Name: fabrication of three-dimensional paper-based microfluidic devices for immunoassays
Uploaded: 2017-03-09T14:00:00
Description: Watch this Scientific Journal Video about Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays at JoVE.com

This work was supported by Tufts University and by a generous gift from Dr. James Kanagy. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship Program under Grant No. (DGE-1325256) that was awarded to S.C.F. D.J.W. was supported by a U.S. Department of Education GAANN fellowship. We thank Dr. Jeremy Schonhorn (JanaCare), Dr. Jason Rolland (Carbon3D), and Rachel Deraney (Brown University) for helping develop the design of the three-dimensional paper-based microfluidic device and immunoassay.

1.基于纸张的微流体器件层的制备<ol><li>准备用于使用图形设计软件程序的纸张，尼龙，和粘合剂层的图案。 6每个层可以具有不同的图案。 注：该图案可以包括被不需要的官能纸质免疫对准孔，但与三维器件的重现性制造协助。如果设备是单独组装，成条，或作为全片这些孔的位置会有所不同。用于设计图形的软件程序可能基于图案化技术（ 例如 ，光刻，蜡染，?...

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确定一个可重复的制造策略是试验发展的一个重要组成部分。 22我们使用顺序，层-层的方法来制造三维纸基微流体装置。相反那些适用折叠或折纸技术来产生从纸23的单个片材的多层器件的方法，24添加剂制造提供了许多优点：（ⅰ）多材料可以并入到单个设备架构无需修改方法印刷，对齐，或层的装配。 （二）图案化粘合膜可以集?...

<html>纸是广泛使用的范围内的制剂或等级，可以官能化以调整其性能，并且可以通过毛细作用或芯吸自主地输送流体。如果纸是用疏水物质图案化（ 例如 ，光致抗蚀剂1或蜡2），流体的芯吸可以在空间上的纸层内的控制。例如，所施加的含水样品可以定向成许多不同的区，以与存储在纸内的化学和生化试剂反应。这些纸基微流体装置已被证明是用于便携式和廉价的分析测定3，4，5，6，7的发展有用的平台。纸基微流体装置的应用包括点的护理诊断EF“&gt; 8，环境污染物9的监测，检测假药10，和离域医疗（或”远程医疗“）的有限资源设置11。 构图纸的多个层可被组装成一个集成装置，其中来自相邻层（ 即 ，高于或低于）亲水区连接，以形成连续的流体网络，其入口和出口可联接或左独立。 12每一层可以包括一个唯一的图案，使试剂和多个测定的空间分离在单个设备上执行。所得三维微流体装置，不仅能够芯吸流体，以使分析测定的（ 例如 ，肝功能试验13和小分子14的电化学检测），但它也可以SUP端口一些复杂的功能（ 例如 ，阀门15和简单机械16）常见的传统微的方法。重要的是，因为纸灯芯通过毛细作用流体，这些设备可以从用户最小的努力工作。 因为试剂可以存储在纸基装置的三维结构中，复杂的协议可以减少到单一的加入含水样品到设备。最近，我们介绍了可用于使用蜡印技术来创建图案化层纸质免疫发展的一般三维器件结构。 17，18，这些研究主要集中在与层叠的层的设备号的设计方面是如何使用的，组合物的层，并且该三维微流体网络控制的总体％的图案免疫测定的性能。最终，我们能够利用这些设计规则，以促进一个多路复用免疫测定19的快速发展。在此手稿，人类绒毛膜促性腺激素（hCG;怀孕激素）一个先前开发的免疫测定17被用作一个例子来说明，我们已对三维纸基免疫测定的装配和制造开发的策略。因此，我们专注于一个设备，而不是化验开发的组装和操作。 在夹心免疫测定，这是一种用于检测hCG的，特定于激素的一个亚基涂覆到固体基质中，然后被阻止，以限制一个样品或任何后续的试剂的非特异性吸附捕获抗体的格式。该基板是最经常的聚苯乙烯微孔板（ 例如 ，酶联免疫吸附测定或ELISA）。然后将样品加入到孔中，温育一段时间。严谨洗涤后，特定的hCG的另一亚基的抗体并使其孵育。该检测抗体可以产生可测量的信号缀合的胶体粒子，酶，或荧光团。该井再次解释的测定的结果（ 例如 ，使用读板器）之前洗涤。而商业试剂盒依靠这种耗时多步过程中，所有这些步骤都可以迅速地在纸基微流体装置以最少的干预来的用户执行。 用于hCG的免疫测定的装置，包括六个有源层，其是，从顶部到底部，用于样品此外，共轭存储，孵化，捕获，洗涤和印迹（ 图1）。试样添加层从定性滤纸制成。它有利于引入液体样品的和保护试剂在缀合物Laye的ř从由用户环境或意外接触污染。缀合物层（定性滤纸）持有用于免疫测定的色剂（ 例如 ，胶体金标记的抗体）。孵化层（定性滤纸）允许样品到纸平面内横向行进到达下一个层，捕获层之前促进与试剂的分析物的结合。所述捕获层（尼龙膜）含有特异性吸附到材料被分析物的配体。在测定完成后，该层显露以使完成的免疫复合体的可视化。洗涤层（定性滤纸）绘制过量液体，包括自由共轭试剂从脸部的捕获层的入印迹层（厚色谱纸）的距离。六层设备是由图案化的，双面粘合剂的五层保持在一起：永久性粘合剂的四层保持ASSEM的完整性放血装置以及可移除的粘合剂的一层便于剥离装置的检查捕获层上的免疫测定的结果。 对于本手稿的目的，我们使用的hCG仅阴性和阳性对照样品（0 MIU / mL和81 MIU /毫升，分别），以提供一纸基免疫分析，其允许之间的关系的一个专门讨论的代表性结果制造方法和设备的性能。除了演示如何制造设备的成功，我们强调的几个制造误差，可能导致设备或不可重复测定结果的失败。在这个手稿详细的协议，并讨论将提供有价值的洞察纸质免疫是如何设计和制造的研究人员。虽然我们集中我们的免疫证明，我们预计这里提出的指导方针将是三地扪制造广泛有用简称AE，下同纸基微流体装置。

<table border="0" cellpadding="0" cellspacing="0" width="1059">
	<tbody>
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			<td height="21" style="height:21px;width:447px;">Illustrator CC</td>
			<td style="width:135px;">Adobe</td>
			<td style="width:175px;"></td>
			<td style="width:304px;">to design patterns for layers of paper and adhesive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Xerox ColorQube 8580 printer</td>
			<td style="width:135px;">Amazon</td>
			<td style="width:175px;">B00R92C9DI</td>
			<td>to print wax patterns onto layers of paper and Nylon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Isotemp General Purpose Heating and Drying Oven</td>
			<td style="width:135px;">Fisher Scientific</td>
			<td style="width:175px;">15-103-0509</td>
			<td>to melt wax into paper</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Artograph LightTracer</td>
			<td style="width:135px;">Amazon</td>
			<td style="width:175px;">B000KNHRH6</td>
			<td>to assist with alignment of layers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Apache AL13P laminator</td>
			<td style="width:135px;">Amazon</td>
			<td style="width:175px;">B00AXHSZU2</td>
			<td>to laminate layers together</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Graphtec CE6000 Cutting Plotter</td>
			<td style="width:135px;">Graphtec America</td>
			<td style="width:175px;">CE6000-40</td>
			<td>to pattern adhesive films</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Swingline paper cutter</td>
			<td style="width:135px;">Amazon</td>
			<td style="width:175px;">B0006VNY4C</td>
			<td>to cut paper or devices</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Epson Perfection V500 photo scanner</td>
			<td style="width:135px;">Amazon</td>
			<td style="width:175px;">B000VG4AY0</td>
			<td>to scan images of readout layer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">economy plier-action hole punch</td>
			<td style="width:135px;">McMaster-Carr</td>
			<td>3488A9</td>
			<td>to remove alignment holes&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Whatman chromatogrpahy paper, Grade 4</td>
			<td style="width:135px;">Sigma Aldrich</td>
			<td>WHA1004917</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Fisherbrand chromatography paper (thick)&#160;</td>
			<td style="width:135px;">Fisher Scientific</td>
			<td>05-714-4</td>
			<td>to function as blot layer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Immunodyne ABC (0.45 &#181;m pore size )</td>
			<td style="width:135px;">Pall Corporation</td>
			<td>NBCHI3R</td>
			<td>to function as material for capture layer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">removable/permanent adhesive-double faced liner</td>
			<td style="width:135px;">FLEXcon</td>
			<td style="width:175px;">DF021621</td>
			<td>to facilitate peeling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">permanent adhesive-double faced liner</td>
			<td style="width:135px;">FLEXcon</td>
			<td style="width:175px;">DF051521</td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:19px;width:447px;">wax liner</td>
			<td style="width:135px;">FLEXcon</td>
			<td style="width:175px;">FLEXMARK 80 D/F PFW LINER</td>
			<td>to assist with patterning adhesive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">acrylic sheet</td>
			<td style="width:135px;">McMaster-Carr</td>
			<td style="width:175px;">8560K266&#160;</td>
			<td>to fabricate frame</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">self-adhesive sheets</td>
			<td style="width:135px;">Fellowes</td>
			<td style="width:175px;">CRC52215</td>
			<td>to use as protective slip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">absolute ethanol</td>
			<td style="width:135px;">VWR</td>
			<td style="width:175px;">89125-172</td>
			<td>to sanitize work area</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">bovine serum albumin</td>
			<td style="width:135px;">AMRESCO</td>
			<td style="width:175px;">0332</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sekisui Diagnostics OSOM hCG Urine Controls</td>
			<td style="width:135px;">Fisher Scientific</td>
			<td>22-071-066</td>
			<td>to use as positive and negative samples</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anti-&#946;-hCG monoclonal antibody colloidal gold conjugate (clone 1)</td>
			<td style="width:135px;">Arista Biologicals&#160;</td>
			<td>CGBCG-0701</td>
			<td>to treat conjugate layer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">goat anti-&#945;-hCG antibody</td>
			<td style="width:135px;">Arista Biologicals&#160;</td>
			<td>ABACG-0500</td>
			<td>to treat capture layer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">10X phosphate buffered saline</td>
			<td style="width:135px;">Fisher Scientific</td>
			<td style="width:175px;">BP3991</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Oxoid skim milk powder</td>
			<td style="width:135px;">Thermo Scientific</td>
			<td style="width:175px;">OXLP0031B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Tween 20</td>
			<td style="width:135px;">AMRESCO</td>
			<td style="width:175px;">M147</td>
			<td></td>
		</tr>
	</tbody>
</table>

fabrication of three-dimensional paper-based microfluidic devices for immunoassays

纸中自主灯芯流体由于毛细管作用。通过用疏水性障碍图案的纸张，流体输送可以控制和一层纸内定向。此外，堆叠构图纸的多层创建复杂的三维微流体网络可以支持的分析和生物分析测定法的发展。纸基微流体装置是便宜的，携带方便，易于使用，并且不需要外部设备进行操作。其结果是，他们拥有巨大潜力，作为点的护理诊断的平台。为了正确评价的效用和纸为基础的设备的分析性能，必须开发合适的方法，以确保它们的制造是可重复的，并在一个尺度适合于实验室环境。在这个手稿中，一种方法来制造，可以用于纸基免疫测定一般器件结构进行说明。我们使用添加剂manufacturin的一种形式克（多层层压）来制备构成图案的纸张和图案化粘合剂的多层器件。除了演示正确使用这些三维纸基微流体装置与人绒毛膜促性腺激素（hCG）的免疫测定，在制造过程中的错误可能导致设备故障进行了讨论。我们预计这种方法制造的纸张为基础的设备会发现专门针对资源有限的设置旨在分析应用的广泛的实用性。

在三维纸基微流体装置获得可再现的测定性能依赖于确保设备之间的一致性的制造方法。为了实现这一目标，我们已经确定了一些制造工艺和材料方面的考虑，并在展示纸质免疫的情况下在这里他们讨论。我们使用的蜡印刷方法，以形成纸基微流体装置（ 图2A）内的疏水性的障碍。 2此方法是理想的，因为它仅依赖于广泛使用的办公设备，需...

Watch this Scientific Journal Video about Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays at JoVE.com

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

我们详细介绍的方法来制造三维纸基微流体装置用于在免疫测定的开发使用。我们对设备组装方式是一种多层，添加剂制造。我们展示了一个夹心免疫提供对这些类型的纸张为基础的设备的代表性结果。

三维纸基微流体装置的制造用于免疫分析

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Research

JoVE Journal

Bioengineering

Planar and Three-Dimensional Printing of Conductive Inks

平面和立体印刷的导电油墨

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

科研

生物工程

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

人体软骨组织制作采用三维喷墨打印技术

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Analysis of Cell Migration within a Three-dimensional Collagen Matrix

在一个三维胶原蛋白矩阵分析细胞迁移

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic ...

Paper-based Devices for Isolation and Characterization of Extracellular Vesicles

基于纸张的设备进行隔离外囊泡与表征

Extracellular vesicles (EVs), membranous particles released from various types of cells, hold a great potential for clinical applications. They contain ...

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization

层与层胶原沉积在微流体装置的Microtissue稳定

Although microfluidics provides exquisite control of the cellular microenvironment, culturing cells within microfluidic devices can be challenging. 3D ...

Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation

娇嫩的组织操纵定制的软机器人气动设备手爪基于杆制造

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations ...

Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

Fabrication of Inverted Colloidal Crystal Polyethylene glycol Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

倒胶体晶体聚（乙二醇）脚手架制作：肝脏组织工程三维细胞培养平台

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing ...

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

多级可变高度的光刻多层带瓣微流体装置

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to ...

A Simple and Scalable Fabrication Method for Organic Electronic Devices on Textiles

关于纺织品有机电子设备的简单和可扩展的制作方法

Today, wearable electronics devices combine a large variety of functional, stretchable, and flexible technologies. However, in many cases, these devices ...

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

三维热塑性材料的印刷, 以创建自动注射器泵的反馈控制微流控应用

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...