatically controlling the valve. Pour 10 milliliters of liquid PDMS and one milliliter of curing agent into a prepared pneumatic valve channel mold, and heat activate at 90 degrees Celsius for 30 minutes. After the PDMS structures are cured, separate the SU8 mold.Punch three 1.5 millimeter pneumatic ports into the pneumatic valve channel using a 1.5 millimeter puncture. Pour 10 milliliters of liquid PDMS and one milliliter of curing agent into a clean SU8 mold. Spin coat for

pneumatically-driven-microfluidic-platform-for-micro-particle

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JoVE Journal

Engineering

A novel microfluidic system has been developed using the phenomenon of passive pumping and a user controlled fluid delivery system. This microfluidic system has the potential to be used in a wide variety of biological applications given its low cost, ease of use, volumetric precision, high speed, repeatability and automation.

High Speed Droplet-based Delivery System for Passive Pumping in Microfluidic Devices

In this article, we present a microfluidic-based method for particle confinement based on hydrodynamic flow.  We demonstrate stable particle trapping at a fluid stagnation point using a feedback control mechanism, thereby enabling confinement and micromanipulation of arbitrary particles in an integrated microdevice.

A Microfluidic-based Hydrodynamic Trap for Single Particles

Droplet-based microfluidic platforms are promising candidates for high throughput experimentation since they are able to generate picoliter, self-compartmentalized vessels inexpensively at kHz rates. Through integration with fast, sensitive and high resolution fluorescence spectroscopic methods, the large amounts of information generated within these systems can be efficiently extracted, harnessed and utilized.

Fluorescence detection methods for microfluidic droplet platforms

This protocol describes a system architecture for performing automated small volume (0.15–1.5 ml) particle separations using a microfluidic device, and discusses methods to optimize acoustofluidic device performance and operation.

A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice

Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets

We propose a simple self-assembly technique of silica colloidal nanoparticles to create a nanofluidic junction between two microchannels in polydimethylsiloxane (PDMS). Using this technique, a nanoporous bead membrane with a pore size down to ~45 nm was built inside a microchannel and applied to electrokinetic preconcentration of DNA samples.

Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles

Lab-in-a-drop reaction systems allow the versatile implementation of complex reactions in a microfluidic scale. An automated actuation platform consisting of a 3 x 3 matrix of electromagnetic coils was developed and successfully used to merge two 10 µL microreactors and thereby initiate an enzymatic reaction in the resulting liquid marbles.

Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation

We demonstrate a microfluidic platform with an integrated surface electrode network that combines resistive pulse sensing (RPS) with code division multiple access (CDMA), to multiplex detection and sizing of particles in multiple microfluidic channels.

Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles

We present a microfluidic system for high throughput studies on complex life machinery, which consists of 1500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. The microfluidic chip allows for the analysis of the highly complex and dynamic micro-environmental conditions in vivo.

Generation of Dynamical Environmental Conditions using a High-Throughput Microfluidic Device

The fabrication of a polydimethylsiloxane (PDMS)-based bilayer device for the production of combinatorial libraries in water-in-oil emulsions (plugs) is presented here. The necessary hardware and software required to automate plug production are detailed in the protocol, and the production of a quantitative library of fluorescent plugs is also demonstrated.

Bilayer Microfluidic Device for Combinatorial Plug Production

This protocol presents a microfluidic system for studying the microbial precipitation of manganese oxide minerals. Biomineral formation is measured in situ with optical microscopy and image analysis. This approach can be adapted to investigate the precipitation of other distinctively colored biogenic minerals in porous environments.

A Microfluidic Platform to Investigate Microbial Precipitation of Metal Oxides in Porous Media

学习气动微流控平台技术，用于高效浓缩微颗粒并防止堵塞与细胞损伤。该方法采用PDMS结构和气动阀门控制，适用于生物医学工程中的生物样本处理与分析。

Pneumatically Driven Microfluidic Platform for Micro-Particle Concentration

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