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Nanoplasmon-enhanced drop-screen for high throughput single-cell nucleocytoplasmic miRNA profiling
Jia Liu1, Guoyun Sun, Shih-Chung Wei
1Department of Biomedical Engineering, National University of Singapore, 4 Engineering Drive 3, #04-08, 117583 Singapore.
Lab on a Chip
|April 18, 2020
Summary
This study introduces a novel nanoplasmon-enhanced droplet screening platform for high-throughput analysis of single-cell microRNA (miRNA) nucleocytoplasmic profiles. The platform achieves sensitive quantification, enabling a deeper understanding of cellular functions.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Cell nucleocytoplasmic profiles of microRNAs (miRNAs) are crucial for cellular functions like transcription and export.
- Assessing nucleocytoplasmic heterogeneity is vital but challenging due to limitations in high-throughput screening and miRNA quantification sensitivity.
- Existing droplet-based methods for single-cell miRNA assays struggle with the sensitivity needed to reveal nucleocytoplasmic heterogeneity.
Purpose of the Study:
- To develop a high-throughput screening platform for quantifying single-cell nucleocytoplasmic miRNA heterogeneity.
- To achieve highly sensitive detection of miRNAs, enabling precise analysis of their distribution within single cells.
- To establish a method for comprehensive quantitative cell biology through continuous-flow measurement of nucleocytoplasmic profiles.
Main Methods:
- Development of a nanoplasmon-enhanced droplet screening platform.
- Synthesis of nucleic acid-functionalized plasmonic nanosensors using silver nanoparticles.
- Utilizing competitive binding assays and fluorescence signal changes for sensitive miRNA quantification.
- Implementing continuous-flow measurement for high-throughput single-cell analysis.
Main Results:
- The platform demonstrates high sensitivity for miRNA quantification, reaching 0.1 nM.
- Synergistic integration of droplet screening and multiplexed plasmonic assays enables precise miRNA profiling.
- The system allows for high-throughput measurement of single-cell nucleocytoplasmic profiles at approximately 100 cells per minute.
Conclusions:
- The nanoplasmon-enhanced droplet screening platform effectively quantifies single-cell nucleocytoplasmic miRNA heterogeneity with high sensitivity.
- This technology addresses the limitations of previous methods, facilitating advanced quantitative cell biology.
- The developed platform offers a powerful tool for studying cellular processes and heterogeneity in a high-throughput manner.
