Building Dynamic Cellular Machineries in Droplet-Based Artificial Cells with Single-Droplet Tracking and Analysis
Building Dynamic Cellular Machineries in Droplet-Based Artificial Cells with Single-Droplet Tracking and Analysis
复制标题
通过单液滴跟踪和分析在基于液滴的人造细胞中构建动态细胞机器
DOI：
10.1021/acs.analchem.9b01481
复制
发表时间：
2019
影响因子：
7.4
通讯作者：
Yang, Qiong
中科院分区：
文献类型：
--
作者：
Sun, Meng;Li, Zhengda;Wang, Shiyuan;Maryu, Gembu;Yang, Qiong
尽管在过去的几十年里，液滴微流体在化学和生物学上的应用呈指数级增长，但用于常规生成和综合分析液滴人工细胞的强大通用平台仍然很少。在这里，我们报告使用微流控液滴在体外人工细胞中复制各种类型的细胞机制。结合独特的基于图像的分析方法，该系统可以全自动跟踪单个液滴，具有高精度、高通量和高灵敏度。这些强大的性能使得我们开发的三个具有代表性的基于液滴的分析原型具有广泛的适用性，用于(i)液滴数字检测，（ii）体外转录和翻译反应，以及（iii）细胞周期振荡的时空动力学。该平台通过实时、长期成像来生成、孵化、跟踪和分析单个微滴的能力释放了其在加速无细胞合成生物学方面的巨大潜力。此外，从数字到模拟再到形态检测的广泛范围使这种液滴分析技术适用于许多其他不同类型的基于液滴的化学和生物分析。
Although the application of droplet microfluidics has grown exponentially in chemistry and biology over the past decades, robust universal platforms for the routine generation and comprehensive analysis of droplet-based artificial cells are still rare. Here we report using microfluidic droplets to reproduce a variety of types of cellular machinery in in vitro artificial cells. In combination with a unique image-based analysis method, the system enables full automation in tracking single droplets with high accuracy, high throughput, and high sensitivity. These powerful performances allow broad applicability evident in three representative droplet-based analytical prototypes that we develop for (i) droplet digital detection, (ii) in vitro transcription and translation reactions, and (iii) spatiotemporal dynamics of cell-cycle oscillations. The capacities of this platform to generate, incubate, track, and analyze individual microdroplets via real-time, long-term imaging unleash its great potential in accelerating cell-free synthetic biology. Moreover, the wide scope covering from digital to analog to morphological detections makes this droplet analysis technique adaptable for many other divergent types of droplet-based chemical and biological assays.