Name: fabrication of refractive-index-matched devices for biomedical microfluidics
Uploaded: 2018-09-10T14:45:00
Description: Watch this Scientific Journal Video about Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics at JoVE.com

Dette arbejde blev støttet af University of Utah office vicepræsident for forskning og af midler i forbindelse med give P30 CA042014 tildelt til jægeren Cancer Institute og CRR Program på jægeren Cancer Institute.

1. fabrikation af Polydimethylsiloxan Negative
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Forberedelse af Polydimethylsiloxan
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<li>Foranstaltning 18 g PDMS silikoneelastomer og 1,8 g hærdning reagens. Hæld den hærdning reagens i en måling båd indeholdende elastomer.</li>
		<li>Bland elastomer og hærdning reagenset kraftigt i 1 min og sæt blandingen i et vakuumkammer i 30 min.</li>
		<li>Fjerne PDMS fra vakuum, hæld 15 g på det negative ved hjælp af en cookie cutter (radius = 3,8 cm) til at holde PDMS fra kører ned af si...

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	<li>Zare, R. N., Kim, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+platforms+for+single-cell+analysis.">Microfluidic platforms for single-cell analysis.</a> Annual Review Biomedical Engineering. 12, 187-201 (2010).</li><li>Neuzi, P., Giselbrecht, S., Lange, K., Huang, T. J., Manz, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revisiting+lab-on-a-chip+technology+for+drug+discovery.">Revisiting lab-on-a-chip technology for drug discovery.</a> Nature Reviews Drug Discovery. 11 (8), 620-632 (2012).</li><li>Xu, W., 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=Cell+stiffness+is+a+biomarker+of+the+metastatic+potential+of+ovarian+cancer+cells.">Cell stiffness is a biomarker of the metastatic potential of ovarian cancer cells.</a> PLoS ONE. 7 (10), 46609 (2012).</li><li>Karakas, H. E., 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=A+microfluidic+chip+for+screening+individual+cancer+cells+via+eavesdropping+on+autophagy-inducing+crosstalk+in+the+stroma+niche.">A microfluidic chip for screening individual cancer cells via eavesdropping on autophagy-inducing crosstalk in the stroma niche.</a> Scientific Reports. 7 (1), 2050 (2017).</li><li>DeBerardinis, R. J., Lum, J. J., Hatzivassiliou, G., Thompson, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biology+of+cancer:+metabolic+reprogramming+fuels+cell+growth+and+proliferation.">The biology of cancer: metabolic reprogramming fuels cell growth and proliferation.</a> Cell Metabolism. 7 (1), 11-20 (2008).</li><li>Yin, H., Marshall, D. <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+single+cell+analysis.">Microfluidics for single cell analysis.</a> Current Opinion in Biotechnology. 23 (1), 110-119 (2012).</li><li>Kemper, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+of+laser+micromanipulated+optically+trapped+cells+by+digital+holographic+microscopy.">Monitoring of laser micromanipulated optically trapped cells by digital holographic microscopy.</a> Journal of Biophotonics. 3 (7), 425-431 (2010).</li><li>Barer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interference+micorscopy+and+mass+determination.">Interference micorscopy and mass determination.</a> Nature. 169 (4296), 366-367 (1952).</li><li>Zangle, T. A., Teitell, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live-cell+mass+profiling:+an+emerging+approach+in+quantitative+biophysics.">Live-cell mass profiling: an emerging approach in quantitative biophysics.</a> Nature Methods. 11 (12), 1221-1228 (2014).</li><li>Chun, J., 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=Rapidly+quantifying+drug+sensitivity+of+dispersed+and+clumped+breast+cancer+cells+by+mass+profiling.">Rapidly quantifying drug sensitivity of dispersed and clumped breast cancer cells by mass profiling.</a> Analyst. 137 (23), 5495-5498 (2012).</li><li>Reed, J., 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=Live+cell+interferometry+reveals+cellular+dynamism+during+force+propagation.">Live cell interferometry reveals cellular dynamism during force propagation.</a> Acs Nano. 2 (5), 841-846 (2011).</li><li>Eldridge, W. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Refractive-index-matched+hydrogel+materials+for+measuring+flow-structure+interactions.">Refractive-index-matched hydrogel materials for measuring flow-structure interactions.</a> Experiments in Fluids. 54 (2), 6 (2013).</li><li>Ogawa, T., Hanada, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfabrication+of+the+UV+transparent+polymer+CYTOP+using+a+conventional+pulsed+green+laser.">Microfabrication of the UV transparent polymer CYTOP using a conventional pulsed green laser.</a> Applied Physics a-Materials Science &amp; Processing. 122 (3), 6 (2016).</li><li>Hanada, Y., Ogawa, T., Koike, K., Sugioka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Making+the+invisible+visible:+a+microfluidic+chip+using+a+low+refractive+index+polymer.">Making the invisible visible: a microfluidic chip using a low refractive index polymer.</a> Lab on a Chip. 16 (13), 2481-2486 (2016).</li><li>Zangle, T. 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MY133-V2000 kan bruges som et alternativ til traditionelle blødt litografi fabrikation materialer såsom PDMS. Tidligere arbejde har vist, at materialer med et højt indeks af brydning, såsom PDMS, indføre betydelige artefakter nær kanalen væggene på grund af forkert indekser i brydning mellem fabrikation materiale og den vandige opløsning inde kanalen 13. MY133-V2000 giver mulighed for matchende brydningsindekset af enheden mikrofluid at de vandige opløsninger almindeligt anvendt i biomed...

Udviklingen af mikrofluid teknologi har muliggjort en lang række nye biomedicinske teknikker, der udnytter den unikke fysik af mikroskala strømme1,2. Dette omfatter de diagnostiske teknikker bygget på mikrofluid platforme at kvantificere klinisk relevante biomarkører, herunder celle stivhed3, celleoverflademarkører4og vækst5. Ved at manipulere enkelt celler, kan mikrofluid enheder også bruges til at måle biomarkør heterogenitet, for eksempel som en indikator for malignitet6. Evnen til at kombinere mikrofluid programmer med mikroskopi steget yderligere nytte af disse platforme ved at tillade enheder, der måler flere biomarkører samtidig7.
QPM er en mikroskopi teknik, der måler faseskift, som lyset passerer gennem og interagerer med sagen inde i gennemsigtig prøver. Massen af individuelle celler kan beregnes ud fra QPM målinger, ved hjælp af det kendte forholdet mellem brydningsindekset og biomasse massefylde8,9. Tidligere arbejde har vist, at QPM er i stand til at måle klinisk relevante parametre såsom celle vækst10,11 og celle mekaniske egenskaber via lidelse styrke12. Når det kombineres med mikrofluidik, kan QPM potentielt bruges til at måle celle adfærd i et stærkt kontrolleret miljø i vitro. En af de primære udfordringer ved at kombinere QPM med mikrofluidik er høj brydningsindekset af de fleste polymerer, der anvendes til at konstruere mikrofluid kanaler via bløde litografi13.
En vigtig udfordring for kombinationen af mikrofluidik med forskellige mikroskopi-teknikker er misforholdet mellem brydningsindekset af enhed materiale i forhold til brydningsindekset for vand14,15. En metode til at løse dette er gennem brug af en lav brydningsindekset polymer som CYTOP16 eller MY133-V200013. Sidstnævnte er en fluorholdige ultraviolet (UV)-helbredes acrylat polymer, der har en refraktivt indeks svarer til vand (n = 1,33) og der er kompatible med bløde litografi teknikker, giver mulighed for en problemfri integration i mange etablerede mikrofluid enheden fabrikation arbejdsprocesser. Det gør MY133-V2000 ikke kun egnet til mikrofluid indretning fabrikation, men giver også mulighed for at være let kombineret med QPM og andre mikroskopi metoder, til at måle celle opførsel både på kolonien og på en enkelt celle skala. MY133-V2000 eliminerer artefakter som følge af fase udpakning ved at producere lidt, hvis nogen, faseskift som lys passerer gennem vand-MY133 interface.
Selv om at fjerne uoverensstemmelsen i brydningsindeks, er en stor udfordring i forbindelse med enheder fremstillet af fluorholdige polymerer, såsom MY133-V2000, lav overholdelsen af andre materialer som glas eller PDMS. Den nuværende arbejde viser fabrikation af en MY133-V2000 mikrofluid enheden ved hjælp af bløde litografi. Ved hjælp af O2 plasma til behandling af overfladen af både kanalen og PDMS sikrer substrat kombineret med en brugerdefineret opdigtet akryl indehaveren, at enheden klæber til underlaget, at skabe en forseglet kanal. Denne enhed er velegnet til cellekultur og QPM til at måle massen af celler i den kanal, som har vigtige applikationer til at måle væksten af levende celler og intracellulær transport af celle biomasse, som begge har klinisk relevans i diagnostiske medicin og narkotika opdagelsen.

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			<td height="21" style="height:21px;width:276px;">MY133-V2000</td>
			<td style="width:183px;">MY Polymers</td>
			<td style="width:203px;">MY133-V2000</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Sylgard 184</td>
			<td style="width:183px;">Ellsworth Adhesives</td>
			<td>184 SIL ELAST KIT 0.5KG</td>
			<td></td>
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			<td height="21" style="height:21px;width:276px;">Fisher Premium microscope slides</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td>12-544-4</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;">.118&#34;(3.0mm) x 12&#34; x 12&#34; Acrylic Sheet</td>
			<td>United States Plastic Corp</td>
			<td>44290</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">.060&#34;(1.5mm) x 12&#34; x 12&#34; Acrylic Sheet</td>
			<td>United States Plastic Corp</td>
			<td>44200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">SCIGRIP 3 Very Fast Set Acrylic Cement</td>
			<td style="width:183px;">United States Plastic Corp</td>
			<td style="width:203px;">45735</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Standard Aluminum Foil (.6 mm thick)</td>
			<td style="width:183px;">VWR</td>
			<td>89107-726</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Kim Wipes</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">06-666</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Insta-Cure+ Super Glue</td>
			<td style="width:183px;">Bob Smith Industries</td>
			<td style="width:203px;">BSI-109</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:276px;">1/8&#34; PVC tubing</td>
			<td style="width:183px;">McMaster Carr</td>
			<td style="width:203px;">5231K55</td>
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			<td height="21" style="height:21px;width:276px;">McCormick Food Coloring</td>
			<td style="width:183px;">Target</td>
			<td style="width:203px;">13353207</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">X-Acto #1 Precision Knife</td>
			<td style="width:183px;">X-Acto</td>
			<td style="width:203px;">X3201</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">X-Acto #18 Heavyweight wood chiseling blade</td>
			<td style="width:183px;">X-Acto</td>
			<td style="width:203px;">X218</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">VWR Razor Blades</td>
			<td style="width:183px;">VWR</td>
			<td style="width:203px;">55411-055</td>
			<td></td>
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			<td height="21" style="height:21px;width:276px;">Surface Treated Cell Culture Dishes</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">FBO12922</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Fibronectin Human Plasma</td>
			<td style="width:183px;">Sigma-Aldritch</td>
			<td style="width:203px;">F0895-1MG</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Trypsin-EDTA 10x</td>
			<td>Fisher Scientific</td>
			<td>15-400-054</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:276px;">Corning Dulbecco&#39;s Phosphate Buffered Saline</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">MT21030CM</td>
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			<td height="21" style="height:21px;width:276px;">Gibco Penicillin-Streptomycin</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">15-140-148</td>
			<td></td>
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			<td height="21" style="height:21px;width:276px;">HyClone Nonessential Amino Acids 100x</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">SH3023801</td>
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			<td height="21" style="height:21px;width:276px;">Fetal Bovine Serum</td>
			<td style="width:183px;">Omega Scientific</td>
			<td style="width:203px;">FB-12</td>
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			<td height="42" style="height:42px;width:276px;">Corning DMEM with L-glutamine and glucose</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">MT10013CV</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:276px;">Trichloro(1H,1H,2H,2H-perfluorooctyl)silane</td>
			<td style="width:183px;">Sigma-Aldritch</td>
			<td style="width:203px;">448931</td>
			<td>Reacts violently with water</td>
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			<td height="21" style="height:21px;width:276px;">Ethanol, 200 proof Decon Labs</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">04-355-223</td>
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			<td height="21" style="height:21px;width:276px;">Acetone</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">A18P-4</td>
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			<td height="21" style="height:21px;width:276px;">Bel-Art 42025 Plastic Dessicator</td>
			<td style="width:183px;">Cole-Parmer</td>
			<td style="width:203px;">EW-06514-30</td>
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			<td height="21" style="height:21px;width:276px;">Epilog Fusion Laser Cutter, 120 W</td>
			<td style="width:183px;">Epilog Laser</td>
			<td>Epilog Fusion M2 32 Laser</td>
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			<td height="21" style="height:21px;width:276px;">Isotemp Stirring Hotplate</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td>SP88850200</td>
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			<td height="21" style="height:21px;width:276px;">Ateco 14111 1.5 inch stainless steel cutter</td>
			<td style="width:183px;">Ateco</td>
			<td style="width:203px;">14111</td>
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			<td height="21" style="height:21px;">Pyrex Glass Cell Culture Dish</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td>08-747B</td>
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			<td height="21" style="height:21px;">Radio Frequency Plasma Cleaner</td>
			<td style="width:183px;">Harrick Plasma</td>
			<td style="width:203px;">PDC-32G</td>
			<td>Used with Oxygen gas</td>
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			<td height="21" style="height:21px;width:276px;">Black Hole Laboratories Digivac</td>
			<td style="width:183px;">Black Hole Laboratories</td>
			<td style="width:203px;">Model 215</td>
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			<td height="21" style="height:21px;width:276px;">Intelli-Ray Ultraviolet Oven</td>
			<td style="width:183px;">Uvitron</td>
			<td style="width:203px;">UVO338</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Compact Spin Coater</td>
			<td style="width:183px;">MTI Corporation</td>
			<td style="width:203px;">VTC-100A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Fisher Brand Isotemp Oven</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td>15-103-0510</td>
			<td>Forced Air Convection</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:276px;">Gilson Positive Displacement Pipette P1000</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">FD10006G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">HeraCell VIOS 160i</td>
			<td style="width:183px;">Fisher Scientific</td>
			<td style="width:203px;">13 998 212PM</td>
			<td></td>
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fabrication of refractive-index-matched devices for biomedical microfluidics

Brugen af mikrofluid enheder er opstået som et afgørende redskab til biomedicinske programmer. Når det kombineres med moderne mikroskopi-teknikker, kan disse enheder gennemføres som en del af en robust platform i stand til at foretage samtidige supplerende målinger. Den primære udfordring lavet af en kombination af disse to teknikker er uoverensstemmelse i brydningsindeks mellem traditionelt materialer til at gøre mikrofluid enheder og de vandige opløsninger anvendes typisk i biomedicin. Denne uoverensstemmelse kan oprette optisk artefakter nær kanal eller enhed kanterne. En løsning er at reducere brydningsindekset af det materiale, der anvendes til at fabrikere enheden ved hjælp af en fluorholdig polymer som MY133-V2000 hvis brydningsindeks er lig i vandet (n = 1,33). Her, opførelsen af en mikrofluid enhed lavet af MY133-V2000 bruger bløde litografi teknikker er påvist, ved hjælp af O2 plasma sammenholdt med en akryl indehaveren for at øge vedhæftning mellem enheden og MY133-V2000 fabrikeret og den Polydimethylsiloxan (PDMS) substrat. Enheden er derefter testet ved at inkubere det fyldt med celle kultur medier til 24 h for at vise enhedens evne til at opretholde celle kultur betingelser i løbet af en typisk imaging eksperiment. Endelig, kvantitative fase mikroskopi (QPM) bruges til at måle fordelingen af masse i levende vedhængende cellerne i microchannel. Denne måde, godtgøres den øgede præcision, aktiveret ved opdigte enheden fra et lavt index of refraktion polymer som MY133-V2000 i stedet for traditionelle blødt litografi materialer såsom PDMS. Samlet set kan denne tilgang for at fabrikere mikrofluid enheder let integreres i eksisterende bløde litografi arbejdsprocesser for at reducere optisk artefakter og øge måling præcision.

Denne protokol beskriver fabrikation af MY133-V2000, en fluorholdig polymer med en lav brydningsindekset matcher det vand. Et centralt element i denne protokol er hvordan man kan overvinde manglen vedhæftning, der er karakteristisk for fluorholdige polymerer, ved hjælp af ilt plasma og ved opdigte enhed inden for en akryl indehaveren til at give den ekstra mekaniske styrke forpligtet til at forsegle kanalen mod PDMS substrat (figur 1). Lav brydningsindekset...

Watch this Scientific Journal Video about Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics at JoVE.com

Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

Denne protokol beskriver fabrikation af mikrofluid enheder fra MY133-V2000 at fjerne artefakter, der ofte opstår i microchannels på grund af de fejlagtige refraktivt indeks mellem microchannel strukturer og en vandig opløsning. Denne protokol bruger en akryl indehaveren til at komprimere den indkapslet enhed, forbedre adhæsion både kemisk og mekanisk.

Fabrikation af refraktive-index-matchede enheder for biomedicinsk mikrofluidik

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices ...

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Engineering

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