Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University , Ithaca, New York 14853, United States.
Langmuir. 2017 Feb 7;33(5):1285-1294. doi: 10.1021/acs.langmuir.6b04385. Epub 2017 Jan 26.
The supported lipid bilayer has been portrayed as a useful model of the cell membrane compatible with many biophysical tools and techniques that demonstrate its appeal in learning about the basic features of the plasma membrane. However, some of its potential has yet to be realized, particularly in the area of bilayer patterning and phase/composition heterogeneity. In this work, we generate contiguous bilayer patterns as a model system that captures the general features of membrane domains and lipid rafts. Micropatterned polymer templates of two types are investigated for generating patterned bilayer formation: polymer blotting and polymer lift-off stenciling. While these approaches have been used previously to create bilayer arrays by corralling bilayers patches with various types of boundaries impenetrable to bilayer diffusion, unique to the methods presented here, there are no physical barriers to diffusion. In this work, interfaces between contiguous lipid phases define the pattern shapes, with continuity between them allowing transfer of membrane-bound biomolecules between the phases. We examine effectors of membrane domain stability including temperature and cholesterol content to investigate domain dynamics. Contiguous patterning of supported bilayers as a model of lipid rafts expands the application of the SLB to an area with current appeal and brings with it a useful toolset for characterization and analysis. These combined tools should be helpful to researchers investigating lipid raft dynamics and function and biomolecule partitioning studies. Additionally, this patterning technique may be useful for applications such as bioseparations that exploit differences in lipid phase partitioning or creation of membranes that bind species like viruses preferentially at lipid phase boundaries, to name a few.
支持的脂质双层已被描绘为一种与许多生物物理工具和技术兼容的细胞膜有用模型，这些工具和技术证明了其在了解质膜基本特征方面的吸引力。然而，其某些潜力尚未得到实现，特别是在双层图案化和相/组成异质性领域。在这项工作中，我们生成了连续的双层图案作为模型系统，该系统捕获了膜域和脂筏的一般特征。研究了两种类型的微图案化聚合物模板，以生成图案化双层形成：聚合物印迹和聚合物剥离模板印刷。虽然这些方法以前曾被用于通过用各种类型的不可渗透双层扩散的边界来围捕双层补丁来创建双层阵列，但是这里提出的方法的独特之处在于，没有扩散的物理障碍。在这项工作中，连续脂质相之间的界面定义了图案形状，它们之间的连续性允许膜结合生物分子在相之间转移。我们研究了膜域稳定性的效应物，包括温度和胆固醇含量，以研究域动力学。作为脂筏模型的支持双层的连续图案化扩展了 SLB 的应用领域，该应用领域具有当前的吸引力，并带来了用于表征和分析的有用工具集。这些组合工具应该有助于研究脂质筏动力学和功能以及生物分子分区研究的研究人员。此外，这种图案化技术可能对诸如利用脂质相分区差异的生物分离或创建优先在脂质相边界处结合病毒等物质的膜等应用有用。
