Name: three-dimensional printing of thermoplastic materials to create automated syringe pumps with feedback control for microfluidic applications
Uploaded: 2018-08-30T12:30:00
Description: Watch this Scientific Journal Video about Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications at JoVE.com

Gli autori riconoscono supporto dall'Office of Naval Research awards N00014-17-12306 e N00014-15-1-2502, come pure da Air Force Office of Scientific Research award FA9550-13-1-0108 e la National Science Foundation Grant n. 1709238.

1. stampa 3D e montaggio della pompa a siringa
<ol><li>
Preparare e 3D-stampa componenti pompa siringa
	<ol>
<li>Scarica il. File di disegno STL dai File supplementari di questa carta. Nota: Ci sono sei. I file STL, intitolati 'JoVE_Syringe_Clamp_10mL_Size.stl', 'JoVE_Syringe_Platform.stl', 'JoVE_Syringe_Plunger_Connectors.stl', 'JoVE_Syringe_Pump_End_Stop.stl', 'JoVE_Syringe_Pump_Motor_Connector.stl', e ' JoVE_Syringe_Pump_Traveler_ Push.STL', nei file supplementari</strong...

<ol>
	<li>Sackmann, E. K., Fulton, A. L., Beebe, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+present+and+future+role+of+microfluidics+in+biomedical+research.">The present and future role of microfluidics in biomedical research.</a> Nature. 507 (7491), 181-189 (2014).</li><li>Duncombe, T. A., Tentori, A. M., Herr, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics:+reframing+biological+enquiry.">Microfluidics: reframing biological enquiry.</a> Nature Reviews Molecular Cell Biology. 16 (9), 554-567 (2015).</li><li>Prakadan, S. M., Shalek, A. K., Weitz, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scaling+by+shrinking:+empowering+single-cell+'omics'+with+microfluidic+devices.">Scaling by shrinking: empowering single-cell 'omics' with microfluidic devices.</a> Nature Reviews Genetics. 18 (6), (2017).</li><li>Kim, Y., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics+in+nanomedicine.">Microfluidics in nanomedicine.</a> Reviews in Cell Biology and Molecular Medicine. 1, 127-152 (2015).</li><li>Rusconi, R., Garren, M., Stocker, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics+expanding+the+frontiers+of+microbial+ecology.">Microfluidics expanding the frontiers of microbial ecology.</a> Annual Review of Biophysics. 43, 65-91 (2014).</li><li>Skafte-Pedersen, P., Sabourin, D., Dufva, M., Snakenborg, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-channel+peristaltic+pump+for+microfluidic+applications+featuring+monolithic+PDMS+inlay.">Multi-channel peristaltic pump for microfluidic applications featuring monolithic PDMS inlay.</a> Lab on a Chip. 9 (20), 3003-3006 (2009).</li><li>Heo, Y. J., Kang, J., Kim, M. J., Chung, W. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tuning-free+controller+to+accurately+regulate+flow+rates+in+a+microfluidic+network.">Tuning-free controller to accurately regulate flow rates in a microfluidic network.</a> Scientific Reports. 6, 23273 (2016).</li><li>Kuczenski, B., LeDuc, P. R., Messner, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pressure-driven+spatiotemporal+control+of+the+laminar+flow+interface+in+a+microfluidic+network.">Pressure-driven spatiotemporal control of the laminar flow interface in a microfluidic network.</a> Lab on a Chip. 7 (5), 647-649 (2007).</li><li>Lake, J. R., Heyde, K. C., Ruder, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-cost+feedback-controlled+syringe+pressure+pumps+for+microfluidics+applications.">Low-cost feedback-controlled syringe pressure pumps for microfluidics applications.</a> PLoS One. 12 (4), (2017).</li><li>Kong, D. S., 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=Open-source,+community-driven+microfluidics+with+metafluidics.">Open-source, community-driven microfluidics with metafluidics.</a> Nature Biotechnology. 35 (6), 523-529 (2017).</li><li>Wijnen, B., Hunt, E. J., Anzalone, G. C., Pearce, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-source+syringe+pump+library.">Open-source syringe pump library.</a> PLoS One. 9 (9), e107216 (2014).</li><li>Ferry, M. S., Razinkov, I. A., Hasty, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics+for+synthetic+biology:+from+design+to+execution.">Microfluidics for synthetic biology: from design to execution.</a> Methods in Enzymology. , 295-372 (2011).</li><li>. Arduino Libraries for Timer.h Available from: <a target="_blank" href="https://github.com/JChristensen/Timer">https://github.com/JChristensen/Timer</a> (2018)</li><li>. Arduino Libraries for AccelStepper.h Available from: <a target="_blank" href="https://github.com/adafruit/AccelStepper">https://github.com/adafruit/AccelStepper</a> (2018)</li><li>Lin, F., 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=Generation+of+dynamic+temporal+and+spatial+concentration+gradients+using+microfluidic+devices.">Generation of dynamic temporal and spatial concentration gradients using microfluidic devices.</a> Lab on a Chip. 4 (3), 164-167 (2004).</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></ol>

Qui, abbiamo presentato un nuovo design per un sistema di pompa a siringa con controllo di pressione del circuito chiuso. Ciò è stata compiuta mediante l'integrazione di una pompa a siringa 3D-stampato con un sensore di pressione piezoresistivo e un microcontrollore open source. Impiegando un regolatore PID, siamo stati in grado controllare la pressione in ingresso e fornire tempi di risposta rapidi mantenendo nel contempo la stabilità su un set point in modo preciso.
Molti esperimenti util...

Strumenti di microfluidica sono diventati utili per scienziati nella ricerca biologica e chimica. Dovuto l'utilizzazione di basso volume, capacità di misurazione rapida, profili di flusso ben definiti, microfluidica ha guadagnato la trazione in genomica e proteomica ricerca, high throughput screening, diagnostica medica, nanotecnologia e cella singola analisi1,2,3,4. Inoltre, la flessibilità del design dei dispositivi microfluidici consente prontamente ricerca di scienza di base, ad esempio indagando la dinamica spazio-temporale di colonie batteriche colta5.
Molti tipi di sistemi di iniezione di fluido sono stati sviluppati per fornire con precisione il flusso di dispositivi microfluidici. Esempi di tali sistemi di iniezione peristaltiche e pompe di ricircolo6, regolatore di pressione sistemi7e pompe a siringa8. Questi sistemi di iniezione, tra cui pompe a siringa, sono spesso composti da componenti costosi precisione costruita. Aumentando questi sistemi con controllo di feedback a circuito chiuso della pressione nel flusso di uscita aumenta il costo di questi sistemi. In risposta, abbiamo precedentemente sviluppato un sistema di pompa siringa robusto, a basso costo che utilizza il controllo di feedback a circuito chiuso per regolare la pressione di flusso di output. Utilizzando il controllo di pressione del circuito chiuso, la necessità di costosi componenti di altissima precisione è abrogato9.
La combinazione di conveniente stampa 3D hardware e una crescita significativa in associato software open-source ha fatto la progettazione e la fabbricazione di dispositivi microfluidici sempre più accessibile ai ricercatori da una varietà di discipline10. Tuttavia, i sistemi utilizzati per fluido in auto attraverso questi dispositivi rimangono costosi. Per soddisfare questa esigenza per un sistema di controllo fluidi di basso costo, abbiamo sviluppato un design che possa essere fabbricato dai ricercatori in laboratorio, che richiedono solo un piccolo numero di fasi dell'assemblaggio. Nonostante il suo basso costo e semplice montaggio, questo sistema può fornire un controllo di flusso preciso e fornisce un'alternativa ai sistemi a pompa siringa commercialmente disponibili, ciclo chiuso, che può essere proibitivo.
Qui, forniamo i protocolli per la costruzione e l'utilizzo del ciclo chiuso controllato siringa sistema pompa abbiamo sviluppato (Figura 1). Il fluido sistema di gestione è composto da una pompa a siringa fisico ispirata da un precedente studio11, un microcontrollore e un sensore di pressione piezoresistivo. Quando montato e programmato con un regolatore di proporzionale-integrale-derivato (PID), il sistema è in grado di erogare un flusso ben regolato, pressione-driven per dispositivi microfluidici. Questo fornisce un'alternativa di basso costo e flessibile ai prodotti commerciali ad alto costo, consentendo un più ampio gruppo di ricercatori di utilizzare microfluidica nel loro lavoro.

<table border="0" cellpadding="0" cellspacing="0" width="1268">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:289px;">Arduino IDE</td>
			<td style="width:245px;">Arduino.org</td>
			<td style="width:169px;"></td>
			<td style="width:565px;">Arduino Uno R3 control software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Header Connector, 2 Positions</td>
			<td>Digi-Key</td>
			<td>WM4000-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Header Connector, 3 Positions</td>
			<td>Digi-Key</td>
			<td>WM4001-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Header Connector, 4 Positions</td>
			<td>Digi-Key</td>
			<td>WM4002-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hook-up Wire, 22 Gauge, Black</td>
			<td>Digi-Key</td>
			<td>1528-1752-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hook-up Wire, 22 Gauge, Blue</td>
			<td>Digi-Key</td>
			<td>1528-1757-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hook-up Wire, 22 Gauge, Red</td>
			<td>Digi-Key</td>
			<td>1528-1750-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hook-up Wire, 22 Gauge, White</td>
			<td>Digi-Key</td>
			<td>1528-1768-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hook-up Wire, 22 Gauge, Yellow</td>
			<td>Digi-Key</td>
			<td>1528-1751-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Instrumentation Amplifier</td>
			<td>Texas Instruments</td>
			<td>INA122P</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microcontroller, Arduino Uno R3</td>
			<td>Arduino.org</td>
			<td>A000066</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mini Breadboard</td>
			<td>Amazon</td>
			<td>B01IMS0II0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Power Supply</td>
			<td>BK Precision</td>
			<td>1550</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pressure Sensor</td>
			<td>PendoTech</td>
			<td>PRESS-S-000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rectangular Connectors, Housings</td>
			<td>Digi-Key</td>
			<td>WM2802-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rectangular Connectors, Male</td>
			<td>Digi-Key</td>
			<td>WM2565CT-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Resistors, 10k Ohm&#160;</td>
			<td>Digi-Key</td>
			<td>1135-1174-1-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Resistors, 330 Ohm&#160;</td>
			<td>Digi-Key</td>
			<td>330ADCT-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stepper Motor Driver, EasyDriver</td>
			<td>Digi-Key</td>
			<td>1568-1108-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">USB 2.0 Cable, A-Male to B-Male</td>
			<td>Amazon</td>
			<td>PC045</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3D Printed Material, Z-ABS&#160;</td>
			<td>Zortrax</td>
			<td></td>
			<td>A variety of colors are available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3D Printer</td>
			<td>Zortrax</td>
			<td>M200</td>
			<td>Printing out the syringe pump components</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ball Bearing, 17x6x6mm</td>
			<td>Amazon</td>
			<td>B008X18NWK</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex Machine Screws, M3x16mm&#160;</td>
			<td>Amazon</td>
			<td>B00W97MTII</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex Machine Screws, M3x35mm&#160;</td>
			<td>Amazon</td>
			<td>B00W97N2UW</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex Nut, M3 0.5&#160;</td>
			<td>Amazon</td>
			<td>B012U6PKMO</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex Nut, M5&#160;</td>
			<td>Amazon</td>
			<td>B012T3C8YQ</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lathe Round Rod</td>
			<td>Amazon</td>
			<td>B00AUB73HW</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Linear Ball Bearing</td>
			<td>Amazon</td>
			<td>B01IDKG1WO</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Linear Flexible Coupler</td>
			<td>Amazon</td>
			<td>B010MZ8SQU</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Steel Lock Nut, M3 0.5</td>
			<td>Amazon</td>
			<td>B000NBKLOQ</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stepper Motor, NEMA-17, 1.8o/step</td>
			<td>Digi-Key</td>
			<td>1568-1105-ND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe, 10mL, Luer-Lok Tip</td>
			<td>BD</td>
			<td>309604</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Threaded Rod</td>
			<td>Amazon</td>
			<td>B01MA5XREY</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1H,1H,2H,2H-Perfluorooctyltrichlorosilane</td>
			<td>FisherScientific</td>
			<td>AAL1660609</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Camera Module</td>
			<td>Raspberry Pi Foundation</td>
			<td>V2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Compact Oven</td>
			<td>FisherScientific</td>
			<td>PR305220G</td>
			<td>Baking PDMS pre-polymer mixture and the device</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dispensing Needle, 22 Gauge</td>
			<td>McMaster-Carr</td>
			<td>75165A682</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dispensing Needle, 23 Gauge</td>
			<td>McMaster-Carr</td>
			<td>75165A684</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fisherbrand Premium Cover Glasses</td>
			<td>FisherScientific</td>
			<td>12-548-5C</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass Culture Petri Dish, 130x25mm</td>
			<td>American Educational Products</td>
			<td>7-1500-5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plasma Cleaner</td>
			<td>Harrick Plasma</td>
			<td>PDC-32G</td>
			<td>Binding the cover glass with the PDMS device</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Razor Blades</td>
			<td>FisherScientific</td>
			<td>7071A141&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scotch Magic Tape</td>
			<td>Amazon</td>
			<td>B00RB1YAL6</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Single-board Computer</td>
			<td>Raspberry Pi Foundation</td>
			<td>Raspberry Pi 2 model B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Smart Spatula</td>
			<td>FisherScientific</td>
			<td>EW-06265-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sylgard 184 Silicone Elastomer Kit</td>
			<td>FisherScientific</td>
			<td>NC9644388</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe Filters</td>
			<td>Thermo Scientific</td>
			<td>7252520</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tygon Tubing</td>
			<td>ColeParmer&#160;</td>
			<td>EW-06419-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vacuum Desiccator</td>
			<td>FisherScientific</td>
			<td>08-594-15C</td>
			<td>Degasing PDMS pre-polymer mixture and coating fluorosilane on the master mold</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weighing Dishes</td>
			<td>FisherScientific</td>
			<td>S67090A</td>
			<td></td>
		</tr>
	</tbody>
</table>

three-dimensional printing of thermoplastic materials to create automated syringe pumps with feedback control for microfluidic applications

Microfluidica è diventato uno strumento fondamentale nella ricerca attraverso, chimica, scienze biologiche e fisiche. Una componente importante della sperimentazione di microfluidica è un sistema in grado di fornire con precisione una portata aspirata o la pressione di ingresso stabile per fluidi. Qui, abbiamo sviluppato un sistema di pompe siringa in grado di controllare e regolare la pressione del fluido di aspirazione consegnato a un dispositivo microfluidico. Questo sistema è stato progettato utilizzando materiali a basso costo e principi di produzione additiva, sfruttando la stampa tridimensionale (3D) di materiali termoplastici e componenti normalizzati quando possibile. Questo sistema è composto da tre componenti principali: una pompa a siringa, un trasduttore di pressione e un microcontrollore programmabile. All'interno di questa carta, abbiamo dettagliatamente una serie di protocolli di fabbricazione, assemblaggio e programmazione di questo sistema di pompa a siringa. Inoltre, abbiamo incluso i risultati rappresentativi che dimostrano ad alta fedeltà, controllo feedback della pressione di ingresso utilizzando questo sistema. Ci aspettiamo che questo protocollo permetterà ai ricercatori di fabbricare sistemi pompa siringa di basso costo, abbassando la barriera di ingresso per l'utilizzo della microfluidica in biomedica, chimica e ricerca dei materiali.

Qui, presentiamo un protocollo per la costruzione di una siringa di feedback controllato sistema di pompaggio e dimostrare i suoi usi potenziali per applicazioni di microfluidica. La figura 1 Mostra il sistema collegato della pompa a siringa, sensore di pressione, dispositivo microfluidico, microcontrollore, circuito del sensore di pressione e driver del motore passo a passo. Dettagliate didascalie per l'assemblaggio di pompa siringa sono mostrate in <strong ...

Watch this Scientific Journal Video about Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications at JoVE.com

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

Qui presentiamo un protocollo per la costruzione di una pompa a siringa a pressione controllata per essere utilizzato nelle applicazioni di microfluidica. Questa pompa a siringa fatta da un additivo fabbricato corpo, hardware COTS ed elettronica open-source. Il sistema risultante è a basso costo, semplice da costruire ed eroga ben regolato flusso del fluido per attivare la ricerca rapida microfluidica.

Tridimensionale di stampa di materiali termoplastici per creare automatizzati pompe a siringa con controllo in retroazione per applicazioni di microfluidica

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...

This method can help enable research in any field requiring precision liquid handling including molecular, cell, and systems biology, chemical engineering and microfabrication. The main advantage of this approach is that we are able to fabricate low-cost pressure-regulated syringe pumps. This technique should save money across a wide range of disciplines.This technique showcases how we can couple additive manufacturing with open-source electronics to make a useful piece of laboratory instrumentation. To begin, download the STL design files from the supplemental files of this publication. Prepare these files for printing by opening them in a software package dedicated to the conversion of STL model files to executable instruction sets for the 3D printer being used.Ensure that the proper software is being used as some printers will require proprietary software whereas others may be able to print directly from the STL file. Now print the plastic components using acrylonitrile butadiene styrene with a high-quality 3D printer setting. If other common 3D printing materials are being used such as polylactic acid or other thermoplastic elastomers, make sure that the finished mechanical properties are comparable.Once complete, detach the printed parts from the printing platform of the 3D printer. Remove the printed supporting structure from the finished parts. Smooth the printed components by sanding any rough edges using sandpaper.For best results, use sandpaper with a grit size below 220. Make sure all components are smooth before assembling and ensure that all seven parts have been printed. Fasten the stepper motor to a threaded rod using a motor shaft z-axis flexible coupler with set screws.Before continuing, make sure that rotating the stepper motor shaft drives the threaded rod without slippage. Connect the syringe platform to the motor connector by firmly pressing the syringe platform's connection pegs into the mating holes on top of the motor connector. Attach the two parts by fastening four 16 millimeter screws through the motor connector.Now insert two linear ball bearings and a 0.8 millimeter hex nut into the openings located on the bottom of the traveler push. Align the threaded rod on the motor connector through the 0.8 millimeter hex nut in the traveler push. Then insert the two linear shafts through the traveler push in the motor connector.Now place two hex nuts in the hexagonal spaces of the motor connector piece and then use two 16 millimeter screws to tighten the connections securing the linear shafts from moving. Insert the ball bearing into the middle opening of the end stop. Connect the end stop with the assembled components.Place two hex nuts in the hexagonal spaces of the end stop piece and then use two 16 millimeter screws to tighten the connections to affix the end stop to the assembly. Next, attach the syringe plunger female connector piece to the traveler push piece using two steel locknuts and two 16 millimeter screws. Place a 10 milliliter syringe on the top of the pump.Ensure the head of the plunger is aligned into the notch of the syringe plunger female connector piece and that the top of the syringe barrel is fixed into the slot of the motor connector. Now insert the syringe plunger male connector piece into the syringe plunger female connector. Connect the syringe clamp to the syringe platform using two hex nuts and two 35 millimeter screws ensuring the syringe barrel is fixed into the slot of the syringe clamp.Ensure that there is a tight fit between the male and female components securing the plunger in place. Proceed to microfluidic device preparation as detailed in the text protocol. To begin the syringe pump system assembly, remove 80%of the length of the wire insulation and shielding from the pressure sensor's electrical cable using a razor.Be gentle when cutting to ensure the wires are not compromised above the desired length. Once the insulation and shielding are removed, connect the wires to male rectangular connectors. Using a similar approach, remove one to two centimeters of the wire insulation from a stepper motor's leads and connect the wires to male rectangular connectors.Now affix the syringe onto the inlet side of the pressure sensor. Connect the 22 gauge dispensing needle onto the outlet side of the pressure sensor. Slide one end of 0.51 centimeter diameter tubing over the 22 gauge dispensing needle attached to the pressure sensor.Slide the other end of the tubing over a 22 gauge dispensing needle that can be connected to the microfluidic device. Then connect the needle to the inlet port of the microfluidic device. Connect the outlet port of the microfluidic device to a waste disposal reservoir using a 22 gauge needle and 0.51 centimeter diameter tubing similar to the inlet port's connection.Next, assemble the electronic circuit on a prototyping breadboard according to the diagram in the text protocol. Connect the wires from the stepper motor with the stepper motor driver. Then connect the wires from the pressure sensor and the stepper motor driver to the breadboard.Now connect the output signal from the breadboard with the analog input pin on the microcontroller. Connect the logic input pins from the stepper motor driver with the digital pins on the microcontroller. The step input on the stepper motor driver is connected with a pulse width modulated port of digital pins on the microcontroller denoted by a tilde sign.Connect the power supply to the breadboard as shown in the text. Then set the power supply to 10 volts for the breadboard and the stepper motor driver. To control the syringe pressure pumps, first open the Integrated Development Environment or IDE for the open-source microcontroller.Download the Timer. h and ExcelStepper. h libraries to the microcontroller's IDE library directory.Download the controller code titled Dual Pump PID Control.INO. This code is used to control the feedback-controlled syringe pump system with two pumps. Program the controller code so that it fits the experiment being conducted.Modify the control parameters for the timing parameters to fit the desired response and duration of the experiment. Compile and upload the code to the microcontroller before running the experiment. Turn on the power supply for the syringe pump system.Finally, set the voltage to 10 volts for the stepper motor power supply. Shown here are representative results showing how control parameters may be tuned. Modifying the proportional coefficients as shown here lead to different response profiles for the pressure signals.Modifying the integral and differential coefficients also lead to changes in the response profiles. Parameters may be optimized for different experimental designs. Shown here are representative results showing the control and modulation of a laminar flow profile in a microfluidic device.By controlling the inlet pressure at two inlet ports of a Y-shaped microfluidic device, the interface position may be precisely controlled downstream. Following this procedure, other techniques such as cell culturing and microscopy can be performed in order to answer questions related to synthetic gene network regulatory dynamics, cell physiology and biological regulatory networks. These syringe pump systems have enabled more rapid development of microfluidic procedures within our lab group.These devices are an inexpensive and time-efficient way for groups to control flow through many different microfluidic devices. We hope that researchers across a wide range of disciplines will be able to use and extend these pressure-regulated syringe pumps for their microfluidic needs.

Research

JoVE Journal

Bioengineering

Planar and Three-Dimensional Printing of Conductive Inks

Stampa planare e tridimensionale di inchiostri conduttivi

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

Ricerca

Bioingegneria

Designing Silk-silk Protein Alloy Materials for Biomedical Applications

Progettazione di materiali in lega di proteine ​​della seta-seta per le applicazioni biomediche

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Umana Cartilagine Tessuto Fabrication Uso tridimensionale Tecnologia Inkjet Printing

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Analysis of Cell Migration within a Three-dimensional Collagen Matrix

Analisi della migrazione delle cellule all&#39;interno di una tridimensionale di collagene Matrix

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

Generazione di una tridimensionale a tutto spessore della pelle Equivalent e Automated Ferimento

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

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

Una piattaforma per l&#39;elaborazione di precisione Microfluidic piccola Volume del campione e il suo uso per Dimensione particelle biologiche separate con un Microdevice Acoustic

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

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

Realizzazione superidrofobiche Materiali Polimerici per applicazioni biomediche

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial ...

Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

Fabrication of Inverted Colloidal Crystal Polyethylene glycol Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

Fabbricazione di invertito colloidale cristallo poli (glicole etilenico) Scaffold: una piattaforma Cell Culture tridimensionale per il fegato Tissue Engineering

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Fabbricazione di dispositivi microfluidici basati su carta tridimensionale per gli immunodosaggi

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

Sciogliere elettrofilatura scrittura di scaffold tridimensionale Poly(ε-caprolactone) con morfologie controllabile per applicazioni di ingegneria del tessuto

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...