Generation of phospholipid vesicle-nanotube networks and transport of molecules therein
Generation of phospholipid vesicle-nanotube networks and transport of molecules therein
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DOI：
10.1038/nprot.2011.321
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发表时间：
2011-05-01
期刊：
影响因子：
14.8
通讯作者：
Orwar, Owe
中科院分区：
文献类型：
--
作者：
Jesorka, Aldo;Stepanyants, Natalia;Orwar, Owe
我们描述了由脂质双分子层纳米管和表面固定化囊泡组成的软物质网络的显微操作和显微注射过程。这些仿生膜系统具有独特的结构灵活性和可扩展性，与自上而下制备的固态微流体和纳米流体设备不同，它们允许对单个容器和互连管道进行动态控制的网络设计。制造是建立在磷脂分子的自组装上，然后是微操作操作，例如膜电穿孔和微注射，以影响膜的形状转换并创建一系列相互连接的隔室。网络的大小和几何形状可以根据其所需的功能来选择。膜的组成主要是在自组装步骤中控制的，而单个容器的内部内容物是通过一系列微针注射来确定的。网络不能用其他目前可用的巨型单层囊泡制备方法（大单层囊泡融合或电形成）制备。详细描述了三种传输模式，它们适用于在网络中移动水溶性或膜结合的小分子、聚合物、DNA、蛋白质和纳米颗粒。如果所有必要的准备工作都提前做好，制作程序大约需要90分钟。运输研究需要额外的60-120分钟，取决于运输制度。
We describe micromanipulation and microinjection procedures for the fabrication of soft-matter networks consisting of lipid bilayer nanotubes and surface-immobilized vesicles. These biomimetic membrane systems feature unique structural flexibility and expandability and, unlike solid-state microfluidic and nanofluidic devices prepared by top-down fabrication, they allow network designs with dynamic control over individual containers and interconnecting conduits. The fabrication is founded on self-assembly of phospholipid molecules, followed by micromanipulation operations, such as membrane electroporation and microinjection, to effect shape transformations of the membrane and create a series of interconnected compartments. Size and geometry of the network can be chosen according to its desired function. Membrane composition is controlled mainly during the self-assembly step, whereas the interior contents of individual containers is defined through a sequence of microneedle injections. Networks cannot be fabricated with other currently available methods of giant unilamellar vesicle preparation (large unilamellar vesicle fusion or electroformation). Described in detail are also three transport modes, which are suitable for moving water-soluble or membrane-bound small molecules, polymers, DNA, proteins and nanoparticles within the networks. The fabrication protocol requires similar to 90 min, provided all necessary preparations are made in advance. The transport studies require an additional 60-120 min, depending on the transport regime.