Name: self-assembly of hybrid lipid membranes doped with hydrophobic organic molecules at the water/air interface
Uploaded: 2020-05-01T14:00:00
Description: Watch this Scientific Journal Video about Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface at JoVE.com

这项工作得到了日本科学技术厅（JPMJCR14F3）的CREST项目和日本科学促进协会（19H00846和18K14120）的赠款援助支持。这项工作部分在东北大学电气通信研究所纳米电子和自旋电子学实验室进行。

1. 准备混合解决方案
<ol><li>在超声波浴中清洗四个 4 mL 一次性玻璃瓶和螺钉盖（带 PTFE 涂层密封件），在蒸馏水中清洗 10 分钟（用过滤系统净化），然后分别清洗乙醇和氯仿。在氮气流中干燥玻璃瓶和瓶盖。</li>
	<li>在厌氧手套箱中，在洗净的玻璃瓶中，通过溶解氯仿中的聚氯聚氯，在洗净的玻璃瓶中准备一个CuPc库存溶液（10mg/mL）。</li>
	<li>通过 0.2 μm 聚四氟乙烯 （PTFE） 膜过滤 CuPc 溶液。</li...

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在杂交膜的前体溶液中，混合有机溶剂（氯仿和六烷）而不是纯氯仿用于溶解脂质和CuPc。如果使用纯氯仿，前体溶液的密度将高于水。因此，溶液极有可能沉入水底，而不是扩散到水面上。将低密度溶剂六烷添加到前体溶液中，可确保溶液漂浮在水面上，并在溶剂蒸发后形成均匀的混合膜。还应当指出，溶液与水面接触后，会立即观察到轻微的表面波，这可能是由于膜和有机溶剂的扩散，这将改...

作为细胞膜的基本框架，细胞内部通过脂质双层系统从外部分离。该系统由两栖磷脂组成，由亲水磷酯"头"和疏水脂肪酸"尾巴"组成。由于脂质双层体在水环境中具有显著的流动性和自组装能力，1、2、人工脂质双层体可以采用简单的方法3、4形成。各种类型的膜蛋白，如离子通道，膜受体和酶，已被纳入人工脂质双层，以模仿和研究细胞膜5，6的功能。最近，脂质双层体掺杂了纳米材料（如金属纳米粒子、石墨烯和碳纳米管），形成功能性混合膜7、8、9、10、11、12、13。形成这种混合膜的广泛使用的方法涉及形成掺杂脂囊泡，其中含有疏水性材料，如改性Au-nano粒子7或碳纳米管11，然后产生的囊泡被融合到平面支持的脂质双层体。然而，这种方法是复杂和耗时的，这限制了这种混合膜的潜在用途。
在这项工作中，脂质膜掺杂有机分子，以产生混合脂质膜，通过自组装在水/空气界面形成。该协议涉及三个步骤：混合溶液的制备，在水/空气界面形成混合膜，以及将膜转移到Si基板上。与以前报告的其他方法相比，此处描述的方法更简单，不需要复杂的检测。使用这种方法，可以在更短的时间内形成空气稳定的混合脂质膜，具有较大的面积。本研究中使用的纳米材料是一种半导体有机分子，铜（II）2，9，16，23-四-tert-丁基-29H，31H-邻苯二甲酸酯（CuPc），广泛应用于许多应用，包括太阳能电池、光探测器、气体传感器和催化14，15。CuPc是一种具有平面结构的小有机分子，其疏水特性对磷脂二重奏的"尾巴"具有很高的亲和力。其他组报告说，CuPc分子可以在单晶表面上自组装，形成高有序的结构16，17。因此，通过自组装，极有可能将CuPc分子并入脂质双层体。
我们详细介绍了用于形成膜的程序，并为顺利实施此程序提供了一些建议。此外，本文还提出了混合脂膜的一些现性结果，并讨论了该方法的潜在应用。

<table>
	<tbody>
		<tr>
			<td>Chloroform</td>
			<td>Wako&#160;Chemicals</td>
			<td>033-08631</td>
			<td></td>
		</tr>
		<tr>
			<td>CuPc</td>
			<td>Sigma-Aldrich</td>
			<td>423165</td>
			<td></td>
		</tr>
		<tr>
			<td>DPhPc</td>
			<td>Avanti Polar Lipids</td>
			<td>850356C</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass vials with screw cap</td>
			<td>Nichiden-Rike Glass Co., Ltd</td>
			<td>6-29801</td>
			<td></td>
		</tr>
		<tr>
			<td>Hexane</td>
			<td>Wako&#160;Chemicals</td>
			<td>084-03421</td>
			<td></td>
		</tr>
		<tr>
			<td>Membrane filters</td>
			<td>Merck Millipore Ltd.</td>
			<td>R8CA42836</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-syringe</td>
			<td>Hamilton</td>
			<td>80530</td>
			<td></td>
		</tr>
		<tr>
			<td>Peristaltic pump</td>
			<td>Tokyo Rikakikai Co., Ltd.</td>
			<td>11914199</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex mixer</td>
			<td>Scientific Industries, Inc.</td>
			<td>SI-0286</td>
			<td></td>
		</tr>
	</tbody>
</table>

self-assembly of hybrid lipid membranes doped with hydrophobic organic molecules at the water/air interface

由于其独特的特性，包括超薄厚度（3-4纳米），超高电阻率，流动性和自组装能力，脂质双层层可以很容易地功能化，并可用于各种应用，如生物传感器和生物器件。在这项研究中，我们引入了一个平面有机分子：铜（II）2，9，16，23-四-tert-丁基-29H，31H-邻苯二甲酸酯（CuPc）掺杂脂膜。CuPc/脂质混合膜通过自组装在水/空气界面形成。在这个膜中，疏水性CuPc分子位于脂质分子的疏水尾之间，形成脂质/CuPc/脂质夹层结构。有趣的是，通过将混合膜转移到Si基板上，可以很容易形成空气稳定的混合脂质双层体。我们报告了将纳米材料整合到脂质双层系统中的直接方法，这是制造生物传感器和生物设备的新方法。

由于存在CuPc分子，成型膜具有均匀的浅蓝色。彩色膜的面积通常为几平方厘米。在图1A和图1B中，我们显示了Si基板上混合脂膜的微观图像和原子力显微镜（AFM）图像（包括高度轮廓）。在 AFM 图像中，左上角的膜很厚，厚度为 79.4 nm，右下角的膜很薄，厚度为 4.9 nm。薄膜显示 Ra = 0.4 nm 的表面粗糙度，接近清洁的 Si 基材。此外，基于多个测量，膜厚度?...

Watch this Scientific Journal Video about Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface at JoVE.com

Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface

我们报告一个协议，通过掺杂脂质双层铜（II）2，9，16，23-四-特-丁基-29H，31H-邻苯二甲酸酯（CuPc）分子，在水/空气界面上生产混合脂质膜。由此产生的混合脂质膜具有脂质/CuPc/脂质夹层结构。该协议也适用于其他功能纳米材料的形成。

在水/空气界面上掺杂与疏水有机分子混合脂膜的自组装

Because of their unique properties, including an ultrathin thickness (3-4 nm), ultrahigh resistivity, fluidity and self-assembly ability, lipid bilayers ...

我们的协议演示了一种制造有机分子掺杂混合脂膜的简单方法。膜的空气稳定性能可将脂质双层结构的应用扩展到固态器件。我们使用自组装工艺形成厚度为几纳米的混合脂质膜。该协议简单易用，不需要复杂的设备。该方法可用于制造其他相同的生物混合脂质膜，并可方便地应用于传感器和其他传感设备。在厌氧手套箱中工作，将铜邻苯二甲酸溶解在洗净玻璃瓶内的氯仿中，以每毫升铜邻苯二甲酸酮库存溶液中准备 10 毫克。通过 0.2 微米 PTFE 膜过滤溶液。在 2，300 RPM 下将 DPHPC 溶液与涡流混合器混合 10 秒。然后用氯仿冲洗玻璃微注射器五次，并用它来将溶液的200微升转移到预洗的玻璃瓶中。用温和的氮气流蒸发小瓶中的溶剂。用氯仿冲洗另一个玻璃微注射器，然后用DPHPC将200、2.6微升氯仿加入玻璃瓶。将过滤的铜邻苯二甲酸盐溶液的47.4微升加入DPHPC溶液中，该溶液应产生10比1的DPHPC与邻苯二甲氨酸的摩尔比。使用另一个干净的注射器向溶液中加入 250 微升六烷。然后在 2，300 RPM 下与涡流混合器混合 10 秒。通过 0.2 微米 PTFE 膜过滤准备好的溶液。从硅晶片上切割三厘米硅基板。然后在超声波浴中用纯净水清洁10分钟，然后用乙醇清洗，然后清洗氯仿。用氧气等离子体处理基材五分钟，去除表面吸附的有机材料，提高亲水性。用流动的纯净水清洗PTFE烧杯三分钟。然后将清洁的硅基板放入烧杯中，以小角度倾斜。将足够数量的纯净水倒入烧杯中，直到整个硅基板被淹没。将准备好的混合溶液从冰柜中取出，让其加热至室温。然后在 2，300 RPM 的涡流混合器中搅拌 15 秒。使用冲洗的 50 微升微注射器将混合溶液的三到五微升滴到水面上，形成浮动混合脂膜。要将膜转移到硅基板上，蒸发有机溶剂，然后用渗透泵将水泵出烧杯，速度为每分钟三毫升。转移过程完成后，将硅基板放在洁净室雨刷器上，让所有残留水蒸发。由于存在铜邻苯二甲酸酮分子，形成混合脂质膜具有均匀的浅蓝色，面积为几平方厘米。此处显示的是硅基板上膜的共和显微镜图像和原子力显微镜图像。在 AFM 图像中，左上角的膜厚度为 79.4 纳米，右下部的膜厚度为 4.9 纳米。薄膜表面粗糙度为 0.4 纳米，接近清洁硅基板。利用能量分散X射线分析进一步研究硅基板上混合膜的组成。代表性元素的原子比，如铜、磷、氮和碳分别是0.33、0.97、4.06和68.56%。铜、磷、氮、碳的理论摩尔比应为1比3，对11比192，接近杂交膜中的测量元素比，表明脂质与铜邻苯二甲酸分子的比例在膜形成和传递过程后保持不变。通过将脂质膜与其他纳米材料（如石墨烯或薄荷纳米颗粒）一起掺杂，可以很容易形成具有各种功能的纳米混合膜。

Research

JoVE Journal

Chemistry

Origami Inspired Self-assembly of Patterned and Reconfigurable Particles

折纸灵感的图案和可重构颗粒自组装

There are numerous techniques such as photolithography, electron-beam lithography and soft-lithography that can be used to precisely pattern two ...

科研

化学

Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria

隔离和脂质A的化学表征革兰氏阴性菌

Lipopolysaccharide (LPS) is the major cell surface molecule of gram-negative bacteria, deposited on the outer leaflet of the outer membrane bilayer. LPS ...

Formation of Ordered Biomolecular Structures by the Self-assembly of Short Peptides

生物分子有序结构由短肽的自组装形成

In nature, complex functional structures are formed by the self-assembly of biomolecules under mild conditions. Understanding the forces that control ...

Preparation of Hydrophobic Metal-Organic Frameworks via Plasma Enhanced Chemical Vapor Deposition of Perfluoroalkanes for the Removal of Ammonia

增强型氟烷烃的化学气相沉积等离子通过疏水金属 - 有机骨架的制备氨的去除

Plasma enhanced chemical vapor deposition (PECVD) of perfluoroalkanes has long been studied for tuning the wetting properties of surfaces. For high ...

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

复杂的二维形状的从单链DNA的瓷砖自组装

Current methods in DNA nano-architecture have successfully engineered a variety of 2D and 3D structures using principles of self-assembly. In this ...

Synthesis, Characterization, and Functionalization of Hybrid Au/CdS and Au/ZnS Core/Shell Nanoparticles

合成，表征和混合金/硫化镉和Au / ZnS核/壳纳米颗粒功能化

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large ...

Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level

环状聚合物的合成及其在熔融状态扩散运动的表征在单分子水平

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single ...

Synthesis and Characterization of Fe-doped Aluminosilicate Nanotubes with Enhanced Electron Conductive Properties

以增强电子导电性能的合成与表征Fe掺杂碳纳米管硅铝的

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe ...

Grafting Multiwalled Carbon Nanotubes with Polystyrene to Enable Self-Assembly and Anisotropic Patchiness

用聚苯乙烯接枝多壁纳米碳纳米管使自组装和各向异性斑块

We demonstrate a straightforward protocol to graft pristine multiwalled carbon nanotubes (MWCNTs) with polystyrene (PS) chains at the sidewalls through a ...

Liquid-cell Transmission Electron Microscopy for Tracking Self-assembly of Nanoparticles

用于跟踪纳米粒子自组装的液体细胞透射电子显微镜

Drying a nanoparticle dispersion is a versatile way to create self-assembled structures of nanoparticles, but the mechanism of this process is not fully ...