perfusable-vascular-network-with-tissue-model-microfluidic

Perfusable Vascular Network with a Tissue Model in a Microfluidic Device

A spheroid (a multicellular aggregate) is regarded as a good model of living tissues in the human body. Despite the significant advancement in the ...

Kyoto University

&#x672C;&#x6587;&#x63D0;&#x51FA;&#x4E86;&#x4E24;&#x4E2A;&#x6052;&#x5B9A;&#x6D41;&#x6295;&#x5165;&#x81EA;&#x4E3B;&#x8F6C;&#x6362;&#x4EA4;&#x66FF;&#x7684;&#x632F;&#x8361;&#x6D41;&#x8F93;&#x51FA;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x8BBE;&#x5907;&#x3002;&#x6211;&#x4EEC;&#x5B8C;&#x6210;&#x6B64;&#x5D4C;&#x5165;&#x5F0F;&#x786C;&#x4EF6;&#x81EA;&#x6211;&#x63A7;&#x5236;&#x7F16;&#x7A0B;&#x4F7F;&#x7528;&#x901A;&#x5E38;&#x5C01;&#x95ED;&#x7684;&#x819C;&#x9600;&#x6709;&#x8FDB;&#x6C14;&#x53E3;&#xFF0C;&#x51FA;&#x6C14;&#x53E3;&#xFF0C;&#x548C;&#x4E00;&#x4E2A;&#x819C;&#x52A0;&#x538B;&#x5206;&#x5EAD;&#x8FDE;&#x63A5;&#x5230;&#x7B2C;&#x4E09;&#x4E2A;&#x7AEF;&#x5B50;&#x3002;&#x8C03;&#x6574;&#x7684;&#x9608;&#x503C;&#x5F00;&#x653E;&#x538B;&#x529B;&#x5F00;&#x5173;&#x9600;&#x8FD9;&#x4E9B; 3 &#x7EC8;&#x7AEF;&#x6D41;&#x4E2D;&#x542F;&#x7528;&#x632F;&#x8361;&#x5468;&#x671F;&#x5230; 57 &#x548C; 360 s &#x4E0E;&#x5DE5;&#x4F5C;&#x5468;&#x671F;&#x7684; 0.2-0.5 &#x4E4B;&#x95F4;&#x7684;&#x8C03;&#x6574;&#x3002;&#x8FD9;&#x4E9B;&#x503C;&#x662F;&#x5728;&#x76F8;&#x5BF9;&#x8F83;&#x597D;&#x7684;&#x534F;&#x8BAE;&#xFF0C;&#x4E0E;&#x7406;&#x8BBA;&#x503C;&#xFF0C;&#x4E3A;&#x4E00;&#x4E2A;&#x66F4;&#x52A0;&#x5E7F;&#x6CDB;&#x7684;&#x4E0D;&#x540C;&#x6CE2;&#x5F62;&#x632F;&#x8361;&#x7684;&#x5408;&#x7406;&#x8BBE;&#x8BA1;&#x63D0;&#x4F9B;&#x65B9;&#x5F0F;&#x3002;&#x6211;&#x4EEC;&#x8FD8;&#x663E;&#x793A;&#x4F7F;&#x7528;&#x8FD9;&#x4E9B;&#x632F;&#x8361;&#x5668;&#x6765;&#x6267;&#x884C;&#x4E16;&#x4FD7;&#x5730;&#x56FE;&#x6848;&#x7684;&#x4EA4;&#x4ED8;&#x6D3B;&#x7EC6;&#x80DE;&#x7684;&#x5316;&#x5B66;&#x7269;&#x8D28;&#x7684;&#x80FD;&#x529B;&#x3002;&#x8BE5;&#x8BBE;&#x5907;&#x53EA;&#x9700;&#x8981;&#x4E00;&#x4E2A;&#x6CE8;&#x5C04;&#x5668;&#x6CF5;&#xFF0C;&#x4ECE;&#x800C;&#x6D88;&#x9664;&#x590D;&#x6742;&#x3001; &#x6602;&#x8D35;&#x7684;&#x5916;&#x90E8;&#x6267;&#x884C;&#x5668;&#x7684;&#x4F7F;&#x7528;&#x3002;&#x8FD9;&#x4E9B;&#x53EF;&#x8C03;&#x8C10;&#x6CE2;&#x5F62;&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x88AB;&#x8BBE;&#x60F3;&#xFF0C;&#x4EE5;&#x4FBF;&#x8FDB;&#x884C;&#x57FA;&#x4E8E;&#x5355;&#x5143;&#x683C;&#x7684;&#x7814;&#x7A76;&#x9700;&#x8981;&#x4E16;&#x4FD7;&#x7684;&#x523A;&#x6FC0;&#x3002;

&#x6052;&#x5B9A;&#x6D41;&#x9A71;&#x52A8;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x7684;&#x4E0D;&#x540C;&#x5DE5;&#x4F5C;&#x5468;&#x671F;&#x3002;

This paper presents microfluidic devices that autonomously convert two constant flow inputs into an alternating oscillatory flow output. We accomplish this hardware embedded self-control programming using normally closed membrane valves that have an inlet, an outlet, and a membrane-pressurization chamber connected to a third terminal. Adjustment of threshold opening pressures in these 3-terminal flow switching valves enabled adjustment of oscillation periods to between 57 and 360 s with duty cycles of 0.2-0.5. These values are in relatively good agreement with theoretical values, providing the way for rational design of an even wider range of different waveform oscillations. We also demonstrate the ability to use these oscillators to perform temporally patterned delivery of chemicals to living cells. The device only needs a syringe pump, thus removing the use of complex, expensive external actuators. These tunable waveform microfluidic oscillators are envisioned to facilitate cell-based studies that require temporal stimulation.

Constant flow-driven microfluidic oscillator for different duty cycles.

Cet article pr&eacute;sente les dispositifs microfluidiques autonome de convertir deux entr&eacute;es de d&eacute;bit constant en une sortie d&#39;&eacute;coulement oscillatoire alternatif. Nous accomplissons cette programmation de ma&icirc;trise de soi de mat&eacute;riel incorpor&eacute; &agrave; l&#39;aide de soupapes de membrane normalement ferm&eacute; disposant d&#39;une entr&eacute;e, une sortie, et une chambre de membrane-pressurisation reli&eacute; &agrave; un troisi&egrave;me terminal. R&eacute;glage du seuil d&#39;ouverture pressions dans ces flux de bornier &agrave; 3 valves de commutation activ&eacute; ajustement des p&eacute;riodes d&#39;oscillation &agrave; entre 57 et 360 s avec des cycles d&#39;utilisation de 0,2 &agrave; 0,5. Ces valeurs sont en assez bon accord avec les valeurs th&eacute;oriques, pr&eacute;voyant la fa&ccedil;on dont la conception rationnelle d&#39;&eacute;largir encore l&#39;&eacute;ventail des oscillations de forme d&#39;onde diff&eacute;rentes. Nous d&eacute;montrons &eacute;galement la possibilit&eacute; d&#39;utiliser ces oscillateurs pour effectuer la livraison dans le temps &agrave; motifs de produits chimiques aux cellules vivantes. L&#39;appareil doit uniquement un pousse-seringue, &eacute;liminant ainsi l&#39;utilisation d&#39;actionneurs externes complexes et co&ucirc;teuses. Ces oscillateurs de microfluidique de forme d&#39;onde accordable sont envisag&eacute;es pour faciliter les &eacute;tudes sur les cellules qui ont besoin de stimulation temporelle.

Oscillateur microfluidique de pilot&eacute;e par le flux constant de cycles d&#39;utilisation diff&eacute;rents.

Questo libro presenta dispositivi microfluidici che convertono autonomamente due ingressi di flusso costante in un&#39;uscita di flusso oscillatorio alternato. Compiamo questa programmazione di autocontrollo hardware embedded utilizzando valvole a membrana normalmente chiuso che dispongono di un ingresso, un&#39;uscita, e una camera a membrana-pressurizzazione collegato ad un terminale di terzo. Regolazione della soglia di apertura pressioni in questi 3-terminale flusso valvole di commutazione attivato regolazione dei periodi di oscillazione a tra 57 e 360 s con duty cycle di 0,2-0,5. Questi valori sono in accordo con i valori teorici, che fornisce il modo per progettazione razionale di una gamma ancora pi&ugrave; ampia delle oscillazioni diverse forme d&#39;onda relativamente. Abbiamo anche dimostrare la capacit&agrave; di utilizzare questi oscillatori per eseguire temporaneamente fantasia consegna di prodotti chimici per le cellule viventi. Il dispositivo ha solo bisogno di una pompa a siringa, eliminando cos&igrave; l&#39;uso di complessi e costosi attuatori esterni. Questi oscillatori di microfluidica onda sintonizzabile sono immaginati per facilitare gli studi basati su celle che richiedono la stimolazione temporale.

Costante flusso-driven microfluidica oscillatore per cicli di lavoro diversi.

&#x81EA;&#x5F8B;&#x7684; 2 &#x3064;&#x306E;&#x4E00;&#x5B9A;&#x306E;&#x6D41;&#x308C;&#x5165;&#x529B;&#x3001;&#x4EA4;&#x4E92;&#x306E;&#x6D41;&#x91CF;&#x51FA;&#x529B;&#x306B;&#x5909;&#x63DB;&#x3059;&#x308B;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x3092;&#x63D0;&#x6848;&#x3059;&#x308B;.&#x6211;&#x3005; &#x306F;&#x30B3;&#x30F3;&#x30BB;&#x30F3;&#x30C8;&#x306B;&#x5165;&#x53E3;&#x304C;&#x3042;&#x308B;&#x901A;&#x5E38;&#x30AF;&#x30ED;&#x30FC;&#x30BA;&#x30C9;&#x819C;&#x5F01;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3053;&#x306E;&#x30CF;&#x30FC;&#x30C9;&#x30A6;&#x30A7;&#x30A2;&#x57CB;&#x3081;&#x8FBC;&#x307E;&#x308C;&#x305F;&#x81EA;&#x5DF1;&#x30D7;&#x30ED;&#x30B0;&#x30E9;&#x30DF;&#x30F3;&#x30B0;&#x3092;&#x9054;&#x6210;&#x3057;&#x3001;&#x819C;&#x52A0;&#x5727;&#x5BA4;&#x7B2C; 3 &#x30BF;&#x30FC;&#x30DF;&#x30CA;&#x30EB;&#x306B;&#x63A5;&#x7D9A;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x8ABF;&#x6574;&#x958B;&#x53E3;&#x5727;&#x529B;&#x30D0;&#x30EB;&#x30D6;&#x3092;&#x5207;&#x308A;&#x66FF;&#x3048;&#x308B;&#x3053;&#x308C;&#x3089; 3 &#x30BF;&#x30FC;&#x30DF;&#x30CA;&#x30EB; &#x30D5;&#x30ED;&#x30FC;&#x306E;&#x3057;&#x304D;&#x3044;&#x5024;&#x306E;&#x767A;&#x632F;&#x671F;&#x9593;&#x306B; 57 &#x3068; 360 s 0.2 &#xFF5E; 0.5 &#x306E;&#x30C7;&#x30E5;&#x30FC;&#x30C6;&#x30A3; &#x30B5;&#x30A4;&#x30AF;&#x30EB;&#x3092;&#x6301;&#x3064;&#x9593;&#x306E;&#x8ABF;&#x6574;&#x304C;&#x6709;&#x52B9;&#x306B;&#x306A;&#x3063;&#x3066;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x5024;&#x306F;&#x7570;&#x306A;&#x308B;&#x6CE2;&#x5F62;&#x632F;&#x52D5;&#x306E;&#x3088;&#x308A;&#x5E83;&#x3044;&#x7BC4;&#x56F2;&#x306E;&#x5408;&#x7406;&#x7684;&#x306A;&#x8A2D;&#x8A08;&#x306E;&#x305F;&#x3081;&#x306E;&#x65B9;&#x6CD5;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x3001;&#x7406;&#x8AD6;&#x5024;&#x3068;&#x6BD4;&#x8F03;&#x7684;&#x3088;&#x304F;&#x4E00;&#x81F4;&#x3067;&#x3059;&#x3002;&#x307E;&#x305F;&#x3053;&#x308C;&#x3089;&#x306E;&#x767A;&#x632F;&#x5668;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x7D30;&#x80DE;&#x3078;&#x5316;&#x5B66;&#x7269;&#x8CEA;&#x306E;&#x6642;&#x9593;&#x7684;&#x30D1;&#x30BF;&#x30FC;&#x30F3;&#x306E;&#x914D;&#x4FE1;&#x3092;&#x5B9F;&#x884C;&#x3059;&#x308B;&#x80FD;&#x529B;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306B;&#x306F;&#x3001;&#x3053;&#x306E;&#x3088;&#x3046;&#x306B;&#x8907;&#x96D1;&#x306A;&#x9AD8;&#x4FA1;&#x306A;&#x5916;&#x90E8;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30E5;&#x30A8;&#x30FC;&#x30BF;&#x306E;&#x4F7F;&#x7528;&#x3092;&#x9664;&#x53BB;&#x3059;&#x308B;&#x30B7;&#x30EA;&#x30F3;&#x30B8; &#x30DD;&#x30F3;&#x30D7;&#x3001;&#x3060;&#x3051;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x53EF;&#x5909;&#x6CE2;&#x5F62;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x306E;&#x6642;&#x9593;&#x7684;&#x306A;&#x523A;&#x6FC0;&#x3092;&#x5FC5;&#x8981;&#x3068;&#x3059;&#x308B;&#x7D30;&#x80DE;&#x306B;&#x57FA;&#x3065;&#x304F;&#x7814;&#x7A76;&#x3092;&#x4FC3;&#x9032;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x60F3;&#x5B9A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;

&#x3055;&#x307E;&#x3056;&#x307E;&#x306A;&#x30C7;&#x30E5;&#x30FC;&#x30C6;&#x30A3; &#x30B5;&#x30A4;&#x30AF;&#x30EB;&#x7528;&#x306E;&#x4E00;&#x5B9A;&#x306E;&#x6D41;&#x308C;&#x3092;&#x99C6;&#x52D5;&#x3055;&#x308C;&#x308B;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x3002;

&#xC774; &#xBB38;&#xC11C;&#xC5D0;&#xC11C;&#xB294; &#xC790;&#xC728;&#xC801;&#xC73C;&#xB85C; 2 &#xAC1C;&#xC758; &#xC77C;&#xC815; &#xD55C; &#xAD50;&#xB958; &#xC785;&#xB825; &#xAD50;&#xB958; &#xCD95;&#xC804;&#xAE30; &#xD750;&#xB984; &#xCD9C;&#xB825;&#xC73C;&#xB85C; &#xBCC0;&#xD658; &#xD558;&#xB294; &#xBBF8;&#xC138; &#xC7A5;&#xCE58; &#xC124;&#xBA85;. &#xC6B0;&#xB9AC;&#xB294; &#xC785;&#xAD6C;, &#xCD9C;&#xAD6C;, &#xD3C9;&#xC0C1;&#xC2DC; &#xB9C9; &#xBC38;&#xBE0C;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC784;&#xBCA0;&#xB514;&#xB4DC; &#xD558;&#xB4DC;&#xC6E8;&#xC5B4; &#xC790;&#xAE30; &#xD504;&#xB85C;&#xADF8;&#xB798;&#xBC0D;&#xC774; &#xC218;&#xD589; &#xD558; &#xACE0; &#xB9C9; &#xAC00;&#xC555; &#xCC54;&#xBC84; &#xC81C; 3 &#xD130;&#xBBF8;&#xB110;&#xC5D0; &#xC5F0;&#xACB0;. &#xC870;&#xC815; &#xC784;&#xACC4;&#xAC12;&#xC774; 3 &#xD130;&#xBBF8;&#xB110; &#xD750;&#xB984; &#xC804;&#xD658; &#xBC38;&#xBE0C;&#xC5D0;&#xC11C;&#xC5D0;&#xC11C; &#xC555;&#xB825; &#xC5EC;&#xC758; 0.2-0.5&#xC758; &#xB4C0;&#xD2F0; &#xC0AC;&#xC774;&#xD074; 57, 360 s &#xC0AC;&#xC774; &#xC9C4;&#xB3D9; &#xAE30;&#xAC04;&#xC758; &#xC870;&#xC815; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xAC12;&#xC740; &#xC774;&#xB860;&#xC801;&#xC778; &#xAC12;, &#xC11C;&#xB85C; &#xB2E4;&#xB978; &#xD30C;&#xD615; &#xC9C4;&#xB3D9;&#xC758; &#xB354; &#xB113;&#xC740; &#xBC94;&#xC704;&#xC758; &#xD569;&#xB9AC;&#xC801;&#xC778; &#xB514;&#xC790;&#xC778;&#xC5D0; &#xB300; &#xD55C; &#xBC29;&#xBC95;&#xC744; &#xC81C;&#xACF5;&#xACFC; &#xC0C1;&#xB300;&#xC801;&#xC73C;&#xB85C; &#xC88B;&#xC740; &#xACC4;&#xC57D;. &#xC6B0;&#xB9AC; &#xB610;&#xD55C;&#xC774; &#xBC1C;&#xC9C4;&#xAE30;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC0B4;&#xC544;&#xC788;&#xB294; &#xC138;&#xD3EC;&#xC5D0; &#xD654;&#xD559; &#xBB3C;&#xC9C8;&#xC758; &#xC77C;&#xC2DC;&#xC801;&#xC73C;&#xB85C; &#xD328;&#xD134;&#xD654; &#xB41C; &#xB0A9;&#xD488;&#xC744; &#xC218;&#xD589; &#xD558;&#xB294; &#xAE30;&#xB2A5;&#xC744; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xC7A5;&#xCE58;&#xB9CC; &#xC8FC;&#xC0AC;&#xAE30; &#xD38C;&#xD504;, &#xBCF5;&#xC7A1; &#xD558; &#xACE0; &#xBE44;&#xC2FC; &#xC678;&#xBD80; &#xC561;&#xCE04;&#xC5D0;&#xC774;&#xD130;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC81C;&#xAC70; &#xD574;&#xC57C; &#xD569;&#xB2C8;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xAC00;&#xBCC0; &#xD30C;&#xD615; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30;&#xB294; &#xC77C;&#xC2DC;&#xC801;&#xC778; &#xC790;&#xADF9;&#xC744; &#xD544;&#xC694;&#xB85C; &#xD558;&#xB294; &#xC138;&#xD3EC; &#xAE30;&#xBC18; &#xC5F0;&#xAD6C;&#xB97C; &#xC6A9;&#xC774; &#xD558; &#xAC8C; &#xC0C1;&#xC0C1;.

&#xC11C;&#xB85C; &#xB2E4;&#xB978; &#xB4C0;&#xD2F0; &#xC0AC;&#xC774;&#xD074;&#xC5D0; &#xB300; &#xD55C; &#xC9C0;&#xC18D;&#xC801;&#xC778; &#xD750;&#xB984; &#xAE30;&#xBC18; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30;.

Este papel presenta dispositivos microflu&iacute;dicos que aut&oacute;nomamente convierten dos entradas de flujo constante en una salida alterna de flujo oscilatorio. Logramos esta programaci&oacute;n de autocontrol de hardware incrustado utilizando v&aacute;lvulas de membrana normalmente cerrado que tienen una entrada, una salida, y una c&aacute;mara de presurizaci&oacute;n de membrana conectado a un terminal tercero. Ajuste del umbral de apertura presiones en estas v&aacute;lvulas de conmutaci&oacute;n de la corriente de 3 terminales hab&iacute;a habilitado ajuste de periodos de oscilaci&oacute;n entre 57 y 360 s con ciclos de trabajo de 0.2-0.5. Estos valores son relativamente buena de acuerdo con los valores te&oacute;ricos, prever la forma de dise&ntilde;o racional de un mayor rango de oscilaciones de onda diferentes. Tambi&eacute;n demostramos la capacidad de utilizar estos osciladores para realizar la entrega temporal con motivos de sustancias qu&iacute;micas a las c&eacute;lulas vivas. El dispositivo s&oacute;lo requiere una bomba de jeringa, eliminando as&iacute; el uso de actuadores externos complejos, costosos. Estos osciladores de onda sintonizable microflu&iacute;dicos se prev&eacute;n para facilitar los estudios celulares que requieren estimulaci&oacute;n temporal.

Oscilador de microfluidos basadas en el flujo constante de ciclos de trabajo diferentes.

A microfluidic device integrated with a Total Internal Reflection (TIR)-based chip for cell observation and analysis was developed. This integrated device enables in situ Total Internal Reflection Fluorescence Microscopy (TIRFM) on adherent cells cultured under continuous medium perfusion. This TIR-based chip, allows TIRFM to be easily performed on cells without the assembly of complicated optical components and cell culture chambers. The integrated device was evaluated by tracking the movement of fluorescent beads and monitoring the location of insulin granules in mouse pancreatic β-cells. This system offers higher signal-to-noise (S/N) ratio than epi-fluorescence microscopy (EPIFM), and comparable image quality to commercial TIRFM systems when imaging insulin granules. We also detected repetitive changes in intracellular Ca(2+) concentration in MIN6-m9 cells stimulated with KCl, which demonstrates quick perfusion for cell analysis while maintaining high S/N ratio.

A perfusable microfluidic device with on-chip total internal reflection fluorescence microscopy (TIRFM) for in situ and real-time monitoring of live cells.

&#x672C;&#x6587;&#x6982;&#x8FF0;&#x4E86;&#x5F39;&#x6027;&#x9600;&#x548C;&#x6DB2;&#x6EF4;-&#x57FA;&#x4E8E;&#x5FAE;&#x6D41;&#x63A7;&#x7CFB;&#x7EDF;&#x65E8;&#x5728;&#x5C3D;&#x91CF;&#x51CF;&#x5C11;&#x5916;&#x90E8;&#x538B;&#x529B;&#x7684;&#x63A7;&#x5236;&#x6D41;&#x4F53;&#x6D41;&#x52A8;&#x7684;&#x5FC5;&#x8981;&#x6027;&#x3002;&#x6B64;&#x6982;&#x5FF5;&#x6587;&#x7AE0;&#x4ECB;&#x7ECD;&#x6709;&#x4EE3;&#x8868;&#x6027;&#x7684;&#x7EC4;&#x6210;&#x90E8;&#x5206;&#xFF0C;&#x5728;&#x8FD9;&#x4E9B;&#x8BBE;&#x5907;&#x4EE5;&#x53CA;&#x76F8;&#x5173;&#x7684;&#x751F;&#x5316;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x7684;&#x5DE5;&#x4F5C;&#x539F;&#x7406;&#x3002;&#x7D27;&#x63A5;&#x7740;&#x7684;&#x662F;&#x63D0;&#x4F9B;&#x89C2;&#x70B9;&#x4E0D;&#x540C;&#x5FAE;&#x6D41;&#x63A7;&#x9600;&#x95E8;&#x548C;&#x7CFB;&#x7EDF;&#x901A;&#x8FC7;&#x6BD4;&#x8F83;&#x5176;&#x5F02;&#x540C;&#x53CA;&#x4E0E;&#x6676;&#x4F53;&#x7BA1; (&#x9600;&#x95E8;) &#x548C;&#x7CFB;&#x7EDF;&#x5728;&#x5FAE;&#x7535;&#x5B50;&#x65B9;&#x9762;&#x7684;&#x4F5C;&#x7528;&#x3002;&#x5C3D;&#x7BA1;&#x4ECE;&#x7535;&#x5B50;&#x7EBF;&#x8DEF;&#x8FDB;&#x884C;&#x7C7B;&#x6BD4;&#x7684;&#x4E00;&#x4E9B;&#x7269;&#x7406;&#x9650;&#x5236;&#xFF0C;&#x81EA;&#x52A8;&#x5316;&#x5FAE;&#x6D41;&#x63A7;&#x7535;&#x8DEF;&#x8BBE;&#x8BA1;&#x53EF;&#x4EE5;&#x83B7;&#x5F97;&#x4ECE;&#x7535;&#x5B50;&#x7EBF;&#x8DEF;&#x5C3D;&#x91CF;&#x51CF;&#x5C11;&#x5916;&#x90E8;&#x63A7;&#x5236;&#x5355;&#x4F4D;&#xFF0C;&#x540C;&#x65F6;&#x6267;&#x884C;&#x9AD8;&#x590D;&#x6742;&#x6027;&#x548C;&#x9AD8;&#x541E;&#x5410;&#x91CF;&#x7684;&#x5206;&#x6790;&#x89C1;&#x89E3;&#x3002;

&#x4F7F;&#x7528;&#x5F39;&#x6027;&#x9600;&#x548C;&#x6DB2;&#x6EF4;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x81EA;&#x52A8;&#x5316;&#xFF1A; &#x964D;&#x4F4E;&#x5BF9;&#x5916;&#x90E8;&#x63A7;&#x5236;&#x5668;&#x7684;&#x4F9D;&#x8D56;&#x3002;

This paper gives an overview of elastomeric valve- and droplet-based microfluidic systems designed to minimize the need of external pressure to control fluid flow. This Concept article introduces the working principle of representative components in these devices along with relevant biochemical applications. This is followed by providing a perspective on the roles of different microfluidic valves and systems through comparison of their similarities and differences with transistors (valves) and systems in microelectronics. Despite some physical limitation of drawing analogies from electronic circuits, automated microfluidic circuit design can gain insights from electronic circuits to minimize external control units, while implementing high-complexity and high-throughput analysis.

Microfluidic automation using elastomeric valves and droplets: reducing reliance on external controllers.

Le pr&eacute;sent document donne un aper&ccedil;u des &eacute;lastom&egrave;res vanne et gouttelettes-microfluidique syst&egrave;mes con&ccedil;us pour minimiser la n&eacute;cessit&eacute; de pressions ext&eacute;rieures pour contr&ocirc;ler l&#39;&eacute;coulement du fluide. Cet article de Concept introduit le principe de fonctionnement des composants repr&eacute;sentatifs dans ces dispositifs ainsi que les applications biochimiques pertinentes. Il est suivi en fournissant un point de vue sur les r&ocirc;les de microfluidique diff&eacute;rentes valves et syst&egrave;mes gr&acirc;ce &agrave; la comparaison de leurs similitudes et diff&eacute;rences avec transistors (vannes) et en micro&eacute;lectronique. Malgr&eacute; une certaine limitation physique des analogies de circuits &eacute;lectroniques de dessin, conception de circuit microfluidique automatis&eacute; peut acqu&eacute;rir des connaissances de circuits &eacute;lectroniques pour r&eacute;duire au minimum les unit&eacute;s de contr&ocirc;le externe, mise en &oelig;uvre des analyses de grande complexit&eacute; et haut d&eacute;bit.

Microfluidique automation &agrave; l&#39;aide de vannes en &eacute;lastom&egrave;re et gouttelettes : r&eacute;duire la d&eacute;pendance &agrave; l&#39;&eacute;gard des contr&ocirc;leurs externes.

Questo documento offre una panoramica dei sistemi basati su valvola e gocciolina elastomerico microfluidica progettato per ridurre al minimo la necessit&agrave; di pressione esterna per controllare il flusso del fluido. Questo articolo di concetto introduce il principio di funzionamento dei componenti rappresentativi in questi dispositivi insieme ad applicazioni di biochimiche. Questa &egrave; seguita da fornire una prospettiva sui ruoli di microfluidica diverse valvole e sistemi nel campo della microelettronica e sistemi attraverso il confronto delle loro somiglianze e differenze con transistor (valvole). Nonostante alcune limitazioni fisiche di analogie da circuiti elettronici di disegno, disegno circuito automatizzato microfluidica pu&ograve; acquisire conoscenze da circuiti elettronici per ridurre al minimo di unit&agrave; di controllo esterna, mentre l&#39;attuazione di analisi ad alta complessit&agrave; e ad alta velocit&agrave;.

Microfluidica automazione utilizzando valvole elastomeriche e gocce: ridurre la dipendenza dai controllori esterni.

&#x672C;&#x30D0;&#x30EB;&#x30D6; &#x30FB;&#x6DB2;&#x6EF4;&#x30D9;&#x30FC;&#x30B9;&#x306E;&#x5F3E;&#x6027;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306E;&#x6D41;&#x4F53;&#x306E;&#x6D41;&#x308C;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x5916;&#x90E8;&#x304B;&#x3089;&#x306E;&#x5727;&#x529B;&#x306E;&#x5FC5;&#x8981;&#x6027;&#x3092;&#x6700;&#x5C0F;&#x9650;&#x306B;&#x3059;&#x308B;&#x3088;&#x3046;&#x306B;&#x8A2D;&#x8A08;&#x306E;&#x6982;&#x8981;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x4EE3;&#x8868;&#x7684;&#x306A;&#x30B3;&#x30F3;&#x30DD;&#x30FC;&#x30CD;&#x30F3;&#x30C8;&#x306B;&#x95A2;&#x9023;&#x3059;&#x308B;&#x751F;&#x5316;&#x5B66;&#x7684;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3068;&#x4E00;&#x7DD2;&#x306B;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306E;&#x52D5;&#x4F5C;&#x539F;&#x7406;&#x306F;&#x3053;&#x306E;&#x30B3;&#x30F3;&#x30BB;&#x30D7;&#x30C8;&#x3092;&#x7D39;&#x4ECB;&#x3057;&#x3066;&#x304F;&#x3060;&#x3055;&#x3044;&#x3002;&#x3053;&#x308C;&#x306F;&#x3001;&#x7570;&#x306A;&#x308B;&#x30DE;&#x30A4;&#x30AF;&#x30ED; &#x30D0;&#x30EB;&#x30D6;&#x3068;&#x306E;&#x985E;&#x4F3C;&#x70B9;&#x3068;&#x76F8;&#x9055;&#x30C8;&#x30E9;&#x30F3;&#x30B8;&#x30B9;&#x30BF; &#xFF08;&#x30D0;&#x30EB;&#x30D6;&#xFF09; &#x306E;&#x6BD4;&#x8F03;&#x3092;&#x901A;&#x3057;&#x3066;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3001;&#x30DE;&#x30A4;&#x30AF;&#x30ED; &#x30A8;&#x30EC;&#x30AF;&#x30C8;&#x30ED;&#x30CB;&#x30AF;&#x30B9;&#x306E;&#x5F79;&#x5272;&#x306B;&#x8996;&#x70B9;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x3053;&#x3068;&#x3067;&#x7D9A;&#x3044;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x96FB;&#x5B50;&#x56DE;&#x8DEF;&#x304B;&#x3089;&#x985E;&#x4F3C;&#x56F3;&#x9762;&#x306E;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x7269;&#x7406;&#x7684;&#x306A;&#x5236;&#x9650;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x3001;&#x81EA;&#x52D5;&#x5316;&#x3055;&#x308C;&#x305F;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x56DE;&#x8DEF;&#x8A2D;&#x8A08;&#x306F;&#x5916;&#x90E8;&#x5236;&#x5FA1;&#x30E6;&#x30CB;&#x30C3;&#x30C8;&#x3001;&#x9AD8;&#x8907;&#x96D1;&#x3055;&#x3068;&#x9AD8;&#x30B9;&#x30EB;&#x30FC;&#x30D7;&#x30C3;&#x30C8;&#x306E;&#x89E3;&#x6790;&#x3092;&#x5B9F;&#x88C5;&#x3059;&#x308B;&#x4E2D;&#x3092;&#x6700;&#x5C0F;&#x9650;&#x306B;&#x6291;&#x3048;&#x308B;&#x3057;&#x3001;&#x96FB;&#x5B50;&#x56DE;&#x8DEF;&#x304B;&#x3089;&#x306E;&#x6D1E;&#x5BDF;&#x529B;&#x3092;&#x5F97;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

&#x30A8;&#x30E9;&#x30B9;&#x30C8;&#x30DE;&#x30FC; &#x30D0;&#x30EB;&#x30D6;&#x3068;&#x6DB2;&#x6EF4;&#x3092;&#x7528;&#x3044;&#x305F;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30AA;&#x30FC;&#x30C8;&#x30E1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;: &#x5916;&#x90E8;&#x30B3;&#x30F3;&#x30C8; &#x30ED;&#x30FC;&#x30E9;&#x30FC;&#x3078;&#x306E;&#x4F9D;&#x5B58;&#x3092;&#x6E1B;&#x3089;&#x3059;&#x3053;&#x3068;&#x3002;

&#xC774; &#xC885;&#xC774; &#xD0C4;&#xC131; &#xBC38;&#xBE0C; &#xBC0F; &#xB4DC;&#xB86D;&#xB9BF; &#xAE30;&#xBC18; &#xBBF8;&#xC138; &#xC720;&#xCCB4; &#xD750;&#xB984;&#xC744; &#xC81C;&#xC5B4; &#xD558;&#xB294; &#xC678;&#xBD80; &#xC555;&#xB825;&#xC758; &#xD544;&#xC694;&#xB97C; &#xCD5C;&#xC18C;&#xD654; &#xD558;&#xB3C4;&#xB85D; &#xC124;&#xACC4; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xAC1C;&#xC694;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xAC1C;&#xB150; &#xAE30;&#xC0AC; &#xAD00;&#xB828; &#xC0DD;&#xD654;&#xD559; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xACFC; &#xD568;&#xAED8;&#xC774; &#xC7A5;&#xCE58;&#xC5D0;&#xC11C; &#xB300;&#xD45C;&#xC801;&#xC778; &#xBD80;&#xD488;&#xC758; &#xC791;&#xB3D9; &#xC6D0;&#xB9AC;&#xB97C; &#xC18C;&#xAC1C;&#xD569;&#xB2C8;&#xB2E4;. &#xC774;&#xAC83;&#xC740; &#xB2E4;&#xB978; &#xBBF8;&#xC138; &#xBC38;&#xBE0C; &#xBC0F; &#xADF8;&#xB4E4;&#xC758; &#xC720;&#xC0AC;&#xC810;&#xACFC; &#xCC28;&#xC774;&#xC810; &#xD2B8;&#xB79C;&#xC9C0;&#xC2A4;&#xD130; (&#xBC38;&#xBE0C;)&#xC640; &#xBE44;&#xAD50;&#xB97C; &#xD1B5;&#xD574; &#xC2DC;&#xC2A4;&#xD15C; &#xBC0F; &#xB9C8;&#xC774;&#xD06C;&#xB85C; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xC5ED;&#xD560;&#xC5D0; &#xB300; &#xD55C; &#xAD00;&#xC810;&#xC744; &#xC81C;&#xACF5; &#xD558; &#xC5EC; &#xB530; &#xB790; &#xB2E4;. &#xC804;&#xC790; &#xD68C;&#xB85C;&#xC5D0;&#xC11C; &#xC720;&#xCD94;&#xB97C; &#xADF8;&#xB9AC;&#xAE30;&#xC758; &#xC77C;&#xBD80; &#xBB3C;&#xB9AC;&#xC801; &#xD55C;&#xACC4;&#xC5D0;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0; &#xC790;&#xB3D9;&#xD654; &#xB41C; &#xBBF8;&#xC138; &#xD68C;&#xB85C; &#xB514;&#xC790;&#xC778; &#xB192;&#xC740; &#xBCF5;&#xC7A1;&#xC131; &#xBC0F; &#xB192;&#xC740; &#xCC98;&#xB9AC;&#xB7C9; &#xBD84;&#xC11D;&#xC744; &#xAD6C;&#xD604; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC678;&#xBD80; &#xC81C;&#xC5B4; &#xC7A5;&#xCE58;&#xB97C; &#xCD5C;&#xC18C;&#xD654; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC804;&#xC790; &#xD68C;&#xB85C;&#xC5D0;&#xC11C; &#xD1B5;&#xCC30;&#xB825;&#xC744; &#xC5BB;&#xC744; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xD0C4;&#xC131; &#xBC38;&#xBE0C;&#xC640; &#xBC29;&#xC6B8;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xBBF8;&#xC138; &#xC790;&#xB3D9;&#xD654;: &#xC678;&#xBD80; &#xCEE8;&#xD2B8;&#xB864;&#xB7EC;&#xC5D0; &#xB300; &#xD55C; &#xC758;&#xC874;&#xB3C4; &#xC904;&#xC774;&#xACE0;.

Este documento da una descripci&oacute;n de sistemas microflu&iacute;dicos elastom&eacute;rico v&aacute;lvula y gotita basada en dise&ntilde;ado para minimizar la necesidad de controlar el flujo de fluido de presi&oacute;n externa. Este art&iacute;culo del concepto introduce el principio de funcionamiento de componentes representativos de estos dispositivos junto con aplicaciones bioqu&iacute;micas pertinentes. Esto es seguido por proporcionar una perspectiva sobre las funciones de microfluidos diferentes v&aacute;lvulas y sistemas en microelectr&oacute;nica y sistemas a trav&eacute;s de la comparaci&oacute;n de sus similitudes y diferencias con los transistores (v&aacute;lvulas). A pesar de algunas limitaciones f&iacute;sicas de dibujo analog&iacute;as de circuitos electr&oacute;nicos, dise&ntilde;o de circuito microflu&iacute;dico automatizado puede hacerse una idea de circuitos electr&oacute;nicos para minimizar las unidades de control externo, ejecuci&oacute;n de an&aacute;lisis de alta complejidad y alto rendimiento.

Automatizaci&oacute;n de microfluidos usando v&aacute;lvulas elastom&eacute;ricas y gotas: reduce la dependencia de controladores externos.

The field of microfluidics has drastically contributed to downscale the size of benchtop experiments to the dimensions of a chip without compromising results. However, further miniaturization and the ability to directly manipulate individual molecules require a platform that permits organized molecular transport. The motor proteins and microtubules that carry out orderly intracellular transport are ideal for driving in vitro nanotransport. Here, we demonstrate that a reconstruction of the cellular kinesin/dynein-microtubule system in nanotracks provides a molecular total analysis system (MTAS) to control massively parallel chemical reactions. The mobility of kinesin and a microtubule dissociation method enable orientation of a microtubule in an array for directed transport of reactive molecules carried by kinesin or dynein. The binding of glutathione S-transferase (GST) to glutathione (GSH) and the binding of streptavidin to biotin are visualized as colocalizations of quantum dots (Q-dots) when motor motilities bring them into contact. The organized nanotransport demonstrated here suggests the feasibility of using our platform to perform parallel biochemical reactions focused at the molecular level.

Colocalization of quantum dots by reactive molecules carried by motor proteins on polarized microtubule arrays.

&#x672C;&#x6587;&#x63ED;&#x793A;&#x4E86;&#x4E00;&#x4E2A;&#x5FAE;&#x6D41;&#x63A7;&#x819C;&#x9600; (&mu;MV) &#x548C;&#x7535;&#x5B50;&#x6676;&#x4F53;&#x7BA1;&#x7684;&#x7535;&#x6DB2;&#x7C7B;&#x6BD4;&#x7684;&#x5173;&#x952E;&#x9650;&#x5236;&#x3002;&#x4E0E;&#x4E0D;&#x540C;&#x7684;&#x6253;&#x5F00;&#x548C;&#x5173;&#x95ED;&#x9608;&#x503C;&#x7535;&#x538B;&#xFF0C;&#x5728;&#x6DB2;&#x538B; &mu;MVs&#xFF0C;&#x6709;&#x7C7B;&#x4F3C;&#x7684;&#x5178;&#x578B;&#x6676;&#x4F53;&#x7BA1;&#x7528;&#x4E8E;&#x6253;&#x5F00;&#x548C;&#x5173;&#x95ED;&#x9608;&#x503C;&#x538B;&#x529B;&#x5927;&#x5927;&#x4E0D;&#x540C;&#xFF0C;&#x53EF;&#x4EE5;&#x66F4;&#x6539;&#xFF0C;&#x5373;&#x4F7F;&#x5BF9;&#x4E8E;&#x540C;&#x4E00; &mu;MVs &#x6839;&#x636E;&#x6574;&#x4F53;&#x7684;&#x7535;&#x8DEF;&#x8BBE;&#x8BA1;&#x548C;&#x64CD;&#x4F5C;&#x6761;&#x4EF6;&#x3002;&#x6211;&#x4EEC;&#x89E3;&#x91CA;&#xFF0C;&#x7279;&#x522B;&#x662F;&#xFF0C;&#x5982;&#x4F55;&#x5173;&#x95ED;&#x9608;&#x503C;&#x7684;&#x8D1F;&#x503C;&#x7684;&#x538B;&#x529B;&#x5927;&#x5927;&#x9650;&#x5236;&#x751A;&#x81F3;&#x7B80;&#x5355;&#x6DB2;&#x538B; &mu;MV &#x7535;&#x8DEF;&#x7B49;&#x81EA;&#x4E3B;&#x5207;&#x6362;&#x4E24;&#x4E2A;&#x9600;&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x7684;&#x64CD;&#x4F5C;&#x3002;&#x8FD9;&#x4E9B;&#x8C05;&#x89E3;&#x6709;&#x91CD;&#x5927;&#x7684;&#x5F71;&#x54CD;&#xFF0C;&#x5728;&#x8BBE;&#x8BA1;&#x81EA;&#x4E3B;&#x5FAE;&#x6D41;&#x63A7;&#x8BBE;&#x5907;&#x3002;

&#x5206;&#x6790;&#x5FAE;&#x6D41;&#x63A7;&#x6676;&#x4F53;&#x7BA1;&#x4E3A;&#x81EA;&#x4E3B;&#x5FAE;&#x6D41;&#x63A7;&#x7535;&#x8DEF;&#x7684;&#x9608;&#x503C;&#x538B;&#x529B;&#x9650;&#x5236;&#x3002;

This paper reveals a critical limitation in the electro-hydraulic analogy between a microfluidic membrane-valve (Î¼MV) and an electronic transistor. Unlike typical transistors that have similar on and off threshold voltages, in hydraulic Î¼MVs, the threshold pressures for opening and closing are significantly different and can change, even for the same Î¼MVs depending on overall circuit design and operation conditions. We explain, in particular, how the negative values of the closing threshold pressures significantly constrain operation of even simple hydraulic Î¼MV circuits such as autonomously switching two-valve microfluidic oscillators. These understandings have significant implications in designing self-regulated microfluidic devices.

Analyzing threshold pressure limitations in microfluidic transistors for self-regulated microfluidic circuits.

Ce document fait appara&icirc;tre une limite critique dans l&#39;analogie &eacute;lectro-hydraulique entre une valve &agrave; membrane microfluidiques (&mu;MV) et un transistor &eacute;lectronique. Contrairement aux transistors typiques qui ont similaires sur et en dehors des tensions de seuil, en &mu;MVs hydrauliques, la pression seuil d&#39;ouverture et de fermeture est significativement diff&eacute;rente et peut changer, m&ecirc;me pour les m&ecirc;mes &mu;MVs selon les conditions de conception et d&#39;exploitation de circuit globales. Nous expliquer, en particulier, comment les valeurs n&eacute;gatives du seuil de fermeture pressions consid&eacute;rablement limiter bon fonctionnement des circuits &mu;MV hydraulique m&ecirc;me simple telle que la commutation autonome deux soupapes microfluidique oscillateurs. Ces accords ont des implications importantes dans la conception de dispositifs microfluidiques autor&eacute;glement&eacute;e.

L&#39;analyse des limites de pression de seuil dans les transistors de microfluidique pour circuits microfluidiques autor&eacute;glement&eacute;e.

Questo libro rivela una limitazione critica nell&#39;analogia elettro-idraulico tra una valvola membrana microfluidica (&mu;MV) e un transistor elettronico. A differenza dei transistor tipiche che hanno simili e spegnere le tensioni di soglia, in &mu;MVs idraulico, le pressioni di soglia di apertura e chiusura sono significativamente differenti e possono cambiare, anche per la stessa &mu;MVs a seconda delle condizioni di progettazione e funzionamento circuito globale. Spieghiamo, in particolare, come i valori negativi della soglia chiusura pressioni notevolmente vincolare funzionamento di circuiti &mu;MV idraulico anche semplici come autonomamente oscillatori microfluidica due valvole di commutazione. Questi accordi hanno implicazioni significative nella progettazione di dispositivi microfluidici auto-regolato.

Analizzare i limiti soglia pressione microfluidica transistori per circuiti microfluidici auto-regolato.

&#x3053;&#x306E;&#x8AD6;&#x6587;&#x3067;&#x306F;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x819C;&#x5F01; &#xFF08;&mu;MV&#xFF09; &#x304A;&#x3088;&#x3073;&#x96FB;&#x5B50;&#x30C8;&#x30E9;&#x30F3;&#x30B8;&#x30B9;&#x30BF;&#x306E;&#x96FB;&#x6C17;&#x6CB9;&#x5727;&#x30A2;&#x30CA;&#x30ED;&#x30B8;&#x30FC;&#x306E;&#x91CD;&#x8981;&#x306A;&#x5236;&#x9650;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3059;&#x3002;&#x3088;&#x3046;&#x306A;&#x30AA;&#x30F3;&#x3068;&#x30AA;&#x30D5;&#x306E;&#x3057;&#x304D;&#x3044;&#x5024;&#x96FB;&#x5727;&#x6CB9;&#x5727; &mu;MVs &#x306E;&#x5178;&#x578B;&#x7684;&#x306A;&#x30C8;&#x30E9;&#x30F3;&#x30B8;&#x30B9;&#x30BF;&#x3068;&#x306F;&#x7570;&#x306A;&#x308A;&#x958B;&#x9589;&#x306E;&#x95BE;&#x5024;&#x5727;&#x529B;&#x306F;&#x5927;&#x5E45;&#x306B;&#x7570;&#x306A;&#x3063;&#x3066;&#x3044;&#x308B;&#x3057;&#x3082;&#x540C;&#x3058; &mu;MVs &#x5168;&#x4F53;&#x306E;&#x56DE;&#x8DEF;&#x306E;&#x8A2D;&#x8A08;&#x3068;&#x904B;&#x7528;&#x6761;&#x4EF6;&#x306B;&#x5FDC;&#x3058;&#x3066;&#x5909;&#x66F4;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x7279;&#x306B;&#x3001;&#x6211;&#x3005; &#x304C;&#x8AAC;&#x660E;&#x7D42;&#x4E86;&#x3057;&#x304D;&#x3044;&#x5024;&#x306E;&#x8CA0;&#x306E;&#x5024;&#x306E;&#x5727;&#x529B;&#x3069;&#x306E;&#x3088;&#x3046;&#x306B;&#x5927;&#x5E45;&#x306B; 2 &#x3064;&#x306E;&#x30D0;&#x30EB;&#x30D6; &#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x306E;&#x81EA;&#x5F8B;&#x7684;&#x5207;&#x308A;&#x66FF;&#x3048;&#x306A;&#x3069;&#x5358;&#x7D14;&#x306A;&#x6CB9;&#x5727; &mu;MV &#x56DE;&#x8DEF;&#x306E;&#x52D5;&#x4F5C;&#x3092;&#x5236;&#x9650;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x7406;&#x89E3;&#x81EA;&#x5DF1;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306E;&#x8A2D;&#x8A08;&#x306B;&#x5927;&#x304D;&#x306A;&#x5F71;&#x97FF;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;

&#x81EA;&#x5DF1;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x56DE;&#x8DEF;&#x7528;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x30C8;&#x30E9;&#x30F3;&#x30B8;&#x30B9;&#x30BF;&#x306E;&#x95BE;&#x5024;&#x5727;&#x529B;&#x5236;&#x9650;&#x306E;&#x5206;&#x6790;&#x3002;

&#xC774; &#xC885;&#xC774; &#xBBF8;&#xC138; &#xB9C9; &#xBC38;&#xBE0C; (&mu;MV)&#xC640; &#xC804;&#xC790; &#xD2B8;&#xB79C;&#xC9C0;&#xC2A4;&#xD130; &#xC0AC;&#xC774; &#xC804;&#xAE30; &#xC720;&#xC555; &#xBE44;&#xC720;&#xC5D0;&#xC11C; &#xC911;&#xC694; &#xD55C; &#xD55C;&#xACC4;&#xB97C; &#xBCF4;&#xC5EC;&#xC900;&#xB2E4;. &#xC784;&#xACC4; &#xC804;&#xC555;, &#xC720;&#xC555; &Mu;mvs&#xC5D0;&#xC11C; &#xC774;&#xB530;&#xAE08; &#xBE44;&#xC2B7;&#xD55C; &#xC788;&#xB294; &#xC77C;&#xBC18;&#xC801;&#xC778; &#xD2B8;&#xB79C;&#xC9C0;&#xC2A4;&#xD130;&#xC640; &#xB2EC;&#xB9AC; &#xAC1C;&#xD3D0;&#xC5D0; &#xB300; &#xD55C; &#xC784;&#xACC4;&#xAC12; &#xC555;&#xB825; &#xD604;&#xC800; &#xD558; &#xAC8C; &#xB2E4;&#xB974;&#xB2E4; &#xBC0F; &#xC804;&#xCCB4; &#xD68C;&#xB85C; &#xB514;&#xC790;&#xC778; &#xBC0F; &#xC791;&#xC5C5; &#xC870;&#xAC74;&#xC5D0; &#xB530;&#xB77C; &#xB3D9;&#xC77C;&#xD55C; &Mu;mvs&#xC5D0;&#xB3C4; &#xBCC0;&#xACBD;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xC124;&#xBA85; &#xD2B9;&#xD788;, &#xC74C;&#xC218; &#xB2EB;&#xB294; &#xC784;&#xACC4;&#xAC12;&#xC758; &#xC555;&#xB825;&#xC744; &#xD06C;&#xAC8C; &#xD558;&#xB294; &#xBC29;&#xBC95; &#xC81C;&#xD55C; 2 &#xBC38;&#xBE0C; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30;&#xB97C; &#xC790;&#xC728;&#xC801;&#xC73C;&#xB85C; &#xC804;&#xD658; &#xB4F1;&#xB3C4; &#xAC04;&#xB2E8;&#xD55C; &#xC720;&#xC555; &mu;MV &#xD68C;&#xB85C;&#xC758; &#xB3D9;&#xC791;. &#xC774;&#xB7EC;&#xD55C; &#xC774;&#xD574; &#xC790;&#xCCB4; &#xADDC;&#xC81C; &#xBBF8;&#xC138; &#xC7A5;&#xCE58; &#xC124;&#xACC4;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xC601;&#xD5A5;&#xC744; &#xBBF8;&#xCE60;.

&#xC790;&#xCCB4; &#xADDC;&#xC81C; &#xBBF8;&#xC138; &#xD68C;&#xB85C; &#xB300; &#xD55C; &#xBBF8;&#xC138; &#xD2B8;&#xB79C;&#xC9C0;&#xC2A4;&#xD130;&#xC5D0;&#xC11C; &#xC784;&#xACC4;&#xAC12; &#xC555;&#xB825; &#xD55C;&#xACC4;&#xB97C; &#xBD84;&#xC11D; &#xD569;&#xB2C8;&#xB2E4;.

Este documento revela una limitaci&oacute;n fundamental en la electro-hidr&aacute;ulica analog&iacute;a entre una v&aacute;lvula de membrana microfluidos (&mu;MV) y un transistor electr&oacute;nico. A diferencia de transistores t&iacute;picos que tienen similares dentro y fuera de los voltajes de umbral, en &mu;MVs hidr&aacute;ulicos, las presiones de umbral de apertura y cierre son significativamente diferentes y pueden cambiar, incluso para los mismos &mu;MVs dependiendo de las condiciones de dise&ntilde;o y operaci&oacute;n del circuito de generales. Explicamos, en particular, c&oacute;mo los valores negativos del umbral de cierre presiones significativamente restringir operaci&oacute;n de circuitos &mu;MV hidr&aacute;ulico incluso simple como aut&oacute;nomo cambiar dos v&aacute;lvulas microflu&iacute;dicos osciladores. Estos acuerdos tienen implicaciones significativas en el dise&ntilde;o de dispositivos microflu&iacute;dicos autorregulado.

Analizar las limitaciones de presi&oacute;n umbral en transistores de microfluidos para circuitos de microfluidos autorregulado.

&#x6211;&#x4EEC;&#x76EE;&#x524D;&#x7684;&#x5B9E;&#x9A8C;&#x4E0E;&#x7406;&#x8BBA;&#x7684;&#x4E00;&#x79CD;&#x6052;&#x5B9A;&#x6D41;&#x9A71;&#x52A8;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x4E0E;&#x5E7F;&#x6CDB;&#x53EF;&#x8C03;&#x8C10;&#x632F;&#x8361;&#x5468;&#x671F;&#x957F;&#x3002;&#x5C06;&#x8FD9;&#x4E2A;&#x632F;&#x8361;&#x5668;&#x4E24;&#x4E2A;&#x5E38;&#x6570;&#x8F93;&#x5165;&#x6D41;&#x52A8;&#x6CE8;&#x5C04;&#x5668;&#x6CF5;&#x4E2D;&#x8F6C;&#x6362;&#x4E3A;&#x4E0E;&#x632F;&#x8361;&#x5468;&#x671F;&#x53EF;&#x4EE5;&#x8C03;&#x6574;&#x5230; 0.3 &#x4E4B;&#x95F4;&#x7684;&#x4EA4;&#x66FF;&#x3001; &#x5B9A;&#x671F;&#x8F93;&#x51FA;-&#x6D41;&#x5230;&#x901A;&#x8FC7;&#x4F18;&#x5316;&#x5916;&#x90E8;&#x819C;&#x7535;&#x5BB9;&#x5668; 4.1 h s&#x3002;&#x6B64;&#x7535;&#x5BB9;&#x5728;&#x7ED9;&#x5B9A;&#x8F93;&#x5165;&#x6D41;&#x91CF;&#xFF0C;&#x4ECE;&#x800C;&#x63D0;&#x4F9B;&#x4E86;&#x6781;&#x5927;&#x7684;&#x7075;&#x6D3B;&#x6027;&#xFF0C;&#x5728;&#x8BBE;&#x5907;&#x64CD;&#x4F5C;&#x4E2D;&#x5141;&#x8BB8;&#x591A;&#x4E2A;&#x53EF;&#x8C03;&#x6574;&#x7684;&#x65F6;&#x671F;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x8868;&#x660E;&#xFF0C;&#x8DB3;&#x591F;&#x5927;&#x7684;&#x5916;&#x90E8;&#x7535;&#x5BB9;&#xFF0C;&#x4E0E;&#x5185;&#x90E8;&#x7535;&#x5BB9;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x9600;&#x95E8;&#x672C;&#x8EAB;&#xFF0C;&#x662F;&#x632F;&#x8361;&#x5173;&#x952E;&#x8981;&#x6C42;&#x3002;&#x8FD9;&#x4E9B;&#x5E7F;&#x6CDB;&#x53EF;&#x8C03;&#x8C10;&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x8BBE;&#x60F3;&#xFF0C;&#x4E3A;&#x4E0A;&#x82AF;&#x7247;&#x8BA1;&#x65F6;&#x6E90;&#x7684;&#x5FAE;&#x6D41;&#x63A7;&#x903B;&#x8F91;&#x7535;&#x8DEF;&#xFF0C;&#x88AB;&#x5E7F;&#x6CDB;&#x7528;&#x4E8E;&#x751F;&#x7269;&#x8282;&#x594F;&#x7684;&#x7814;&#x7A76;&#x548C;&#x5176;&#x4ED6;&#x9700;&#x8981;&#x7684;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x7684;&#x53D8;&#x5316;&#x5B9A;&#x65F6;&#x6D41;&#x4EA4;&#x6362;&#x3002;

&#x5FAE;&#x6D41;&#x63A7;&#x632F;&#x8361;&#x5668;&#x4E0E;&#x5E7F;&#x6CDB;&#x53EF;&#x8C03;&#x8C10;&#x671F;&#x3002;

We present experiments and theory of a constant flow-driven microfluidic oscillator with widely tunable oscillation periods. This oscillator converts two constant input-flows from a syringe pump into an alternating, periodic output-flow with oscillation periods that can be adjusted to between 0.3 s to 4.1 h by tuning an external membrane capacitor. This capacitor allows multiple adjustable periods at a given input flow-rate, thus providing great flexibility in device operation. Also, we show that a sufficiently large external capacitance, relative to the internal capacitance of the microfluidic valve itself, is a critical requirement for oscillation. These widely tunable microfluidic oscillators are envisioned to be broadly useful for the study of biological rhythms, as on-chip timing sources for microfluidic logic circuits, and other applications that require variation in timed flow switching.

Microfluidic oscillators with widely tunable periods.

Nous pr&eacute;sentons des exp&eacute;riences et la th&eacute;orie d&#39;un oscillateur de microfluidique de pilot&eacute;e par le flux constant avec des p&eacute;riodes d&#39;oscillation largement accordable. Cet oscillateur convertit deux flux d&#39;entr&eacute;e constantes d&#39;un pousse-seringue en un flux de sortie alternatif, p&eacute;riodique avec des p&eacute;riodes d&#39;oscillation r&eacute;glable entre 0,3 &agrave; s &agrave; 4,1 h en ajustant un condensateur de la membrane externe. Ce condensateur permet de multiples p&eacute;riodes r&eacute;glables &agrave; un donn&eacute;e d&#39;entr&eacute;e d&eacute;bit, offrant ainsi une grande flexibilit&eacute; dans le fonctionnement du dispositif. En outre, nous montrent qu&#39;une capacit&eacute; d&#39;entr&eacute;e externe suffisamment grande, par rapport &agrave; la capacit&eacute; interne de la vanne de microfluidique elle-m&ecirc;me, est une exigence cruciale &agrave; l&#39;oscillation. Ces oscillateurs largement accordable microfluidiques sont envisag&eacute;es pour &ecirc;tre largement utile pour l&#39;&eacute;tude des rythmes biologiques, comme sources de chronom&eacute;trage sur puce pour circuits logiques microfluidique, et autres applications n&eacute;cessitant une variation chronom&eacute;tr&eacute; commutation de flux.

Microfluidique oscillateurs avec p&eacute;riodes largement accordables.

Vi presentiamo esperimenti e la teoria di un oscillatore di microfluidica basata sul flusso costante con periodi di oscillazione ampiamente sintonizzabile. Questo oscillatore converte due flussi di input costanti da una pompa a siringa in un flusso di uscita alternato, periodico con periodi di oscillazione che pu&ograve; essere regolato a tra 0,3 s a 4,1 h sintonizzando un condensatore di membrana esterna. Questo condensatore permette molteplici periodi regolabili ad una determinato input-portata, fornendo cos&igrave; grande flessibilit&agrave; nel funzionamento del dispositivo. Inoltre, che ci mostra una capacit&agrave; esterna sufficientemente grande, rispetto la capacit&agrave; interna di microfluidica valvola stessa, &egrave; un requisito fondamentale per oscillazione. Questi oscillatori ampiamente sintonizzabile microfluidica sono immaginati essere ampiamente utile per lo studio dei ritmi biologici, come fonti di on-chip timing per circuiti logici microfluidica, e altre applicazioni che richiedono la variazione nel tempo flusso di commutazione.

Microfluidica oscillatori con periodi ampiamente sintonizzabili.

&#x6CE2;&#x9577;&#x53EF;&#x5909;&#x767A;&#x632F;&#x671F;&#x9593;&#x3068;&#x672C;&#x5B9F;&#x9A13;&#x3068;&#x7406;&#x8AD6;&#x306E;&#x4E00;&#x5B9A;&#x306E;&#x6D41;&#x308C;&#x3092;&#x99C6;&#x52D5;&#x3055;&#x308C;&#x308B;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x3002;&#x3053;&#x306E;&#x767A;&#x632F;&#x5668; 2 &#x3064;&#x306E;&#x5B9A;&#x6570;&#x306E;&#x5165;&#x529B;&#x30D5;&#x30ED;&#x30FC; &#x30B7;&#x30EA;&#x30F3;&#x30B8; &#x30DD;&#x30F3;&#x30D7;&#x304B;&#x3089;&#x3001;&#x4EA4;&#x4E92;&#x3001;&#x5B9A;&#x671F;&#x7684;&#x306A;&#x51FA;&#x529B;&#x30D5;&#x30ED;&#x30FC;&#x306B; 0.3 &#x306E;&#x9593;&#x3092;&#x8ABF;&#x6574;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x632F;&#x52D5;&#x5468;&#x671F;&#x3092;&#x5909;&#x63DB;&#x819C;&#x306E;&#x5916;&#x90E8;&#x30B3;&#x30F3;&#x30C7;&#x30F3;&#x30B5;&#x30FC; &#x30C1;&#x30E5;&#x30FC;&#x30CB;&#x30F3;&#x30B0;&#x306B;&#x3088;&#x3063;&#x3066; 4.1 h s&#x3002;&#x3053;&#x306E;&#x30B3;&#x30F3;&#x30C7;&#x30F3;&#x30B5;&#x30FC;&#x8ABF;&#x6574;&#x53EF;&#x80FD;&#x306A;&#x8907;&#x6570;&#x306E;&#x671F;&#x9593;&#x3092;&#x4E0E;&#x3048;&#x3089;&#x308C;&#x305F;&#x5165;&#x529B; - &#x3053;&#x306E;&#x3088;&#x3046;&#x306B;&#x30C7;&#x30D0;&#x30A4;&#x30B9;&#x306E;&#x64CD;&#x4F5C;&#x3001;&#x512A;&#x308C;&#x305F;&#x67D4;&#x8EDF;&#x6027;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x6D41;&#x91CF;&#x3067;&#x3001;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x307E;&#x305F;&#x3001;&#x6211;&#x3005; &#x306F;&#x30DE;&#x30A4;&#x30AF;&#x30ED; &#x30D0;&#x30EB;&#x30D6;&#x81EA;&#x4F53;&#x306E;&#x5185;&#x90E8;&#x5BB9;&#x91CF;&#x306B;&#x5BFE;&#x3057;&#x3066;&#x306E;&#x5341;&#x5206;&#x306A;&#x5927;&#x304D;&#x3055;&#x306E;&#x5916;&#x90E8;&#x5BB9;&#x91CF;&#x3092;&#x793A;&#x3059;&#x632F;&#x52D5;&#x306E;&#x91CD;&#x8981;&#x306A;&#x8981;&#x4EF6;&#x3067;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x5E83;&#x304F;&#x53EF;&#x5909;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x306F;&#x3001;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x8AD6;&#x7406;&#x56DE;&#x8DEF;&#x5185;&#x8535;&#x306E;&#x30BF;&#x30A4;&#x30DF;&#x30F3;&#x30B0; &#x30BD;&#x30FC;&#x30B9;&#x3068;&#x3057;&#x3066;&#x5E83;&#x304F;&#x751F;&#x7269;&#x30EA;&#x30BA;&#x30E0;&#x306E;&#x691C;&#x8A0E;&#x306E;&#x305F;&#x3081;&#x306B;&#x6709;&#x7528;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x60F3;&#x5B9A;&#x3055;&#x308C;&#x3066;&#x3001;&#x30D5;&#x30ED;&#x30FC;&#x306E;&#x5909;&#x5316;&#x3092;&#x5FC5;&#x8981;&#x3068;&#x3059;&#x308B;&#x4ED6;&#x306E;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x304C;&#x30BF;&#x30A4;&#x30E0;&#x30A2;&#x30A6;&#x30C8;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;

&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x6D41;&#x4F53;&#x767A;&#x632F;&#x5668;&#x5468;&#x6CE2;&#x6570;&#x53EF;&#x5909;&#x671F;&#x9593;&#x3002;

&#xC120;&#xBB3C;&#xC774; &#xB110;&#xB9AC; &#xC870;&#xC815;&#xD560; &#xC218; &#xC788;&#xB294; &#xC9C4;&#xB3D9; &#xAE30;&#xAC04;&#xACFC; &#xC77C;&#xC815; &#xD750;&#xB984; &#xAE30;&#xBC18; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30;&#xC758; &#xC774;&#xB860;&#xACFC; &#xC2E4;&#xD5D8;. &#xC774; &#xBC1C;&#xC9C4;&#xAE30;&#xB294; &#xBC88;&#xAC08;&#xC544;&#xAC00; &#xBA70;, &#xC815;&#xAE30;&#xC801;&#xC778; &#xCD9C;&#xB825; &#xD750;&#xB984; 0.3 &#xC0AC;&#xC774; &#xC870;&#xC815;&#xD560; &#xC218; &#xC788;&#xB294; &#xC9C4;&#xB3D9; &#xAE30;&#xAC04;&#xACFC; &#xC8FC;&#xC0AC;&#xAE30; &#xD38C;&#xD504;&#xC5D0;&#xC11C; &#xB450; &#xC0C1;&#xC218; &#xC785;&#xB825; &#xD750;&#xB984; &#xBCC0;&#xD658; &#xC678;&#xBD80; &#xB9C9; &#xCEE4;&#xD328;&#xC2DC;&#xD130; &#xC870;&#xC815; 4.1 h s. &#xC774; &#xCEE4;&#xD328;&#xC2DC;&#xD130;&#xB294; &#xC8FC;&#xC5B4;&#xC9C4;&#xB41C; &#xC785;&#xB825;&#xB41C; &#xC720;&#xB7C9;, &#xC7A5;&#xCE58; &#xC791;&#xC5C5;&#xC5D0; &#xC720;&#xC5F0;&#xC131; &#xC81C;&#xACF5;&#xC5D0; &#xC5EC;&#xB7EC; &#xC870;&#xC815; &#xAE30;&#xAC04;&#xC744; &#xD5C8;&#xC6A9; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC;&#xB294; &#xBBF8;&#xC138; &#xBC38;&#xBE0C; &#xC790;&#xCCB4;&#xC758; &#xB0B4;&#xBD80; &#xCEE4;&#xD328;&#xC2DC;&#xD134;&#xC2A4;&#xB97C; &#xAE30;&#xC900;&#xC73C;&#xB85C; &#xCDA9;&#xBD84;&#xD788; &#xD070; &#xC678;&#xBD80; &#xCEE4;&#xD328;&#xC2DC;&#xD134;&#xC2A4; &#xD45C;&#xC2DC; &#xC9C4;&#xB3D9;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD55C; &#xC694;&#xAD6C; &#xC0AC;&#xD56D;&#xC774;&#xC785;&#xB2C8;&#xB2E4;. &#xC774; &#xAD11;&#xBC94;&#xC704; &#xD558; &#xAC8C; &#xC870;&#xC815;&#xD560; &#xC218; &#xC788;&#xB294; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30; &#xCE69;&#xC5D0; &#xD0C0;&#xC774;&#xBC0D; &#xC18C;&#xC2A4; &#xBBF8;&#xC138; &#xB17C;&#xB9AC; &#xD68C;&#xB85C; &#xB300; &#xD55C; &#xC0DD;&#xBB3C; &#xD559;&#xC801; &#xB9AC;&#xB4EC;&#xC758; &#xC5F0;&#xAD6C;&#xC5D0; &#xB300; &#xD55C; &#xAD11;&#xBC94;&#xC704; &#xD558; &#xAC8C; &#xC720;&#xC6A9; &#xD558; &#xAC8C; &#xAD6C;&#xC0C1; &#xD558; &#xACE0; &#xB2E4;&#xB978; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC5D0;&#xC11C; &#xBCC0;&#xD654;&#xB97C; &#xC694;&#xAD6C; &#xD558;&#xB294; &#xD750;&#xB984; &#xC2A4;&#xC704;&#xCE6D; &#xD0C0;&#xC784;.

&#xAD11;&#xBC94;&#xC704; &#xD558; &#xAC8C; &#xC870;&#xC815;&#xD560; &#xC218; &#xC788;&#xB294; &#xAE30;&#xAC04;&#xC73C;&#xB85C; &#xBBF8;&#xC138; &#xBC1C;&#xC9C4;&#xAE30;&#xC785;&#xB2C8;&#xB2E4;.

Presentamos experimentos y teor&iacute;a de un oscilador de microfluidos basadas en flujo constante con per&iacute;odos de oscilaci&oacute;n ampliamente regulables. Este oscilador convierte dos flujos de entrada constantes de una bomba de jeringa en un flujo salida peri&oacute;dico, alternando con per&iacute;odos de oscilaci&oacute;n que puede ajustarse entre 0.3 s a h 4,1 templando un capacitor de la membrana externa. Este condensador permite m&uacute;ltiples per&iacute;odos ajustables a dado entrado caudal, permitiendo gran flexibilidad en la operaci&oacute;n del dispositivo. Adem&aacute;s, demostramos que una capacitancia externa suficientemente grande, en relaci&oacute;n con la capacidad interna de la v&aacute;lvula de microfluidos, es un requisito cr&iacute;tico para oscilaci&oacute;n. Estos osciladores ampliamente sintonizables microflu&iacute;dicos son previstos para ser ampliamente &uacute;til para el estudio de los ritmos biol&oacute;gicos, como fuentes de sincronizaci&oacute;n de la en-viruta para circuitos de l&oacute;gica de microfluidos, y otras aplicaciones que requieren variaci&oacute;n en tiempo de conmutaci&oacute;n de flujo.

Osciladores de microfluidos con per&iacute;odos muy armoniosas.

Intracellular cargo is transported by multiple motor proteins. Because of the force balance of motors with mixed polarities, cargo moves bidirectionally to achieve biological functions. Here, we propose a microtubule gliding assay for a tug-of-war study of kinesin and dynein. A boundary of the two motor groups is created by photolithographically patterning gold to selectively attach kinesin to the glass and dynein to the gold surface using a self-assembled monolayer. The relationship between the ratio of two antagonistic motor numbers and the velocity is derived from a force-velocity relationship for each motor to calculate the detachment force and motor backward velocity. Although the tug-of-war involves >100 motors, values are calculated for a single molecule and reflect the collective dynein and non-collective kinesin functions when they work as a team. This assay would be useful for detailed in vitro analysis of intracellular motility, e.g., mitosis, where a large number of motors with mixed polarities are involved.

Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface.

One of challenges for using microtubules (MTs) driven by kinesin motors in microfluidic environments is to control their direction of movement. Although applying physical biases to rectify MTs is prevalent, it has not been established as a design methodology in conjunction with microfluidic devices. In the future, the methodology is expected to achieve functional motor-driven nanosystems. Here, we propose a method to guide kinesin-propelled MTs in multiple directions under an electric field by designing a charged surface of MT minus ends labeled with dsDNA via a streptavidin-biotin interaction. MTs labeled with 20-bp or 50-bp dsDNA molecules showed significantly different trajectories according to the DNA length, which were in good agreement with values predicted from electrophoretic mobilities measured for their minus ends. Since the effective charge of labeled DNA molecules was equal to that of freely dispersed DNA molecules in a buffer solution, MT trajectory could be estimated by selecting labeling molecules with known charges. Moreover, the estimated trajectory enables to define geometrical sizes of a microfluidic device. This rational molecular design and prediction methodology allows MTs to be guided in multiple directions, demonstrating the feasibility of using molecular sorters driven by motor proteins.

Control of microtubule trajectory within an electric field by altering surface charge density.

Microtubules driven by kinesin motors have been utilised as "molecular shuttles" in microfluidic environments with potential applications in autonomous nanoscale manipulations such as capturing, separating, and/or concentrating biomolecules. However, the conventional flow cell-based assay has difficulty in separating bound target molecules from free ones even with buffer flushing because molecular manipulations by molecular shuttles take place on a glass surface and molecular binding occurs stochastically; this makes it difficult to determine whether molecules are carried by molecular shuttles or by diffusion. To address this issue, we developed a microtubule-based transport system between two compartments connected by a single-micrometre-scale channel array that forms dynamically via pneumatic actuation of a polydimethylsiloxane membrane. The device comprises three layers-a control channel layer (top), a microfluidic channel layer (middle), and a channel array layer (bottom)-that enable selective injection of assay solutions into a target compartment and dynamic formation of the microchannel array. The pneumatic channel also serves as a nitrogen supply path to the assay area, which reduces photobleaching of fluorescently labelled microtubules and deactivation of kinesin by oxygen radicals. The channel array suppresses cross-contamination of molecules caused by diffusion or pressure-driven flow between compartments, facilitating unidirectional transport of molecular shuttles from one compartment to another. The method demonstrates, for the first time, efficient and unidirectional microtubule transport by eliminating diffusion of target molecules on a chip and thus may constitute one of the key aspects of motor-driven nanosystems.

Dynamic formation of a microchannel array enabling kinesin-driven microtubule transport between separate compartments on a chip.

Self-switching microfluidic circuits that are able to perform biochemical experiments in a parallel and autonomous manner, similar to instruction-embedded electronics, are rarely implemented. Here, we present design principles and demonstrations for gravity-driven, integrated, microfluidic pulsatile flow circuits. With a common gravity head as the only driving force, these fluidic oscillator arrays realize a wide range of periods (0.4 s-2 h) and flow rates (0.10-63 μl min(-1)) with completely independent timing between the multiple oscillator sub-circuits connected in parallel. As a model application, we perform systematic, parallel analysis of endothelial cell elongation response to different fluidic shearing patterns generated by the autonomous microfluidic pulsed flow generation system.

Multiple independent autonomous hydraulic oscillators driven by a common gravity head.

The Escherichia coli RuvB hexameric ring motor proteins, together with RuvAs, promote branch migration of Holliday junction DNA. Zero mode waveguides (ZMWs) constitute of nanosized holes and enable the visualization of a single fluorescent molecule under micromolar order of the molecules, which is applicable to characterize the formation of RuvA-RuvB-Holliday junction DNA complex. In this study, we used ZMWs and counted the number of RuvBs binding to RuvA-Holliday junction DNA complex. Our data demonstrated that different nucleotide analogs increased the amount of Cy5-RuvBs binding to RuvA-Holliday junction DNA complex in the following order: no nucleotide, ADP, ATPγS, and mixture of ADP and ATPγS. These results suggest that not only ATP binding to RuvB but also ATP hydrolysis by RuvB facilitates a stable RuvA-RuvB-Holliday junction DNA complex formation.

Synergistic effect of ATP for RuvA-RuvB-Holliday junction DNA complex formation.

Tau protein is a well-established biomarker for a group of neurodegenerative diseases collectively called tauopathies. So far, clinically relevant detection of tau species in cerebrospinal fluid (CSF) cannot be achieved without immunological methods. Recently, it was shown that different tau isoforms including the ones carrying various types of mutations affect microtubule (MT)-kinesin binding and velocity in an isoform specific manner. Here, based on these observations, we developed a microfluidic device to analyze tau mutations, isoforms and their ratios. The assay device consists of three regions: a MT reservoir which captures MTs from a solution to a kinesin-coated surface, a microchannel which guides gliding MTs, and an arrowhead-shaped collector which concentrates MTs. Tau-bound fluorescently labeled MTs (tau-MTs) were assayed, and the increase in fluorescence intensity (FI) corresponding to the total number of MTs accumulated was measured at the collector. We show that our device is capable of differentiating 3R and 4R tau isoform ratios and effects of point mutations within 5 minutes. Furthermore, radially oriented collector regions enable simultaneous FI measurements for six independent assays. Performing parallel assays in the proposed device with minimal image processing provides a cost-efficient, easy-to-use and fast tau detection platform.

On-chip microtubule gliding assay for parallel measurement of tau protein species.

Organ culture systems are used to elucidate the mechanisms of pattern formation in developmental biology. Various organ culture techniques have been used, but the lack of microcirculation in such cultures impedes the long-term maintenance of larger tissues. Recent advances in microfluidic devices now enable us to utilize self-organized perfusable capillary networks in organ cultures. In this review, we will overview past approaches to organ culture and current technical advances in microfluidic devices, and discuss possible applications of microfluidics towards the study of developmental biology.

Tissue culture on a chip: Developmental biology applications of self-organized capillary networks in microfluidic devices.

Creating vascular networks in tissues is crucial for tissue engineering. Although recent studies have demonstrated the formation of vessel-like structures in a tissue model, long-term culture is still challenging due to the lack of active perfusion in vascular networks. Here, we present a method to create a three-dimensional cellular spheroid with a perfusable vascular network in a microfluidic device. By the definition of the cellular interaction between human lung fibroblasts (hLFs) in a spheroid and human umbilical vein endothelial cells (HUVECs) in microchannels, angiogenic sprouts were induced from microchannels toward the spheroid; the sprouts reached the vessel-like structures in a spheroid to form a continuous lumen. We demonstrated that the vascular network could administer biological substances to the interior of the spheroid. As cell density in the spheroid is similar to that of a tissue, the perfusable vasculature model opens up new possibilities for a long-term tissue culture in vitro.

Integrating perfusable vascular networks with a three-dimensional tissue in a microfluidic device.

Vasculogenesis is the earliest process in development for spontaneous formation of a primitive capillary network from endothelial progenitor cells. When human umbilical vein endothelial cells (HUVECs) are cultured on Matrigel, they spontaneously form a network structure which is widely used as an in vitro model of vasculogenesis. Previous studies indicated that chemotaxis or gel deformation was involved in spontaneous pattern formation. In our study, we analyzed the mechanism of vascular pattern formation using a different system, meshwork formation by HUVECs embedded in fibrin gels. Unlike the others, this experimental system resulted in a perfusable endothelial network in vitro. We quantitatively observed the dynamics of endothelial cell protrusion and developed a mathematical model for one-dimensional dynamics. We then extended the one-dimensional model to two-dimensions. The model showed that random searching by endothelial cells was sufficient to generate the observed network structure in fibrin gels.

Mathematical modeling for meshwork formation of endothelial cells in fibrin gels.

Intracellular transport is affected by the filament network in the densely packed cytoplasm. Biophysical studies focusing on intracellular transport based on microtubule-kinesin system frequently use in vitro motility assays, which are performed either on individual microtubules or on random (or simple) microtubule networks. Assembling intricate networks with high flexibility requires the manipulation of 25 nm diameter microtubules individually, which can be achieved through the use of pick-and-place assembly. Although widely used to assemble tiny objects, pick-and-place is not a common practice for the manipulation of biological materials. Using the high-level handling capabilities of microelectromechanical systems (MEMS) technology, tweezers are designed and fabricated to pick and place single microtubule filaments. Repeated picking and placing cycles provide a multilayered and multidirectional microtubule network even for different surface topographies. On-demand assembly of microtubules forms crossings at desired angles for biophysical studies as well as complex networks that can be used as nanotransport systems.

Article	Year
Constant flow-driven microfluidic oscillator for different duty cycles. Analytical chemistry\| PubMed ID: 22206453	2012
A perfusable microfluidic device with on-chip total internal reflection fluorescence microscopy (TIRFM) for in situ and real-time monitoring of live cells. Biomedical microdevices\| PubMed ID: 22585136	2012
Microfluidic automation using elastomeric valves and droplets: reducing reliance on external controllers. Small (Weinheim an der Bergstrasse, Germany)\| PubMed ID: 22761019	2012
Colocalization of quantum dots by reactive molecules carried by motor proteins on polarized microtubule arrays. ACS nano\| PubMed ID: 23230973	2013
Analyzing threshold pressure limitations in microfluidic transistors for self-regulated microfluidic circuits. Applied physics letters\| PubMed ID: 23284181	2012
Microfluidic oscillators with widely tunable periods. Lab on a chip\| PubMed ID: 23429765	2013
Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface. Scientific reports\| PubMed ID: 24923426	2014
Control of microtubule trajectory within an electric field by altering surface charge density. Scientific reports\| PubMed ID: 25567007	2015
Dynamic formation of a microchannel array enabling kinesin-driven microtubule transport between separate compartments on a chip. Lab on a chip\| PubMed ID: 25805147	2015
Multiple independent autonomous hydraulic oscillators driven by a common gravity head. Nature communications\| PubMed ID: 26073884	2015
Synergistic effect of ATP for RuvA-RuvB-Holliday junction DNA complex formation. Scientific reports\| PubMed ID: 26658024	2015
On-chip microtubule gliding assay for parallel measurement of tau protein species. Lab on a chip\| PubMed ID: 27056640	2016
Tissue culture on a chip: Developmental biology applications of self-organized capillary networks in microfluidic devices. Development, growth & differentiation\| PubMed ID: 27272910	2016
Integrating perfusable vascular networks with a three-dimensional tissue in a microfluidic device. Integrative biology : quantitative biosciences from nano to macro\| PubMed ID: 28561127	2017
Mathematical modeling for meshwork formation of endothelial cells in fibrin gels. Journal of theoretical biology\| PubMed ID: 28648563	2017
Pick-and-Place Assembly of Single Microtubules. Small (Weinheim an der Bergstrasse, Germany)\| PubMed ID: 28692749	2017

Ryuji Yokokawa

Biography

JoVE Journal Publications

Other Publications