Bu çalışma NIH R21 EB008177-01A2 ve New York Üniversitesi Abu Dabi (NYUAD) Araştırma Geliştirme Fonu 2013 Biz mikrofabrikasyon sırasında verdikleri destek nedeniyle MİT MTL teknik personele teşekkürlerimi ifade ve James Weston ve onların için NYUAD Nikolas Giakoumidis tarafından desteklenen SEM fotoğraf çekmek ve sırasıyla bir gerilim bölücü bina desteği. PDMS cihaz imalat NYUAD ve mikroimalat çekirdek tesisi yapılmıştır. Son olarak, biz video çekimi ve düzenleme için dijital Burs NYUAD Merkezi&#39;nden Rebecca Pittam teşekkür etmek istiyorum.

Silika Kolloidal Boncuk Cezalar 1. Hazırlık <ol><li> 300 nm ve 500 nm silika kordon süspansiyonların hazırlanması <ol><li> Vorteks 30 sn için silika kordon stok süspansiyonu (% 10 ağırlık / su hac). Homojen bir süspansiyon elde edildi. 1 dakika için 2600 x g&#39;de, 1.5 ml tüp ve santrifüj içine 600 ul stok süspansiyonu toplam Pipet. </li><li> 1 mM sodyum fosfat tampon maddesi (PB, pH 7.0), 400 ul süpernatant değiştirin. </li><li> vorteks yoluyla pH 7.0&#39;da 1 mM sodyum fosfat çözeltisi içinde% 15 nihai konsantrasyonu silis boncuklar askıya alınması. </li></ol></li><li> Poli yüzey işlevselleştirilmesi 500 nm silika karboksil boncukları (allilamin hidroklorür, PAH) ve poli (sodyum stiren sülfonat, PSS) polielektrolitler <ol><li> % 1 10 mi, 1 M NaCl (pH 7.0) (w / v) boncuk süspansiyonu ile karboksil grubu ile 500 nm silis boncuklar 0.1 g süspanse edin. </li><li> 1 M nac 0.4% PAH (MW 65K) hazırlayın1 M NaCl, 15 ml stok çözeltisi 300 ul (% 20 ağırlık / su içinde hac) çözülmesiyle l. 20 mi, 1 M NaCl çözeltisi içinde 0.18 g PSS çözülmesiyle, 1 M NaCl çözeltisi içinde% 0.9 PSS (MW 70K) hazırlayın. Vortex hem 1 dakika için çözümler. tamamen polielektrolit çözmek için. </li><li> karboksil fonksiyonel grup ile silis boncuklar üzerinde pozitif yüklü polielektrolit tabakası biriktirilmesi için 15 ml&#39;lik bir tüp içinde% 1 silis karboksil boncuk 9.8 ml PAH çözeltisi 200 ul ekle. 1 dakika için boncuk süspansiyonu vorteksleyin. ve 60 dakika boyunca bir tüp rotator üzerinde inkübe edin. Oda sıcaklığında karıştırıldı. </li><li> 1 dakika boyunca 1.801 xg&#39;de boncuk süspansiyonu santrifüj. ve 10 ml DI su ile ilişkisiz PAH beş kez yıkayın. Her bir santrifüj ve süpernatan çıkarılmasından sonra boncuklar yoğun tüpün altındaki sıkıştırıldı. Boncuklar yeniden süspanse edildi ve bir sonraki santrifüj aşamasından önce yıkanabilir ve böylece DI, 8 ml su ilave edilmeden önce DI 2 ml su ile kuvvetli bir şekilde pipetleme kordon yığın bozabilir. </li><li> takipPSS kaplama 1.2.3 ve 1.2.4 adımları boncuklar üzerinde, negatif yüklü bir tabaka yatırmak. PSS birikimi öncesinde 1.2.4 5 inci yıkama adımından DI su süpernatant çıkardıktan sonra 1 M NaCl 9.8 ml boncuk yeniden askıya. <ol><li> 15 ml tüp altındaki köşe yığın parçalamak ve daha sonra 1 M NaCl, 8 ml eklemek için 1 M NaCl 2 ml kullanılarak alan güçlü pipetleme adımı kullanın. Tek PAH tabaka tevdi silis boncuklar 9.8 ml PSS çözeltisi 200 ul ekle. 1 dakika boyunca vorteks sonra. ve 60 dakika boyunca kuluçkalama. Tüp rotator, DI su ile 5 yıkama adımları tekrarlayın. </li><li> doğru gerçekleştirilmiştir polielektrolit yerleştirme işlemini kontrol etmek için, üreticinin protokolüne uygun olarak, bir dinamik ışık saçma sistemi kullanılarak her bir polielektrolit kaplama öncesi ve sonrası boncukların zeta potansiyelinin ölçülmesi (Tablo 1 e bakınız). </li></ol></li><li> Tek PSS katmanda aşağıdaki DI suyla beş yıkama adımları tekrarlayınyerleştirme ve% 0.05 Tween 20 öncesinde akışkanlık arttırmak için mikroakışkan cihaz kullanmak (a / h% 15) ile 1 mM sodyum fosfat tampon 650 ul boncuk yeniden askıya. </li></ol></li><li> PSS tek bir tabakası bırakmak üzere özellikle amin fonksiyonel grubu ile 500 nm silis boncuklar 1.2.6 etmek 1.2.5 yukarıda açıklanan prosedürü izleyin. </li></ol> PDMS mikroakışkan Chip 2. Fabrikasyon <ol><li> Silikon master mikroüretim <ol><li> aşağıdaki gibi mikroüretim teknikleri kullanılarak PDMS kalıplama için silikon ana imal. <ol><li> Spin kat silikon gofret 4,000 rpm&#39;de 1 mikron ince fotorezist. Desen projeksiyon litografi (pozlama süresi 170 milisaniye.) Ve 700 nm derin ve 2 mikron genişliğinde düzlemsel nanochannels etch kullanarak katman reaktif iyon aşındırma ile (silis boncuklar nanotraps olarak hareket). </li><li> CHF 3 (45 sccm: 3.5 nm / s aşındırma oranı elde etmek için aşağıdaki aşındırma parametrelerini kullanın), CF 4 (15 sccm), Ar (100 sccm), basınç 100 mTorr&#39;luk, RF güç 200 W </li></ol></li><li> Spin kat, ikinci 1 mikron kalınlığında fotorezist 2.000 rpm&#39;de katman ve daha önce desenli nanotraps bir hizalama gerçekleştirin. mikrokanalların iletişim litografi yoluyla ve silikon derin reaktif iyon aşındırma (DRIE) tarafından Desen. DRIE Tablo 2&#39;de 26 parametreleri kullanın. </li></ol></li><li> PDMS kalıp imalatı <ol><li> Bir vakum kavanoz O / N trichlorosilane (50 ul) ile silikon ustası Silanize. DİKKAT: Tricholorosilane zehirli ve korozif bir malzemedir. Her zaman uygun kişisel koruyucu ekipman ile kimyasal bir kaput kullanabilirsiniz. </li><li> 10 de sertleştirme maddesi başlangıç ​​karışımı: silanize silikon ana 1 oranı ve döküm PDMS ve bir konveksiyon fırınında 2 saat boyunca 70 ° C &#39;de tedavi. </li><li> Bir pl sonra plazma temizleyici kullanarak boş bir gofret bir bıçak ve plazma bağı onunla silikon ustadan PDMS levha çıkarın1 dakika boyunca plazma temizleyici asma tedavisi. adımı döküm aşağıdaki PDMS için bir bölüm çizgi işaretlemek için kenarı boyunca kasetleri takın. </li><li> triklorosilan (50 ul), O / N oranı ile bir vakum kavanoza PDMS kalıp Silanize. </li><li> PDMS döküm (baz:: 1 oranında 10 sertleştirici) silanize PDMS kalıp ve bir konveksiyon fırınında 2 saat boyunca 70 ° C &#39;de tedavi. </li></ol></li><li> PDMS cihazın imalatı <ol><li> bant ile işaretlenmiş bölüm hattı boyunca PDMS kalıp tedavi PDMS levha soyun. </li><li> Bir bant ile temiz 1,5 mm biyopsi yumruk ile Punch rezervuar delikleri, izopropil alkol (IPA) ve azot ile kuru ile durulayın. </li><li> Plazma bağı, 1 dakika için bir plazma temizleme bir plazma tedavi sonrası 25 mm x 75 mm mikroskop cam slayt PDMS cihaz. </li></ol></li><li> Ultrasonicate 60 dakika boyunca boncuk süspansiyonu. Dolum için ultrasonik banyoda önce. 10 ul boncuk süspansiyonu pipetle (300 nm olmayan işlevselleştirilmiş silika olmakreklamlar veya PAH-PSS tabakaları ile 500 nm, silika karboksil boncuklar ya da girişler 4 ve 6 bir PSS tabakası ile 500 nm silika amin boncuk) her biri için (Şekil 1 A, B) hemen PDMS çip plazma bağ sonra bkz cam alt-tabaka. boncuk ambalaj geliştirmek için bir pipet ile PDMS çip üzerinde hafifçe dokunun. <ol><li> boncuk dağıtım kanallarının doldurduktan sonra, bant ile 1 ve 9 hariç tüm girişleri kapsamaktadır. . Kullanımdan önce 4 ° C&#39;de 3 saat ve mağaza cihazı Hava kuruması 2 kolloidal kendinden montaj işleminin adım adım şematik verir Şekil. </li></ol></li></ol> DNA&#39;nın Elektrokinetik Konsantrasyon 3. Deney <ol><li> hazneleri 3, 7, bir tampon çözeltisi ile (1 mM PB 10 | il) ve hazne 5, bir DNA örneği ile (1 mM PB 10 nM 10 ul) doldurun ve rezervuar 2 ters pipet ucu ile hafifçe negatif bir basınç uygulamak 8 ve 10 (bkz kabarcıkları olmadan çözümleri ile kanal doldurmak için <strong&gt; Şekil 1 B). </li><li> basıncı dengelemek ve 5 dakika beklemek 10 Rezervuara rezervuar 2 ve 8 ve 10 nM DNA 10 ul 1 mM PB 10 ul ekle. dengeye ulaşmak için. </li><li> rezervuar 3, 5, 7, 10 içine Pt teller yerleştirin. </li><li> Bir kaynak metre ve Pt tel bağlı bir gerilim bölücü kullanılarak nanofluidic kavşak arasındaki gerilimi uygulayın. İlk rezervuarların 5, rezervuarlar 3, 7, 10 ve GND üzerinde 30 V uygulayın. </li><li> ~ 30 saniye sonra rezervuar 10 25 V gerilim azaltın. </li><li> DNA floresan sinyalleri kaydederken numunenin photobleaching en aza indirmek için, her 5 saniyede bir aralıklarla açıklık ile mekanik bir deklanşör kullanın. </li></ol>

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</ol>

The authors have nothing to disclose.

NANO AKIŞKAN incelemek için ortak cihaz tasarım düzeni takiben, nanochannels bir dizi desenlendirme lithographically kolloidal nanoparçacıkların buharlaşma odaklı kendini takımı kullanılarak yerine iki mikroakışkan kanallar arasında bir nanofluidic kavşak fabrikasyon. topuk kısmı dağıtım kanalı, 700 nm lik bir derinliği ve 100 nm&#39;lik toplam genişlikte kordon dağıtım kanalının her iki tarafta 2 um genişliğinde nanotraps bir diziye kolloidal parçacıklar akan zaman akmasını boncuk sü...

10 2 nm - NANO AKIŞKAN 10 1 uzunluğu ölçeğinde biyolojik süreçleri ya da iyonların taşınım olaylarını ve molekülleri incelemek için μ TMS ortaya çıkan bir araştırma alanı (Micro Toplam Analiz Sistemleri) &#39;dir. Gerekirse bu tür nanochannels olarak nanofluidic araçları gelişine, moleküllerin ve iyonların taşıma işlemleri sadece ayrılma ve tespiti için bu uzunluk ölçekte kullanılabilen özellikleri istismar ederek, benzeri görülmemiş bir hassasiyetle takip ve manipüle edilebilir. 1,2 Bir bu karakteristik nano ölçekli özellikleri bir yük dengesizliğe neden ve nanochannel ve Mikrokanallı arasındaki iyon konsantrasyonu kutuplaşma (ICP) başlatabilir nanochannels toplu ücret (veya Dukhin numarasına) yüzeyinin yüksek bir orandır. 3 Nanofluidic süreçleri için bir ortak cihaz platformu bir birleşme olarak nanochannels dizisi ile bağlı bir, iki mikro sisteminden oluşur. 4-6 </sup&gt; Bir nanofluidic cihazı oluşturmak için tercih edilen malzeme, çünkü bağlama işlemleri sırasında çökmesini kanal engelleyen yüksek sertlik silikondur. 7 Bununla birlikte, silisyum cihaz fabrikasyonu pahalı maskeleri ve temiz oda tesiste işleme önemli gerektirir. 8- nedeniyle kalıplama ve plazma yapıştırma, polidimetilsiloksan (PDMS) aracılığıyla cihaz imalat kolaylık 10. yaygın Mikroakiskan için bir yapı malzemesi olarak kabul edilmiştir ve bunun yanı sıra NANO AKIŞKAN için ideal bir malzemedir olacaktır. Ancak, düşük Young modülü 360-870 KPa etrafında, plazma yapıştırılması sırasında PDMS kanal kolayca katlanabilir hale getirir. nanochannel derinliği 100 nm altında olması olan, standart fotolitografi vasıtasıyla PDMS cihazların imalat, bir kanal genişliği gerektiren, son derece zorlu hale geleceği anlamına gelir: 1 nanochannel (derinliğine genişliği) minimum en-boy oranı en az 10 olması gerekir photolith mevcut sınırından daha azcivarında 1 mikron coğrafyayı. Bu sınırlamayı aşmak için, böyle bir plazma tedaviden sonra 78 nm 11 veya oluşturmak için kırışıklıkların ortalama derinliği ile çatlaklar başlatmak için germe olmayan lithographical yöntemlerle PDMS nanochannels oluşturmak için girişimleri olmuştur. 12 mekanik basınç ile PDMS kanal izin çöken bir 60 nm gibi düşük nanochannel yüksekliği. 13 bu son derece yaratıcı olmayan taşbaskı yöntemler derinlemesine 100 nm altında bina nanochannels izin olsa da, nanochannel imalat boyutlu kontrol edilebilirlik hala nanofluidic cihazlar için bir yapı malzemesi olarak PDMS geniş bir kabul bir engel teşkil etmektedir. nanochannels bir diğer önemli bir sorun, ister silikon veya PDMS, iyonlar veya moleküller manipülasyonu için bir kanal duvarı yüzey yükünün değiştirilmesi için bir ihtiyaç vardır durumunda yüzey işlevselleştirilmesi. bağlanması aracılığıyla cihaz grubu sonra nanochannels zor olannedeniyle difüzyon ile sınırlı taşıma yüzey fonksiyonlandırmalar için ulaşır. Yüksek boyutlu sadakat ve kolay bir yüzey fonksiyonlandırmalar bir nano kanalı oluşturmak için, mikroakışkan cihazlar buharlaşma 14-16 tarafından uyarılan kolloidal parçacıkların öz-montaj yöntemi umut verici yaklaşımlardan biri olabilir. Gözenek boyutu ve yüzey özelliği kontrol edilebilirliği yanı sıra, ayarlamak için bir olasılık sıcaklık, 17 pH, 18,19 ve iyonik kuvvete kontrol ederek polielektrolitler ile kaplanmış koloidal parçacıklar kullanılarak in-situ gözenek boyutu da vardır. Çünkü bu 18 elektrokromatografi, 20 biyosensörler, 21 protein konsantrasyonu 22 ve Mikroakiskan proteinlerin ve DNA&#39;nın ayrılması. 14,23 Bu çalışmada için avantajlar, kolloidal parçacıkların öz-montaj yöntemi zaten bulduğu uygulamaları, biz bir inşa etmek için bu öz-montaj yöntemi konuşlandırılmış elektrokinetik zenginleştirme cihazıİki mikro arasındaki nanofluidic birleşim gerektirir PDMS. 24 elektrokinetik konsantrasyonda arkasındaki temel mekanizma, iyon konsantrasyon kutuplaşma (ICP) dayanır. Imalat ve montaj adımları 25 ayrıntılı bir açıklaması aşağıdaki protokole dahil edilir.

<table><tbody><tr><td>Poly(Styrenesulfonic Acid) Sodium Salt</td><td>Polysciences&amp;nbsp;</td><td>08772</td><td></td></tr><tr><td>Poly(allylamine) Solution</td><td>Sigma Aldrich</td><td>479144-5G</td><td></td></tr><tr><td>Silica Microsphere - 300 nm</td><td>Polysciences&amp;nbsp;</td><td>24321</td><td></td></tr><tr><td>Silica Microsphere - 500 nm</td><td>Polysciences&amp;nbsp;</td><td>24323</td><td></td></tr><tr><td>Silica Microsphere Carboxyl Functional - 500 nm</td><td>Polysciences&amp;nbsp;</td><td>24753</td><td></td></tr><tr><td>Silica Microsphere Amine Functional - 500 nm</td><td>Polysciences&amp;nbsp;</td><td>24756</td><td></td></tr><tr><td>Sylgard 184 Silicone Elastomer kit</td><td>Dow Corning</td><td></td><td></td></tr><tr><td>Trichlorosilane</td><td>Sigma Aldrich</td><td>175552</td><td></td></tr><tr><td>Ultrasonic Cleaner</td><td>Branson</td><td>3510</td><td></td></tr><tr><td>Tube Rotator&amp;nbsp;</td><td>VWR</td><td>10136-084</td><td></td></tr><tr><td>Vortex Mixer</td><td>WiseMix</td><td>VM-10</td><td></td></tr><tr><td>Microcentrifuge</td><td>VWR</td><td>Micro 1207</td><td></td></tr><tr><td>Plasma Cleaner</td><td>Harrick Plasma</td><td>PDC-001-HP</td><td></td></tr><tr><td>PDMS Mixer</td><td>Thinky</td><td>ARE-250</td><td></td></tr><tr><td>Oven</td><td>Thermo Scientific</td><td>PR305220M</td><td></td></tr><tr><td>Epi-fluorescence Microscope</td><td>Nikon</td><td>Eclipse Ti</td><td></td></tr><tr><td>CCD Camera</td><td>Andor</td><td>Clara</td><td></td></tr><tr><td>Platinum Electrodes</td><td>Alfa Aesar</td><td>43014</td><td></td></tr><tr><td>Source Meter</td><td>Keithley</td><td>2400</td><td></td></tr><tr><td>Digital Multimeter&amp;nbsp;</td><td>Extech</td><td>410</td><td></td></tr><tr><td>Microscopy Glass Slides</td><td>Thermo Scientific</td><td>2951-001</td><td></td></tr><tr><td>Tween 20</td><td>Merck Millipore</td><td>822184</td><td></td></tr><tr><td>Sodium chloride</td><td>Fisher Scientific</td><td>7646-14-5</td><td></td></tr><tr><td>Sodium phosphate monobasic</td><td>Sigma Aldrich</td><td>71505</td><td></td></tr><tr><td>Sodium phosphate dibasic</td><td>Sigma Aldrich</td><td>S3264</td><td></td></tr><tr><td>DNA</td><td>IDT</td><td></td><td>CAA CCG ATG CCA CAT CAT TAG CTA C</td></tr><tr><td>B-Phycoerythrin</td><td>Life Technologies</td><td>P-800</td><td></td></tr><tr><td>Dynamic light scattering system for Zeta Potential Measurement</td><td>Malvern</td><td>Zetasizer Nano S</td><td></td></tr><tr><td>Photoresist&amp;nbsp;</td><td>Shipley</td><td>SPR700-1.0</td><td></td></tr><tr><td>Projection lithography</td><td>Nikon</td><td>NSR2005i9</td><td></td></tr><tr><td>Reactive Ion Etcher</td><td>Applied Materials</td><td>AME P5000</td><td></td></tr><tr><td>ICP deep reactive ion etcher</td><td>STS</td><td>STS-6&quot;</td><td></td></tr><tr><td>Contact lithography</td><td>Electronic Visions</td><td>EV620</td><td></td></tr><tr><td>Photoresist Coater Developer</td><td>SSI</td><td>SSI 150</td><td></td></tr><tr><td>Non-contact surface profiler</td><td>Wyko</td><td>NT 9800</td><td></td></tr></tbody></table>

creating sub-50 nm nanofluidic junctions in pdms microfluidic chip via self-assembly process of colloidal particles

Polidimetilsiloksan (PDMS), kalıplama ve bağlama kolaylığı ve saydamlığı mikroakışkan cihazları olmak için geçerli olan yapı malzemesidir. Nedeniyle PDMS malzemenin yumuşak olması, ancak, nanochannels oluşturmak için PDMS kullanmak zordur. kanallar plazma yapıştırılması sırasında çöker kolayca eğilimindedir. Bu yazıda alt-50, iki mikrokanallar arasında gözenekler nm nanofluidic kavşaklar oluşturmak için silika koloidal nanoparçacıkların bir buharlaşma odaklı kendini montaj yöntem mevcut. gözenek boyutu yanı sıra nanofluidic kavşak yüzey yükü sadece öz-montaj işleminden önce bir şişeye monte mikroakışkan cihazın dışında koloidal silis boncuk boyutu ve yüzey fonksiyonelleşmesine değiştirerek ayarlanabilir olduğunu. 300 nm, 500 nm ve 900 nm arasında bir kordon boyutu Nanopartiküllerin kendini düzenlemesi kullanılarak, sırasıyla, bir gözenek 45 nm ~ büyüklüğü, ~ 75 nm ve ~ 135 nm gözenekli bir membranın imal etmek mümkün olmuştur. elektrik altındaEl potansiyeli, bir katyon seçici membran gibi bu nano-gözenekli membran başlatılan iyon konsantrasyon kutuplaşma (ICP) oyunculuk 15 dakika içinde ~ 1.700 kat DNA konsantre. Bu sigara litografik nanofabrikasyon süreci PDMS mikroakışkan çip içinde iyonların ve moleküllerin nano taşıma süreçlerinin çalışma için ayarlanabilir bir nanofluidic kavşak inşa etmek için yeni bir fırsat açılır.

İki Mikrokanallarda arasında kendi kendine monte nanofluidic kavşak içeren PDMS bir elektrokinetik yoğunlaştırıcı çip) Şekil 1A gösterilmiştir. Cihazın ortasında kanal 50 mikron genişliğinde boncuk dağıtım kanalı (Şekil 1B) üzerinden bir DNA numune çözeltisi ile doldurulmuş ve her iki tarafta iki tampon çözelti kanallar ile çevrili. silika koloidal süspansiyon numune ve tampon çözelti kanalı arasında bir nanofluidic kav?...

Watch this Scientific Journal Video about Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles at JoVE.com

Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles

We propose a simple self-assembly technique of silica colloidal nanoparticles to create a nanofluidic junction between two microchannels in polydimethylsiloxane (PDMS). Using this technique, a nanoporous bead membrane with a pore size down to ~45 nm was built inside a microchannel and applied to electrokinetic preconcentration of DNA samples.

Kolloidal Parçacıkların Öz-Montaj Süreci yoluyla PDMS mikroakışkan Chip Sub-50 Nm Nanofluidic Junctions oluşturma

Polydimethylsiloxane (PDMS) is the prevailing building material to make microfluidic devices due to its ease of molding and bonding as well as its ...

creating-sub-50-nm-nanofluidic-junctions-pdms-microfluidic-chip-via

Research

JoVE Journal

Engineering

10.6K Görüntüleme.  New York University Abu Dhabi (NYUAD). We propose a simple self-assembly technique of silica colloidal nanoparticles to create a nanofluidic junction between two microchannels in polydimethylsiloxane (PDMS). Using this technique, a nanoporous bead membrane with a pore size down to ~45 nm was built inside a microchannel and applied to electrokinetic preconcentration of DNA samples.

Video: Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles

A Microfluidic-based Hydrodynamic Trap for Single Particles

The ability to confine and manipulate single particles in free solution is a key enabling technology for fundamental and applied science. Methods for particle trapping based on optical, magnetic, electrokinetic, and acoustic techniques have led to major advancements in physics and biology ranging from the molecular to cellular level. In this article, we introduce a new microfluidic-based technique for particle trapping and manipulation based solely on hydrodynamic fluid flow. Using this method, we demonstrate trapping of micro- and nano-scale particles in aqueous solutions for long time scales. The hydrodynamic trap consists of an integrated microfluidic device with a cross-slot channel geometry where two opposing laminar streams converge, thereby generating a planar extensional flow with a fluid stagnation point (zero-velocity point). In this device, particles are confined at the trap center by active control of the flow field to maintain particle position at the fluid stagnation point. In this manner, particles are effectively trapped in free solution using a feedback control algorithm implemented with a custom-built LabVIEW code. The control algorithm consists of image acquisition for a particle in the microfluidic device, followed by particle tracking, determination of particle centroid position, and active adjustment of fluid flow by regulating the pressure applied to an on-chip pneumatic valve using a pressure regulator. In this way, the on-chip dynamic metering valve functions to regulate the relative flow rates in the outlet channels, thereby enabling fine-scale control of stagnation point position and particle trapping. The microfluidic-based hydrodynamic trap exhibits several advantages as a method for particle trapping. Hydrodynamic trapping is possible for any arbitrary particle without specific requirements on the physical or chemical properties of the trapped object. In addition, hydrodynamic trapping enables confinement of a "single" target object in concentrated or crowded particle suspensions, which is difficult using alternative force field-based trapping methods. The hydrodynamic trap is user-friendly, straightforward to implement and may be added to existing microfluidic devices to facilitate trapping and long-time analysis of particles. Overall, the hydrodynamic trap is a new platform for confinement, micromanipulation, and observation of particles without surface immobilization and eliminates the need for potentially perturbative optical, magnetic, and electric fields in the free-solution trapping of small particles.

In this article, we present a microfluidic-based method for particle confinement based on hydrodynamic flow. We demonstrate stable particle trapping at a fluid stagnation point using a feedback control mechanism, thereby enabling confinement and micromanipulation of arbitrary particles in an integrated microdevice.

Tek Parçacıklar için mikroakışkan tabanlı Hidrodinamik Tuzak

The ability to confine and manipulate single particles in free solution is a key enabling technology for fundamental and applied science.  Methods for ...

Biyomühendislik

Fluorescence detection methods for microfluidic droplet platforms

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that accompany system miniaturisation. Advantages include the ability to efficiently process nano- to femoto- liter volumes of sample, facile integration of functional components, an intrinsic predisposition towards large-scale multiplexing, enhanced analytical throughput, improved control and reduced instrumental footprints.1
In recent years much interest has focussed on the development of droplet-based (or segmented flow) microfluidic systems and their potential as platforms in high-throughput experimentation.2-4 Here water-in-oil emulsions are made to spontaneously form in microfluidic channels as a result of capillary instabilities between the two immiscible phases. Importantly, microdroplets of precisely defined volumes and compositions can be generated at frequencies of several kHz. Furthermore, by encapsulating reagents of interest within isolated compartments separated by a continuous immiscible phase, both sample cross-talk and dispersion (diffusion- and Taylor-based) can be eliminated, which leads to minimal cross-contamination and the ability to time analytical processes with great accuracy. Additionally, since there is no contact between the contents of the droplets and the channel walls (which are wetted by the continuous phase) absorption and loss of reagents on the channel walls is prevented.
Once droplets of this kind have been generated and processed, it is necessary to extract the required analytical information. In this respect the detection method of choice should be rapid, provide high-sensitivity and low limits of detection, be applicable to a range of molecular species, be non-destructive and be able to be integrated with microfluidic devices in a facile manner. To address this need we have developed a suite of experimental tools and protocols that enable the extraction of large amounts of photophysical information from small-volume environments, and are applicable to the analysis of a wide range of physical, chemical and biological parameters. Herein two examples of these methods are presented and applied to the detection of single cells and the mapping of mixing processes inside picoliter-volume droplets. We report the entire experimental process including microfluidic chip fabrication, the optical setup and the process of droplet generation and detection.

Droplet-based microfluidic platforms are promising candidates for high throughput experimentation since they are able to generate picoliter, self-compartmentalized vessels inexpensively at kHz rates. Through integration with fast, sensitive and high resolution fluorescence spectroscopic methods, the large amounts of information generated within these systems can be efficiently extracted, harnessed and utilized.

Mikroakışkan damlacık platformları için Floresan algılama yöntemleri

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

Microfluidic Chips Controlled with Elastomeric Microvalve Arrays

Miniaturized microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays. Robust flow control and precise fluidic volumes are two critical requirements for these applications. We have developed microfluidic chips featuring elastomeric polydimethylsiloxane (PDMS) microvalve arrays that: 1) need no extra energy source to close the fluidic path, hence the loaded device is highly portable; and 2) allow for microfabricating deep (up to 1 mm) channels with vertical sidewalls and resulting in very precise features.

The PDMS microvalves-based devices consist of three layers: a fluidic layer containing fluidic paths and microchambers of various sizes, a control layer containing the microchannels necessary to actuate the fluidic path with microvalves, and a middle thin PDMS membrane that is bound to the control layer. Fluidic layer and control layers are made by replica molding of PDMS from SU-8 photoresist masters, and the thin PDMS membrane is made by spinning PDMS at specified heights. The control layer is bonded to the thin PDMS membrane after oxygen activation of both, and then assembled with the fluidic layer. The microvalves are closed at rest and can be opened by applying negative pressure (e.g., house vacuum). Microvalve closure and opening are automated via solenoid valves controlled by computer software.

Here, we demonstrate two microvalve-based microfluidic chips for two different applications. The first chip allows for storing and mixing precise sub-nanoliter volumes of aqueous solutions at various mixing ratios. The second chip allows for computer-controlled perfusion of microfluidic cell cultures.

The devices are easy to fabricate and simple to control. Due to the biocompatibility of PDMS, these microchips could have broad applications in miniaturized diagnostic assays as well as basic cell biology studies.

We demonstrate protocols for manufacturing and automating elastomeric polydimethylsiloxane (PDMS)-based microvalve arrays that need no extra energy to close and feature photolithographically defined precise volumes. A parallel subnanoliter-volume mixer and an integrated microfluidic perfusion system are presented.

Elastomerik Mikrovalf Diziler Kontrollü mikroakışkan Chips

Miniaturized microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays.  ...

Biyoloji

Non-plasma Bonding of PDMS for Inexpensive Fabrication of Microfluidic Devices

In this video, we demonstrate how to use the neuron microfluidic device without plasma bonding.  In some cases it may be desirable to reversibly bond devices to the Corning No. 1 cover glass.  This could be due, perhaps, to a plasma cleaner not being available.  In other instances, it may be desirable to remove the device from the glass after the culturing of neurons for certain types of microscopy or for immunostaining, though it is not necessary to remove the device for immunostaining since the neurons can be stained in the device.  Some researchers, however, still prefer to remove the device.  In this case, reversible bonding of the device to the cover glass makes that possible.  There are some disadvantages to non-plasma bonding of the devices in that not as tight of a seal is formed.  In some cases axons may grow under the grooves rather than through them.  Also, because the glass and PDMS are hydrophobic, liquids do not readily enter the device making it necessary at times to force media and other reagents into the device.  Liquids will enter the device via capillary action, but it takes significantly longer as compared to devices that have been plasma bonded.  The plasma cleaner creates temporary hydrophilic charges on the glass and device that facilitate the flow of liquids through the device after bonding within seconds.  For non-plasma bound devices, liquid flow through the devices takes several minutes.  It is also important to note that the devices to be used with non-plasma bonding need to be sterilized first, whereas plasma treated devices do not need to be sterilized prior to use because the plasma cleaner will sterilize them.

In this video we demonstrate how to use the neuron microfluidic device without plasma bonding.

Ucuz mikroakışkan Cihazlar İmalat PDMS Non-plazma Yapışma

In this video, we demonstrate how to use the neuron microfluidic device without plasma bonding.  In some cases it may be desirable to reversibly bond ...

Microfluidic Pneumatic Cages: A Novel Approach for In-chip Crystal Trapping, Manipulation and Controlled Chemical Treatment

The precise localization and controlled chemical treatment of structures on a surface are significant challenges for common laboratory technologies. Herein, we introduce a microfluidic-based technology, employing a double-layer microfluidic device, which can trap and localize in situ and ex situ synthesized structures on microfluidic channel surfaces. Crucially, we show how such a device can be used to conduct controlled chemical reactions onto on-chip trapped structures and we demonstrate how the synthetic pathway of a crystalline molecular material and its positioning inside a microfluidic channel can be precisely modified with this technology. This approach provides new opportunities for the controlled assembly of structures on surface and for their subsequent treatment.

Herein, we describe the fabrication and operation of a double-layer microfluidic system made of polydimethylsiloxane (PDMS). We demonstrate the potential of this device for trapping, directing the coordination pathway of a crystalline molecular material and controlling chemical reactions onto on-chip trapped structures.

Mikroakışkan Pnömatik Kafesler: Bir In-yonga Kristal Bindirme, Manipülasyon için Yeni Yaklaşım ve Kontrollü Kimyasal Arıtma

The precise localization and controlled chemical treatment of structures on a surface are significant challenges for common laboratory technologies. ...

Kimya

High Throughput Microfluidic Rapid and Low Cost Prototyping Packaging Methods

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging techniques.
The one-time use packaging technique employs UV-based and temperature curing epoxies to connect microtubes to access holes, wire-bonding for integrated circuit connections, and silver epoxy for electrical connections. This method is based on a robust assembly technique that can support relatively high pressure close to 1 psi and does not need any support to strengthen the microfluidic architecture.
Reusable packaging techniques consist of PDMS-based microtube interconnectors and anisotropic adhesive films for electrical connections. These devices are more sensitive and fragile. Consequently, Plexiglas support is added to the microfluidic structure to improve the electrical contact when anisotropic adhesive films are used, and also to strengthen the microfluidic architecture. In addition, a micromanipulator is needed to maintain tubes while using a thin PDMS layer to connect them to the access holes. Different PDMS layer thicknesses, ranging from 0.45-3 mm, are tested to compare the best adherence versus injection rates. Applied injection rates are varied from 50-300 &#956;l/hr for 0.45-3 mm PDMS layers, respectively. These techniques are mainly applicable for low-pressure applications. However, they can be extended for high-pressure ones through plasma-oxygen process to permanently seal the PDMS to glass substrates. The main advantage of this technique, besides the fact that it is reusable, consists of keeping the device observable when the microchannel length is very short (in the range of 3 mm or lower).

In this article we describe different techniques for microfluidic rapid prototyping platforms. The proposed techniques are based on ultraviolet (UV) sensitive and temperature curing epoxies, polydimethylsiloxane (PDMS) based tubing, wire-bonding, and anisotropic adhesive films. The assembling procedures presented are developed for both one-time use devices as well as reusable microfluidic systems.

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging ...

Patterning of Microorganisms and Microparticles through Sequential Capillarity-assisted Assembly

Controlled patterning of microorganisms into defined spatial arrangements offers unique possibilities for a broad range of biological applications, including studies of microbial physiology and interactions. At the simplest level, accurate spatial patterning of microorganisms would enable reliable, long-term imaging of large numbers of individual cells and transform the ability to quantitatively study distance-dependent microbe-microbe interactions. More uniquely, coupling accurate spatial patterning and full control over environmental conditions, as offered by microfluidic technology, would provide a powerful and versatile platform for single-cell studies in microbial ecology. 
This paper presents a microfluidic platform to produce versatile and user-defined patterns of microorganisms within a microfluidic channel, allowing complete optical access for long-term, high-throughput monitoring. This new microfluidic technology is based on capillarity-assisted particle assembly and exploits the capillary forces arising from the controlled motion of an evaporating suspension inside a microfluidic channel to deposit individual microsized objects in an array of traps microfabricated onto a polydimethylsiloxane (PDMS) substrate. Sequential depositions generate the desired spatial layout of single or multiple types of micro-sized objects, dictated solely by the geometry of the traps and the filling sequence. 
The platform has been calibrated using colloidal particles of different dimensions and materials: it has proven to be a powerful tool to generate diverse colloidal patterns and perform surface functionalization of trapped particles. Furthermore, the platform was tested on microbial cells, using Escherichia coli cells as a model bacterium. Thousands of individual cells were patterned on the surface, and their growth was monitored over time. In this platform, the coupling of single-cell deposition and microfluidic technology allows both geometric patterning of microorganisms and precise control of environmental conditions. It thus opens a window into the physiology of single microbes and the ecology of microbe-microbe interactions, as shown by preliminary experiments.

We present a technology that uses capillarity-assisted assembly in a microfluidic platform to pattern micro-sized objects suspended in a liquid, such as bacteria and colloids, into prescribed arrays on a polydimethylsiloxane substrate.

Sıralı Capillarity destekli Montaj yoluyla Mikroorganizmaların ve Mikropartiküllerin Deseni

Controlled patterning of microorganisms into defined spatial arrangements offers unique possibilities for a broad range of biological applications, ...

Mühendislik

Pneumatically Driven Microfluidic Platform for Micro-Particle Concentration

The present article introduces a method for fabricating and operating a pneumatic valve to control particle concentration using a microfluidic platform. This platform has a three-dimensional (3D) network with curved fluid channels and three pneumatic valves, which create networks, channels, and spaces through duplex replication with polydimethylsiloxane (PDMS). The device operates based on the transient response of a fluid flow rate controlled by a pneumatic valve in the following order: (1) sample loading, (2) sample blocking, (3) sample concentration, and (4) sample release. The particles are blocked by thin diaphragm layer deformation of the sieve valve (Vs) plate and accumulate in the curved microfluidic channel. The working fluid is discharged by the actuation of two on/off valves. As a result of the operation, all particles of various magnifications were successfully intercepted and disengaged. When this technology is applied, the operating pressure, the time required for concentration, and the concentration rate may vary depending on the device dimensions and particle size magnification.

The present protocol describes a pneumatic microfluidic platform that can be used for efficient microparticle concentration.

Mikro Partikül Konsantrasyonu için Pnömatik Tahrikli Mikroakışkan Platform

The present article introduces a method for fabricating and operating a pneumatic valve to control particle concentration using a microfluidic platform. ...

Kombinatoryal Plug Üretimi için Mikroakışkan Bir Cihazın Kurulumu

Bilayer Microfluidic Device for Combinatorial Plug Production

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such systems have been used to encapsulate a variety of biochemical reactions - from incubation of single cells to implementation of PCR reactions, from genomics to chemical synthesis. Coupling the microfluidic channels with regulatory valves allows control over their opening and closing, thereby enabling the rapid production of large-scale combinatorial libraries consisting of a population of droplets with unique compositions. In this paper, protocols for the fabrication and operation of a pressure-driven, PDMS-based bilayer microfluidic device that can be utilized to generate combinatorial libraries of water-in-oil emulsions called plugs are presented. By incorporating software programs and microfluidic hardware, the flow of desired fluids in the device can be controlled and manipulated to generate combinatorial plug libraries and to control the composition and quantity of constituent plug populations. These protocols will expedite the process of generating combinatorial screens, particularly to study drug response in cells from cancer patient biopsies.

Yazar Spotlight: Hesaplamalı ve Deneysel Yaklaşımları Hassas Onkolojiye Entegre Etme

The fabrication of a polydimethylsiloxane (PDMS)-based bilayer device for the production of combinatorial libraries in water-in-oil emulsions (plugs) is presented here. The necessary hardware and software required to automate plug production are detailed in the protocol, and the production of a quantitative library of fluorescent plugs is also demonstrated.

Kombinatoryal Fiş Üretimi için İki Katmanlı Mikroakışkan Cihaz

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such ...

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Kombinatoryal Fiş Üretimi için İki Katmanlı Mikroakışkan Cihaz

Kombinatoryal Fiş Üretimi için İki Katmanlı Mikroakışkan Cihaz