Raman-Activated Droplet Sorting (RADS) for Label-Free High-Throughput Screening of Microalgal Single-Cells,Analytical Chemistry

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Raman-Activated Droplet Sorting (RADS) for Label-Free High-Throughput Screening of Microalgal Single-Cells
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-09 00:00:00 , DOI: 10.1021/acs.analchem.7b03884
Xixian Wang _{1,

2} , Lihui Ren _{1,

2} , Yetian Su ₁ , Yuetong Ji _{1,

2} , Yaoping Liu ₃ , Chunyu Li _{1,

2} , Xunrong Li _{1,

2} , Yi Zhang ₁ , Wei Wang ₃ , Qiang Hu ₄ , Danxiang Han ₄ , Jian Xu _{1,

2} , Bo Ma _{1,

2}

Affiliation

Raman-activated cell sorting (RACS) has attracted increasing interest, yet throughput remains one major factor limiting its broader application. Here we present an integrated Raman-activated droplet sorting (RADS) microfluidic system for functional screening of live cells in a label-free and high-throughput manner, by employing AXT-synthetic industrial microalga Haematococcus pluvialis (H. pluvialis) as a model. Raman microspectroscopy analysis of individual cells is carried out prior to their microdroplet encapsulation, which is then directly coupled to DEP-based droplet sorting. To validate the system, H. pluvialis cells containing different levels of AXT were mixed and underwent RADS. Those AXT-hyperproducing cells were sorted with an accuracy of 98.3%, an enrichment ratio of eight folds, and a throughput of ∼260 cells/min. Of the RADS-sorted cells, 92.7% remained alive and able to proliferate, which is equivalent to the unsorted cells. Thus, the RADS achieves a much higher throughput than existing RACS systems, preserves the vitality of cells, and facilitates seamless coupling with downstream manipulations such as single-cell sequencing and cultivation.

中文翻译：

拉曼激活的液滴分选（RADS）用于微藻单细胞的无标签高通量筛选

拉曼激活细胞分选术（RACS）引起了越来越多的兴趣，但通量仍然是限制其广泛应用的主要因素。在这里，我们介绍了一个集成的拉曼活化液滴分选（RADS）微流控系统，以无标记和高通量的方式，通过使用AXT合成的工业微藻生血红球菌（H. pluvialis）作为模型，对活细胞进行功能筛选。在单个细胞进行微滴封装之前，先进行单个细胞的拉曼光谱分析，然后将其直接与基于DEP的液滴分选结合。为了验证系统，幽门螺杆菌将含有不同水平AXT的细胞混合并进行RADS。这些AXT高产细胞的分选精度为98.3％，富集率是8倍，吞吐率约为260个细胞/分钟。在RADS分选的细胞中，仍有92.7％存活并能够增殖，这与未分选的细胞相当。因此，与现有的RACS系统相比，RADS可获得更高的通量，保留了细胞的活力，并促进了与下游操作（如单细胞测序和培养）的无缝结合。

更新日期：2017-11-09

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