Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Enzyme-linked immunosorbent assay (ELISA) is a promising method to detect small amount of proteins in biological samples. The devices providing a platform ...

School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), UNIST-gil 50, Ulsan, Republic of Korea

We have developed new surface to ensure a proper spacing between immobilized biomolecules. While DNA microarray on this surface provided each probe DNA with ample space for hybridization with incoming target DNAs, the microarray showed enhanced discrimination efficiency for various types of single nucleotide polymorphism. The high discrimination efficiency holds for all tested cases (100:<1 for internal mismatched cases; 100:<28 for terminal mismatched ones). In addition, by investigating influence of hybridization temperature and washing condition on the fluorescence intensity and the discrimination efficiency with and without controlled mesospacing, it was observed that the nanoscale-controlled surface showed good discrimination efficiency in a wide range of temperature (37-50 degrees C), and hybridization behavior on the surface was in agreement with the solution one. Intriguingly, it was found that washing process after the hybridization was critical for the high discrimination efficiency. For the particular case, washing process was so efficient that only 30 s washing was sufficient to reach the optimal discrimination ratio.

DNA microarrays on nanoscale-controlled surface.

High throughput analysis of DNA in low concentration and small volume is an important issue and a continuing challenge in the field of DNA microarray and sensor. Recently, we have demonstrated that the DNA microarray on nano-scale controlled surface provides ample space for hybridization resulting in the best discrimination efficiency for SNP analysis. Here, we report the utility of the nano-scale controlled surface in conjunction with a multiply tagged protein. Application of streptavidin-fluorophore conjugates in combination with the highly controlled surface that suppresses non-specific binding of DNA allows highly sensitive detection of DNA while maintaining superior SNP discrimination efficiency comparable to our earlier results. The sensitivity of DNA microarray on the mesospaced surface is two orders of magnitude higher than that of the generic surface when a streptavidin-fluorophore conjugate was employed, and the detection limit on the former surface was found to be 50 fM of 15-mer target DNA. Various streptavidin-fluorophore conjugates including streptavidin-Cy3, streptavidin-Cy5, streptavidin-Alexa Flour 555 and streptavidin-phycoerythrin were examined.

Sensitivity enhancement of DNA microarray on nano-scale controlled surface by using a streptavidin-fluorophore conjugate.

Noncovalent functionalization of carbon nanotubes (CNTs) by dendrons was demonstrated. Certain types of dendrons successfully functionalized CNT surfaces through the noncovalent interactions between the peripheries of the dendrons and the sidewalls of CNTs. Dendrons have a unique anisotropic shape and an orthogonal functional group at their apex, and thus can generate a certain spacing between the functional groups upon immobilization on surfaces. Atomic force microscope (AFM) imaging, dispersion experiments, and MicroRaman spectroscopy were employed for the characterization of the functionalization. The binding was found to be governed by the chemical nature of the terminal groups, namely, the "fingertips", through a comparison study on the adsorption efficiency of the dendron analogs. Functional groups such as the carboxylic acid group and the benzyl amide group were effective for the cooperative binding. AFM analysis showed that the average spacing generated by the dendrons was 14-15 nm at a particular adsorption condition. Assembling streptavidin on the tubes through the dendrons and biotin confirmed the realization of the regulated spacing as well as the elimination of unwanted aggregation. The noncovalent functionalization of CNTs by a dendron can be a new approach toward sensible nanobiodevices, not only by introducing biomolecular probes on CNTs without disruption of the electronic network of the tubes, but also by providing the immobilized probe molecules with a space ample enough to minimize steric hindrance for the unhindered interaction with their target species.

"Fingertip"-guided noncovalent functionalization of carbon nanotubes by dendrons.

We describe a simple and versatile method for bonding thermoplastics to elastomeric polydimethylsiloxane (PDMS) at room temperature. The bonding of various thermoplastics including polycarbonate (PC), cyclic olefin copolymer (COC), polymethylmethacrylate (PMMA), and polystyrene (PS), to PDMS has been demonstrated at room temperature. An irreversible bonding was formed instantaneously when the thermoplastics, activated by oxygen plasma followed by aminopropyltriethoxysilane modification, were brought into contact with the plasma treated PDMS. The surface modified thermoplastics were characterized by water contact angle measurements and X-ray photoelectron spectroscopy. The tensile strength of the bonded hybrid devices fabricated with PC, COC, PMMA, and PS was found to be 430, 432, 385, and 388 kPa, respectively. The assembled devices showed high burst resistance at a maximum channel pressure achievable by an in-house built syringe pump, 528 kPa. Furthermore, they displayed very high hydrolytic stability; no significant change was observed even after the storage in water at 37 °C over a period of three weeks. In addition, this thermoplastic-to-PDMS bonding technique has been successfully employed to fabricate a relatively large sized device. For example, a lab-on-a-disc with a diameter of 12 cm showed no leakage when it spins for centrifugal fluidic pumping at a very high rotating speed of 6000 rpm.

Simple room temperature bonding of thermoplastics and poly(dimethylsiloxane).

&#x96C7;&#x7528;&#x7EB3;&#x7C73;&#x7ED3;&#x6784;&#x7684;&#x6D41;&#x63A7;&#x8BBE;&#x5907; &#xFF08;&lt; 100 &#x6BEB;&#x5FAE;&#x7C73;&#x5728;&#x4E00;&#x4E2A;&#x6216;&#x4E24;&#x4E2A;&#x7EF4;&#x5EA6;&#x3001; &#x7F1D;&#x6216;&#x7535;&#x89C6;&#x9891;&#x9053;&#xFF0C;&#x5206;&#x522B;&#xFF09; &#x6B63;&#x5728;&#x4EA7;&#x751F;&#x6781;&#x5927;&#x7684;&#x5174;&#x8DA3;&#xFF0C;&#x56E0;&#x4E3A;&#x6B64;&#x5927;&#x5C0F;&#x7684;&#x57DF;&#x76F8;&#x6BD4;&#xFF0C;&#x4ED6;&#x4EEC;&#x7684;&#x5FAE;&#x5C3A;&#x5EA6;&#x540C;&#x884C;&#x6240;&#x63D0;&#x4F9B;&#x7684;&#x72EC;&#x7279;&#x5C5E;&#x6027;&#x3002;&#x6709;&#x8DA3;&#x7684;&#x7EB3;&#x7C73;&#x5C3A;&#x5EA6;&#x73B0;&#x8C61;&#x7684;&#x4F8B;&#x5B50;&#x5305;&#x62EC; preconcentrate &#x79BB;&#x5B50;&#x7269;&#x79CD;&#x7531;&#x4E8E;&#x79BB;&#x5B50;&#x9009;&#x62E9;&#x6027;&#x8FC1;&#x79FB;&#xFF0C;&#x72EC;&#x7279;&#x7684;&#x5206;&#x5B50;&#x5206;&#x79BB;&#x65B9;&#x5F0F;&#xFF0C;&#x5BC6;&#x95ED;&#x73AF;&#x5883;&#x5141;&#x8BB8;&#x751F;&#x7269;&#x805A;&#x5408;&#x7269;&#x62C9;&#x4F38;&#x548C;&#x65AD;&#x88C2;&#x4F38;&#x957F;&#x7387;&#x548C;&#x4E0D;&#x53D7;&#x7EA6;&#x675F;&#x7684;&#x56FA;&#x76F8; bioreactions &#x5927;&#x89C4;&#x6A21;&#x8FD0;&#x8F93;&#x6587;&#x7269;&#x6781;&#x9AD8;&#x4E00;&#x7EA7;&#x7684;&#x80FD;&#x529B;&#x3002;&#x4E8B;&#x5B9E;&#x4E0A;&#xFF0C;&#x5728;&#x6587;&#x732E;&#x4E2D;&#x7684;&#x8BB8;&#x591A;&#x4F8B;&#x5B50;&#x8868;&#x660E;&#x8FD9;&#x4E9B;&#x72EC;&#x7279;&#x7684;&#x673A;&#x4F1A;&#xFF0C;&#x867D;&#x7136;&#x7EDD;&#x5927;&#x591A;&#x6570;&#x4F7F;&#x7528;&#x73BB;&#x7483;&#x7194;&#x5316;&#x7845;&#x4F5C;&#x4E3A;&#x886C;&#x5E95;&#x6750;&#x6599;&#x3002;&#x805A;&#x5408;&#x7269;&#x5FAE;&#x6D41;&#x4F53;&#x5DF2;&#x786E;&#x7ACB;&#x4F5C;&#x4E3A;&#x73BB;&#x7483;&#x3001; &#x77F3;&#x82F1;&#x3001; &#x6216;&#x57FA;&#x4E8E;&#x7845;&#x7684;&#x6D41;&#x63A7;&#x8BBE;&#x5907;&#x7684;&#x66FF;&#x4EE3;&#x65B9;&#x6CD5;&#x3002;&#x805A;&#x5408;&#x7269;&#x7684;&#x4F7F;&#x7528;&#x6240;&#x5F15;&#x8D77;&#x7684;&#x4E3B;&#x8981;&#x4F18;&#x52BF;&#x662F;&#x53EF;&#x4EE5;&#x7528;&#x4E8E;&#x751F;&#x4EA7;&#x6240;&#x9700;&#x7684;&#x7ED3;&#x6784;&#x3001; &#x7406;&#x5316;&#x7279;&#x6027;&#x4E0E;&#x4E0D;&#x540C;&#x7684;&#x9AD8;&#x5206;&#x5B50;&#x6750;&#x6599;&#xFF0C;&#x5E76;&#x53EF;&#x4EE5;&#x7528;&#x6765;&#x6539;&#x53D8;&#x7684;&#x57FA;&#x6750;&#x8868;&#x9762;&#x5316;&#x5B66;&#x7684;&#x7B80;&#x5355;&#x548C;&#x9C81;&#x68D2;&#x7684;&#x4FEE;&#x6539;&#x6218;&#x7565;&#x76F8;&#x5173;&#x7684;&#x5E7F;&#x6CDB;&#x7684;&#x591A;&#x6837;&#x52A0;&#x5DE5;&#x534F;&#x8BAE;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x867D;&#x7136;&#x76EE;&#x524D;&#x5B9E;&#x73B0;&#x805A;&#x5408;&#x7269;&#x5FAE;&#x6D41;&#x4F53;&#x7684;&#x4F18;&#x52BF;&#xFF0C;&#x57FA;&#x4E8E;&#x805A;&#x5408;&#x7269;&#x7684;&#x5948;&#x7C73;&#x6D41;&#x4F53;&#x5DE5;&#x5B66;&#x7684;&#x6F14;&#x53D8;&#x53EA;&#x6700;&#x8FD1;&#x62A5;&#x544A;&#x4E86;&#x3002;&#x5728;&#x8FD9;&#x5173;&#x952E;&#x7684;&#x5BA1;&#x67E5;&#x4E2D;&#xFF0C;&#x5C06;&#x8BA8;&#x8BBA;&#x4E86;&#x4F7F;&#x7528;&#x5F39;&#x6027;&#x548C;&#x70ED;&#x5851;&#x6027;&#x6750;&#x6599;&#x7684;&#x805A;&#x5408;&#x7269;&#x57FA;&#x5948;&#x7C73;&#x6D41;&#x4F53;&#x5DE5;&#x5B66;&#x6240;&#x63D0;&#x4F9B;&#x7684;&#x673A;&#x4F1A;&#x3002;&#x5C24;&#x5176;&#x662F;&#xFF0C;&#x4EE5;&#x53CA;&#x4ED6;&#x4EEC;&#x53EF;&#x4EE5;&#x5B9E;&#x73B0;&#x4E3A;&#x5F39;&#x6027;&#x4F53;&#x548C;&#x70ED;&#x5851;&#x6027;&#x6750;&#x6599;&#x7684;&#x7EB3;&#x7C73;&#x5927;&#x5C0F;&#x57DF;&#x5C06;&#x8BA8;&#x8BBA;&#x5236;&#x4F5C;&#x7684;&#x5404;&#x79CD;&#x65B9;&#x5F0F;&#x3002;&#x53EF;&#x4EE5;&#x7528;&#x4E8E;&#x5948;&#x7C73;&#x6D41;&#x4F53;&#x5DE5;&#x5B66;&#x7684;&#x4E0D;&#x540C;&#x805A;&#x5408;&#x7269;&#x57FA;&#x4F53;&#x5C06;&#x4ECB;&#x7ECD;&#x4EE5;&#x53CA;&#x4E0E;&#x65E0;&#x673A;&#x7EB3;&#x7C73;&#x5668;&#x4EF6;&#x7684;&#x6BD4;&#x8F83;&#x548C;&#x6280;&#x672F;&#x8BBE;&#x5907; &#xFF08;257 &#x5F15;&#x7528;&#xFF09; &#x7684;&#x64CD;&#x4F5C;&#x548C;&#x5236;&#x4F5C;&#x6750;&#x6599;&#x5DEE;&#x5F02;&#x7684;&#x540E;&#x679C;&#x3002;

&#x7075;&#x6D3B;&#x7684;&#x5236;&#x4F5C;&#x4E0E;&#x5E94;&#x7528;&#x805A;&#x5408;&#x7269;&#x6ED1;&#x79FB;&#x548C; nanoslits&#x3002;

Fluidic devices that employ nanoscale structures (<100 nm in one or two dimensions, slits or channels, respectively) are generating great interest due to the unique properties afforded by this size domain compared to their micro-scale counterparts. Examples of interesting nanoscale phenomena include the ability to preconcentrate ionic species at extremely high levels due to ion selective migration, unique molecular separation modalities, confined environments to allow biopolymer stretching and elongation and solid-phase bioreactions that are not constrained by mass transport artifacts. Indeed, many examples in the literature have demonstrated these unique opportunities, although predominately using glass, fused silica or silicon as the substrate material. Polymer microfluidics has established itself as an alternative to glass, fused silica, or silicon-based fluidic devices. The primary advantages arising from the use of polymers are the diverse fabrication protocols that can be used to produce the desired structures, the extensive array of physiochemical properties associated with different polymeric materials, and the simple and robust modification strategies that can be employed to alter the substrate's surface chemistry. However, while the strengths of polymer microfluidics is currently being realized, the evolution of polymer-based nanofluidics has only recently been reported. In this critical review, the opportunities afforded by polymer-based nanofluidics will be discussed using both elastomeric and thermoplastic materials. In particular, various fabrication modalities will be discussed along with the nanometre size domains that they can achieve for both elastomer and thermoplastic materials. Different polymer substrates that can be used for nanofluidics will be presented along with comparisons to inorganic nanodevices and the consequences of material differences on the fabrication and operation of nanofluidic devices (257 references).

Flexible fabrication and applications of polymer nanochannels and nanoslits.

Dispositifs fluidiques qui emploient des structures nanom&eacute;triques (&lt; 100 nm dans une ou deux dimensions, fentes ou canaux, respectivement) suscitent beaucoup d&#39;int&eacute;r&ecirc;t en raison des propri&eacute;t&eacute;s uniques offertes par ce domaine de taille par rapport &agrave; leurs homologues de micro-&eacute;chelle. Exemples de ph&eacute;nom&egrave;nes int&eacute;ressants de nanoscale incluent la capacit&eacute; de preconcentrate des esp&egrave;ces ioniques &agrave; des niveaux extr&ecirc;mement &eacute;lev&eacute;s en raison de l&#39;ion s&eacute;lective de la migration, les modalit&eacute;s de s&eacute;paration mol&eacute;culaire unique, confin&eacute; environnements pour permettre le biopolym&egrave;re &eacute;tirement et &eacute;longation et artefacts de transport de masse bioreactions de phase solide qui ne sont pas contraints par. En effet, beaucoup d&#39;exemples dans la litt&eacute;rature ont d&eacute;montr&eacute; ces occasions uniques, bien que principalement &agrave; l&#39;aide de verre, fondue silice ou silicium comme mat&eacute;riau de substrat. Microfluidique de polym&egrave;re s&#39;est impos&eacute; comme une alternative aux dispositifs fluidiques &agrave; base de silicium, en verre ou en silice fondue. Les principaux avantages d&eacute;coulant de l&#39;utilisation de polym&egrave;res sont les protocoles de fabrication diversifi&eacute;e qui peuvent &ecirc;tre utilis&eacute;s pour produire les structures d&eacute;sir&eacute;es, la vaste gamme de propri&eacute;t&eacute;s physico-chimiques associ&eacute;s &agrave; diff&eacute;rents mat&eacute;riaux polym&eacute;riques et les strat&eacute;gies de modification simple et robuste qui peuvent &ecirc;tre utilis&eacute;es pour modifier la chimie de surface du substrat. Cependant, alors que les points forts de la microfluidique de polym&egrave;re est actuellement r&eacute;alis&eacute;, l&#39;&eacute;volution des nanofluidics &agrave; base de polym&egrave;res que r&eacute;cemment a &eacute;t&eacute; signal&eacute;e. Dans cette revue critique, les possibilit&eacute;s offertes par nanofluidics &agrave; base de polym&egrave;res seront examin&eacute;es &agrave; l&#39;aide de mat&eacute;riaux les &eacute;lastom&egrave;res et thermoplastiques. En particulier, diverses modalit&eacute;s de fabrication seront discut&eacute;es avec les domaines de taille nanom&eacute;trique qui ils peuvent r&eacute;aliser pour les &eacute;lastom&egrave;res et thermoplastiques. Substrats diff&eacute;rents polym&egrave;re qui peuvent &ecirc;tre utilis&eacute;s pour nanofluidics seront pr&eacute;sent&eacute;s ainsi que des comparaisons avec des nanodispositifs inorganiques et les cons&eacute;quences des diff&eacute;rences mat&eacute;rielles sur la fabrication et le fonctionnement des dispositifs de nanofluidiques (257 r&eacute;f&eacute;rences).

Fabrication flexible et applications des polym&egrave;res NANOCANAUX et nanoslits.

Dispositivi idraulici che impiegano strutture su scala nanometrica (&lt; 100 nm in una o due dimensioni, fessure o canali, rispettivamente) stanno generando grande interesse a causa delle propriet&agrave; uniche di questo dominio di dimensioni rispetto ai loro omologhi di micro-scala. Esempi di fenomeni su scala nanometrica interessanti includono la possibilit&agrave; di preconcentrate specie ioniche a livelli estremamente elevati a causa dello ione selettivo migrazione, modalit&agrave; unica separazione molecolare, confinati gli ambienti per consentire biopolimero stretching e allungamento e fase solida bioreactions che non sono vincolati da artefatti di trasporto di massa. Infatti, molti esempi in letteratura hanno dimostrato queste opportunit&agrave; uniche, anche se prevalentemente utilizzando vetro, fuso silice o silicio come materiale di substrato. Microfluidics polimero si &egrave; affermata come alternativa al vetro, fused silica, dispositivi idraulici basati sul silicio. I vantaggi principali derivanti dall&#39;uso di polimeri sono i protocolli di fabbricazione diversi che possono essere utilizzati per produrre le strutture desiderate, la vasta gamma di propriet&agrave; fisiochimiche associate a diversi materiali polimerici e le strategie di modifica semplice e robusto che possono essere impiegate per alterare la chimica di superficie del substrato. Tuttavia, mentre i punti di forza del polimero microfluidica &egrave; attualmente essendo realizzato, l&#39;evoluzione della base di polimeri nanofluidica ha solo recentemente stato segnalato. In questa revisione critica, saranno discusse le opportunit&agrave; offerte da nanofluidica polimerici utilizzando materiali sia elastomerici e termoplastici. In particolare, diverse modalit&agrave; di fabbricazione sar&agrave; discusso insieme con i domini di dimensioni nanometro che possono ottenere sia per elastomeri e materiali termoplastici. Substrati differenti polimeri che possono essere utilizzati per la nanofluidica saranno presentati insieme a confronti di nanodispositivi inorganici e le conseguenze delle differenze di materiale sulla fabbricazione e il funzionamento di dispositivi nanofluidiche (257 riferimenti).

Fabbricazione flessibile e applicazioni del polimero nanochannels e nanofessure.

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&#x30A2;&#x30FC;&#x30C6;&#x30A3;&#x30D5;&#x30A1;&#x30AF;&#x30C8;&#x8CEA;&#x91CF;&#x9078;&#x629E;&#x30A4;&#x30AA;&#x30F3;&#x306B;&#x3088;&#x308B;&#x30A4;&#x30AA;&#x30F3;&#x7A2E;&#x3092; preconcentrate &#x3059;&#x308B;&#x6A5F;&#x80FD;&#x306A;&#x3069;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x4E3B;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x30AC;&#x30E9;&#x30B9;&#x304C;&#x3001;&#x78BA;&#x304B;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x30E6;&#x30CB;&#x30FC;&#x30AF;&#x306A;&#x6A5F;&#x4F1A;&#x3001;&#x6587;&#x5B66;&#x306E;&#x591A;&#x304F;&#x306E;&#x4F8B;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x308B;&#x77F3;&#x82F1;&#x307E;&#x305F;&#x306F;&#x30B7;&#x30EA;&#x30B3;&#x30F3;&#x57FA;&#x677F;&#x6750;&#x6599;&#x3068;&#x3057;&#x3066;&#x878D;&#x5408;&#x3002;&#x9AD8;&#x5206;&#x5B50;&#x30DE;&#x30A4;&#x30AF;&#x30ED; &#x30AC;&#x30E9;&#x30B9;&#x3001;&#x77F3;&#x82F1;&#x307E;&#x305F;&#x306F;&#x30B7;&#x30EA;&#x30B3;&#x30F3; &#x30D9;&#x30FC;&#x30B9;&#x6D41;&#x4F53;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306B;&#x4EE3;&#x308F;&#x308B;&#x3082;&#x306E;&#x3068;&#x3057;&#x3066;&#x5B9A;&#x7740;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x9AD8;&#x5206;&#x5B50;&#x6750;&#x6599;&#x306E;&#x4F7F;&#x7528;&#x306B;&#x8D77;&#x56E0;&#x3059;&#x308B;&#x4E3B;&#x306A;&#x5229;&#x70B9;&#x306F;&#x3001;&#x6240;&#x671B;&#x306E;&#x69CB;&#x9020;&#x306E;&#x7570;&#x306A;&#x308B;&#x9AD8;&#x5206;&#x5B50;&#x6750;&#x6599;&#x3068;&#x57FA;&#x677F;&#x306E;&#x754C;&#x9762;&#x5316;&#x5B66;&#x3092;&#x5909;&#x66F4;&#x3059;&#x308B;&#x306B;&#x306F;&#x63A1;&#x7528;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x30B7;&#x30F3;&#x30D7;&#x30EB;&#x304B;&#x3064;&#x5805;&#x7262;&#x306A;&#x5909;&#x66F4;&#x6226;&#x7565;&#x3092;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x7269;&#x7406;&#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x306E;&#x5E83;&#x7BC4;&#x306A;&#x914D;&#x5217;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x591A;&#x69D8;&#x306A;&#x88FD;&#x9020;&#x30D7;&#x30ED;&#x30C8;&#x30B3;&#x30EB;&#x3067;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;&#x9AD8;&#x5206;&#x5B50;&#x6D41;&#x4F53;&#x306E;&#x5F37;&#x307F;&#x306F;&#x73FE;&#x5728;&#x5B9F;&#x73FE;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x30DD;&#x30EA;&#x30DE;&#x30FC; &#x30D9;&#x30FC;&#x30B9;&#x6D41;&#x4F53;&#x7D20;&#x5B50;&#x306E;&#x9032;&#x5316;&#x306F;&#x3001;&#x6700;&#x8FD1;&#x5831;&#x544A;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x91CD;&#x8981;&#x306A;&#x30EC;&#x30D3;&#x30E5;&#x30FC;&#x3067;&#x306F;&#x3001;&#x30A8;&#x30E9;&#x30B9;&#x30C8;&#x30DE;&#x30FC;&#x3068;&#x71B1;&#x53EF;&#x5851;&#x6027;&#x6750;&#x6599;&#x3092;&#x7528;&#x3044;&#x305F;&#x30DD;&#x30EA;&#x30DE;&#x30FC; &#x30D9;&#x30FC;&#x30B9;&#x6D41;&#x4F53;&#x7D20;&#x5B50;&#x306B;&#x3088;&#x3063;&#x3066;&#x4E0E;&#x3048;&#x3089;&#x308C;&#x308B;&#x6A5F;&#x4F1A;&#x3092;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3059;&#x3002;&#x7279;&#x306B;&#x3001;&#x30A8;&#x30E9;&#x30B9;&#x30C8;&#x30DE;&#x30FC;&#x3068;&#x71B1;&#x53EF;&#x5851;&#x6027;&#x6A39;&#x8102;&#x6750;&#x6599;&#x306E;&#x4E21;&#x65B9;&#x3092;&#x9054;&#x6210;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x30CA;&#x30CE;&#x30E1;&#x30FC;&#x30C8;&#x30EB; &#x30B5;&#x30A4;&#x30BA; &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x3068;&#x5171;&#x306B;&#x69D8;&#x3005; &#x306A;&#x4F5C;&#x88FD;&#x65B9;&#x6CD5;&#x3092;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3059;&#x3002;&#x6D41;&#x4F53;&#x7D20;&#x5B50;&#x3092;&#x4F7F;&#x7528;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x7570;&#x306A;&#x308B;&#x30DD;&#x30EA;&#x30DE;&#x30FC;&#x57FA;&#x677F;&#x7121;&#x6A5F;&#x30CA;&#x30CE;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306E;&#x6BD4;&#x8F03;&#x3068;&#x4F5C;&#x88FD;&#x306B;&#x95A2;&#x3059;&#x308B;&#x6750;&#x6599;&#x306E;&#x9055;&#x3044;&#x306E;&#x7D50;&#x679C;&#x3068;&#x5316;&#x5B66; # &#x5206;&#x5B50;&#x30C7;&#x30D0;&#x30A4;&#x30B9; (257 &#x53C2;&#x7167;) &#x306E;&#x52D5;&#x4F5C;&#x304C;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;

&#x67D4;&#x8EDF;&#x306A;&#x52A0;&#x5DE5;&#x3068;&#x30DD;&#x30EA;&#x30DE;&#x30FC;&#x65BD;&#x3057;&#x305F;&#x3068; nanoslits &#x306E;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3002;

&#xB098;&#xB178; &#xAD6C;&#xC870;&#xB97C; &#xC0AC;&#xC6A9; &#xD558;&#xB294; &#xC720;&#xCCB4; &#xC7A5;&#xCE58; (&lt; 100 nm&#xC5D0;&#xC11C; &#xD558;&#xB098; &#xB610;&#xB294; &#xB450; &#xAC1C;&#xC758; &#xCC28;&#xC6D0;, &#xC2AC;&#xB9BF; &#xB610;&#xB294; &#xCC44;&#xB110; &#xAC01;&#xAC01;) &#xB9C8;&#xC774;&#xD06C;&#xB85C; &#xC2A4;&#xCF00;&#xC77C; &#xB4E4;&#xC5D0; &#xBE44;&#xD574;&#xC774; &#xD06C;&#xAE30; &#xB3C4;&#xBA54;&#xC778;&#xC5D0;&#xC11C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xB3C5;&#xD2B9;&#xD55C; &#xD2B9;&#xC131;&#xC73C;&#xB85C; &#xC778;&#xD574; &#xD070; &#xAD00;&#xC2EC;&#xC744; &#xC0DD;&#xC131; &#xD558;&#xB294;. &#xC7AC;&#xBBF8; &#xC788;&#xB294; &#xB098;&#xB178; &#xC2A4;&#xCF00;&#xC77C; &#xD604;&#xC0C1;&#xC758; &#xC608;&#xB85C; &#xC774;&#xC628; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;, &#xB3C5;&#xD2B9;&#xD55C; &#xBD84;&#xC790; &#xBD84;&#xB9AC; modalities &#xAD6D;&#xD55C; biopolymer &#xC2A4;&#xD2B8;&#xB808;&#xCE6D; &#xBC0F; &#xC2E0;&#xC7A5; &#xC218; &#xC788;&#xB3C4;&#xB85D; &#xD658;&#xACBD; &#xBC0F;&#xC5D0; &#xC758;&#xD574; &#xC81C;&#xD55C; &#xB418;&#xC9C0; &#xACE0;&#xCCB4; &#xC0C1; bioreactions &#xB300;&#xB7C9; &#xC804;&#xC1A1; &#xC544;&#xD2F0;&#xD329;&#xD2B8; &#xC120;&#xD0DD;&#xC73C;&#xB85C; &#xC778;&#xD574; &#xB9E4;&#xC6B0; &#xB192;&#xC740; &#xC218;&#xC900;&#xC5D0;&#xC11C; &#xC774;&#xC628; &#xC885; preconcentrate &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC2E4;&#xC81C;&#xB85C;, &#xBB38;&#xD559;&#xC5D0; &#xB9CE;&#xC740; &#xC608;&#xC81C; &#xC774;&#xB7EC;&#xD55C; &#xB3C5;&#xD2B9;&#xD55C; &#xAE30;&#xD68C;&#xB97C; &#xBCF4;&#xC5EC;&#xC900; &#xC720;&#xB9AC; &#xC8FC;&#xB85C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xBE44;&#xB85D; &#xC735;&#xD569; &#xC2E4;&#xB9AC; &#xCE74; &#xB610;&#xB294; &#xC2E4;&#xB9AC;&#xCF58; &#xAE30;&#xD310; &#xC18C;&#xC7AC;&#xB85C;. &#xD3F4;&#xB9AC;&#xBA38; microfluidics &#xC720;&#xB9AC;, &#xC6A9;&#xC735; &#xC2E4;&#xB9AC; &#xCE74; &#xB610;&#xB294; &#xC2E4;&#xB9AC;&#xCF58; &#xC720;&#xCCB4; &#xC7A5;&#xCE58; &#xB300;&#xC2E0; &#xC790;&#xCCB4;&#xB97C; &#xC124;&#xB9BD; &#xD588;&#xB2E4;. &#xACE0;&#xBD84;&#xC790;&#xC758; &#xC0AC;&#xC6A9;&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC8FC;&#xC694; &#xC7A5;&#xC810;&#xC740; &#xB2E4;&#xC591; &#xD55C; &#xC81C;&#xC791; &#xD504;&#xB85C;&#xD1A0;&#xCF5C; &#xC6D0;&#xD558;&#xB294; &#xAD6C;&#xC870;, &#xB2E4;&#xB978; &#xACE0;&#xBD84;&#xC790; &#xC7AC;&#xB8CC;&#xC640; &#xAE30;&#xD310;&#xC758; &#xD45C;&#xBA74; &#xD654;&#xD559;&#xC744; &#xBCC0;&#xACBD; &#xD558;&#xB824;&#xBA74; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xB294; &#xAC04;&#xB2E8; &#xD558; &#xACE0; &#xAC15;&#xB825;&#xD55C; &#xC218;&#xC815; &#xC804;&#xB7B5;&#xC5D0; &#xC5F0;&#xACB0; &#xD558;&#xB294; physiochemical &#xC18D;&#xC131;&#xC758; &#xAD11;&#xBC94;&#xC704; &#xD55C; &#xBC30;&#xC5F4;&#xC744; &#xC0DD;&#xC131; &#xD558;&#xB294; &#xB370; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xD3F4;&#xB9AC;&#xBA38; microfluidics &#xAC15;&#xC810;&#xC744; &#xD604;&#xC7AC; &#xC2E4;&#xD604; &#xB418; &#xACE0;, &#xD558;&#xB294; &#xB3D9;&#xC548; &#xD3F4;&#xB9AC;&#xBA38; &#xAE30;&#xBC18; nanofluidics&#xC758; &#xC9C4;&#xD654; &#xCD5C;&#xADFC;&#xC5D0; &#xBCF4;&#xACE0; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xC911;&#xC694; &#xD55C; &#xAC80;&#xD1A0;&#xC5D0;&#xC11C; &#xD3F4;&#xB9AC;&#xBA38; &#xAE30;&#xBC18; nanofluidics&#xC5D0;&#xC11C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xAE30;&#xD68C; &#xD0C4;&#xC131;&#xACFC; &#xC5F4;&#xAC00; &#xC18C;&#xC131; &#xC7AC;&#xB8CC;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xB17C;&#xC758; &#xB41C;&#xB2E4;. &#xD2B9;&#xD788;, &#xC81C;&#xC791; modalities &#xB2E4;&#xC591; &#xD55C; &#xADF8;&#xB4E4;&#xC740; &#xD0C4;&#xC131;&#xACFC; &#xC5F4;&#xAC00; &#xC18C;&#xC131; &#xC7AC;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xC5BB;&#xC744; &#xC218; &#xC788;&#xB294; &#xB098;&#xB178;&#xBBF8;&#xD130; &#xD06C;&#xAE30; &#xB3C4;&#xBA54;&#xC778; &#xD568;&#xAED8; &#xB17C;&#xC758; &#xD569;&#xB2C8;&#xB2E4;. Nanofluidics&#xC5D0; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xB294; &#xB2E4;&#xB978; &#xD3F4;&#xB9AC;&#xBA38; &#xAE30;&#xD310; &#xBB34;&#xAE30; &#xB098;&#xB178;&#xC5D0; &#xB300; &#xD55C; &#xBE44;&#xAD50; &#xBC0F; nanofluidic &#xC7A5;&#xCE58; (257 &#xCC38;&#xC870;)&#xC758; &#xC791;&#xC5C5; &#xADF8;&#xB9AC;&#xACE0; &#xC81C;&#xC870;&#xC5D0; &#xC18C;&#xC7AC; &#xCC28;&#xC774;&#xC758; &#xACB0;&#xACFC; &#xD568;&#xAED8; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4;.

&#xC720;&#xC5F0;&#xD55C; &#xC81C;&#xC791; &#xBC0F; &#xD3F4;&#xB9AC;&#xBA38; nanochannels&#xACFC; nanoslits&#xC758; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;.

Dispositivos flu&iacute;dicos que emplean estructuras de nanoescala (&lt; 100 nm en uno o dos dimensiones, ranuras o canales, respectivamente) est&aacute;n generando gran inter&eacute;s debido a las propiedades &uacute;nicas de este dominio de tama&ntilde;o en comparaci&oacute;n con sus contrapartes de micro escala. Ejemplos de fen&oacute;menos de nanoescala interesantes incluyen la capacidad de preconcentrate de especies i&oacute;nicas en niveles extremadamente altos por ion selectivo, migraci&oacute;n, modalidades de separaci&oacute;n molecular &uacute;nica, confinado entornos para permitir biopol&iacute;mero estiramiento y elongaci&oacute;n y bioreactions de la fase s&oacute;lida que no est&aacute;n limitados por artefactos de transporte en masa. De hecho, muchos ejemplos en la literatura han demostrado estas oportunidades &uacute;nicas, aunque predominantemente utilizando cristal, fundi&oacute; s&iacute;lice o silicio como el material del sustrato. Pol&iacute;mero microflu&iacute;dica se ha consolidado como una alternativa al vidrio, S&iacute;lice fundida o dispositivos flu&iacute;dicos basados en el silicio. Las principales ventajas derivadas de la utilizaci&oacute;n de pol&iacute;meros son los protocolos de fabricaci&oacute;n diversa que pueden ser utilizados para producir las estructuras deseadas, la amplia gama de propiedades fisicoqu&iacute;micas asociadas con diferentes materiales polim&eacute;ricos y las estrategias de modificaci&oacute;n sencilla y robusta que pueden emplearse para alterar la qu&iacute;mica superficial del sustrato. Sin embargo, mientras que actualmente, se est&aacute; llevando a cabo las fortalezas de la microflu&iacute;dica de pol&iacute;mero, la evoluci&oacute;n de nanoflu&iacute;dica basados en pol&iacute;meros ha recientemente divulgada. En esta revisi&oacute;n cr&iacute;tica, se discutir&aacute;n las oportunidades que ofrece nanoflu&iacute;dica con pol&iacute;meros materiales elast&oacute;meros y termopl&aacute;sticos. En particular, se discutir&aacute;n diversas modalidades de fabricaci&oacute;n junto con los dominios de tama&ntilde;o nan&oacute;metro que ellos puedan alcanzar para elast&oacute;meros y termopl&aacute;sticos. Sustratos de pol&iacute;meros diferentes que pueden utilizarse para nanoflu&iacute;dica se presentar&aacute; junto con comparaciones a nanodispositivos inorg&aacute;nicos y las consecuencias de las diferencias materiales en la fabricaci&oacute;n y operaci&oacute;n de dispositivos de editoriales (257 referencias).

Fabricaci&oacute;n flexible y aplicaciones de pol&iacute;meros nanocanales y nanoslits.

This paper presents a cost-effective, rapid, and fully automated lab-on-a-disc for simultaneous detection of multiple protein biomarkers in raw samples such as whole blood or whole saliva. For the diagnosis of cardiovascular disease, here, a novel centrifugal microfluidic layout was designed to conduct the simultaneous detection of high sensitivity C-reactive protein, cardiac troponin I, and N-terminal pro-B type natriuretic peptide based on a bead-based sandwich type enzyme-linked immunosorbent assay (ELISA). Three reaction chambers are initially interconnected for the common processes such as sample injection, incubation, and washing and then isolated on-demand for the independent processes such as substrate incubation and final detection. The assay performances such as the limit of detection and the dynamic range were comparable with those of the conventional ELISA despite the significant reduction of the minimum sample volume (200 Î¼L), the amount of washing buffer (700 Î¼L), and the total process time (20 min).

Lab-on-a-Disc for Fully Integrated Multiplex Immunoassays.

&#x672C;&#x6587;&#x63D0;&#x51FA;&#x4E86;&#x539F;&#x59CB;&#x6837;&#x672C;&#x5982;&#x5168;&#x8840;&#x6216;&#x553E;&#x6DB2;&#x4E2D;&#x7684;&#x7ECF;&#x6D4E;&#x9AD8;&#x6548;&#x3001; &#x5FEB;&#x901F;&#x548C;&#x5B8C;&#x5168;&#x81EA;&#x52A8;&#x5316;&#x5B9E;&#x9A8C;&#x5BA4;--&#x4E00;&#x76D8;&#x540C;&#x65F6;&#x68C0;&#x6D4B;&#x591A;&#x4E2A;&#x86CB;&#x767D;&#x8D28;&#x751F;&#x7269;&#x6807;&#x5FD7;&#x7269;&#x3002;&#x5FC3;&#x8840;&#x7BA1;&#x75BE;&#x75C5;&#x7684;&#x8BCA;&#x65AD;&#xFF0C;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x4E00;&#x79CD;&#x65B0;&#x578B;&#x79BB;&#x5FC3;&#x5F0F;&#x5FAE;&#x6D41;&#x63A7;&#x5E03;&#x5C40;&#x4E3A;&#x4E86;&#x8FDB;&#x884C;&#x540C;&#x65F6;&#x68C0;&#x6D4B;&#x7684;&#x9AD8;&#x7075;&#x654F;&#x5EA6; C-&#x53CD;&#x5E94;&#x86CB;&#x767D;&#x3001; &#x5FC3;&#x808C;&#x808C;&#x9499;&#x86CB;&#x767D;&#xFF0C;&#x4E0E; N &#x7EC8;&#x7AEF; pro-B &#x578B;&#x94A0;&#x5C3F;&#x80BD;&#x57FA;&#x4E8E;&#x57FA;&#x4E8E;&#x73E0;&#x7684;&#x4E09;&#x660E;&#x6CBB;&#x7C7B;&#x578B;&#x9176;&#x8054;&#x9176;&#x8054;&#x514D;&#x75AB;&#x5438;&#x9644;&#x6CD5; (ELISA)&#x3002;&#x4E09;&#x4E2A;&#x53CD;&#x5E94;&#x5BA4;&#x8FDB;&#x6837;&#x3001; &#x5B75;&#x5316;&#x548C;&#x6D17;&#x6DA4;&#x7B49;&#x5E38;&#x89C1;&#x8FDB;&#x7A0B;&#x6700;&#x521D;&#x76F8;&#x4E92;&#x8FDE;&#x63A5;&#xFF0C;&#x7136;&#x540E;&#x9694;&#x79BB;&#x70B9;&#x64AD;&#x57FA;&#x677F;&#x5B75;&#x5316;&#x548C;&#x6700;&#x540E;&#x68C0;&#x6D4B;&#x7B49;&#x7684;&#x72EC;&#x7ACB;&#x8FC7;&#x7A0B;&#x3002;&#x5AB2;&#x7F8E;&#x5C3D;&#x7BA1;&#x5927;&#x5E45;&#x51CF;&#x5C11;&#x7684;&#x6700;&#x5C0F;&#x6837;&#x672C;&#x91CF; &#xFF08;200 &mu;L)&#x3001; &#x6D17;&#x6DA4;&#x7F13;&#x51B2;&#x533A; &#xFF08;700 &mu;L&#xFF09;&#xFF0C;&#x91D1;&#x989D;&#x548C;&#x8FC7;&#x7A0B;&#x7684;&#x603B;&#x65F6;&#x95F4; (20 &#x5206;&#x949F;) &#x7684;&#x5E38;&#x89C4;&#x9176;&#x8054;&#x514D;&#x75AB;&#x5438;&#x9644;&#x6D4B;&#x5B9A;&#x8868;&#x6F14;&#x7B49;&#x7684;&#x68C0;&#x6D4B;&#x548C;&#x52A8;&#x6001;&#x8303;&#x56F4;&#x7684;&#x9650;&#x5236;&#x3002;

&#x5B9E;&#x9A8C;&#x5BA4;--&#x4E00;&#x76D8;&#x7684;&#x5B8C;&#x5168;&#x96C6;&#x6210;&#x7684;&#x591A;&#x8DEF;&#x590D;&#x7528;&#x514D;&#x75AB;&#x3002;

Cet article pr&eacute;sente un rentable, rapid et enti&egrave;rement automatis&eacute; lab-sur-un-disque pour la d&eacute;tection simultan&eacute;e de diff&eacute;rents biomarqueurs de prot&eacute;ines dans des &eacute;chantillons bruts tels que le sang ou la salive enti&egrave;re. Pour le diagnostic des maladies cardiovasculaires, ici, une mise en page du roman microfluidique centrifuge a &eacute;t&eacute; con&ccedil;u pour effectuer la d&eacute;tection simultan&eacute;e de haute sensibilit&eacute; de prot&eacute;ine C - r&eacute;active, troponine cardiaque I et N-terminale pro-B tapez peptide natriur&eacute;tique based on un &quot;sandwich&quot; ax&eacute;e sur la perle type enzyme-linked immunosorbent assay (ELISA). Trois chambres de r&eacute;action sont initialement interconnect&eacute;s pour des proc&eacute;d&eacute;s tels que l&#39;injection de l&#39;&eacute;chantillon, l&#39;incubation et laver et ensuite isolement &agrave; la demande des processus ind&eacute;pendants tels que l&#39;incubation du substrat et d&eacute;tection finale. Les performances du test comme la limite de d&eacute;tection et de la plage dynamique ont &eacute;t&eacute; comparables &agrave; celles du test ELISA classique malgr&eacute; la r&eacute;duction importante du volume minimale de l&#39;&eacute;chantillon (200 &mu;L), la quantit&eacute; de m&eacute;moire tampon de lavage (700 &mu;L) et le temps d&#39;ex&eacute;cution total (20 min).

Lab-sur-un-disque pour les immunoessais multiplexes enti&egrave;rement int&eacute;gr&eacute;s.

Dieser Beitrag stellt eine kosteng&uuml;nstige, schnelle und vollautomatische Lab-am-a-Disc f&uuml;r den gleichzeitigen Nachweis von mehreren Protein-Biomarkern in rohe Proben wie Vollblut oder ganze Speichel. F&uuml;r die Diagnose von Herz-Kreislauf-Erkrankungen hier eine neuartige zentrifugaler Microfluidic-Layout wurde entwickelt, um den gleichzeitigen Nachweis von hochempfindlichen C - reaktives Protein f&uuml;hren, Herztroponin I und N-Terminal Pro-B-Typ natriuretisches Peptid, basierend auf einer Bead-basierter Sandwich Typ Enzyme-linked Immunosorbent Assay (ELISA). Drei Reaktion Kammern sind f&uuml;r die gemeinsame Prozesse wie Beispiel Injektion, Inkubation und waschen zun&auml;chst miteinander verbunden und dann isoliert auf Abruf f&uuml;r die unabh&auml;ngige Prozesse wie Substrat Inkubation und endg&uuml;ltige Erkennung. Der Assay-Performances wie die Begrenzung der Aufdeckung und der dynamische Bereich waren vergleichbar mit denen des konventionellen ELISA trotz der erheblichen Reduktion der minimale Probenvolumen (200 &mu;L), die H&ouml;he der Wasch-Puffer (700 &mu;L) und die Prozess-Gesamtzeit (20 min.).

Lab-am-a-Disc f&uuml;r voll integrierte Multiplex-Immunoassays.

Questo libro presenta un conveniente, rapido e completamente automatizzato lab-su-un disco per la rilevazione simultanea di biomarcatori multipli di proteine in campioni di raw come sangue intero o saliva intera. Per la diagnosi di malattie cardiovascolari, qui, un romanzo microfluidica centrifughe layout &egrave; stato progettato per condurre la rilevazione simultanea di alta sensibilit&agrave; proteina C - reattiva, troponina cardiaca I e pro-B N-terminale tipo peptide natriuretico basato su un panino basato su tallone tipo enzyme-linked immunosorbent assay (ELISA). Tre camere di reazione sono inizialmente interconnessione per i comuni processi come iniezione del campione, incubazione e lavaggio e quindi isolato su richiesta per i processi indipendenti come incubazione del substrato e la rivelazione finale. Le prestazioni di analisi come il limite di rilevamento e la gamma dinamica erano comparabili con quelli del Elisa convenzionale nonostante la significativa riduzione del volume campione minimo (200 &mu;L), la quantit&agrave; di tampone di lavaggio (700 &micro; l) e il tempo di processo totale (20 min).

Laboratorio-su-un-disco per immunodosaggi Multiplex completamente integrati.

Raw &#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x5168;&#x4F53;&#x306E;&#x8840;&#x6DB2;&#x3084;&#x553E;&#x6DB2;&#x306A;&#x3069;&#x306B;&#x30B3;&#x30B9;&#x30C8;&#x52B9;&#x7387;&#x306E;&#x6025;&#x901F;&#x306A;&#x3068;&#x5B8C;&#x5168;&#x306B;&#x81EA;&#x52D5;&#x5316;&#x3055;&#x308C;&#x305F;&#x30E9;&#x30DC;-&#x306B;-&#x306F;-&#x30C7;&#x30A3;&#x30B9;&#x30AF;&#x306E;&#x8907;&#x6570;&#x306E;&#x86CB;&#x767D;&#x8CEA;&#x30D0;&#x30A4;&#x30AA; &#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x306E;&#x540C;&#x6642;&#x691C;&#x51FA;&#x306B;&#x3064;&#x3044;&#x3066;&#x8FF0;&#x3079;&#x308B;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x5FC3;&#x8840;&#x7BA1;&#x75BE;&#x60A3;&#x306E;&#x8A3A;&#x65AD;, &#x65B0;&#x898F;&#x9060;&#x5FC3;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30EC;&#x30A4;&#x30A2;&#x30A6;&#x30C8;&#x9AD8;&#x611F;&#x5EA6; C &#x53CD;&#x5FDC;&#x6027;&#x86CB;&#x767D;&#x306E;&#x540C;&#x6642;&#x691C;&#x51FA;&#x3092;&#x884C;&#x3046;&#x305F;&#x3081;&#x306B;&#x8A2D;&#x8A08;&#x3055;&#x308C;&#x305F;&#x3001;&#x5FC3;&#x7B4B;&#x30C8;&#x30ED;&#x30DD;&#x30CB;&#x30F3; I&#x3001;&#x304A;&#x3088;&#x3073; N &#x672B;&#x7AEF;&#x30D7;&#x30ED; B &#x578B;&#x30CA;&#x30C8;&#x30EA;&#x30A6;&#x30E0;&#x5229;&#x5C3F;&#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x3001;&#x30D3;&#x30FC;&#x30C9; &#x30D9;&#x30FC;&#x30B9; &#x30B5;&#x30F3;&#x30C9;&#x30A4;&#x30C3;&#x30C1;&#x578B; elisa &#x30A2;&#x30C3;&#x30BB;&#x30A4; (elisa &#x6CD5;) &#x306B;&#x57FA;&#x3065;&#x304D;&#x307E;&#x3059;&#x3002;3 &#x3064;&#x306E;&#x53CD;&#x5FDC;&#x30C1;&#x30E3;&#x30F3;&#x30D0;&#x30FC;&#x306F;&#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x6CE8;&#x5165;&#x3001;&#x30A4;&#x30F3;&#x30AD;&#x30E5;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3001;&#x6D17;&#x6FEF;&#x306A;&#x3069;&#x306E;&#x4E00;&#x822C;&#x7684;&#x306A;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#x5F53;&#x521D;&#x76F8;&#x4E92;&#x63A5;&#x7D9A;&#x3055;&#x308C;&#x3001;&#x57FA;&#x677F;&#x30A4;&#x30F3;&#x30AD;&#x30E5;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3068;&#x6700;&#x7D42;&#x7684;&#x306A;&#x691C;&#x51FA;&#x306A;&#x3069;&#x306E;&#x72EC;&#x7ACB;&#x3057;&#x305F;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#x30AA;&#x30F3; &#x30C7;&#x30DE;&#x30F3;&#x30C9;&#x3092;&#x5206;&#x96E2;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x6027;&#x80FD;&#x306E;&#x691C;&#x51FA;&#x3068;&#x30C0;&#x30A4;&#x30CA;&#x30DF;&#x30C3;&#x30AF; &#x30EC;&#x30F3;&#x30B8;&#x306E;&#x5236;&#x9650;&#x306A;&#x3069;&#x306F;&#x6700;&#x5C0F;&#x30B5;&#x30F3;&#x30D7;&#x30EB; &#x30DC;&#x30EA;&#x30E5;&#x30FC;&#x30E0; &#xFF08;200 &mu;&#xFF09;&#x3001;&#x6D17;&#x6D44;&#x30D0;&#x30C3;&#x30D5;&#x30A1;&#x30FC; (700 &mu; L) &#x306E;&#x91CF;&#x3001;&#x5408;&#x8A08;&#x51E6;&#x7406;&#x6642;&#x9593; (20 &#x5206;) &#x306E;&#x5927;&#x5E45;&#x306A;&#x524A;&#x6E1B;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x5F93;&#x6765;&#x306E; ELISA &#x306E;&#x305D;&#x308C;&#x3089;&#x3068;&#x540C;&#x7B49;&#x3060;&#x3063;&#x305F;&#x3002;

&#x30E9;&#x30DC;-&#x306B;-&#x306F;-&#x30C7;&#x30A3;&#x30B9;&#x30AF;&#x3092;&#x5B8C;&#x5168;&#x306B;&#x7D71;&#x5408;&#x3055;&#x308C;&#x305F;&#x591A;&#x91CD;&#x30A4;&#x30E0;&#x30CE;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x3002;

&#xC774; &#xC2E0;&#xBB38;&#xC740; &#xC804;&#xCCB4; &#xD608;&#xC561; &#xB610;&#xB294; &#xC804;&#xCCB4; &#xD0C0; &#xC561; &#xAC19;&#xC740; &#xC6D0;&#xC2DC; &#xC0D8;&#xD50C; &#xBE44;&#xC6A9; &#xD6A8;&#xC728;&#xC801;&#xC778; &#xBE60;&#xB974;&#xACE0; &#xC644;&#xBCBD; &#xD558; &#xAC8C; &#xC790;&#xB3D9;&#xD654; &#xB41C; &#xC5F0;&#xAD6C;&#xC18C;-&#xC5D0;-&#xD55C;-&#xB514;&#xC2A4;&#xD06C; &#xC5EC;&#xB7EC; &#xB2E8;&#xBC31;&#xC9C8; &#xBC14;&#xC774;&#xC624; &#xB9C8;&#xCEE4;&#xC758; &#xB3D9;&#xC2DC; &#xAC80;&#xCD9C;&#xC744; &#xC704;&#xD55C; &#xC120;&#xBB3C; &#xD55C;&#xB2E4;. &#xC2EC;&#xD608; &#xAD00; &#xC9C8;&#xD658;&#xC758; &#xC9C4;&#xB2E8;&#xC5D0; &#xB300; &#xD55C; &#xC5EC;&#xAE30;, &#xC18C;&#xC124; &#xC6D0;&#xC2EC; &#xBBF8;&#xC138; &#xB808;&#xC774;&#xC544;&#xC6C3; &#xACE0;&#xAC10;&#xB3C4; C-&#xBC18;&#xC751;&#xC131; &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xB3D9;&#xC2DC; &#xAC80;&#xCD9C;&#xC744; &#xC218;&#xD589; &#xD558;&#xB3C4;&#xB85D; &#xC124;&#xACC4; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;, &#xADF8;&#xB9AC;&#xACE0; &#xC2EC;&#xC7A5; troponin I &#xBC0F; N-&#xD130;&#xBBF8;&#xB110; &#xD504;&#xB85C; B &#xC785;&#xB825; natriuretic &#xD3A9; &#xD2F0; &#xB4DC; &#xBE44;&#xB4DC;&#xC5D0; &#xAE30;&#xCD08;&#xB97C; &#xB454; &#xC0CC;&#xB4DC;&#xC704;&#xCE58; &#xC885;&#xB958; &#xD6A8;&#xC18C; &#xC5F0;&#xACB0; &#xB41C; immunosorbent &#xC2DC;&#xD5D8; (ELISA)&#xC5D0; &#xB530;&#xB77C;. 3 &#xAC1C;&#xC758; &#xBC18;&#xC751; &#xCC54;&#xBC84; &#xCC98;&#xC74C; &#xC0D8;&#xD50C; &#xC8FC;&#xC785;, &#xBD80; &#xD654;, &#xC138;&#xCC99; &#xB4F1; &#xC77C;&#xBC18;&#xC801;&#xC778; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC5D0; &#xB300; &#xD55C; &#xC0C1;&#xD638; &#xC5F0;&#xACB0; &#xB41C; &#xC788;&#xC73C;&#xBA70; &#xB2E4;&#xC74C; &#xC808;&#xC5F0; &#xAE30;&#xD310; &#xC778;&#xD050;&#xBCA0;&#xC774;&#xC158; &#xCD5C;&#xC885; &#xAC80;&#xC0C9; &#xB4F1; &#xB3C5;&#xB9BD; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC5D0; &#xB300; &#xD55C; &#xC218;&#xC694;. &#xAC80;&#xC0C9; &#xBC0F; &#xB3D9;&#xC801; &#xBC94;&#xC704; &#xC81C;&#xD55C; &#xB4F1; &#xBD84;&#xC11D; &#xACB0;&#xACFC; &#xACF5;&#xC5F0; &#xCD5C;&#xC18C; &#xC0D8;&#xD50C; &#xBCFC;&#xB968; (200 &mu; L), &#xC138;&#xCC99; &#xBC84;&#xD37C; (700 &mu; L), &#xAE08;&#xC561; &#xBC0F; &#xCD1D; &#xD504;&#xB85C;&#xC138;&#xC2A4; &#xC2DC;&#xAC04; (20 &#xBD84;)&#xC758; &#xB300;&#xD3ED; &#xAC10;&#xC18C;&#xC5D0;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0; &#xAE30;&#xC874;&#xC758; ELISA&#xC640; &#xBE44;&#xAD50; &#xD588;&#xB2E4;.

&#xC2E4;&#xD5D8;&#xC2E4;-&#xC5D0;-&#xD55C;-&#xB514;&#xC2A4;&#xD06C;&#xB97C; &#xC644;&#xBCBD; &#xD558; &#xAC8C; &#xD1B5;&#xD569; &#xB41C; &#xBA40;&#xD2F0;&#xD50C;&#xB809;&#xC2A4; Immunoassays.

Este papel presenta un rentable, r&aacute;pido y totalmente automatizado laboratorio-en-un-disco para la detecci&oacute;n simult&aacute;nea de m&uacute;ltiples biomarcadores de prote&iacute;nas en muestras crudas como sangre o saliva todo. Para el diagn&oacute;stico de las enfermedades cardiovasculares, aqu&iacute;, un dise&ntilde;o de novela microflu&iacute;dicos centr&iacute;fugo fue dise&ntilde;ado para llevar a cabo la detecci&oacute;n simult&aacute;nea de prote&iacute;na C reactiva de alta sensibilidad, troponina I y N-terminal pro-B tipo p&eacute;ptido natriur&eacute;tico basado en un emparedado basadas en grano tipo enzima-linked immunosorbent assay (ELISA). Tres c&aacute;maras de reacci&oacute;n son inicialmente interconectados para los procesos comunes como la inyecci&oacute;n de la muestra, incubaci&oacute;n y lavado y entonces aislaron on-demand para los procesos independientes como sustrato incubaci&oacute;n y detecci&oacute;n final. Las actuaciones de ensayo como el l&iacute;mite de detecci&oacute;n y el rango din&aacute;mico eran comparables con los del ELISA convencional a pesar de la significativa reducci&oacute;n de la cantidad de tamp&oacute;n de lavado (700 &mu;L), el volumen m&iacute;nimo de la muestra (200 &mu;L) y el tiempo total de proceso (20 min).

Laboratorio-en-un-disco para inmunoensayos m&uacute;ltiplexes completamente integrados.

A possible mechanism for the aminosilane-mediated room-temperature (RT) bonding of thermoplastics and poly(dimethylsiloxane) (PDMS) is presented. The plasma-activated thermoplastic or PDMS substrates were modified with alkoxy silanes having different organo functional groups, and their bonding characteristics were studied. Manual peeling tests revealed that strong bonding was realized only when the silane had a free amino group and at least two alkoxy groups on the silicon. Silanization was carried out in both aqueous and anhydrous conditions; bonding occurred readily at RT in the former case, but a longer incubation time or a higher temperature was needed for the latter. The presence of the silane on the surface was confirmed by contact-angle measurements and UV spectrophotometric, attenuated total reflectance infrared spectroscopic (ATR-IR) and X-ray photoelectron spectroscopic (XPS) analyses. In the case where the aminosilane was deposited from aqueous solution, the amino functionality of the silane-catalyzed siloxane bond formation between the silanol on the modified thermoplastic surface and the silanol of the plasma-activated PDMS. In the case of anhydrous phase deposition, the aminosilane first catalyzed the hydrolysis of the ethoxy groups on the silicon, and then, catalyzed the condensation between the silanol groups of both materials. Shelf life tests of the modified thermoplastics showed that the aminosilane was stable over 2 weeks, and that bonding occurred at RT when the substrates were soaked in water before bonding.

Investigation on the mechanism of aminosilane-mediated bonding of thermoplastics and poly(dimethylsiloxane).

We present a novel fully integrated centrifugal microfluidic device with features for target antigen capture from biological samples, via a bead-based enzyme-linked immune-sorbent assay, and flow-enhanced electrochemical detection. The limit of detection (LOD) of our device for the C-reactive protein (CRP) was determined to be 4.9 pg mL(-1), a 17-fold improvement over quantification by optical density. The complete sample-to-answer protocol of our device is fully automated and takes less than 20 min. Overall, the presented microfluidic disc adds to the comparatively small number of fully integrated microfluidic-based platforms that utilize electrochemical detection and exemplifies how electrochemical detection can be enhanced by flow to successfully detect very low levels of biomarkers (e.g. pg mL(-1)).

Flow-enhanced electrochemical immunosensors on centrifugal microfluidic platforms.

The chemistry and the structure of aminosilane layer on the plasma activated thermoplastic substrates, e.g., polycarbonate (PC), polystyrene (PS), poly(methyl methacrylate) (PMMA), and cyclic olefin co-polymer (COC) were investigated at the molecular level. The nature of the surface functional groups of the silane layers prepared by solution phase deposition in aqueous and anhydrous solvents were studied using various techniques including ellipsometry, goniometry, atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and attenuated total reflectance infrared spectroscopy (ATR-IR). The XPS analyses revealed the presence of various oxygen functionalities on the plasma activated thermoplastics. Considerable differences were observed for the structure of aminosilane depending on the solvent used for the reaction. Deposition from aqueous solution resulted in relatively flat and smooth surfaces with consistent thickness compared to the anhydrous solution deposition. In the former case, 33% of the total nitrogen accounted for protonated amine and 16% for the free amino groups. In the latter, only 6% accounted for the protonated amine. The point of zero charge (pzc), on the aminosilane modified PC was found to be around 7, indicated that the surface is positively charged below pH 7 and negatively charged above pH 7. The surface analysis data suggested that various interactions are possible between the plasma activated thermoplastic surface and the aminosilane. In general, they are bound to the surface through covalent bond formation between the oxygen functionalities on the thermoplastic surface and the amino or the silanol groups of the aminosilane.

Aminosilane layers on the plasma activated thermoplastics: influence of solvent on its structure and morphology.

Circulating tumor cells (CTCs) have gained increasing attention owing to their roles in cancer recurrence and progression. Due to the rarity of CTCs in the bloodstream, an enrichment process is essential for effective target cell characterization. However, in a typical pressure-driven microfluidic system, the enrichment process generally requires complicated equipment and long processing times. Furthermore, the commonly used immunoaffinity-based positive selection method is limited, as its recovery rate relies on EpCAM expression of target CTCs, which shows heterogeneity among cell types. Here, we propose a centrifugal-force-based size-selective CTC isolation platform that can isolate and enumerate CTCs from whole blood within 30 s with high purity. The device was validated using the MCF-7 breast cancer cell line spiked in phosphate-buffered saline and whole blood, and an average capture efficiency of 61% was achieved, which is typical for size-based filtration. The capture efficiency for whole blood samples varied from 44% to 84% under various flow conditions and dilution factors. Under the optimized operating conditions, a few hundred white blood cells per 1 mL of whole blood were captured, representing a 20-fold decrease compared to those obtained using a commercialized size-based CTC isolation device. In clinical validation, normalized CTC counts varied from 10 to 60 per 7.5 mL of blood from gastric and lung cancer patients, yielding a detection rate of 50% and 38%, respectively. Overall, our CTC isolation device enables rapid and label-free isolation of CTCs with high purity, which should greatly improve downstream molecular analyses of captured CTCs.

All-in-one centrifugal microfluidic device for size-selective circulating tumor cell isolation with high purity.

ELISA-based devices are promising tools for the detection of low abundant proteins in biological samples. Reductions of the sample volume and assay time as well as full automation are required for their potential use in point-of-care diagnostic applications. Here, we present a highly efficient lab-on-a-disc composed of a TiO2 nanofibrous mat for sensitive detection of serum proteins with a broad dynamic range, with only 10 μL of whole blood within 30 min. The TiO2 nanofibers provide high specific surface area as well as active functional groups to capture large amounts of antibodies on the surface. In addition, the device offers efficient mixing and washing for improving the signal-to-noise ratio, thus enhancing the overall detection sensitivity. We employ the device for the detection of cardiac biomarkers, C-reactive protein (CRP) and cardiac troponin I (cTnI), spiked in phosphate-buffered saline (PBS) as well as in serum or whole blood. The device exhibited a wide dynamic range of six orders of magnitude from 1 pg mL(-1) (~8 fM) to 100 ng mL(-1) (~0.8 pM) and a low detection limit of 0.8 pg mL(-1) (~6 fM) for CRP spiked in CRP-free serum and a dynamic range of 10 pg mL(-1) (~0.4 pM) to 100 ng mL(-1) (~4 nM) with a detection limit of 37 pg mL(-1) (~1.5 pM) for cTnI spiked in whole blood.

Electrospun TiO2 nanofiber integrated lab-on-a-disc for ultrasensitive protein detection from whole blood.

Extracellular vesicles (EVs) are cell-derived nanovesicles, present in almost all types of body fluids, which play an important role in intercellular communication and are involved in the transport of biological signals for regulating diverse cellular functions. Due to the increasing clinical interest in the role of EVs in tumor promotion, various techniques for their isolation, detection, and characterization are being developed. In this review, we present an overview of the current EV isolation and characterization methods in addition to their applications and limitations. Furthermore, EVs as the potential emerging biomarkers in cancer management and their clinical implementation are briefly discussed.

Vijaya Sunkara

Department of Biomedical Engineering

School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), UNIST-gil 50, Ulsan, Republic of Korea

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Vijaya Sunkara

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biomedical Engineering

School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), UNIST-gil 50, Ulsan, Republic of Korea

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Department of Biomedical Engineering