Highly Multiplexed, Efficient, and Automated Single-Cell MicroRNA Sequencing with Digital Microfluidics.,Small Methods

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Highly Multiplexed, Efficient, and Automated Single-Cell MicroRNA Sequencing with Digital Microfluidics.
Small Methods ( IF 10.7 ) Pub Date : 2023-11-28 , DOI: 10.1002/smtd.202301250
Yingwen Chen ₁ , Xuanqun Wang ₁ , Xing Na ₁ , Yingkun Zhang ₁ , Zan Li ₂ , Xiaohui Chen _{3,

4} , Linfeng Cai ₁ , Jia Song ₅ , Ren Xu _{3,

4} , Chaoyong Yang _{1,

5}

Affiliation

Single-cell microRNA (miRNA) sequencing has allowed for comprehensively studying the abundance and complex networks of miRNAs, which provides insights beyond single-cell heterogeneity into the dynamic regulation of cellular events. Current benchtop-based technologies for single-cell miRNA sequencing are low throughput, limited reaction efficiency, tedious manual operations, and high reagent costs. Here, a highly multiplexed, efficient, integrated, and automated sample preparation platform is introduced for single-cell miRNA sequencing based on digital microfluidics (DMF), named Hiper-seq. The platform integrates major steps and automates the iterative operations of miRNA sequencing library construction by digital control of addressable droplets on the DMF chip. Based on the design of hydrophilic micro-structures and the capability of handling droplets of DMF, multiple single cells can be selectively isolated and subject to sample processing in a highly parallel way, thus increasing the throughput and efficiency for single-cell miRNA measurement. The nanoliter reaction volume of this platform enables a much higher miRNA detection ability and lower reagent cost compared to benchtop methods. It is further applied Hiper-seq to explore miRNAs involved in the ossification of mouse skeletal stem cells after bone fracture and discovered unreported miRNAs that regulate bone repairing.

中文翻译：

使用数字微流体进行高度多重、高效和自动化的单细胞 MicroRNA 测序。

单细胞 microRNA (miRNA) 测序可以全面研究 miRNA 的丰度和复杂网络，从而提供超越单细胞异质性的见解，深入了解细胞事件的动态调节。目前基于台式的单细胞 miRNA 测序技术通量低、反应效率有限、手动操作繁琐且试剂成本高。这里，引入了一个高度多重、高效、集成和自动化的样本制备平台，用于基于数字微流控（DMF）的单细胞miRNA测序，名为Hiper-seq。该平台通过对DMF芯片上可寻址液滴的数字控制，集成了主要步骤并自动化了miRNA测序文库构建的迭代操作。基于亲水微结构的设计和处理DMF液滴的能力，可以选择性地分离多个单细胞并以高度并行的方式进行样品处理，从而提高单细胞miRNA测量的通量和效率。与台式方法相比，该平台的纳升反应体积可实现更高的 miRNA 检测能力和更低的试剂成本。进一步应用Hiper-seq探索参与小鼠骨骼干细胞骨折后骨化的miRNA，并发现了未报道的调节骨修复的miRNA。

更新日期：2023-11-28

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