Name: a performance-testing platform for a conduction micropump with an fr-4 copper-clad electrode plate
Uploaded: 2017-10-09T13:00:00
Description: Watch this Scientific Journal Video about A Performance-testing Platform for a Conduction Micropump with an FR-4 Copper-clad Electrode Plate at JoVE.com

This work was sponsored by the National Natural Science Foundation of China (51375176); the Guangdong Provincial Natural Science Foundation of China (2014A030313264); and the Science and Technology Planning Project of Guangdong Province, China (2014B010126003).

Caution: Please consult all relevant material safety data sheets (MSDS) before use. Acetone is highly flammable and can cause irritation to the eyes and respiratory tract. The voltage involved is as high as several thousand volts; hence, electrical sparks are expected when conducting the experiment. Carry out the experiments in a room with good ventilation to avoid explosions and fire from the sparks.
1. Fabrication of the Plates and Holder
NOTE: In this work, the ele...

<ol>
	<li>Kazemi, P. Z., Selvaganapathy, P. R., Ching, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+micropillar+electrode+spacing+on+the+performance+of+electrohydrodynamic+micropumps.">Effect of micropillar electrode spacing on the performance of electrohydrodynamic micropumps.</a> J Electrostat. 68 (4), 376-383 (2010).</li><li>Kano, I., Nishina, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+electrode+arrangements+on+EHD+conduction+pumping.">Effect of electrode arrangements on EHD conduction pumping.</a> IEEE Trans Ind Appl. 49 (2), 679-684 (2013).</li><li>Laser, D. J., Santiago, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+micropumps.">A review of micropumps.</a> J Micromech Microeng. 14 (6), R35 (2004).</li><li>Fylladitakis, E. D., Theodoridis, M. P., Moronis, A. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+on+the+history,+research,+and+applications+of+electrohydrodynamics.">Review on the history, research, and applications of electrohydrodynamics.</a> IEEE Trans Plasma Sci. 42 (2), 358-375 (2014).</li><li>Yazdani, M., Seyed-Yagoobi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electrostatics Joint Conf. , (2009).</li><li>Gharraei, R., Esmaeilzadeh, E., Hemayatkhah, M., Danaeefar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+investigation+of+electrohydrodynamic+conduction+pumping+of+various+liquids+film+using+flush+electrodes.">Experimental investigation of electrohydrodynamic conduction pumping of various liquids film using flush electrodes.</a> J Electrostat. 69 (1), 43-53 (2011).</li><li>Gharraei, R., Esmaeilzadeh, E., Nobari, M. R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Numerical+investigation+of+conduction+pumping+of+dielectric+liquid+film+using+flush-mounted+electrodes.">Numerical investigation of conduction pumping of dielectric liquid film using flush-mounted electrodes.</a> Theor Comp Fluid Dyn. 28 (1), 89 (2014).</li><li>Jeong, S. -. I., Seyed-Yagoobi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+study+of+electrohydrodynamic+pumping+through+conduction+phenomenon.">Experimental study of electrohydrodynamic pumping through conduction phenomenon.</a> J Electrostat. 56 (2), 123-133 (2002).</li><li>Seyed-Yagoobi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrohydrodynamic+pumping+of+dielectric+liquids.">Electrohydrodynamic pumping of dielectric liquids.</a> J Electrostat. 63 (6), 861-869 (2005).</li><li>Hojjati, M., Esmaeilzadeh, E., Sadri, B., Gharraei, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrohydrodynamic+conduction+pumps+with+cylindrical+electrodes+for+pumping+of+dielectric+liquid+film+in+an+open+channel.">Electrohydrodynamic conduction pumps with cylindrical electrodes for pumping of dielectric liquid film in an open channel.</a> Colloid Surface A. 392 (1), 294-299 (2011).</li><li>Yazdani, M., Seyed-Yagoobi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Numerical+investigation+of+electrohydrodynamic-conduction+pumping+of+liquid+film+in+the+presence+of+evaporation.">Numerical investigation of electrohydrodynamic-conduction pumping of liquid film in the presence of evaporation.</a> J Heat Trans-T ASME. 131 (1), 011602 (2009).</li><li>Vafaie, R. H., Ghavifekr, H. B., Lintel, H., Brugger, J., Renaud, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bi-directional+AC+electrothermal+micropump+for+on-chip+biological+applications.">Bi-directional AC electrothermal micropump for on-chip biological applications.</a> Electrophoresis. 37 (5-6), 719-726 (2016).</li><li>Pearson, M. R., Seyed-Yagoobi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Study+of+Linear+and+Radial+Two-Phase+Heat+Transport+Devices+Driven+by+Electrohydrodynamic+Conduction+Pumping.">Experimental Study of Linear and Radial Two-Phase Heat Transport Devices Driven by Electrohydrodynamic Conduction Pumping.</a> J Heat Trans-T ASME. 137 (2), 022901 (2015).</li></ol>

One of the critical steps within the protocol is to inspect the electrode plate carefully. Small burrs on the edge of an electrode can result in a short-circuit, and surface integrity can greatly affect pump performance. The cleaning of the electrode plate and holder is also very important. The electrode chamber height is less than 1 mm, so small dust particles may block the working liquid flow and cause a short-circuit. Before the test, injecting acetone into the chamber can remove the bubbles outside of the chamber.</p...

Micropumps can drive liquid flow on a much smaller scale than most pumps. In recent years, various driving schemes have been applied successfully to microfluidic systems1,2,3,4,5. The electrohydrodynamic (EHD) pump can exert forces directly on the liquid, without any moving parts, which makes it simpler and easier to fabricate6. According to the charge types, EHD pumps can be classified as injection pumps, induction pumps, or conduction pumps. Induction pumps do not work on isothermal liquids, while injection pumps change the liquid conductivity. Because they lack such problems, conduction pumps are more stable and have a wider application.
The conduction pump is based on the mismatch of the dissociation and recombination rates of liquid molecules. Normally, the dissociation and recombination process can be expressed as follows7,8: 
<img alt="Equation" src="/files/ftp_upload/55867/55867eq1.jpg" /> 
where the recombination rate kr is constant while the dissociation rate kd is a function of the electric field strength. When the electric field strength reaches a certain value, the dissociation rate will exceed the recombination rate. Then, more and more free charges travel to the two electrodes of opposite polarity, and heterocharge layers form. These heterocharge layers are the key to the pump, as the movement of the charges pushes the liquid molecules forward. Therefore, net body force can be generated in the liquid within the chamber using asymmetric electrodes or the mismatch of the mobility of positive and negative ions9,10,11,12.
This work introduces a new way of fabricating a symmetric planar electrode plate for a conduction pump. The electrode plate is prepared on FR-4 CCL, and the pump chamber is prepared by micromachining. The fabrication processes are relatively simpler and more convenient than those of other manufacturing methods, such as nanolithography. A testing platform is set up to investigate the performance of the conduction micropump under different conditions. Furthermore, the reliability of the conduction micropump is also investigated under different circumstances.

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			<td height="21" style="width:207px;height:21px;">Amperemeter</td>
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			<td height="21" style="width:207px;height:21px;">Fuse</td>
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			<td height="21" style="width:207px;height:21px;">Ultrasonic cleaner</td>
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			<td height="21" style="width:207px;height:21px;">Soldering iron</td>
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</table>

a performance-testing platform for a conduction micropump with an fr-4 copper-clad electrode plate

Here, a conduction micropump with symmetric planar electrode pairs prepared on flame-retardant glass-reinforced epoxy (FR-4) copper-clad laminate (CCL) is fabricated. It is used to investigate the influence of chamber dimensions on the performance of a conduction micropump and to determine the reliability of the conduction pump when acetone is used as the working fluid. A testing platform is set up to evaluate conduction micropump performance under different conditions. When the chamber height is 0.2 mm, the pump pressure reaches its peak value.

As shown in Figure 11, the pump pressure and its increasing rate rise when the voltage increases. When the voltage reaches 500 V, the pump pressure reaches 1,100 Pa.
The pump static pressure rises with the pump chamber height increasing when the chamber height is under 0.2 mm. The pump performance reaches its highest point when the chamber height is 0.2 mm. Then, the static pressure drops when the c...

Watch this Scientific Journal Video about A Performance-testing Platform for a Conduction Micropump with an FR-4 Copper-clad Electrode Plate at JoVE.com

A Performance-testing Platform for a Conduction Micropump with an FR-4 Copper-clad Electrode Plate

This paper presents a protocol for the fabrication of a conduction micropump using symmetric planar electrodes on flame-retardant glass-reinforced epoxy (FR-4) copper-clad laminate (CCL) to test the influence of chamber dimensions on the performance of a conduction micropump.

Here, a conduction micropump with symmetric planar electrode pairs prepared on flame-retardant glass-reinforced epoxy (FR-4) copper-clad laminate (CCL) is ...

The overall goal of this experiment is to fabricate a conduction micropump using symmetric planar electrodes on flame-retardant, glass-reinforced expoxy copper-clad laminate in order to test the influence of chamber dimensions on the performance of a conduction micropump. The main advantage of this technique is the voltage involved is a low several hundred volts. And the fabrication cost is relatively low.Though this method can provide insight into the mechanism of the conduction pump, it can also be applied to many other systems such as trunk delivering systems and micro-cooling systems. To begin, place the plate inside the beaker with the electrode surface facing down to help remove small particles like dust from the plate surface. Also, place the holder, the inlet and outlet tubes and other tools used in the experiments inside the beaker.Then pour enough 99.5 per cent acetone to immerse the items. Place the beaker inside the ultrasonic washer, and turn it on for five minutes. Next insert the inlet and outlet stainless steel tubes into the two holes on the cover plate.Place a chamber plate made of silicone membrane on the electrode plate, and then cover it with a cover plate. Stack and align the cover plate, the chamber plate and the electrode plate from top to bottom and insert the aligned plates into the holder. Use an M2 bolt to fix the plates inside the holder.Press the plates together by tightening the bolts. Next use two polyurethane hoses with external diameters of four millimeters and internal diameters of two millimeters to connect the inlet and outlet stainless steel tubes. Connect an amperemeter, a 500 volt DC power source and the micropump in series.Then insert a one milliampere fuse between the amperemeter and the power source to protect the amperemeter in case the micropump is shorted. Finally insert the inlet hose into a 50 milliliter beaker with 20 to 30 milliliters of acetone inside. To prepare for the experiment, use a cylinder to extract acetone to fill up the micropump.After the liquid level reaches the outlet hose, continue to extract ten milliliters of acetone inside until all bubbles are pushed away from the chamber. Repeatedly aspirate the acetone out of the pump and then re-inject it without bringing any air inside the pump to help remove bubbles. To perform the static pressure test, attach the outlet hose to a small frame so that the hose can remain straight and vertical.Put a ruler alongside the outlet hose to measure the liquid level. Connect the micropump to the power source. Start the test by raising the voltage to 500 volts and then mark down the initial liquid level.After the liquid level becomes stable, record the time, the final liquid level and the electric current. Continue to record the liquid level and the current every ten seconds until the micropump breaks down. To perform the flow rate test, use a large measuring cylinder to collect the liquid coming out of the outlet hose.Be sure to fix the outlet hose so that the end remains at the same altitude as the liquid level in the beaker. Switch on the power source. Start the test by raising the voltage to 500 volts and then mark down the initial liquid level.As the liquid starts to flow out of the outlet hose, record the volume of acetone inside the measuring cylinder every ten seconds. As the experiment goes on, add acetone to the beaker to maintain the liquid level. Shown here is the increasing rate of pump pressure with an increase in voltage.When the voltage reaches 500 volts, the pump pressure reaches 1, 100 Pascal. The pump static pressure rises with the pump chamber height. The pump performance reaches its highest point when the chamber height is 0.2 millimeters.Then the static pressure drops when the chamber height continues to increase. Based on these results, 0.2 millimeters is considered the best value for the chamber height. By increasing the chamber length, the static pressure rises, with a big slope until a chamber length of 23 millimeters.Then it continues to rise with a smaller slope, indicating a gradual rise. There is a slight decrease in the static pressure versus chamber width curve when the chamber width increases from two millimeters to three millimeters. Afterwards, the static pressure remains at a certain level as the chamber width increases.Once mastered, this technique can be done in half an hour if it is performed properly. After watching this video, you should have a good understanding of how to set up a platform and test the performance of a conduction micropump. Don't forget that working with acetone can be hazardous and precautions such as wearing rubber gloves should always be taken while performing this procedure.We showed demonstration of this method is critical. A short circuit may occur during the experiment if the washing and inspecting steps are not conducted carefully.

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Bioengineering

Environmentally-controlled Microtensile Testing of Mechanically-adaptive Polymer Nanocomposites for ex vivo Characterization

Implantable microdevices are gaining significant attention for several biomedical applications1-4. Such devices have been made from a range of materials, each offering its own advantages and shortcomings5,6. Most prominently, due to the microscale device dimensions, a high modulus is required to facilitate implantation into living tissue. Conversely, the stiffness of the device should match the surrounding tissue to minimize induced local strain7-9. Therefore, we recently developed a new class of bio-inspired materials to meet these requirements by responding to environmental stimuli with a change in mechanical properties10-14. Specifically, our poly(vinyl acetate)-based nanocomposite (PVAc-NC) displays a reduction in stiffness when exposed to water and elevated temperatures (e.g. body temperature). Unfortunately, few methods exist to quantify the stiffness of materials in vivo15, and mechanical testing outside of the physiological environment often requires large samples inappropriate for implantation. Further, stimuli-responsive materials may quickly recover their initial stiffness after explantation. Therefore, we have developed a method by which the mechanical properties of implanted microsamples can be measured ex vivo, with simulated physiological conditions maintained using moisture and temperature control13,16,17. 
To this end, a custom microtensile tester was designed to accommodate microscale samples13,17 with widely-varying Young's moduli (range of 10 MPa to 5 GPa). As our interests are in the application of PVAc-NC as a biologically-adaptable neural probe substrate, a tool capable of mechanical characterization of samples at the microscale was necessary. This tool was adapted to provide humidity and temperature control, which minimized sample drying and cooling17. As a result, the mechanical characteristics of the explanted sample closely reflect those of the sample just prior to explantation.
The overall goal of this method is to quantitatively assess the in vivo mechanical properties, specifically the Young's modulus, of stimuli-responsive, mechanically-adaptive polymer-based materials. This is accomplished by first establishing the environmental conditions that will minimize a change in sample mechanical properties after explantation without contributing to a reduction in stiffness independent of that resulting from implantation. Samples are then prepared for implantation, handling, and testing (Figure 1A). Each sample is implanted into the cerebral cortex of rats, which is represented here as an explanted rat brain, for a specified duration (Figure 1B). At this point, the sample is explanted and immediately loaded into the microtensile tester, and then subjected to tensile testing (Figure 1C). Subsequent data analysis provides insight into the mechanical behavior of these innovative materials in the environment of the cerebral cortex.

Environmentally-controlled Microtensile Testing of Mechanically-adaptive Polymer Nanocomposites for ex vivo Characterization

A method is discussed by which the in vivo mechanical behavior of stimuli-responsive materials is monitored as a function of time. Samples are tested ex vivo using a microtensile tester with environmental controls to simulate the physiological environment. This work further promotes understanding the in vivo behavior of our material.

Implantable microdevices are gaining significant attention for several biomedical applications1-4. Such devices have been made from a range of materials, ...

Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This technology has shown potential in addressing previously challenging applications; including, delivery to primary immune cells, cell reprogramming, carbon nanotube, and quantum dot delivery. This vector-free microfluidic platform relies on mechanical disruption of the cell membrane to facilitate cytosolic delivery of the target material. Herein, we describe the detailed method of use for these microfluidic devices including, device assembly, cell preparation, and system operation. This delivery approach requires a brief optimization of device type and operating conditions for previously unreported applications. The provided instructions are generalizable to most cell types and delivery materials as this system does not require specialized buffers or chemical modification/conjugation steps. This work also provides recommendations on how to improve device performance and trouble-shoot potential issues related to clogging, low delivery efficiencies, and cell viability.

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This protocol provides detailed steps on how to use the system for a broad range of applications.

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This ...

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

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. However, to achieve robust sample manipulation it is necessary to address device integration with the macroscale environment. To realize repeatable, sensitive particle separation with microfluidic devices, this protocol presents a complete automated and integrated microfluidic platform that enables precise processing of 0.15&#8211;1.5 ml samples using microfluidic devices. Important aspects of this system include modular device layout and robust fixtures resulting in reliable and flexible world to chip connections, and fully-automated fluid handling which accomplishes closed-loop sample collection, system cleaning and priming steps to ensure repeatable operation. Different microfluidic devices can be used interchangeably with this architecture. Here we incorporate an acoustofluidic device, detail its characterization, performance optimization, and demonstrate its use for size-separation of biological samples. By using real-time feedback during separation experiments, sample collection is optimized to conserve and concentrate sample. Although requiring the integration of multiple pieces of equipment, advantages of this architecture include the ability to process unknown samples with no additional system optimization, ease of device replacement, and precise, robust sample processing.

This protocol describes a system architecture for performing automated small volume (0.15&#8211;1.5 ml) particle separations using a microfluidic device, and discusses methods to optimize acoustofluidic device performance and operation.

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. ...

Three-Dimensionally Printed Microfluidic Cross-flow System for Ultrafiltration/Nanofiltration Membrane Performance Testing

Minimization and management of membrane fouling is a formidable challenge in diverse industrial processes and other practices that utilize membrane technology. Understanding the fouling process could lead to optimization and higher efficiency of membrane based filtration. Here we show the design and fabrication of an automated three-dimensionally (3-D) printed microfluidic cross-flow filtration system that can test up to 4 membranes in parallel. The microfluidic cells were printed using multi-material photopolymer 3-D printing technology, which used a transparent hard polymer for the microfluidic cell body and incorporated a thin rubber-like polymer layer, which prevents leakages during operation. The performance of ultrafiltration (UF), and nanofiltration (NF) membranes were tested and membrane fouling could be observed with a model foulant bovine serum albumin (BSA). Feed solutions containing BSA showed flux decline of the membrane. This protocol may be extended to measure fouling or biofouling with many other organic, inorganic or microbial containing solutions. The microfluidic design is especially advantageous for testing materials that are costly or only available in small quantities, for example polysaccharides, proteins, or lipids due to the small surface area of the membrane being tested. This modular system may also be easily expanded for high throughput testing of membranes.&#160;

Design and fabrication of a three-dimensionally (3-D) printed microfluidic cross-flow filtration system is demonstrated. The system is used to test performance and observe fouling of ultrafiltration and nanofiltration (thin film composite) membranes.

Minimization and management of membrane fouling is a formidable challenge in diverse industrial processes and other practices that utilize membrane ...

Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles

Microfluidic processing of biological samples typically involves differential manipulations of suspended particles under various force fields in order to spatially fractionate the sample based on a biological property of interest. For the resultant spatial distribution to be used as the assay readout, microfluidic devices are often subjected to microscopic analysis requiring complex instrumentation with higher cost and reduced portability. To address this limitation, we have developed an integrated electronic sensing technology for multiplexed detection of particles at different locations on a microfluidic chip. Our technology, called Microfluidic CODES, combines Resistive Pulse Sensing with Code Division Multiple Access to compress 2D spatial information into a 1D electrical signal. In this paper, we present a practical demonstration of the Microfluidic CODES technology to detect and size cultured cancer cells distributed over multiple microfluidic channels. As validated by the high-speed microscopy, our technology can accurately analyze dense cell populations all electronically without the need for an external instrument. As such, the Microfluidic CODES can potentially enable low-cost integrated lab-on-a-chip devices that are well suited for the point-of-care testing of biological samples.

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 processing of biological samples typically involves differential manipulations of suspended particles under various force fields in order to ...

An Ultrasonic Tool for Nerve Conduction Block in Diabetic Rat Models

Nerve conduction block with a high intensity-focused ultrasound (HIFU) transducer has been performed in normal and diabetic animal models recently. HIFU can reversibly block the conduction of peripheral nerves without damaging the nerves while using an appropriate ultrasonic parameter. Temporary and partial block of the action potentials of nerves shows that HIFU has the potential to be a useful clinical treatment for pain relief. This work demonstrates the procedures for suppressing the action potentials of neuropathic nerves in diabetic rats in vivo using an HIFU transducer. The first step is to generate adult male diabetic neuropathic rats by streptozotocin (STZ) injection. The second step is to evaluate the peripheral diabetic neuropathy in STZ-induced diabetic rats by an electronic von Frey probe and a hot plate. The final step is to record in vivo extracellular action potentials of the nerve exposed to HIFU sonication. The method showed here may benefit the study of ultrasound analgesic applications.

This work presents the methodology of applying high intensity-focused ultrasound to block the action potentials of diabetic neuropathic nerves.

Nerve conduction block with a high intensity-focused ultrasound (HIFU) transducer has been performed in normal and diabetic animal models recently. HIFU ...

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved useful as a convenient live imaging platform providing capabilities for precise control of experimental conditions in real time. In this article, we demonstrate live imaging of individual worms employing WormSpa, a previously-published custom microfluidic device. In the device, multiple worms are individually confined to separate chambers, allowing multiplexed longitudinal surveillance of various biological processes. To illustrate the capability, we performed proof-of-principle experiments in which worms were infected in the device with pathogenic bacteria, and the dynamics of expression of immune response genes and egg laying were monitored continuously in individual animals. The simple design and operation of this device make it suitable for users with no previous experience with microfluidic-based experiments. We propose that this approach will be useful for many researchers interested in longitudinal observations of biological processes under well-defined conditions.

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

In this article, we demonstrate live imaging of individual worms employing a custom microfluidic device. In the device, multiple worms are individually confined to separate chambers, allowing multiplexed longitudinal surveillance of various biological processes.

In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression

The limitations of cell-based synthetic biology are becoming increasingly apparent as researchers aim to develop larger and more complex synthetic genetic regulatory circuits. The analysis of synthetic genetic regulatory networks in vivo is time consuming and suffers from a lack of environmental control, with exogenous synthetic components interacting with host processes resulting in undesired behavior. To overcome these issues, cell-free characterization of novel circuitry is becoming more prevalent. In vitro transcription and translation (IVTT) mixtures allow the regulation of the experimental environment and can be optimized for each unique system. The protocols presented here detail the fabrication of a multilayer microfluidic device that can be utilized to sustain IVTT reactions for prolonged durations. In contrast to batch reactions, where resources are depleted over time and (by-) products accumulate, the use of microfluidic devices allows the replenishment of resources as well as the removal of reaction products. In this manner, the cellular environment is emulated by maintaining an out-of-equilibrium environment in which the dynamic behavior of gene circuits can be investigated over extended periods of time. To fully exploit the multilayer microfluidic device, hardware and software have been integrated to automate the IVTT reactions. By combining IVTT reactions with the microfluidic platform presented here, it becomes possible to comprehensively analyze complex network behaviors, furthering our understanding of the mechanisms that regulate cellular processes.

The fabrication process of a PDMS-based, multilayer, microfluidic device that allows in vitro transcription and translation (IVTT) reactions to be performed over prolonged periods is described. Furthermore, a comprehensive overview of the hardware and software required to automate and maintain these reactions for prolonged durations is provided.

The limitations of cell-based synthetic biology are becoming increasingly apparent as researchers aim to develop larger and more complex synthetic genetic ...

A Microfluidic Platform for Stimulating Chondrocytes with Dynamic Compression

Mechanical stimuli are known to modulate biological functions of cells and tissues. Recent studies have suggested that compressive stress alters growth plate cartilage architecture and results in growth modulation of long bones of children. To determine the role of compressive stress in bone growth, we created a microfluidic device actuated by pneumatic pressure, to dynamically (or statically) compress growth plate chondrocytes embedded in alginate hydrogel cylinders. In this article, we describe detailed methods for fabricating and characterizing this device. The advantages of our protocol are: 1) Five different magnitudes of compressive stress can be generated on five technical replicates in a single platform, 2) It is easy to visualize cell morphology via a conventional light microscope, 3) Cells can be rapidly isolated from the device after compression to facilitate downstream assays, and 4) The platform can be applied to study mechanobiology of any cell type that can grow in hydrogels.

This article provides detailed methods for fabricating and characterizing a pneumatically actuating microfluidic device for chondrocyte compression.

Mechanical stimuli are known to modulate biological functions of cells and tissues. Recent studies have suggested that compressive stress alters growth ...