这项工作是由美国国立卫生研究院R21 EB008177-01A2和纽约大学阿布扎比（NYUAD）加强研究2013年基金微细加工，我们在表示感谢MIT的MTL的鼎力支持技术人员和詹姆斯·韦斯顿和NYUAD的尼古拉Giakoumidis他们的支持支持采取SEM照片分别建立一个分压器。在PDMS的设备制造在NYUAD的微细核心设施进行。最后，我们想从NYUAD中心感谢丽贝卡Pittam为视频拍摄和编辑数字奖学金。

1.胶体二氧化硅悬浮液珠的制备<ol><li> 为300nm和500nm的硅石珠粒悬浮液制剂 <ol><li>涡旋二氧化硅珠纸浆悬浮液（10％w / v的水溶液）30秒。以获得均匀悬浮液。共600微升储备悬浮液吸取到1.5ml试管离心它2600 xg离心1分钟。 </li><li>替换上清液用400μl1 mM磷酸钠缓冲液（PB，pH7.0）中。 </li><li>暂停硅珠成的15％，以1 mM磷酸钠溶液在pH 7.0经涡旋的最终浓度。 </li></ol></li><li> 聚表面官能化500纳米的二氧化硅羧基珠子（烯丙胺盐酸盐，PAH），并与聚（苯乙烯磺酸钠，PSS）聚电解质 <ol><li>暂停0.1用10ml 1M NaCl的（pH 7.0）中为1％（重量/体积）珠悬浮克与羧基500nm的二氧化硅珠粒。 </li><li>在1M NAC准备0.4％PAH（MW 65K）升通过在15ml 1M NaCl的溶解原液的300微升（20％w / v的水溶液）。通过在20毫升的1M NaCl溶液0.18克PSS溶解在1M NaCl的溶液制备0.9％的PSS（MW 70K）。涡旋都为1分钟的解决方案。以完全溶解聚电解质。 </li><li>加入200微升的PAH溶液9.8毫升1％的二氧化硅羧基珠子在15ml管以沉积具有羧基官能团上的二氧化硅珠粒带正电荷的聚电解质层。 VORTEX珠悬挂1分钟。孵育它在管旋转60分钟。在室温。 </li><li>离心珠悬浮液在1801 XG 1分钟。并用10毫升去离子水洗掉未结合的PAH五次。每个离心和除去上清液后，被致密地填充在管底部的小珠。加入8毫升DI水，以使珠可重悬于和到下一个离心机步骤洗掉现有之前破坏通过用2ml DI水剧烈移液珠丛。 </li><li>跟随对于PSS涂敷在1.2.3和1.2.4的步骤在珠子上沉积带负电荷的层。从1.2.4第5 次洗涤步骤除去DI水上清液之前的PSS的沉积后重新悬浮珠子9.8毫升1- M氯化钠的。 <ol><li>使用使用2-毫升1- M氯化钠的打破了珠丛在15ml试管的底部，然后加入8毫升1- M氯化钠的相同的蓬勃移液步骤。加入200μl的PSS的溶液9.8毫升沉积有单一的PAH层的二氧化硅珠粒。涡旋1分钟后和温育60分钟。在试管旋转器中，重复用DI水5洗涤步骤。 </li><li>使用根据制造商的方案，动态光散射系统，以验证已正确执行了聚电解质沉积过程测量之前和之后的每个聚电解质涂层珠的ζ电位（见表1）。 </li></ol></li><li>重复用DI水的单一的PSS层以下五个洗涤步骤用0.05％吐温20（15％重量/体积）中的微流体装置在使用之前，以提高其流动性的沉积和再悬浮于650微升1mM的磷酸钠缓冲液中的珠。 </li></ol></li><li>按照从1.2.5如上所述1.2.6为与胺官能团为500nm的二氧化硅珠粒沉积的PSS的单层的步骤。 </li></ol> 2.将PDMS微流控芯片的制备<ol><li> 硅主的微细 <ol><li>制造用的微细加工技术如下PDMS成型硅主。 <ol><li>旋涂1μm的薄光致抗蚀剂以4000rpm在硅片上。图案使用投影光刻（曝光时间170毫秒）和蚀刻700nm的深和2微米宽的平面纳米通道的层（作为nanotraps为二氧化硅珠）用反应性离子蚀刻。 </li><li>使用以下蚀刻参数达到3.5纳米/秒的刻蚀速率:CHF 3（45 SCCM），CF 4（15标准立方厘米），Ar（气体为100sccm），压力100毫乇，RF功率200W。 </li></ol></li><li>旋转涂布，以2000rpm的转速第二个为1微米厚的光刻胶层和执行对准先前图案nanotraps。通过接触光刻和硅的深反应离子蚀刻（DRIE）的微通道图案。使用DRIE参数26在表2中。 </li></ol></li><li> PDMS模具的制作 <ol><li> Silanize在真空罐O / N三氯硅烷（50微升）的硅主人。 注意:Tricholorosilane是有毒和腐蚀性的材料。总是在化学罩与适当的个人防护装备使用。 </li><li>在10混合基与固化剂:在硅烷化硅母1的比例，并投PDMS和在70℃下固化它用于在对流烘箱2小时。 </li><li>用后一个PL的等离子净化器取下，用小刀和等离子债券它的空白晶圆上的芯片主的PDMS板阿斯玛治疗在1分钟的等离子体清洁器。连接带沿边缘标记以下PDMS铸造步骤分割线。 </li><li>在真空罐子三氯硅烷（50微升）O / N Silanize的PDMS模具。 </li><li>施放的PDMS（碱:1的比例:在10固化剂）的硅烷化的PDMS模具，并在70℃下固化它用于在对流烘箱2小时。 </li></ol></li><li> 与PDMS器件的制造 <ol><li>剥去一起打上胶带分割线的PDMS模具硫化硅橡胶板。 </li><li>为1.5mm活检穿孔，清洁用胶带冲贮存器孔，冲洗用异丙醇（IPA）中，用氮气干燥。 </li><li>等离子体键上在等离子体清洁器的等离子体处理1分钟后25毫米×75毫米的显微镜载玻片的PDMS设备。 </li></ol></li><li> Ultrasonicate珠停牌60分钟。在超声浴中之前填充。吸管10微升悬浮微珠（300 nm的非功能化的二氧化硅是广告，或500纳米的二氧化硅羧基珠PAH-PSS层，或500与PSS层纳米二氧化硅胺珠）进入入口4和6的每个（参见图1的A，B）的PDMS芯片的等离子体接合到后，立即玻璃衬底。轻轻拍打PDMS芯片用枪头上提升珠包装。 <ol><li>填充珠的供货渠道后，覆盖除了1和9，用胶带所有的入口。空气干燥设备3小时并储存在+ 4℃下在使用前， 图2给出了胶体自组装过程的一个步骤的分步示意图。 </li></ol></li></ol> 3.试验研究DNA的浓度电动<ol><li>填充储层3,7用缓冲溶液（10微升1mM的PB）和储存器5用DNA样品（10微升在1mM的PB为10nM），并应用一个轻柔的负压与储2倒置枪头，8和10来填充用的溶液中的信道没有气泡（见<strong&gt;的图1B）。 </li><li>加10微升1mM的的PB到水库2和8和10μl10nM的DNA的储层10来平衡压力，并等待5分钟。以达到平衡。 </li><li>将铂针入储层3，5，7，10。 </li><li>横跨使用连接到源米和Pt线的分压器纳米流体结施加电压。先上水库5座，10和GND上水库3,7适用30 V。 </li><li>约30秒后，减少对水库10的电压25 V。 </li><li>使用机械快门，在每5秒一周期开口记录从DNA的荧光信号时，以减少样品的光漂白。 </li></ol>

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The authors have nothing to disclose.

继通用设备设计方案，研究纳米流体，我们用胶体纳米粒子的蒸发驱动的自组装，而不是光刻图案纳米通道阵列制造的两微流体通道之间的纳米流体连接处。当流动的胶体粒子进入珠输送通道，nanotraps的具有700nm的深度和2微米的小珠传送通道的两侧，在100微米的总宽度的宽度的阵列防止珠悬浮液流入缓冲液和样品通道由于在nanotraps的表面张力。一旦捕获，装在胎圈输送通道迅速胶体粒子和形成在?...

纳流控是μTAS的一个新兴的研究领域（微全分析系统）在10 1的长度尺度来研究生物过程或离子的迁移现象和分子- 10 2纳米。用的纳米流体的工具，如纳米通道的到来，分子和离子的输运过程可以以前所未有的精度进行监测和操纵，如果需要的话，通过利用只在该长度尺度对分离和检测是可用的功能。1,2之一这些特征纳米级特征是表面以在纳米通道散装电荷（或Dukhin数）的高比例，可能会导致电荷不平衡，并启动纳米通道和微通道之间的离子浓度极化（ICP）。3 对于纳米流体现象的研究一种常见的设备平台包括由纳米通道作为结的阵列连接的两个微通道系统。4-6 </sup&gt;用于构建这样的纳米流体装置的选择的材料是硅，因为它的高刚度，防止信道从期间接合工艺倒塌7然而，硅器件制造需要昂贵的掩模和在洁净室设备处理的大量。8- 10由于装置制造的，通过模制和等离子体结合，聚二甲基硅氧烷（PDMS）的便利性已被广泛接受的作为建筑材料为微流体，这将是对纳米流体的理想材料为好。然而，它的杨氏模量低周围360-870千帕，使得等离子体接合时与PDMS通道容易可折叠的。纳米通道（宽深）的最小长宽比必须小于10:1，这意味着通过标准光刻PDMS器件的制造会变得极具挑战性的，如果纳通道深度必须小于100纳米，需要一个信道宽度小于photolith的电流限制地理学在约1微米。为了克服这一限制，已经尝试来创建使用非光刻方法在PDMS纳通道如拉伸以引发裂纹等离子处理后的78纳米11或形成皱纹平均深度12塌陷允许用机械压力将PDMS通道一纳通道的高度低至60纳米。13 即使这些高度发明的非光刻方法允许低于100nm建筑纳通道中的深度，纳米通道制造的尺寸可控性仍对阻碍一个广泛接受的PDMS作为建筑材料为纳米流体装置。纳米通道的另一个关键问题，无论是在硅或PDMS，是在壳体的表面官能有必要改变对离子或分子的操纵通道壁的表面电荷。通过连接装置组装后，纳米通道是非常困难达到表面官能由于扩散限制运输。创建具有高尺寸精确度的和容易的表面官能纳米级信道，在微流体装置引起的蒸发14-16胶体粒子的自组装法可以是有前途的方法之一。除了 ​​孔尺寸和表面特性的可控性，甚至有可能以调谐使用涂有聚电解质的胶体粒子时在原位通过控制温度，17的pH值，18,19和离子强度的细孔的尺寸。18由于这些对于毛细管电，20生物传感器，21蛋白浓度22以及微流体蛋白质和DNA的分离。14,23在这项研究中的优势，胶体粒子的自组装方法已经找到的应用程序，我们部署这种自组装方法来构建在电动富集装置PDMS需要两个微通道之间的纳米流体结24的电动浓度背后的基本机制是基于离子浓度极化（ICP）。包括在以下方案25的制造和装配步骤的详细描述。

<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

聚二甲基硅氧烷（PDMS）是普遍的建筑材料，使微流体装置，因为它易于成型和粘合的以及它的透明度。由于PDMS材料的柔软性，但是，它是具有挑战性的使用PDMS用于构建纳米通道。渠道往往等离子体结合过程中容易崩溃。在本文中，我们提出了二氧化硅胶体纳米粒子的蒸发驱动的自组装方法与子50 2微之间50nm孔纳流控创建路口。孔尺寸以及表面电荷的纳米流体交界的是可调简单地由自组装过程之前，改变组装微流体装置的外胶体二氧化硅珠尺寸和表面官能在小瓶中。使用具有300纳米，500纳米，900纳米的珠粒度的纳米颗粒的自组装，有可能分别与〜45纳米的孔隙尺寸，〜75 nm和〜135毫微米，制作多孔膜。在电人的潜力，这种纳米膜发起离子浓差极化（ICP）作为阳离子选择性膜15分钟内集中精力靠〜1700倍的DNA。这种非光刻纳米制造工艺开辟了建立一个PDMS微流控芯片内的离子和分子纳米运输过程的研究可调谐纳米流体结了新的契机。

在PDMS一个电动集中器芯片包含两个微通道之间的自组装纳米流体结在图1A中示出）。在该装置的中部的通道是通过一个50微米的宽珠输送通道（ 图1B）填充有DNA样品溶液并通过在每侧的两个缓冲溶液的通道两侧。二氧化硅胶体悬浮液流入到珠输送通道等离子体接合后立即创建样品和缓冲液通道之间的纳米流体交界处。由700毫微米深和2微米宽的纳通...

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.

通过胶体粒子自组装过程创建在PDMS微流控芯片分50纳米的纳米流体结

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 次观看.  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.

视频: 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.

一个单粒子为基础的微流控液力陷阱

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

科研

生物工程

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.

荧光检测方法，为微流体液滴平台

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.

微流控芯片控制，弹性微阀阵列

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

生物学

Endothelialized Microfluidics for Studying Microvascular Interactions in Hematologic Diseases

Advances in microfabrication techniques have enabled the production of inexpensive and reproducible microfluidic systems for conducting biological and biochemical experiments at the micro- and nanoscales 1,2. In addition, microfluidics have also been specifically used to quantitatively analyze hematologic and microvascular processes, because of their ability to easily control the dynamic fluidic environment and biological conditions3-6. As such, researchers have more recently used microfluidic systems to study blood cell deformability, blood cell aggregation, microvascular blood flow, and blood cell-endothelial cell interactions6-13.However, these microfluidic systems either did not include cultured endothelial cells or were larger than the sizescale relevant to microvascular pathologic processes. A microfluidic platform with cultured endothelial cells that accurately recapitulates the cellular, physical, and hemodynamic environment of the microcirculation is needed to further our understanding of the underlying biophysical pathophysiology of hematologic diseases that involve the microvasculature.
Here, we report a method to create an "endothelialized" in vitro model of the microvasculature, using a simple, single mask microfabrication process in conjunction with standard endothelial cell culture techniques, to study pathologic biophysical microvascular interactions that occur in hematologic disease. This "microvasculature-on-a-chip" provides the researcher with a robust assay that tightly controls biological as well as biophysical conditions and is operated using a standard syringe pump and brightfield/fluorescence microscopy. Parameters such as microcirculatory hemodynamic conditions, endothelial cell type, blood cell type(s) and concentration(s), drug/inhibitory concentration etc., can all be easily controlled. As such, our microsystem provides a method to quantitatively investigate disease processes in which microvascular flow is impaired due to alterations in cell adhesion, aggregation, and deformability, a capability unavailable with existing assays.

A method to culture an endothelial cell monolayer throughout the entire inner 3D surface of a microfluidic device with microvascular-sized channels (&lt;30 &mu;m) is described. This in vitro microvasculature model enables the study of biophysical interactions between blood cells, endothelial cells, and soluble factors in hematologic diseases.

内皮化微流体研究在血液病的微血管相互作用

Advances in  microfabrication techniques have enabled the production of inexpensive and  reproducible microfluidic systems for conducting biological and ...

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.

PDMS微流体装置的廉价制造的非等离子键合

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.

微流控气动笼:在芯片水晶捕获，处理的新方法和控制化学处理

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

化学

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.

通过顺序毛细管辅助组装对微生物和微粒进行图案化

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

工程学

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.

气动驱动的微流体平台，用于微颗粒浓缩

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

用于组合插头生产的微流体装置的设置

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.

作者聚焦：在精准肿瘤学中整合计算和实验方法

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.

用于组合塞生产的双层微流控装置

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