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Multiplexed detection of micro-RNAs based on microfluidic multi-color fluorescence droplets

Analytical and Bioanalytical Chemistry volume 412, pages 647–655 (2020)Cite this article

Abstract

In this work, simple, rapid, and low-cost multiplexed detection of tumor-related micro-RNAs (miRNAs) was achieved based on multi-color fluorescence on a microfluidic droplet chip, which simplified the complexity of light path to a half. A four-T-junction structure was fabricated to form uniform nano-volume droplet arrays with customized contents. Multi-color quantum dots (QDs) used as the fluorescence labels were encapsulated into droplets to develop the multi-path fluorescence detection module. We designed an integrated multiplex fluorescence resonance energy transfer system assisted by multiple QDs (four colors) and one quencher to detect four tumor-related miRNAs (miRNA-20a, miRNA-21, miRNA-155, and miRNA-221). The qualitative analysis of miRNAs was realized by the color identification of QDs, while the quantitative detection of miRNAs was achieved based on the linear relationship between the quenching efficiency of QDs and the concentration of miRNAs. The practicability of the multiplex detection device was further confirmed by detecting four tumor-related miRNAs in real human serum samples. The detection limits of four miRNAs ranged from 35 to 39 pmol/L was achieved without any target amplification. And the linear range was from 0.1 nmol/L to 1 μmol/L using 10 nL detection volume (one droplet) under the detection speed of 320 droplets per minute. The multiple detection system for miRNAs is simple, fast, and low-cost and will be a powerful platform for clinical diagnostic analysis.

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Funding

This study received financial support from the National Natural Science Foundation of China (21874015 and 21675020).

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  1. Wen-Qi Ye and Yi-Xuan Wei contributed equally to this work.

Affiliations

  1. Research Center for Analytical Sciences, Northeastern University, 3-11 Wenhua Road, Shenyang, 110819, Liaoning, China

    Wen-Qi Ye, Yi-Xuan Wei, Ying-Zhi Zhang, Chun-Guang Yang & Zhang-Run Xu

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  1. Wen-Qi Ye
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  2. Yi-Xuan Wei
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Correspondence to Chun-Guang Yang.

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This study was approved by the Ethics Review Committee of Northeastern University Hospital (Shenyang, China). Informed consent of all volunteers involved in serum experiments.

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Ye, WQ., Wei, YX., Zhang, YZ. et al. Multiplexed detection of micro-RNAs based on microfluidic multi-color fluorescence droplets. Anal Bioanal Chem 412, 647–655 (2020). https://doi.org/10.1007/s00216-019-02266-3

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  • DOI: https://doi.org/10.1007/s00216-019-02266-3

Keywords

  • Microfluidic droplet
  • Multiplex detection
  • Micro-RNAs
  • Multi-color QDs
  • Multi-path fluorescence detection