ligand-nano-cluster-arrays-in-a-supported-lipid-bilayer

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Currently there is considerable interest in creating ordered arrays of adhesive protein islands in a sea of passivated surface for cell biological ...

CNRS, UMR 7325, CINaM

Surface chemistry of topographically patterned grooved samples with ridges of 150 nm width, adsorbed with self-assembled monolayers (SAMs) of alkanethiols on gold, have been characterized by near edge X-ray absorption fine structure (NEXAFS) spectroscopy. Analysis reveals that NEXAFS may discriminate between different chemistries adsorbed to the tops, sidewalls and grooves of the patterns.

A new method to characterize chemically and topographically nanopatterned surfaces.

A label-free two-dimensional colorimetric DNA sensor is reported. This sensor is based on the 2D aggregation of oligonucleotide-modified gold nanoparticle probes induced by the molecular hybridization of single-stranded oligonucleotide probes and their complementary single-stranded DNA targets. To detect the aggregation, we have developed a new detection method based on the selective desorption of nonaggregated nanoparticles. We will show here that this detection method is highly specific and allows the quantification of the DNA targets.

DNA detection method based on the two-dimensional aggregation and selective desorption of nanoparticle probes.

A 2D colorimetric DNA sensor is reported based on the 2D aggregation of oligonucleotide-modified gold nanoparticle probes resulting from the molecular hybridization between these latest and their complementary single stranded DNA targets. To increase their mobility the nanoparticles are adsorbed on a fluid lipid bilayer, itself supported on a substrate. The hybridization between the target and the mobile nanoparticle probes creates links between the nanoparticles resulting in the formation of nanoparticle aggregates in the plane of the substrate. This aggregation is detected using a new method based on the selective desorption of non-aggregated nanoparticles. The addition of dextran sulfate induces the substitution of non-aggregated gold nanoparticles while aggregated ones are stable on the substrate. We show that this detection method is highly specific and allows the detection of DNA mismatches and damages.

2D aggregation and selective desorption of nanoparticle probes: a new method to probe DNA mismatches and damages.

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Interface devices such as integrated planar patch-clamp chips are being developed to study the electrophysiological activity of neuronal networks grown in vitro. The utility of such devices will be dependent upon the ability to align neurons with interface features on the chip by controlling neuronal placement and by guiding cell connectivity. In this paper, we present a strategy to accomplish this goal. Patterned chemical modification of SiN surfaces with poly-d-lysine transferred from PDMS stamps was used to promote adhesion and guidance of cryo-preserved primary rat cortical neurons. We demonstrate that these neurons can be positioned and grown over microhole features which will ultimately serve as patch-clamp interfaces on the chip.

Cell placement and guidance on substrates for neurochip interfaces.

Dispositifs d&#x26;#39;interface tels que planaires int&#xE9;gr&#xE9;s de patch-clamp puces sont d&#xE9;velopp&#xE9;es pour &#xE9;tudier l&#x26;#39;activit&#xE9; &#xE9;lectrophysiologique des r&#xE9;seaux neuronaux cultiv&#xE9;s in vitro. L&#x26;#39;utilit&#xE9; de ces dispositifs sera tributaire de la capacit&#xE9; d&#x26;#39;aligner les neurones avec des fonctionnalit&#xE9;s de l&#x26;#39;interface sur la puce en contr&#xF4;lant le placement neuronale et en guidant la connectivit&#xE9; cellulaire. Dans cet article, nous pr&#xE9;sentons une strat&#xE9;gie pour atteindre cet objectif. Patterned modification chimique des surfaces SiN avec des poly-D-lysine transf&#xE9;r&#xE9;s de timbres PDMS a &#xE9;t&#xE9; utilis&#xE9; pour favoriser l&#x26;#39;adh&#xE9;rence et l&#x26;#39;orientation de la cryo-conserv&#xE9;s neurones corticaux primaires de rat. Nous d&#xE9;montrons que ces neurones peuvent &#xEA;tre positionn&#xE9;s et grandi au fil caract&#xE9;ristiques microtrou qui finira par servir de patch-clamp interfaces sur la puce.

Placement de cellules et de l&#x26;#39;orientation sur des substrats pour les interfaces neuropuces

Interface-Ger&auml;te wie z. B. integrierte planare Patch-Clamp-Chips werden entwickelt, um die elektrophysiologische Aktivit&auml;t neuronaler Netzwerke gewachsen in-vitro-Studie. Das Dienstprogramm solcher Ger&auml;te werden abh&auml;ngig von der F&auml;higkeit, Neuronen mit Features f&uuml;r die Benutzeroberfl&auml;che auf dem Chip durch die Kontrolle der neuronalen Platzierung und durch leitende Verbindungen der Zelle ausrichten. In diesem Beitrag pr&auml;sentieren wir eine Strategie, um dieses Ziel zu erreichen. Gemusterte chemische Modifikation von S&uuml;nde Oberfl&auml;chen mit Poly-d-Lysin von PDMS Briefmarken &uuml;bertragen wurde verwendet, um die Adh&auml;sion und Leitung von Cryo erhaltenen prim&auml;ren Ratte kortikale Neuronen zu f&ouml;rdern. Wir zeigen, dass diese Neuronen positioniert und in Microhole-Features, die letztlich als Patch-Clamp-Schnittstellen auf dem Chip dient gewachsen werden k&ouml;nnen.

Zelle Platzierung und Leitlinien f&uuml;r die Substrate f&uuml;r Neurochip-Schnittstellen.

Dispositivi di interfaccia come chip integrato planar patch-clamp sono stati sviluppati per studiare l&#39;attivit&agrave; elettrofisiologica delle reti neuronali, coltivate in vitro. La capacit&agrave; di allineare i neuroni con funzioni di interfaccia sul chip controllando il posizionamento neuronale e di connettivit&agrave; cellulare guida dipender&agrave; l&#39;utilit&agrave; di tali dispositivi. In questo articolo, vi presentiamo una strategia per realizzare questo obiettivo. Fantasia modificazione chimica delle superfici SiN con poli-d-lisina trasferito da francobolli PDMS &egrave; stato utilizzato per promuovere l&#39;adesione e l&#39;orientamento dei neuroni corticali del ratto primaria crioconservazione. Dimostreremo che questi neuroni possono essere posizionati e cresciuti sulle caratteristiche microhole che serviranno alla fine come interfacce di patch-clamp sul chip.

Cella posizionamento e orientamento su substrati per le interfacce neurochip.

&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x7D71;&#x5408;&#x3055;&#x308C;&#x305F;&#x5E73;&#x9762;&#x30D1;&#x30C3;&#x30C1;&#x30AF;&#x30E9;&#x30F3;&#x30D7;&#x30C1;&#x30C3;&#x30D7;&#x3068;&#x3057;&#x3066;&#x30A4;&#x30F3;&#x30BF;&#x30D5;&#x30A7;&#x30FC;&#x30B9;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306F;&#x3001;in vitro&#x3067;&#x5897;&#x6B96;&#x3055;&#x305B;&#x305F;&#x795E;&#x7D4C;&#x56DE;&#x8DEF;&#x7DB2;&#x306E;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x6D3B;&#x6027;&#x3092;&#x7814;&#x7A76;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x958B;&#x767A;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306E;&#x30E6;&#x30FC;&#x30C6;&#x30A3;&#x30EA;&#x30C6;&#x30A3;&#x3067;&#x306F;&#x3001;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x914D;&#x7F6E;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x308A;&#x3001;&#x30BB;&#x30EB;&#x306E;&#x63A5;&#x7D9A;&#x6027;&#x3092;&#x5C0E;&#x304F;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x30C1;&#x30C3;&#x30D7;&#x4E0A;&#x306E;&#x30A4;&#x30F3;&#x30BF;&#x30D5;&#x30A7;&#x30FC;&#x30B9;&#x6A5F;&#x80FD;&#x3092;&#x6301;&#x3064;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x3092;&#x6574;&#x5217;&#x3055;&#x305B;&#x308B;&#x80FD;&#x529B;&#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x3067;&#x3042;&#x308D;&#x3046;&#x3002;&#x672C;&#x7A3F;&#x3067;&#x306F;&#x3001;&#x3053;&#x306E;&#x76EE;&#x6A19;&#x3092;&#x9054;&#x6210;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x6226;&#x7565;&#x3092;&#x63D0;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002; PDMS&#x30B9;&#x30BF;&#x30F3;&#x30D7;&#x304B;&#x3089;&#x8EE2;&#x9001;&#x3055;&#x308C;&#x305F;&#x30DD;&#x30EA;-D-&#x30EA;&#x30B8;&#x30F3;&#x306E;SiN&#x8868;&#x9762;&#x306E;&#x6A21;&#x69D8;&#x306E;&#x5316;&#x5B66;&#x4FEE;&#x98FE;&#x306F;&#x3001;&#x51CD;&#x7D50;&#x4FDD;&#x5B58;&#x3055;&#x308C;&#x305F;&#x521D;&#x4EE3;&#x30E9;&#x30C3;&#x30C8;&#x76AE;&#x8CEA;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x63A5;&#x7740;&#x3084;&#x30AC;&#x30A4;&#x30C0;&#x30F3;&#x30B9;&#x3092;&#x4FC3;&#x9032;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x304C;&#x914D;&#x7F6E;&#x3055;&#x308C;&#x3001;&#x6700;&#x7D42;&#x7684;&#x306B;&#x30C1;&#x30C3;&#x30D7;&#x4E0A;&#x306B;&#x30D1;&#x30C3;&#x30C1;&#x30AF;&#x30E9;&#x30F3;&#x30D7;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30D5;&#x30A7;&#x30A4;&#x30B9;&#x3068;&#x3057;&#x3066;&#x6A5F;&#x80FD;&#x3057;&#x307E;&#x3059;microhole&#x6A5F;&#x80FD;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x6210;&#x9577;&#x3055;&#x305B;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x308B;&#x3002;

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&#xD1B5;&#xD569;&#xB41C; &#xD3C9;&#xBA74; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xCE69; &#xAC19;&#xC740; &#xC778;&#xD130;&#xD398;&#xC774;&#xC2A4; &#xC7A5;&#xCE58; &#xCCB4; &#xC678; &#xC131;&#xC7A5; &#xC2E0;&#xACBD; &#xB124;&#xD2B8;&#xC6CC;&#xD06C;&#xC758; electrophysiological &#xD65C;&#xB3D9; &#xC5F0;&#xAD6C; &#xAC1C;&#xBC1C; &#xB418; &#xACE0; &#xC788;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xC7A5;&#xCE58;&#xC758; &#xC720;&#xD2F8;&#xB9AC;&#xD2F0; &#xC2E0;&#xACBD; &#xBC30;&#xCE58;&#xB97C; &#xC81C;&#xC5B4; &#xD558; &#xACE0; &#xC140; &#xC5F0;&#xACB0; &#xC9C0;&#xCE68;&#xC5D0; &#xC758;&#xD574; &#xB274;&#xB7F0; &#xCE69; &#xC778;&#xD130;&#xD398;&#xC774;&#xC2A4; &#xAE30;&#xB2A5;&#xC744; &#xC815;&#xB82C; &#xD558;&#xB294; &#xAE30;&#xB2A5;&#xC5D0; &#xB530;&#xB77C; &#xB2EC;&#xB77C; &#xC9D1;&#xB2C8;&#xB2E4; &#xAC83;&#xC785;&#xB2C8;&#xB2E4;. &#xC774; &#xC885;&#xC774;&#xC5D0;&#xC11C;&#xB294; &#xC774;&#xB7EC;&#xD55C; &#xBAA9;&#xD45C;&#xB97C; &#xB2EC;&#xC131; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC804;&#xB7B5; &#xC18C;&#xAC1C;. &#xD3F4; &#xB9AC; d lysine PDMS &#xC2A4;&#xD0EC;&#xD504;&#xC5D0;&#xC11C; &#xC804;&#xC1A1; &#xB41C; &#xC8C4; &#xD45C;&#xBA74; &#xD328;&#xD134;&#xD654; &#xD654;&#xD559; &#xC218;&#xC815; &#xC811;&#xCC29; &#xBC0F; &#xACF3;&#xC744; &#xC54C;&#xC544;&#xB0B4;&#xB294; &#xBCF4;&#xC874; &#xAE30;&#xBCF8; &#xC950; &#xB300;&#xB1CC; &#xD53C; &#xC9C8;&#xC758; &#xB274;&#xB7F0;&#xC758; &#xC9C0;&#xB3C4; &#xD558;&#xC5D0; &#xC0AC;&#xC6A9; &#xB418;&#xC5C8;&#xB2E4;. &#xC774; &#xC704;&#xCE58; &#xBC0F; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xC778;&#xD130;&#xD398;&#xC774;&#xC2A4; &#xCE69;&#xC5D0; &#xAD81;&#xADF9;&#xC801;&#xC73C;&#xB85C; &#xB420; &#xAC83;&#xC785;&#xB2C8;&#xB2E4; microhole &#xAE30;&#xB2A5;&#xC744; &#xD1B5;&#xD574; &#xC131;&#xC7A5; &#xC218; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;.

&#xC140; &#xBC30;&#xCE58; &#xBC0F; neurochip &#xC778;&#xD130;&#xD398;&#xC774;&#xC2A4;&#xC5D0; &#xB300; &#xD55C; &#xAE30;&#xD310;&#xC5D0; &#xB300; &#xD55C; &#xC9C0;&#xCE68;.

Dispositivos de interfaz, tales como planos integrados patch-clamp fichas se est&#xE1;n desarrollando para estudiar la actividad electrofisiol&#xF3;gica de las redes neuronales cultivadas in vitro. La utilidad de tales dispositivos ser&#xE1; dependiente de la capacidad para alinear las neuronas con caracter&#xED;sticas de interfaz en el chip mediante el control de la colocaci&#xF3;n neuronal y guiando la conectividad celular. En este art&#xED;culo se presenta una estrategia para lograr este objetivo. Estampado modificaci&#xF3;n qu&#xED;mica de superficies SiN con poli-D-lisina transferidos de sellos PDMS se utiliz&#xF3; para promover la adhesi&#xF3;n y la orientaci&#xF3;n de la crio-preservados neuronas corticales de rata primarios. Se demuestra que estas neuronas se puede colocar y crecido a trav&#xE9;s de caracter&#xED;sticas MicroHole que en &#xFA;ltima instancia sirven como interfaces de patch-clamp en el chip.

De la c&#xE9;lula de colocaci&#xF3;n y orientaci&#xF3;n sobre sustratos para interfaces neurochip

This Article describes a strategy to stabilize a phospholipid monolayer directly on the surface of a H-terminated silicon substrate in order to provide a useful platform for silicon based biosensors. The stabilization of an acrylated phospholipid monolayer is obtained by two-dimensional chain polymerization. As the formation of the lipid monolayer in aqueous solution competes with the oxidation of the silicon surface, several cycles of oxide removal and lipid exposure are necessary to densify the lipid layer. Lipid monolayer formation is followed by Fourier transform infrared spectroscopy. The resulting monolayer is denser than corresponding alkyl monolayers formed on H-terminated silicon via photochemical or thermally initiated reactions.

Direct stabilization of a phospholipid monolayer on H-terminated silicon.

The electrical performance of stabilized lipid monolayers on H-terminated silicon is reported for the first time. We show that these 2.7 nm thick only ultrathin layers present extremely low current leakage at high electric field and high breakdown voltage that both compare favorably with the best data reported on organic thin film dielectrics. We demonstrate a very unique property of autonomic self-healing of the layer at room temperature with the total recovery of its performance after electrical breakdown. The mechanisms involved in breakdown and self-healing are described.

Autonomic self-healing lipid monolayer: a new class of ultrathin dielectric.

&#x6240;&#x6709;&#x53EF;&#x5174;&#x594B;&#x6027;&#x7EC6;&#x80DE;&#x529F;&#x80FD;&#x4F9D;&#x8D56;&#x4E8E;&#x5D4C;&#x5165;&#x5728;&#x5176;&#x7EC6;&#x80DE;&#x819C;&#x7684;&#x79BB;&#x5B50;&#x901A;&#x9053;&#x3002;&#x79BB;&#x5B50;&#x901A;&#x9053;&#x7ED3;&#x6784;&#x6216;&#x529F;&#x80FD;&#x7684;&#x6270;&#x52A8;&#x5BFC;&#x81F4;&#x5FC3;&#x529F;&#x80FD;&#x4E0D;&#x5168;&#xFF0C;&#x4EE5;&#x81F3;&#x795E;&#x7ECF;&#x9000;&#x884C;&#x6027;&#x75BE;&#x75C5;&#x7684;&#x75C5;&#x7406;&#x73B0;&#x8C61;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x82E5;&#x8981;&#x7406;&#x89E3;&#x53EF;&#x5174;&#x594B;&#x6027;&#x7EC6;&#x80DE;&#x7684;&#x529F;&#x80FD;&#xFF0C;&#x5E76;&#x7EA0;&#x6B63;&#x5176;&#x75C5;&#x7406;&#x751F;&#x7406;&#x5B66;&#xFF0C;&#x5B83;&#x662F;&#x91CD;&#x8981;&#x7684;&#x662F;&#x8981;&#x4E86;&#x89E3;&#x5404;&#x79CD;&#x8BD5;&#x9A8C;&#x6761;&#x4EF6;&#x4E0B;-&#x5305;&#x62EC;&#x66B4;&#x9732;&#x4E8E;&#x65B0;&#x578B;&#x836F;&#x7269;&#x9776;&#x6807;&#x7684;&#x79BB;&#x5B50;&#x901A;&#x9053;&#x529F;&#x80FD;&#x3002;&#x73BB;&#x7483;&#x5438;&#x7BA1;&#x819C;&#x7247;&#x94B3;&#x662F;&#x8981;&#x76D1;&#x89C6;&#x7684;&#x5185;&#x5728;&#x548C;&#x7A81;&#x89E6;&#x5C5E;&#x6027;&#x7684;&#x795E;&#x7ECF;&#x5143;&#x7684;&#x5148;&#x8FDB;&#x7684;&#x6280;&#x672F;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x8FD9;&#x79CD;&#x6280;&#x672F;&#x662F;&#x52B3;&#x52A8;&#x5BC6;&#x96C6;&#x578B;&#x548C;&#x5177;&#x6709;&#x4F4E;&#x7684;&#x6570;&#x636E;&#x541E;&#x5410;&#x91CF;&#x3002;&#x5E73;&#x9762;&#x819C;&#x7247;&#x94B3;&#x82AF;&#x7247;&#xFF0C;&#x96C6;&#x6210;&#x5230;&#x81EA;&#x52A8;&#x5316;&#x7CFB;&#x7EDF;&#xFF0C;&#x63D0;&#x4F9B;&#x9AD8;&#x541E;&#x5410;&#x91CF;&#x4F46;&#x53EA;&#x9650;&#x4E8E;&#x79BB;&#x7EC6;&#x80DE;&#x4ECE;&#x60AC;&#x6D6E;&#x6DB2;&#xFF0C;&#x56E0;&#x800C;&#x9650;&#x5236;&#x4E86;&#x5176;&#x4F7F;&#x7528;&#x4E2D;&#x5EFA;&#x6A21;&#x7684;&#x751F;&#x7406;&#x529F;&#x80FD;&#x3002;&#x8FD9;&#x4E9B;&#x82AF;&#x7247;&#xFF0C;&#x56E0;&#x6B64;&#x5E76;&#x4E0D;&#x6D89;&#x53CA;&#x795E;&#x7ECF;&#x5143;&#x901A;&#x4FE1;&#x7814;&#x7A76;&#x6700;&#x9002;&#x5408;&#x3002;&#x591A;&#x7535;&#x6781;&#x9635;&#x5217; &#xFF08;&#x591A;&#x8FB9;&#x73AF;&#x5883;&#x534F;&#x5B9A;&#xFF09;&#xFF0C;&#x76F8;&#x6BD4;&#x4E4B;&#x4E0B;&#xFF0C;&#x6709;&#x80FD;&#x529B;&#x901A;&#x8FC7;&#x6D4B;&#x91CF;&#x5C40;&#x90E8;&#x573A;&#x7535;&#x4F4D;&#x4ECE;&#x80DE;&#x5916;&#x7684;&#x591A;&#x4E2A;&#x7AD9;&#x70B9;&#xFF0C;&#x76D1;&#x89C6;&#x7F51;&#x7EDC;&#x6D3B;&#x52A8;&#xFF0C;&#x4F46;&#x7279;&#x5B9A;&#x79BB;&#x5B50;&#x901A;&#x9053;&#x6D3B;&#x52A8;&#x4E00;&#x9879;&#x6311;&#x6218;&#x4ECE;&#x8FD9;&#x4E9B;&#x591A;&#x8DEF;&#x590D;&#x7528;&#x4FE1;&#x53F7;&#x4E2D;&#x63D0;&#x53D6;&#x3002;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x4ECB;&#x7ECD;&#x4E00;&#x79CD;&#x65B0;&#x578B;&#x5E73;&#x9762;&#x819C;&#x7247;&#x94B3;&#x82AF;&#x7247;&#x6280;&#x672F;&#xFF0C;&#x4F7F;&#x5355;&#x4E2A;&#x795E;&#x7ECF;&#x5143; synaptically &#x8FDE;&#x63A5;&#x795E;&#x7ECF;&#x5143;&#x7F51;&#x7EDC;&#xFF0C;&#x5728;&#x591A;&#x4E2A;&#x5730;&#x70B9;&#x540C;&#x65F6;&#x9AD8;&#x5206;&#x8FA8;&#x7387;&#x7535;&#x751F;&#x7406;&#x5BA1;&#x8BAF;&#x4ECE;&#x800C;&#x5C06;&#x591A;&#x8FB9;&#x73AF;&#x5883;&#x534F;&#x5B9A;&#x4E0E;&#x819C;&#x7247;&#x94B3;&#x6280;&#x672F;&#x7684;&#x4F18;&#x70B9;&#x7ED3;&#x5408;&#x8D77;&#x6765;&#x3002;&#x6BCF;&#x4E2A;&#x795E;&#x7ECF;&#x5143;&#x53EF;&#x63A2;&#x6D4B;&#x901A;&#x8FC7;&#x8FDE;&#x63A5;&#x5230;&#x4E13;&#x7528;&#x5730;&#x4E0B;&#x5FAE;&#x6D41;&#x63A7;&#x901A;&#x9053;&#x7684;&#x5149;&#x5708;&#x3002;&#x6210;&#x957F;&#x5728;&#x7F51;&#x7EDC;&#x4E2D;&#x7684;&#x795E;&#x7ECF;&#x5143;&#x6309; physisorbed &#x6216; chemisorbed &#x7684;&#x5316;&#x5B66;&#x7EBF;&#x7D22;&#x5230;&#x5B54;&#x5BF9;&#x9F50;&#x3002;&#x5728;&#x8FD9;&#x6B21;&#x5BA1;&#x67E5;&#xFF0C;&#x6211;&#x4EEC;&#x63CF;&#x8FF0;&#x8FD9;&#x4E9B;&#x82AF;&#x7247;&#xFF0C;&#x4ECE;&#x795E;&#x7ECF;&#x5143;&#x7EC6;&#x80DE;&#x5316;&#x5B66;&#x9635;&#x5217;&#x7684;&#x5355;&#x5143;&#x683C;&#x4F4D;&#x7F6E;&#x548C;&#x76EE;&#x524D;&#x7684;&#x751F;&#x7406;&#x6570;&#x636E;&#x65B9;&#x6CD5;&#x7684;&#x8BBE;&#x8BA1;&#x548C;&#x5236;&#x4F5C;&#x8FC7;&#x7A0B;&#x3002;

&#x4ECE;&#x4E86;&#x89E3;&#x5230;&#x65B0;&#x578B;&#x836F;&#x7269;&#x53D1;&#x73B0;&#x7EC6;&#x80DE;&#x529F;&#x80FD;&#xFF1A; &#x5E73;&#x9762;&#x819C;&#x7247;&#x94B3;&#x9635;&#x5217;&#x82AF;&#x7247;&#x6280;&#x672F;&#x7684;&#x4F5C;&#x7528;&#x3002;

All excitable cell functions rely upon ion channels that are embedded in their plasma membrane. Perturbations of ion channel structure or function result in pathologies ranging from cardiac dysfunction to neurodegenerative disorders. Consequently, to understand the functions of excitable cells and to remedy their pathophysiology, it is important to understand the ion channel functions under various experimental conditions - including exposure to novel drug targets. Glass pipette patch-clamp is the state of the art technique to monitor the intrinsic and synaptic properties of neurons. However, this technique is labor intensive and has low data throughput. Planar patch-clamp chips, integrated into automated systems, offer high throughputs but are limited to isolated cells from suspensions, thus limiting their use in modeling physiological function. These chips are therefore not most suitable for studies involving neuronal communication. Multielectrode arrays (MEAs), in contrast, have the ability to monitor network activity by measuring local field potentials from multiple extracellular sites, but specific ion channel activity is challenging to extract from these multiplexed signals. Here we describe a novel planar patch-clamp chip technology that enables the simultaneous high-resolution electrophysiological interrogation of individual neurons at multiple sites in synaptically connected neuronal networks, thereby combining the advantages of MEA and patch-clamp techniques. Each neuron can be probed through an aperture that connects to a dedicated subterranean microfluidic channel. Neurons growing in networks are aligned to the apertures by physisorbed or chemisorbed chemical cues. In this review, we describe the design and fabrication process of these chips, approaches to chemical patterning for cell placement, and present physiological data from cultured neuronal cells.

From understanding cellular function to novel drug discovery: the role of planar patch-clamp array chip technology.

Toutes les fonctions des cellules excitables se fondent sur des canaux ioniques qui sont incorpor&eacute;s dans leur membrane plasmique. Les perturbations de la fonction ou la structure de canal d&#39;ion entra&icirc;ner des pathologies allant de dysfonction cardiaque aux maladies neurod&eacute;g&eacute;n&eacute;ratives. Par cons&eacute;quent, pour comprendre les fonctions des cellules excitables et rem&eacute;dier &agrave; leur physiopathologie, il est important de comprendre les fonctions de canal d&#39;ion dans diverses conditions exp&eacute;rimentales - y compris l&#39;exposition &agrave; des cibles de nouveaux m&eacute;dicaments. Patch-clamp de pipette de verre est la technique de pointe pour surveiller les propri&eacute;t&eacute;s intrins&egrave;ques et synaptiques des neurones. Cependant, cette technique est beaucoup de travail et a donn&eacute;es &agrave; faible d&eacute;bit. Puces Planar patch-clamp, int&eacute;gr&eacute;s dans des syst&egrave;mes automatis&eacute;s, d&#39;offrir des d&eacute;bits &eacute;lev&eacute;s mais sont limit&eacute;es &agrave; des cellules isol&eacute;es des suspensions, ce qui limite leur utilisation dans la mod&eacute;lisation d&#39;une fonction physiologique. Ces puces ne sont donc pas plus adapt&eacute;s aux &eacute;tudes portant sur la communication neuronale. Multi&eacute;lectrodes tableaux (AME), en revanche, avoir la possibilit&eacute; de surveiller l&#39;activit&eacute; r&eacute;seau en mesurant les potentiels de champs locaux de plusieurs sites extracellulaires, mais l&#39;activit&eacute; des canaux ioniques sp&eacute;cifiques est difficile &agrave; extraire de ces signaux multiplex&eacute;s. Nous d&eacute;crivons ici une technologie de puce de nouveaux planar patch clamp qui permet l&#39;interrogatoire &eacute;lectrophysiologique haute r&eacute;solution simultan&eacute;e de neurones individuels sur des sites multiples dans les r&eacute;seaux neurones synapses connect&eacute;s, combinant ainsi les avantages des techniques MEA et patch clamp. Chaque neurone peut &ecirc;tre sond&eacute;e par une ouverture qui se connecte &agrave; un canal microfluidique souterraine d&eacute;di&eacute;e. De plus en plus dans les r&eacute;seaux de neurones sont align&eacute;s sur les ouvertures par physisorb&eacute;es ou chimisorb&eacute;s des signaux chimiques. Dans cette revue, nous d&eacute;crivons le processus de conception et de fabrication de ces puces, approches de structuration chimique pour le placement de la cellule et pr&eacute;senter des donn&eacute;es physiologiques des cellules neuronales cultiv&eacute;es.

De comprendre la fonction cellulaire &agrave; la d&eacute;couverte de nouveaux m&eacute;dicaments : le r&ocirc;le de la technologie de puce planar patch-clamp.

Alle erregbar Zelle Funktionen st&uuml;tzen sich auf Ionenkan&auml;le, die in ihrer Plasmamembran eingebettet sind. St&ouml;rungen von Ionen-Kanal-Struktur oder Funktion f&uuml;hren Pathologien von kardialen Dysfunktion bis hin zu neurodegenerativen Erkrankungen. Um die Funktionen von excitable Zellen zu verstehen und ihre Pathophysiologie zu beseitigen, ist es daher wichtig zu verstehen, die Ion-Kanalfunktionen unter verschiedenen experimentellen Bedingungen - einschlie&szlig;lich der Gef&auml;hrdung durch neuartige Droge-Ziele. Glas-Pipette Patch-Clamp ist die modernste Technik, die intrinsischen und synaptischen Eigenschaften der Neuronen zu &uuml;berwachen. Aber diese Technik ist arbeitsintensiv und hat geringen Datendurchsatz. Planare Patch-Clamp-Chips, Automationen, integriert bieten hohe Durchs&auml;tze aber begrenzt an isolierten Zellen von Suspensionen, ihre Verwendung bei der Modellierung physiologische Funktion eingeschr&auml;nkt sind. Diese Chips sind daher nicht f&uuml;r Studien mit neuronalen Kommunikation am geeignetsten. Multielectrode Arrays (MEAs), im Gegensatz dazu haben die Netzwerk-Aktivit&auml;t &uuml;berwachen durch Messung lokales Feld Potenziale mehrerer extrazellul&auml;re Standorte, aber spezifische Ion Channel-Aktivit&auml;ten ist eine Herausforderung, um aus dieser Multiplex-Signale zu extrahieren. Hier beschreiben wir eine neuartige planare Patch-Clamp-Chip-Technologie, die es erm&ouml;glicht die gleichzeitige hochaufl&ouml;sende elektrophysiologische Befragung des einzelnen Neuronen an mehreren Standorten in synaptisch neuronale Netzwerke, damit kombinieren die Vorteile von MEA und Patch-Clamp-Technik. Jedes Neuron kann durch eine Blende sondiert werden, die einen dedizierten unterirdischen Microfluidic-Kanal verbindet. Neuronen wachsen in Netzwerken werden die &Ouml;ffnungen von Physisorbed oder chemisorbed chemische Signale ausgerichtet. In diesem Bericht beschreiben wir den Prozess-Design und Fertigung dieser Chips, Ans&auml;tze zur chemischen Musterung f&uuml;r Zelle Platzierung und physiologische Daten von kultivierten neuronalen Zellen.

Vom Verst&auml;ndnis der Zellfunktion zu neuartigen Drogeentdeckung: die Rolle der planare Patch-Clamp-Array-Chip-Technologie.

Tutte le funzioni delle cellule eccitabili si basano su canali ionici che sono incorporati nella loro membrana plasmatica. Perturbazioni della struttura di canale ionico o funzione causare patologie che vanno dalla disfunzione cardiaca a patologie neurodegenerative. Di conseguenza, per capire le funzioni delle cellule eccitabili e per porre rimedio a loro Fisiopatologia, &egrave; importante comprendere le funzioni di canale ionico in varie condizioni sperimentali - tra cui l&#39;esposizione a obiettivi farmacologici romanzo. Pipetta vetro patch-clamp &egrave; la tecnica d&#39;avanguardia per monitorare le propriet&agrave; intrinseche e sinaptiche dei neuroni. Tuttavia, questa tecnica &egrave; laborioso e ha velocit&agrave; di trasmissione dati a bassa. Chip planare patch-clamp, integrati in sistemi automatizzati, offrono alta velocit&agrave; ma sono limitati a cellule isolate da sospensioni, limitando il loro uso in funzione fisiologica di modellazione. Questi chip non sono quindi pi&ugrave; adatti per gli studi che coinvolgono la comunicazione neuronale. Matrici multielectrode (AAM), al contrario, hanno la capacit&agrave; di monitorare l&#39;attivit&agrave; di rete misurando i potenziali di campo locale da pi&ugrave; siti extracellulari, ma &egrave; difficile attivit&agrave; del canale ionico specifico per estrarre da questi segnali multiplexati. Qui descriviamo una tecnologia chip romanzo patch-clamp planare che consente la simultanea interrogatorio elettrofisiologico ad alta risoluzione di singoli neuroni in pi&ugrave; siti in reti neuronali sinapticamente connesse, quindi combinando i vantaggi delle tecniche di MEA e patch-clamp. Ogni neurone pu&ograve; essere sondato attraverso un&#39;apertura che si connette a un canale dedicato microfluidica sotterraneo. Crescendo in reti di neuroni sono allineati per le aperture di fisisorbito o chemisorbite segnali chimici. In questa recensione, si descrive il processo di progettazione e fabbricazione di questi chip, approcci di patterning chimico per il posizionamento delle cellule e presentano i dati fisiologici da cellule neuronali in coltura.

Da capire la funzione cellulare alla scoperta di nuovi farmaci: il ruolo della tecnologia di chip array planari patch-clamp.

&#x3059;&#x3079;&#x3066;&#x306E;&#x8208;&#x596E;&#x6027;&#x306E;&#x7D30;&#x80DE;&#x6A5F;&#x80FD;&#x306E;&#x539F;&#x5F62;&#x8CEA;&#x819C;&#x306B;&#x57CB;&#x3081;&#x8FBC;&#x307E;&#x308C;&#x3066;&#x3044;&#x308B;&#x30A4;&#x30AA;&#x30F3; &#x30C1;&#x30E3;&#x30CD;&#x30EB;&#x306B;&#x4F9D;&#x5B58;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30A4;&#x30AA;&#x30F3; &#x30C1;&#x30E3;&#x30CD;&#x30EB;&#x306E;&#x69CB;&#x9020;&#x3084;&#x6A5F;&#x80FD;&#x306E;&#x6442;&#x52D5;&#x304B;&#x3089;&#x306E;&#x5FC3;&#x6A5F;&#x80FD;&#x969C;&#x5BB3;&#x795E;&#x7D4C;&#x5909;&#x6027;&#x75BE;&#x60A3;&#x306B;&#x81F3;&#x308B;&#x307E;&#x3067;&#x306E;&#x75C5;&#x7406;&#x7D50;&#x679C;&#x3057;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E;&#x7D50;&#x679C;&#x3001;&#x8208;&#x596E;&#x6027;&#x7D30;&#x80DE;&#x306E;&#x6A5F;&#x80FD;&#x3092;&#x7406;&#x89E3;&#x3057;&#x3001;&#x305D;&#x306E;&#x75C5;&#x614B;&#x3092;&#x6539;&#x5584;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x65B0;&#x898F;&#x5275;&#x85AC;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3078;&#x306E;&#x66DD;&#x9732;&#x3092;&#x542B;&#x3080;&#x3001;&#x7A2E;&#x3005; &#x306E;&#x5B9F;&#x9A13;&#x6761;&#x4EF6; - &#x4E0B;&#x306E;&#x30A4;&#x30AA;&#x30F3; &#x30C1;&#x30E3;&#x30CD;&#x30EB;&#x6A5F;&#x80FD;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;&#x30AC;&#x30E9;&#x30B9; &#x30D4;&#x30DA;&#x30C3;&#x30C8; &#x30D1;&#x30C3;&#x30C1; &#x30FB; &#x30AF;&#x30E9;&#x30F3;&#x30D7;&#x306E;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x7D44;&#x307F;&#x8FBC;&#x307F;&#x304A;&#x3088;&#x3073;&#x30B7;&#x30CA;&#x30D7;&#x30B9; &#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x3092;&#x76E3;&#x8996;&#x3059;&#x308B;&#x6700;&#x5148;&#x7AEF;&#x306E;&#x6280;&#x8853;&#x3067;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x3053;&#x306E;&#x624B;&#x6CD5;&#x3067;&#x52B4;&#x50CD;&#x96C6;&#x7D04;&#x7684;&#x3001;&#x4F4E;&#x30C7;&#x30FC;&#x30BF; &#x30B9;&#x30EB;&#x30FC;&#x30D7;&#x30C3;&#x30C8;&#x3002;&#x5E73;&#x9762;&#x30D1;&#x30C3;&#x30C1; &#x30FB; &#x30AF;&#x30E9;&#x30F3;&#x30D7;&#x306E;&#x30C1;&#x30C3;&#x30D7;&#x306F;&#x3001;&#x81EA;&#x52D5;&#x5316;&#x3055;&#x308C;&#x305F;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x7D71;&#x5408;&#x3055;&#x308C;&#x305F;&#x9AD8;&#x30B9;&#x30EB;&#x30FC;&#x30D7;&#x30C3;&#x30C8;&#x3067;&#x3059;&#x306B;&#x9694;&#x96E2;&#x3055;&#x308C;&#x305F;&#x30BB;&#x30EB;&#x304B;&#x3089;&#x3053;&#x306E;&#x3088;&#x3046;&#x306B;&#x751F;&#x7406;&#x6A5F;&#x80FD;&#x306E;&#x30E2;&#x30C7;&#x30EA;&#x30F3;&#x30B0;&#x306E;&#x4F7F;&#x7528;&#x3092;&#x5236;&#x9650;&#x3059;&#x308B;&#x61F8;&#x6FC1;&#x6DB2;&#x3001;&#x9650;&#x3089;&#x308C;&#x305F;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C1;&#x30C3;&#x30D7;&#x306F;&#x795E;&#x7D4C;&#x7D30;&#x80DE;&#x306E;&#x30B3;&#x30DF;&#x30E5;&#x30CB;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x542B;&#x3080;&#x8ABF;&#x67FB;&#x306E;&#x6700;&#x3082;&#x9069;&#x3057;&#x305F;&#x3055;&#x308C;&#x307E;&#x305B;&#x3093;&#x3002;&#x591A;&#x96FB;&#x6975;&#x30A2;&#x30EC;&#x30A4; (Mea) &#x306F;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x3001;&#x8907;&#x6570;&#x306E;&#x7D30;&#x80DE;&#x5916;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x304B;&#x3089;&#x306E;&#x5C40;&#x6240;&#x96FB;&#x754C;&#x96FB;&#x4F4D;&#x3092;&#x6E2C;&#x5B9A;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x30CD;&#x30C3;&#x30C8;&#x30EF;&#x30FC;&#x30AF;&#x6D3B;&#x52D5;&#x3092;&#x76E3;&#x8996;&#x3059;&#x308B;&#x80FD;&#x529B;&#x304C;&#x3053;&#x308C;&#x3089;&#x591A;&#x91CD;&#x4FE1;&#x53F7;&#x304B;&#x3089;&#x62BD;&#x51FA;&#x3059;&#x308B;&#x7279;&#x5B9A;&#x306E;&#x30A4;&#x30AA;&#x30F3; &#x30C1;&#x30E3;&#x30CD;&#x30EB;&#x6D3B;&#x6027;&#x306B;&#x6311;&#x6226;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x305D;&#x308C;&#x306B;&#x3088;&#x3063;&#x3066; MEA &#x3068;&#x30D1;&#x30C3;&#x30C1; &#x30FB; &#x30AF;&#x30E9;&#x30F3;&#x30D7;&#x306E;&#x6280;&#x8853;&#x306E;&#x5229;&#x70B9;&#x3092;&#x7D44;&#x307F;&#x5408;&#x308F;&#x305B;&#x3066;&#x6709;&#x52B9;&#x306B;&#x500B;&#x3005; &#x306E;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x63A5;&#x7D9A;&#x306E;&#x795E;&#x7D4C;&#x30CD;&#x30C3;&#x30C8;&#x30EF;&#x30FC;&#x30AF;&#x306B;&#x8907;&#x6570;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x3067;&#x540C;&#x6642;&#x306E;&#x9AD8;&#x89E3;&#x50CF;&#x5EA6;&#x306E;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x5C0B;&#x554F;&#x5C0F;&#x8AAC;&#x5E73;&#x9762;&#x30D1;&#x30C3;&#x30C1;&#x30AF; &#x30C1;&#x30C3;&#x30D7;&#x6280;&#x8853;&#x3092;&#x8A18;&#x8FF0;&#x3057;&#x307E;&#x3059;&#x3002;&#x5404;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306F;&#x5C02;&#x7528;&#x5730;&#x4E0B;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30C1;&#x30E3;&#x30F3;&#x30CD;&#x30EB;&#x306B;&#x63A5;&#x7D9A;&#x3059;&#x308B;&#x958B;&#x53E3;&#x30D7;&#x30ED;&#x30FC;&#x30D6;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3; &#x30CD;&#x30C3;&#x30C8;&#x30EF;&#x30FC;&#x30AF;&#x306E;&#x6210;&#x9577;&#x306B;&#x958B;&#x53E3;&#x90E8;&#x7269;&#x7406;&#x5438;&#x7740;&#x307E;&#x305F;&#x306F;&#x5316;&#x5B66;&#x5438;&#x7740;&#x5206;&#x5B50;&#x306E;&#x5316;&#x5B66;&#x7684;&#x624B;&#x304C;&#x304B;&#x308A;&#x306B;&#x3088;&#x308B;&#x6574;&#x5217;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x7DCF;&#x8AAC;&#x3067;&#x306F;, &#x3053;&#x308C;&#x3089;&#x306E;&#x30C1;&#x30C3;&#x30D7;&#x306F;&#x3001;&#x795E;&#x7D4C;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3059;&#x308B;&#x5316;&#x5B66;&#x30D1;&#x30BF;&#x30FC;&#x30CB;&#x30F3;&#x30B0; &#x30BB;&#x30EB;&#x914D;&#x7F6E;&#x3068;&#x73FE;&#x5728;&#x306E;&#x751F;&#x7406;&#x30C7;&#x30FC;&#x30BF;&#x306E;&#x305F;&#x3081;&#x306E;&#x8A2D;&#x8A08;&#x30FB;&#x4F5C;&#x88FD;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306B;&#x3064;&#x3044;&#x3066;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3059;&#x3002;

&#x65B0;&#x898F;&#x5275;&#x85AC;&#x3078;&#x306E;&#x7D30;&#x80DE;&#x306E;&#x6A5F;&#x80FD;&#x7406;&#x89E3;&#x304B;&#x3089;: &#x5E73;&#x9762;&#x30D1;&#x30C3;&#x30C1;&#x30AF; &#x30A2;&#x30EC;&#x30A4; &#x30C1;&#x30C3;&#x30D7;&#x6280;&#x8853;&#x306E;&#x5F79;&#x5272;&#x3002;

&#xBAA8;&#xB4E0; &#xACE0;&#xB974;&#xAE30;&#xAC00; &#xC140; &#xAE30;&#xB2A5; &#xADF8;&#xB4E4;&#xC758; &#xC6D0;&#xD615;&#xC9C8; &#xB9C9;&#xC5D0; &#xD3EC;&#xD568; &#xB41C; &#xC774;&#xC628; &#xCC44;&#xB110;&#xC5D0; &#xC758;&#xC874;. &#xC774;&#xC628; &#xCC44;&#xB110; &#xAD6C;&#xC870; &#xB610;&#xB294; &#xD568;&#xC218; &#xBB3C;&#xACB0; &#xC2EC;&#xC7A5; &#xAE30;&#xB2A5; &#xC7A5;&#xC560;&#xC5D0;&#xC11C; &#xC2E0;&#xACBD; &#xD1F4;&#xD589; &#xC131; &#xC9C8;&#xD658;&#xC5D0; &#xC18D;&#xD558;&#xB294; &#xBCD1; &#xB9AC; &#xACB0;&#xACFC;. &#xB530;&#xB77C;&#xC11C;, &#xACE0;&#xB974;&#xAE30;&#xAC00; &#xC138;&#xD3EC;&#xC758; &#xAE30;&#xB2A5;&#xC744; &#xC774;&#xD574; &#xD558; &#xACE0; &#xADF8;&#xB4E4;&#xC758; &#xC774;&#xC0C1; &#xD574;&#xACB0; &#xD558;&#xAE30;-&#xC0C8;&#xB85C;&#xC6B4; &#xC57D;&#xBB3C; &#xD45C;&#xC801;&#xC5D0; &#xB300; &#xD55C; &#xB178;&#xCD9C;&#xC744; &#xD3EC;&#xD568; &#xD558; &#xC5EC; &#xB2E4;&#xC591; &#xD55C; &#xC2E4;&#xD5D8; &#xC870;&#xAC74;&#xC5D0;&#xC11C; &#xC774;&#xC628; &#xCC44;&#xB110; &#xAE30;&#xB2A5;&#xC744; &#xC774;&#xD574; &#xD558;&#xB294; &#xAC83;&#xC774; &#xC911;&#xC694; &#xD558;&#xB2E4;. &#xC720;&#xB9AC; &#xD53C; &#xD3AB; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xB274;&#xB7F0;&#xC758; &#xAE30;&#xBCF8; &#xBC0F; &#xC2DC; &#xB0C5; &#xC2A4; &#xC18D;&#xC131;&#xC744; &#xBAA8;&#xB2C8;&#xD130;&#xB9C1; &#xD558;&#xB824;&#xBA74; &#xCCA8;&#xB2E8; &#xAE30;&#xC220;&#xC785;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;,&#xC774; &#xBC29;&#xBC95;&#xC740; &#xB178;&#xB3D9; &#xC9D1;&#xC57D;&#xC801;&#xC778; &#xD558;&#xBA70; &#xB0AE;&#xC740; &#xB370;&#xC774;&#xD130; &#xCC98;&#xB9AC;&#xB7C9;&#xC774;. &#xB192;&#xC740; &#xCC98;&#xB9AC;&#xB7C9; &#xD558;&#xC9C0;&#xB9CC; &#xC81C;&#xD55C; &#xB429;&#xB2C8;&#xB2E4; &#xACA9;&#xB9AC; &#xB41C; &#xC140;&#xC5D0; &#xB530;&#xB77C;&#xC11C; &#xC0DD;&#xB9AC; &#xAE30;&#xB2A5; &#xBAA8;&#xB378;&#xB9C1;&#xC5D0; &#xADF8;&#xB4E4;&#xC758; &#xC0AC;&#xC6A9;&#xC744; &#xC81C;&#xD55C;, &#xC815;&#xC9C0;&#xC5D0;&#xC11C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xD3C9;&#xBA74; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xCE69;, &#xC790;&#xB3D9;&#xD654; &#xC2DC;&#xC2A4;&#xD15C;&#xC5D0; &#xD1B5;&#xD569;. &#xC774;&#xB7EC;&#xD55C; &#xCE69;&#xC740; &#xB530;&#xB77C;&#xC11C; &#xAC00;&#xC7A5; &#xC2E0;&#xACBD; &#xD1B5;&#xC2E0; &#xAD00;&#xB828; &#xC5F0;&#xAD6C;&#xC5D0; &#xC801;&#xD569; &#xD558;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. Multielectrode &#xBC30;&#xC5F4; (MEAs) &#xBC18;&#xBA74;, &#xC5EC;&#xB7EC; &#xC138;&#xD3EC; &#xC678; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0;&#xC11C; &#xB85C;&#xCEEC; &#xD544;&#xB4DC; &#xC804;&#xC704;&#xB97C; &#xCE21;&#xC815; &#xD558; &#xC5EC; &#xB124;&#xD2B8;&#xC6CC;&#xD06C; &#xD65C;&#xB3D9;&#xC744; &#xBAA8;&#xB2C8;&#xD130;&#xB9C1; &#xD558;&#xB294; &#xAE30;&#xB2A5;&#xC744;&#xAC00;&#xC9C0;&#xACE0; &#xD558;&#xC9C0;&#xB9CC; &#xD2B9;&#xC815; &#xC774;&#xC628; &#xCC44;&#xB110; &#xD65C;&#xB3D9;&#xC740; &#xC774;&#xB7EC;&#xD55C; &#xB2E4;&#xC911;&#xD654; &#xB41C; &#xC2E0;&#xD638;&#xC5D0;&#xC11C; &#xCD94;&#xCD9C;&#xD560; &#xB3C4;&#xC804; &#xC774;&#xB2E4;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xAC00; &#xB530;&#xB77C;&#xC11C; MEA &#xBC0F; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xAE30;&#xBC95;&#xC758; &#xC7A5;&#xC810;&#xC774; &#xACB0;&#xD569; synaptically &#xC5F0;&#xACB0; &#xB41C; &#xC2E0;&#xACBD; &#xB124;&#xD2B8;&#xC6CC;&#xD06C;&#xC5D0;&#xC11C; &#xC5EC;&#xB7EC; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0;&#xC11C; &#xAC1C;&#xBCC4; &#xB274;&#xB7F0;&#xC758; &#xB3D9;&#xC2DC; &#xACE0;&#xD574;&#xC0C1;&#xB3C4; electrophysiological &#xC2EC;&#xBB38; &#xC218; &#xC788;&#xB294; &#xC0C8;&#xB85C;&#xC6B4; &#xD3C9;&#xBA74; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xCE69; &#xAE30;&#xC220;&#xC744; &#xC124;&#xBA85; &#xD569;&#xB2C8;&#xB2E4;. &#xAC01; &#xB274;&#xB7F0; &#xC804;&#xC6A9;&#xB41C; &#xC9C0; &#xD558; &#xBBF8;&#xC138; &#xCC44;&#xB110;&#xC5D0; &#xC5F0;&#xACB0; &#xD558;&#xB294; &#xC870;&#xB9AC;&#xAC1C;&#xB97C; &#xD1B5;&#xD574; &#xD0D0;&#xC0C9;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB274;&#xB7F0; &#xB124;&#xD2B8;&#xC6CC;&#xD06C;&#xC5D0;&#xC11C; &#xC131;&#xC7A5; &#xD558; &#xC5EC; apertures physisorbed &#xB610;&#xB294; chemisorbed &#xD654;&#xD559; &#xC2E0;&#xD638;&#xC5D0; &#xC758;&#xD574; &#xC815;&#xB82C; &#xB429;&#xB2C8;&#xB2E4;. &#xC774; &#xAC80;&#xD1A0; &#xC774;&#xB7EC;&#xD55C; &#xCE69;, &#xACBD;&#xC791;&#xB41C; &#xD55C; &#xC2E0;&#xACBD; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xD654;&#xD559; &#xD328;&#xD134;&#xD654; &#xC140; &#xBC30;&#xCE58;&#xC640; &#xD604;&#xC7AC;&#xC758; &#xC0DD;&#xB9AC; &#xC801; &#xB370;&#xC774;&#xD130;&#xC5D0; &#xB300; &#xD55C; &#xC811;&#xADFC;&#xC758; &#xC124;&#xACC4; &#xBC0F; &#xC81C;&#xC870; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC5D0; &#xC124;&#xBA85; &#xD569;&#xB2C8;&#xB2E4;.

&#xC18C;&#xC124; &#xC57D;&#xBB3C; &#xBC1C;&#xACAC;&#xC5D0; &#xD734;&#xB300; &#xC804;&#xD654; &#xAE30;&#xB2A5;&#xC744; &#xC774;&#xD574;: &#xD3C9;&#xBA74; &#xD328;&#xCE58; &#xD074;&#xB7A8;&#xD504; &#xBC30;&#xC5F4; &#xCE69; &#xAE30;&#xC220;&#xC758; &#xC5ED;&#xD560;.

Todas las funciones de la c&eacute;lula excitable dependen de los canales i&oacute;nicos que se encajan en su membrana plasm&aacute;tica. Las perturbaciones de la estructura del canal del ion o funci&oacute;n producir patolog&iacute;as que van desde la disfunci&oacute;n cardiaca a trastornos neurodegenerativos. Por lo tanto, para entender las funciones de las c&eacute;lulas excitables y para remediar su patofisiolog&iacute;a, es importante entender las funciones de canales de iones bajo diferentes condiciones experimentales - incluida la exposici&oacute;n a los objetivos de la nueva droga. Pipeta de vidrio patch-clamp es la t&eacute;cnica de vanguardia para controlar las propiedades intr&iacute;nsecas y sin&aacute;pticas de las neuronas. Sin embargo, esta t&eacute;cnica es laborioso y tiene bajo rendimiento. Fichas de patch-clamp planar, integrados en sistemas automatizados, ofrecen altos rendimientos pero se limitan a c&eacute;lulas aisladas de suspensiones, lo que limita su uso en la funci&oacute;n fisiol&oacute;gica de modelado. Estos chips no son, por lo tanto, m&aacute;s convenientes para estudios que incluyeron la comunicaci&oacute;n neuronal. Micropozo-microcanal arrays (MEAs), por el contrario, tener la capacidad para monitorear la actividad de la red mediante la medici&oacute;n de potenciales de campo local desde m&uacute;ltiples sitios extracelulares, pero la actividad del canal del ion espec&iacute;fico es dif&iacute;cil extraer de estas se&ntilde;ales multiplexadas. Aqu&iacute; describimos una tecnolog&iacute;a de chip de novela planar de la abrazadera del remiendo-que permite el interrogatorio electrofisiol&oacute;gico alta resoluci&oacute;n simult&aacute;neo de neuronas individuales en m&uacute;ltiples sitios en redes neuronales CCIS conectadas, combinando as&iacute; las ventajas de t&eacute;cnicas MEA y patch-clamp. Cada neurona puede ser explorado a trav&eacute;s de una abertura que se conecta a un canal dedicado microflu&iacute;dicos subterr&aacute;neo. Creciendo en redes de neuronas est&aacute;n alineadas con las aberturas por physisorbed o chemisorbed se&ntilde;ales qu&iacute;micas. En esta revisi&oacute;n, se describe el proceso de dise&ntilde;o y fabricaci&oacute;n de estos chips, enfoques patrones qu&iacute;micos para la colocaci&oacute;n de la c&eacute;lula y presentes datos fisiol&oacute;gicos de los cultivos de c&eacute;lulas neuronales.

Comprender la funci&oacute;n celular al descubrimiento de nuevos f&aacute;rmacos: el papel de la tecnolog&iacute;a de chip de matriz plana patch-clamp.

We study the effect of polymerization on the nanomechanical stability of supported lipid monolayers consisting of 1,2-di-(10Z,12Z-tricosadiynoyl)-sn-glycero-3-phosphocholine by means of force mapping using an atomic force microscope. For both nonpolymerized and polymerized lipid monolayers, we investigate the break-through forces required to rupture the monolayers for a whole range of loading velocities. We show that the average break-through force exerted by the tip and required to penetrate the monolayer has a logarithmic dependence on the loading rate. Both Young moduli and intrinsic Gibbs energies have been determined for the nonpolymerized and polymerized lipid monolayers, and we show a drastic effect of polymerization on the nanomechanical stability of the monolayer with an increase by a factor of ∼100 for the young modulus and ∼3 for the intrinsic Gibbs activation energy.

Supported lipid monolayer with improved nanomechanical stability: effect of polymerization.

The purposes of the present study were to (1) assess the effect of the addition of benzalkonium chloride to sodium hypochlorite on its wetting properties, contact angle, and surface energy; (2) determine the critical micellar concentration of benzalkonium chloride in sodium hypochlorite; and (3) investigate the influence of addition of benzalkonium chloride on the free chlorine level, cytotoxicity, and antiseptic properties of the mixture.

Wetting properties and critical micellar concentration of benzalkonium chloride mixed in sodium hypochlorite.

We present a simple cost-effective benchtop protocol to functionalize glass and polydimethylsiloxane (PDMS) with nanometric protein patches for cell adhesion studies. Evaporation masks, covering macroscopic areas on glass, were made using improved strategies for self-assembly of colloidal microbeads which then served as templates for creating the protein patch arrays via the intermediate steps of organo-aminosilane deposition and polyethylene-glycol grafting. The diameter of the patches could be varied down to about 80 nm. The glass substrates were used for advanced optical imaging of T-lymphocytes to explore adhesion by reflection interference contrast microscopy and the possible colocalization of T-cell receptor microclusters and the activating protein patches by total internal reflection fluorescence microscopy. The selectively functionalized glass could also serve as template for transferring the protein nanopatches to the surface of a soft elastomer. We demonstrated successful reverse contact printing onto the surface of thin layers of PDMS with stiffness ranging from 30 KPa to 3 MPa.

Nanometric protein-patch arrays on glass and polydimethylsiloxane for cell adhesion studies.

Field effect transistors have risen as one of the most promising techniques in the development of biomedical diagnosis and monitoring. In such devices, the sensitivity and specificity of the sensor rely on the properties of the active sensing layer (gate dielectric and probe layer). We propose here a new type of transistor developed for the detection of Fe(3+) ions in which this sensing layer is made of a monolayer of lipids, engineered in such a way that it is not sensitive to pH in the acidic range, therefore making the device perfectly suitable for biomedical diagnosis. Probes are γ-pyrone derivatives that have been grafted to the lipid headgroups. Affinity constants derived for the chelator/Fe(3+) complexation as well as for other ions demonstrate very high sensitivity and specificity towards ferric ions with values as high as 5.10(10) M and a detected concentration as low as 50 fM.

A field effect transistor biosensor with a γ-pyrone derivative engineered lipid-sensing layer for ultrasensitive Fe3+ ion detection with low pH interference.

Controlling the deformation of microcapsules and capsules is essential in numerous biomedical applications. The mechanical properties of the membrane of microcapsules made of cross-linked human serum albumin (HSA) are revealed by two complementary experiments in the linear elastic regime. The first provides the surfacic shear elastic modulus Gs by the study of small deformations of a single capsule trapped in an elongational flow: Gs varies from 0.002 to 5 N m(-1). The second gives the volumic Young's modulus E of the membrane by shallow and local indentations of the membrane with an AFM probe: E varies from 20 kPa to 1 MPa. The surfacic and volumic elastic moduli increase with the size of the capsule up to three orders of magnitude and with the protein concentration of the membrane. The membrane thickness is evaluated from these two membrane mechanical characteristics and increases with the size and the initial HSA concentration from 2 to 20 μm.

Mechanical characterization of cross-linked serum albumin microcapsules.

Arrays of protein nanodots with dot-size tuned independently of spacing (e.g., ∼100 to 600 nm diameter for 900 nm spacing) are fabricated. The mechanism of size control is demonstrated, by numerical simulations, to arise from shadow effects during deposition of a sacrificial metal mask. We functionalize the nanodots with antibodies and embed them in a polymer-cushion or in lipid-bilayers or transfer them to soft elastomers. Their ability to influence cell architecture and local membrane organization is demonstrated in T-lymphocytes, using reflection interference contrast and total internal reflection fluorescence microscopy.

Size-Tunable Organic Nanodot Arrays: A Versatile Platform for Manipulating and Imaging Cells.

We report herein the fabrication of novel microarrays based on air-stable functional lipid monolayers over silicon using a combination of e-beam lithography and lift-off. We demonstrate these microarrays can be use as ultrasensitive platform for Kelvin probe force microscopy in sensing experiments. Specificity of the detection is given by the functional group grafted at the lipid headgroup. The arrays developed for the detection of ferric ions, Fe(3+), using a γ-pyrone derivative chelator, demonstrate subpicomolar limit of detection with high specificity. In addition, the technique takes advantage of the structure of the array with the silicon areas playing the role of reference for the measurement, and we determine critical pattern dimensions below which the probe size/shape impacts the measured results.

Anne Charrier

Aix-Marseille Université

CNRS, UMR 7325, CINaM

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Anne Charrier

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Aix-Marseille Université

CNRS, UMR 7325, CINaM

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Aix-Marseille Université