A diffusion-based microfluidic device for single-cell RNA-seq.
A diffusion-based microfluidic device for single-cell RNA-seq.
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DOI：
10.1039/c8lc00967h
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发表时间：
2019-03
期刊：
影响因子：
6.1
通讯作者：
Mimosa Sarma;Jiyoung Lee;Sai Ma;Song Li;Chang Lu
中科院分区：
文献类型：
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作者：
Mimosa Sarma;Jiyoung Lee;Sai Ma;Song Li;Chang Lu
微流体装置为单细胞RNA-seq（scRNA-seq）提供了低输入和高效的平台。现有的微流体装置具有复杂的多室结构，用于处理RNA-seq中涉及的多步骤过程，并且步骤之间的稀释用于消除试剂之间的抑制作用。这使得该装置难以制造和操作。在这里，我们提出了基于微流体扩散的RNA-seq（MID-RNA-seq），用于使用基于扩散的试剂交换方案进行scRNA-seq。该装置将细胞捕获、裂解、逆转录和PCR扩增全部整合在一个简单的微流体装置中。MID-RNA-seq提供与现有scRNA-seq方法相当的高数据质量，同时实现允许多路复用的简单设备设计。MID-RNA-seq设备的鲁棒性和可扩展性对于稀缺细胞样本的转录组学研究将是重要的。
Microfluidic devices provide a low-input and efficient platform for single-cell RNA-seq (scRNA-seq). Existing microfluidic devices have a complicated multi-chambered structure for handling the multi-step process involved in RNA-seq and dilution between steps is used to negate the inhibitory effects among reagents. This makes the device difficult to fabricate and operate. Here we present microfluidic diffusion-based RNA-seq (MID-RNA-seq) for conducting scRNA-seq with a diffusion-based reagent swapping scheme. This device incorporates cell trapping, lysis, reverse transcription and PCR amplification all in one simple microfluidic device. MID-RNA-seq provides high data quality that is comparable to existing scRNA-seq methods while implementing a simple device design that permits multiplexing. The robustness and scalability of the MID-RNA-seq device will be important for transcriptomic studies of scarce cell samples.