Name: aqueous droplets used as enzymatic microreactors and their electromagnetic actuation
Uploaded: 2017-08-28T13:00:00
Description: Watch this Scientific Journal Video about Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation at JoVE.com

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1. 磁性纳米粒子的水化 <ol> <li> 用于合成疏水性磁性微粒, 添加 0.85 g FeCl 3 六水合物 (3.14 摩尔) 和 0.30 g FeCl 2 四水合物 (1.51 摩尔) 至200毫升水/乙醇溶液 (4:1 伏/伏). </li>
	<li> 对此混合物, 增加0.20 毫升 1 h, 1 h, 2 h, 2 h Perfluorooctyltriethoxysilane (PFOTES) (5.23 摩尔) 与剧烈搅拌由磁性搅拌器 (500 rpm)。在惰性气体气氛中 (N 2 ) 进行合成, 使用带有软木塞的圆底烧瓶, 以防止磁铁矿微?...

<ol>
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为了成功地利用微流控技术, 必须将反应量与生物科技合成和分析的要求相对应。在这里提出的驱动平台, 可以移动微流控液滴的磁力。通过将液滴与磁性疏壳封闭在反应平台的平面表面上, 可以在两个维度上自由地进行运动。因此, 在传统的微流控系统中, 采用具有复杂流量控制装置的预定义微流控通道系统来传递溶剂和样品。因此, 小型反应液滴的自动驱动是对已知的实验室芯片平台的显著简化?...

微型反应的技术应用主要是在预定义微通道芯片中进行的。这些系统在文献中得到广泛的建立和全面的描述 (特别是1,2,3)。在 2011年, 微流控技术的营业额在全球总计62亿欧元4。相比之下, 自由移动的微型反应堆舱室的使用以前只在有限的范围内进行过审查和出版。最常见的移动水下微滴的方法是润湿5。其他方法为下落在表面上是基于电场6, 磁力力量7或声学驱动8。由于其不利的表面体积比, 这些 droplet-based 微系统暴露在强烈的蒸发效应。因此, 下落运动通常建立作为液体 two-phase 系统, 其中上部阶段有一个高沸点保护水相从蒸发。然而, 这种方法有很大的风险, 污染的反应液滴不受控制的扩散。这是对上述系统的技术建立的一个重大障碍。
最近的工作涉及换药液固相转变。一种高效的方法是使用疏表面, 从而形成球形水滴。这个反应概念的引伸是使用微反应隔间与疏表面或壳, 例如可能包括聚四氟乙烯 (PTFE) 微粒9。他们的接触角度在表面上通常是在160范围内 (取决于表面粗糙度)。因此, 球形隔间提供了对表面运动的最小阻力, 同时提供了防止水蒸发的保护措施。
含微 PTFE 颗粒的水性水滴可以保持其球形形状, 直径可达2毫米左右。在更高的容量, 疏水壳通常不是完全地闭合的10。利用离子液体12, 对其它壳体材料的影响以及液体大理石在非极性溶剂中的应用范围进行了研究。为形成疏水性基于壳, 到目前为止微粒直径在大小的10毫微米-30 µm 被描述了11,14,16。新的研究表明, 疏水性纳米粒子作为壳体材料, 比微粒13的使用更为有效。第一个稳定性研究证实, 当粒子大小从 ca 减少时, 稳定性提高了.600 nm 到ca 100 nm。这可能是由于在水球体周围的密度更大的粒子分布15。
水的反应隔间的保护由疏水壳和他们的指定作为液体大理石在2001年首先被描述了由 Aussillous et al.和 Mahadevan et al.17,18. 从那以后, 这些定义的反应隔间的应用很少被描述。例如, 基于液体弹珠19的气体传感器和基于光学定性基础的水污染检测方法已被开发成20。作者区分了高反应率和低消耗化学物质的微反应系统的优势。最近的出版物涉及生产 pH 敏感的液体弹珠16或表示 "两面" 粒子与两种不同的功能涂层。例如, Bormashenko et al.可以合成一个微, 其外壳由聚四氟乙烯和半导体炭黑21组成。此外, 通过渗透气液界面24, 反应器能高效、方便地吸收外氧作为单体的合成 polyperoxides。在另一种方法中, 二氧化硅-基于液弹的壳体提供反应基板表面来调节经典的银镜反应26。亲水-核-疏水壳液滴的研究和开发目前存在的问题是粒度调整、单分散液滴的可再生生产、表面润湿性和第二次的效果亲水壳在微反应舱室22, 以及更好地控制液滴轨迹,例如用于开发连续 microPCR 系统4。
这些反应器的磁性驱动提供了相对地高运动范围的好处和一个好选择力量, 当工作在生物化学的系统时。在使用疏水磁铁矿粒子时, 它们既实现了磁力传递到反应器运动的功能, 也满足了疏水壳的作用。在 2006年, 莱曼et al.第一次被假定为液滴内磁性微粒的磁性运动。23和田et al.25, 谁使用手动移动永久磁铁作为执行器的一个单一的水滴的动员。另一种移动少量液体的方法是由赵et al.实现的, 后者使用疏水 Fe3O4粒子作为磁壳。磁性液体大理石的壳在下落的上部被打开了由垂直的反向磁场27。基于这一概念, 薛et al.能够开发出形成微的粒子, 其表面张力为20.1 达因 cm− 1 28。Lin et al.捏造了新的纤维素微/纳米分层球体与 superparamagnetism 和棉织, 提供了神的稳定性磁液滴运输和操作31。到目前为止, 这只是作为一个原理的研究, 而不是用于任何应用程序。液体弹珠的磁性和电控目前在第一种方法中进行。赵et al.在 2010年15和张et al. 2012 29中, 可以通过手动操作在核心壳液滴下的永磁体运动来进行液滴操作。Bormashenko et al.11通过接近钕磁铁, 实现了将铁磁性液体大理石的加速度提高到 25 cm s-1的速度。上述原则研究完全是由一个小型永磁体的手动运动进行的。作为下一步的发展步骤, 赵et al.最近能够通过改变永磁体的距离来估计磁性液体大理石运动所需的磁通密度30。对于与普通实验室芯片系统相媲美的反应控制, 提供自动控制离散液体 v 的方法似乎是不可避免的。olumes为了满足这一需要, 我们开发了一种新的基于变场梯度的控制系统, 以固定、移动和打开磁反应器。

<table border="0" cellpadding="0" cellspacing="0" width="903">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">3D-printer</td>
			<td style="width:163px;">FelixPrinters</td>
			<td style="width:211px;">Pro1</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red)</td>
			<td>Life Technologies</td>
			<td>A12222</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ammonium hydroxide</td>
			<td style="width:163px;">TU-KL</td>
			<td>1072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CAD software</td>
			<td style="width:163px;">Siemens</td>
			<td style="width:211px;">Soled edge</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Contact angle measuring device</td>
			<td style="width:163px;">Dataphysics</td>
			<td style="width:211px;">OCA 20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Cylinder magnet</td>
			<td style="width:163px;">Webcraft GmbH</td>
			<td style="width:211px;">S-04-13-N</td>
			<td>https://www.supermagnete.de/stabmagnete-neodym-rund/stabmagnet-durchmesser-4mm-hoehe-12.5mm-neodym-n42-vernickelt_S-04-13-N</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dipotassium phosphate</td>
			<td>Bernd Kraft</td>
			<td>7758-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Drying oven</td>
			<td>Binder</td>
			<td>FD 115</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Sigma-Aldrich</td>
			<td>68-17-5</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">FeCl2 tetrahydrate</td>
			<td style="width:163px;">TU-KL</td>
			<td>1625</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">FeCl3 hexahydrate</td>
			<td style="width:163px;">TU-KL</td>
			<td>1622</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluorescence probe</td>
			<td style="width:163px;">PerkinElmer</td>
			<td style="width:211px;">LS 55</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Horseradish peroxidase</td>
			<td>Carl Roth</td>
			<td>9003-99-0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hydrogen peroxide</td>
			<td>Th.Geyer GmbH &#38; Co</td>
			<td>7722-84-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monopotassium phosphate</td>
			<td>Bernd Kraft</td>
			<td>7778-77-0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Peltier element</td>
			<td style="width:163px;">Conrad&#160;</td>
			<td style="width:211px;">193569</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Perfluoroctyltriethoxysilane</td>
			<td>Sigma-Aldrich</td>
			<td>51851-37-7</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scanning Electron Microscope</td>
			<td style="width:163px;">FEI</td>
			<td style="width:211px;">Helios NanoLab 650 DualBeam</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Separation bar magnet</td>
			<td style="width:163px;">Webcraft GmbH</td>
			<td style="width:211px;">Q-40-20-10-N,&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Winding machine</td>
			<td style="width:163px;">IWT GmbH</td>
			<td style="width:211px;">FW122</td>
			<td></td>
		</tr>
	</tbody>
</table>

aqueous droplets used as enzymatic microreactors and their electromagnetic actuation

为了成功地实现微流体反应系统, 如 PCR 和电泳, 小液体体积的运动是必不可少的。在传统的实验室芯片平台中, 溶剂和样品通过具有复杂流量控制装置的微流控通道传递。在这里提出的液滴驱动平台是一个有前途的选择。有了它, 它是可能移动液体下落 (微) 在一个反应平台的平面表面 (lab-in-a-下落)。反应器在平台疏水表面的驱动是基于对液滴外层的磁力作用, 它是由一层薄薄的疏磁铁矿颗粒制成的。平台的疏水表面是需要的, 以避免任何接触之间的液体核心和表面, 使微的平滑移动。在平台上, 可以同时定位和移动一个或多个带有10µL 卷的反应器。该平台本身包括一个 3 x 3 矩阵的电气双线圈, 其中既容纳钕或铁芯。磁场梯度是自动控制的。通过磁场梯度的变化, 可以自动操纵反应器的磁性疏水壳, 以使微或打开壳体可逆地运动。基质和相应的酶的反应可以通过合并反应器或使它们与表面固定化催化剂接触来启动。

贝壳颗粒的直径约640纳米。在这个 fluorosilane 壳颗粒内的磁化纳米粒子的直径在22纳米和 37 nm 之间。一个5µL 微与水作为一个液体核心有一个接触角的周围160。
如上所述, 移动10µL 微所需的力是1.34 ±0.08 µN.图 1显示了铜线线圈的电磁力和4500毫安驱动的内铁芯。通过有限元模型 (FEM) 估计力分布, 以确定所需?...

Watch this Scientific Journal Video about Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation at JoVE.com

Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation

Lab-in-a 滴反应系统允许在微流控尺度上实现复杂反应的多用途。一个由 3 x 3 电磁线圈组成的自动驱动平台被开发并成功地用于合并两个10µL 反应器, 从而在产生的液体弹珠中启动酶反应。

用作酶反应器的水滴及其电磁驱动

For the successful implementation of microfluidic reaction systems, such as PCR and electrophoresis, the movement of small liquid volumes is essential. In ...

Research

JoVE Journal

Bioengineering

Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets

固相Submonomer Peptoid聚合物及其自组装成高度有序的纳米片的合成

Peptoids are a novel class of biomimetic, non-natural, sequence-specific heteropolymers that resist proteolysis, exhibit potent biological activity, and ...

科研

生物工程

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

一个工具包，使水环境中的烃类转化

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The ...

Cell Co-culture Patterning Using Aqueous Two-phase Systems

利用双水相系统的细胞共培养图案

Cell patterning  technologies that are fast, easy to use and affordable will be required for the  future development of high throughput cell assays, ...

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

多尺度孔梯度，聚电解质和间接监测改性金属植入物<em&gt;在体内</em

Metallic implants, especially titanium implants, are widely used in  clinical applications. Tissue in-growth and integration to these implants in  the ...

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

测微探针进行远程磁驱动<em&gt;原位</em&gt; 3D细菌生物膜物理性质的映射

Bacterial adhesion and growth on interfaces lead to the formation of three-dimensional heterogeneous structures so-called biofilms. The cells dwelling in ...

Isolation of Cellular Lipid Droplets: Two Purification Techniques Starting from Yeast Cells and Human Placentas

隔离细胞脂滴：两个纯化技术从酵母细胞与人类的胎盘开始

Lipid droplets are dynamic organelles that can be found in most eukaryotic and certain prokaryotic cells. Structurally, the droplets consist of a core of ...

An In Vitro Enzymatic Assay to Measure Transcription Inhibition by Gallium(III) and H3 5,10,15-tris(pentafluorophenyl)corroles

An In Vitro Enzymatic Assay to Measure Transcription Inhibition by GalliumIII and H3 5,10,15-trispentafluorophenylcorroles

一个<em&gt;在体外</em&gt;酶测定来测量转录抑制作用镓（Ⅲ）和H<sub&gt; 3</sub&gt; -5,10,15-三（五氟苯基）corroles

Chemotherapy often involves broad-spectrum cytotoxic agents with many side effects and limited targeting. Corroles are a class of tetrapyrrolic ...

Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing

癌症细胞球体机器人生产与药物测试的双水相系统

Cancer cell spheroids present a relevant in vitro model of avascular tumors for anti-cancer drug testing applications. A detailed protocol for producing ...

Photopatterning Proteins and Cells in Aqueous Environment Using TiO2 Photocatalysis

Photopatterning Proteins and Cells in Aqueous Environment Using TiO2 Photocatalysis

光图案蛋白质和细胞中的水环境中使用二氧化钛<sub&gt; 2</sub&gt;光催化

Organic contaminants adsorbed on the surface of titanium dioxide (TiO2) can be decomposed by photocatalysis under ultraviolet (UV) light. Here we describe ...

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor

蛋白质的MS检测快速酶法处理与流通型微反应器

The vast majority of mass spectrometry (MS)-based protein analysis methods involve an enzymatic digestion step prior to detection, typically with trypsin. ...