Cai Linfeng, Lin Li, Lin Shiyan, Wang Xuanqun, Chen Yingwen, Zhu Huanghuang, Zhu Zhi, Yang Liu, Xu Xing, Yang Chaoyong
The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, Department of Chemical Biology, Department of Chemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
Institute of Molecular Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China.
Small Methods. 2024 Dec;8(12):e2400375. doi: 10.1002/smtd.202400375. Epub 2024 Apr 12.
Proteins as crucial components of cells are responsible for the majority of cellular processes. Sensitive and efficient protein detection enables a more accurate and comprehensive investigation of cellular phenotypes and life activities. Here, a protein sequencing method with high multiplexing, high throughput, high cell utilization, and integration based on digital microfluidics (DMF-Protein-seq) is proposed, which transforms protein information into DNA sequencing readout via DNA-tagged antibodies and labels single cells with unique cell barcodes. In a 184-electrode DMF-Protein-seq system, ≈1800 cells are simultaneously detected per experimental run. The digital microfluidics device harnessing low-adsorbed hydrophobic surface and contaminants-isolated reaction space supports high cell utilization (>90%) and high mapping reads (>90%) with the input cells ranging from 140 to 2000. This system leverages split&pool strategy on the DMF chip for the first time to overcome DMF platform restriction in cell analysis throughput and replace the traditionally tedious bench-top combinatorial barcoding. With the benefits of high efficiency and sensitivity in protein analysis, the system offers great potential for cell classification and drug monitoring based on protein expression at the single-cell level.
蛋白质作为细胞的关键组成部分，负责大多数细胞过程。灵敏且高效的蛋白质检测能够对细胞表型和生命活动进行更准确、全面的研究。在此，我们提出了一种基于数字微流控的具有高复用性、高通量、高细胞利用率和集成性的蛋白质测序方法（DMF-Protein-seq），该方法通过DNA标记抗体将蛋白质信息转化为DNA测序读数，并用独特的细胞条形码标记单个细胞。在一个184电极的DMF-Protein-seq系统中，每次实验运行可同时检测约1800个细胞。利用低吸附疏水表面和污染物隔离反应空间的数字微流控装置，在输入细胞数为140至2000时，支持高细胞利用率（>90%）和高映射读数（>90%）。该系统首次在DMF芯片上利用拆分与合并策略，克服了DMF平台在细胞分析通量方面的限制，并取代了传统繁琐的台式组合条形码技术。凭借蛋白质分析的高效性和灵敏性，该系统在基于单细胞水平的蛋白质表达进行细胞分类和药物监测方面具有巨大潜力。
