Name: multi-analyte biochip (mab) based on all-solid-state ion-selective electrodes (assise) for physiological research
Uploaded: 2013-04-18T08:00:00
Description: Watch this Scientific Journal Video about Multi-analyte Biochip MAB Based on All-solid-state Ion-selective Electrodes ASSISE for Physiological Research at JoVE.com

Vi vil gerne takke NASA Astrobiology Videnskab og Teknologi Instrument Development (ASTID) Program til finansiering support (tilskud numrene 103.498 og 103.692), Gale Lockwood for Birck Nantechnology Center på Purdue University for wirebonding af MAB enheder og Joon Hyeong Park for CAD-tegning af strømmen-celle kammer.

1. Fremstilling af poly (3,4-ethylenedioxythiophene): Poly (natrium-4-styrensulfonat) (PEDOT: PSS) elektropolymerisation Solution for H + og CO 3 2 - Ioner <ol><li> Tilføj 70 mg poly (natrium-4-styrensulfonat) (Na + PSS -) til 10 ml deioniseret (DI) vand og vortex indtil den er helt spredt (ca. 10 sek.) </li><li> Tilføj 10.7 pi 3,4-ethlyenedioxythiophene (EDOT) til opløsning i 1.1 og vortex indtil opløsningen er helt blandet. </li></ol> 2...

<ol>
	<li>Migdalski, J., Bas, B., Blaz, T., Golimowski, J., Lewenstam, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Miniaturized+and+Integrated+Galvanic+Cell+for+the+Potentiometric+Measurement+of+Ions+in+Biological+Liquids.">A Miniaturized and Integrated Galvanic Cell for the Potentiometric Measurement of Ions in Biological Liquids.</a> J. Solid State Electrochem. 13, 149-155 (2009).</li><li>Buehler, M. G., Kounaves, S. P., Martin, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Designing+a+Water-quality+Monitor+with+Ion-selective-electrodes.">Designing a Water-quality Monitor with Ion-selective-electrodes.</a> 1, 331-338 (2001).</li><li>Adamchuk, V. I., Lund, E. D., Sethuramasamyraja, B., Morgan, M. T., Doberman, A., Marx, D. 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Solid State Electrochem. 8, 381-389 (2004).</li><li>Bard, A. J., Faulkner, L. R., ed, 2. n. d. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electrochemical Methods: Fundamentals and Applications. , (2000).</li><li>Claussen, J. C., Artiles, M. S., McLamore, E. S., Mohanty, S., Shi, J., Rickus, J., Fisher, T. S., Porterfield, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrochemical+Glutamate+Biosensing+with+Naanocube+and+Nanosphere+Augmented+Single-walled+Carbon+Nanotube+Networks:+A+Comparative+Study.">Electrochemical Glutamate Biosensing with Naanocube and Nanosphere Augmented Single-walled Carbon Nanotube Networks: A Comparative Study.</a> J. Mater. 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MAB biochip består af Assises der er konstrueret fra en ISM oven på en PEDOT-baserede CP konjugat transduktion lag på en Pt-elektrode, hvis kombination transducerer ionkoncentrationen af ​​interesse til et måleligt elektrisk signal. En stabil elektrode potentiale defineres af både CP lag og ISM lag. Begge lag også bestemme bearbejdning levetid MAB og kvaliteten (støj, fremdrift) af det målte elektriske signal. PEDOT er især attraktivt som en transduktion lag grund både sine ion...

<table border="1">
		<tbody>
		 <tr>
			<td>Name of the items</td>
			<td>Company</td>
			<td>Catalog number</td>
			<td>Comments </td>
		 </tr>
		 <tr>
			<td>3,4-Ethylenedioxythiophene</td>
			<td>Sigma-Aldrich</td>
			<td>483028</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Poly(sodium 4-styrenesulfonate)</td>
			<td>Sigma-Aldrich</td>
			<td>243051</td>
			<td></td>
		 </tr>
		 <tr>
			<td>EC epsilon galvanostat/potentiostat</td>
			<td>Bioanalytical Systems Inc.</td>
			<td>e2P</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Saturated Ag/AgCl reference electrode</td>
			<td>Bioanalytical Systems Inc.</td>
			<td>MF-2052</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Pt gauze</td>
			<td>Alfa Aesar</td>
			<td>10283</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Potassium ferricyanide</td>
			<td>Sigma-Aldrich</td>
			<td>P-8131</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Potassium nitrate</td>
			<td>J.T. Baker</td>
			<td>3190-01</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Sodium bicarbonate</td>
			<td>Mallinckrodt/ Macron</td>
			<td>7412-12</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Sodium carbonate</td>
			<td>Sigma-Aldrich</td>
			<td>S-7127</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Calcium chloride</td>
			<td>J.T. Baker</td>
			<td>1311-01</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Potassium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>P9541</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Calcium sulphate</td>
			<td>Sigma-Aldrich</td>
			<td>237132</td>
			<td></td>
		 </tr>
		 <tr>
			<td>C3 cell stand</td>
			<td>Bioanalytical Systems Inc.</td>
			<td>EF-1085</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Flow-cell chip holder</td>
			<td></td>
			<td></td>
			<td>Custom, courtesy of NASA Ames</td>
		 </tr>
		 <tr>
			<td>Flow-cell electrical fixture</td>
			<td></td>
			<td></td>
			<td>Custom, courtesy of NASA Ames </td>
		 </tr>
		 <tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Table 2. Specific reagents and equipment.</td>
		 </tr>
		</tbody>
 </table>

multi-analyte biochip (mab) based on all-solid-state ion-selective electrodes (assise) for physiological research

Lab-on-a-chip (LOC) applikationer i miljø-, biomedicinske, landbrugs-, biologiske og rumfart forskning kræver en ion-selektiv elektrode (ISE), der kan tåle længere tids opbevaring i komplekse biologiske medier 1-4. En all-solid-state ion-selektive elektroder (Assise) er særligt attraktivt for de førnævnte programmer. Elektroden skal have følgende gunstige karakteristika: let konstruktion, lav vedligeholdelse og (potentiale for) miniaturisering, der giver mulighed for batch-behandling. En mikrofabrikerede Assise beregnet til kvantificering H +, Ca 2 +, og CO 3 2 - ioner blev bygget. Den består af en ædel-metal elektrode lag (dvs. Pt), en transduktion lag, og en ion-selektiv membran (ISM) lag. Transduktion lag fungerer til at transducere koncentrations-afhængig kemisk potentiale ionselektive membran ind i en målbar elektrisk signal. Than levetid af en Assise er fundet at afhænge fastholde potentiale på det ledende lag / membran-grænsefladen 5-7. For at forlænge Assise arbejde levetid og dermed opretholde stabile potentialer ved grænsefladespændinger lag, vi udnyttet den ledende polymer (CP) poly (3,4-ethylenedioxythiophene) (PEDOT) 7-9 i stedet for sølv / sølvchlorid (Ag / AgCl) som transduceren lag. Vi konstruerede den Assise i en lab-on-a-chip-format, som vi kaldte multi-analyt biochip (MAB) (Figur 1). Kalibreringer i testopløsninger påvist, at MAB kan overvåge pH (driftsområdet pH 4-9), CO 3 2 - (målt på 0,01 mM - 1 mM), og Ca2 + (loglineær området 0,01 mM til 1 mM). MAb til pH giver en nær-Nernstian skråning respons efter næsten en måned opbevaring i algernes medium. Karbonatholdige biochips viser en potentiometrisk profil ligner en konventionel ionselektiv elektrode. Physiological målinger blev anvendt til at overvåge den biologiske aktivitet af modelsystem makroalgen Chlorella vulgaris. MAB formidler en fordel i størrelse, alsidighed og multiplekset Analytregistreringsindretning kapacitet, hvilket gør den anvendelig til mange begrænset overvågning situationer på Jorden eller i rummet. Biochip Design og eksperimentelle metoder Den biochip er 10 x 11 mm i dimension og har 9 Assises udpeget som arbejder elektroder (WES) og 5 Ag / AgCl reference-elektroder (RES). Hver arbejdsgruppe elektrode (WE) er 240 um i diameter og er jævnt fordelt på 1,4 mm fra RES, som er 480 um i diameter. Disse elektroder er forbundet til elektriske kontaktpuder med en dimension på 0,5 mm x 0,5 mm. Den skematiske er vist i figur 2.. Cyklisk voltammetri (CV) og galvanostatisk deposition metoder anvendes til at electropolymerize de PEDOT film ved hjælp af en Bioanalytical Systems Inc. (BASI) C3-celle fod (Figur 3). Modionen for PEDOT film er skræddersyet til den analyt-ion af interesse. En PEDOT med poly (styrensulfonat) modion (PEDOT / PSS) er udnyttet til H + og CO 3 2 -, mens en med sulfat (tilsat til opløsningen som CaSO4) er udnyttet til Ca2 +. De elektrokemiske egenskaber af PEDOT-belagt WE analyseres ved hjælp CV&#39;er i redox-aktiv opløsning (dvs. 2 mM kaliumferricyanid (K3Fe (CN) 6)). Baseret på CV-profil, blev Randles-Sevcik analyse bruges til at bestemme det effektive overfladeareal 10.. Spin-coating ved 1.500 rpm anvendes til at kaste ~ 2 um tyk ionselektive membraner (ISM) på MAB arbejdende elektroder (WES). MAB er indeholdt i en mikrofluid flow-celle kammer fyldt med en 150 gl volumen alge medium kontaktpuderne er elektrisk forbundet til BASI systemet (FigURE 4). Den fotosyntetiske aktivitet Chlorella vulgaris overvåges i omgivende lys og mørke forhold.

<html> Et eksempel på en cyklisk voltammogram (CV) resultatet af PEDOT: PSS og dets tilsvarende katodisk spidsstrøm (i p) vs scanningshastighed (v 1/2) er vist i figur 5a og 5b henholdsvis. PEDOT: CaSO4 ved forskellige scan satser og dens katodisk spidsstrøm er ikke vist. Brug Randles-Sevcik analyse 10, de effektive overfladearealer af den faste kontakt PEDOT: PSS og PEDOT: CaSO4 uden ion-selektiv membran blev fu...

Watch this Scientific Journal Video about Multi-analyte Biochip MAB Based on All-solid-state Ion-selective Electrodes ASSISE for Physiological Research at JoVE.com

Multi-analyte Biochip MAB Based on All-solid-state Ion-selective Electrodes ASSISE for Physiological Research

All-solid-state ion-selektive elektroder (Assises) konstrueret ud fra et ledende polymer (CP) transducer giver flere måneders funktionel levetid i flydende medier. Her beskriver vi fabrikation og kalibrering proces Assises i en lab-on-a-chip-format. Den Assise påvises at have opretholdt en nær-Nernstian hældning profil efter længere tids opbevaring i komplekse biologiske medier.

Multi-analyt Biochip (MAB) Baseret på All-solid-state Ion-selektive elektroder (Assise) for Fysiologisk Research

Lab-on-a-chip (LOC) applications in environmental, biomedical, agricultural, biological, and spaceflight research require an ion-selective electrode (ISE) ...

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Multi-analyte Biochip (MAB) Based on All-solid-state Ion-selective Electrodes (ASSISE) for Physiological Research

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