Department of Theory & Bio-Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.
Lab Chip. 2019 Feb 12;19(4):626-633. doi: 10.1039/c8lc01275j.
Biomimetic systems such as model lipid membranes are vital to many research fields including synthetic biology, drug discovery and membrane biophysics. One of the most commonly used are giant unilamellar vesicles (GUVs) due to their size similarity with biological cells and their ease of production. Typical methods for handling such delicate objects are low-throughput and do not allow solution exchange or long-term observations, all of which limits the experimental options. Herein, we present a new device designed to confine large assemblies of GUVs in microfluidic traps but is still able to perform precise and fast solution exchanges. An optimised design allows efficient filling with as many as 114 GUVs per trap and over 23 000 GUVs per device. This allows high-throughput dataset acquisitions which we demonstrate with two proof-of-concept experiments: (i) end-point measurements of vesicle interior pH and (ii) membrane transport kinetics. Moreover, we show that the design is able to selectively trap sub-populations of specific vesicle sizes and assemble them in different layers. The device can easily be applied to other high-throughput membrane studies and will pave the way for future applications using vesicle assemblies to model cellular tissues or even prototissues.
仿生系统,如模型脂质膜,对于包括合成生物学、药物发现和膜生物物理学在内的许多研究领域都至关重要。由于其大小与生物细胞相似,且易于制备,其中最常用的是巨大的单分子层囊泡(GUV)。处理此类脆弱物体的典型方法是低通量的,不允许进行溶液交换或长期观察,所有这些都限制了实验选择。在此,我们提出了一种新的装置,用于将大量 GUV 限制在微流控陷阱中,但仍能够进行精确和快速的溶液交换。优化的设计允许高效填充,每个陷阱中可容纳多达 114 个 GUV,每个装置中可容纳超过 23000 个 GUV。这使得能够进行高通量数据集采集,我们通过两个概念验证实验证明了这一点:(i)囊泡内部 pH 值的终点测量,(ii)膜转运动力学。此外,我们表明该设计能够选择性地捕获特定囊泡大小的亚群,并将它们组装在不同的层中。该装置可以很容易地应用于其他高通量膜研究,并为未来使用囊泡组装来模拟细胞组织甚至原组织的应用铺平道路。