Yandrapalli Naresh, Seemann Tina, Robinson Tom
Department of Theory & Bio-Systems, Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14424 Potsdam, Germany.
Micromachines (Basel). 2020 Mar 10;11(3):285. doi: 10.3390/mi11030285.
Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 ± 4 µm in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions.
脂质体，尤其是巨型单层囊泡（GUVs），是构建人工细胞的理想隔室。传统上，使用它们需要在实验台上进行囊泡生长，然后在显微镜下进行实验。这些步骤耗时且在将囊泡转移到观察室时可能导致囊泡损失。为克服这些问题，我们展示了一种集成微流控芯片，该芯片在同一设备中结合了GUV形成、捕获和多个单独实验。首先，我们优化了缓冲条件，以最大化囊泡在微柱中的产量和后续捕获率。捕获的GUVs直径为18±4µm，具有单分散性。接下来，我们引入了一种带有集成阀的双层设计，可在不到20秒的时间内对捕获囊泡的不同亚群进行快速溶液交换。我们证明，通过膜运输和通透化分析，可以在单个芯片上进行多个实验。总之，我们开发了一种多功能一体化微流控芯片，能够使用少量样品对单批囊泡进行生产并进行多个实验。我们预计该设备对于自下而上的合成生物学将具有高度优势，因为酶促反应需要快速封装和可视化。
