Related Experiment Video
Updated: Apr 13, 2026
09:15
Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
Published on: July 12, 2022
5.8K
A digital microfluidics-based single-cell ddPCR platform for high-throughput gene copy number analysis applied to
Yicheng Sun1, Jiancong Liang2, Hui Li3
1Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, 510515, China.
Analytica Chimica Acta
|April 11, 2026
Summary
A new digital microfluidic platform enables precise single-cell analysis using digital polymerase chain reaction (dPCR). This high-throughput system accurately quantifies nucleic acids and gene copy number variations for advanced genetic studies.
Area of Science:
- Biotechnology
- Microfluidics
- Molecular Biology
Background:
- Digital polymerase chain reaction (dPCR) excels at nucleic acid quantification but faces limitations in single-cell analysis due to droplet control issues.
- Existing dPCR platforms struggle with sensitivity and resolution for detecting gene copy number variations (CNVs) at the single-cell level.
Purpose of the Study:
- To develop an integrated digital microfluidic (DMF) platform for high-throughput, single-cell dPCR analysis.
- To overcome the limitations of current dPCR systems in droplet manipulation and visualization for precise genetic quantification.
Main Methods:
- Development of a DMF chip with 16,384 addressable electrodes for precise nanoliter droplet manipulation.
- Integration of on-chip cell lysis, droplet merging/splitting, and nucleic acid amplification within the DMF workflow.
- Validation using cell lines for quantitative PCR (qPCR) and dPCR, followed by application to primary CAR-T cells for vector copy number (VCN) quantification.
Main Results:
- Achieved single-cell resolution in both qPCR and dPCR applications using the DMF platform.
- Successfully quantified vector copy numbers (VCNs) at the single-cell level in primary CAR-T cells.
- Demonstrated performance comparable to commercial dPCR systems with minimized sample loss and contamination.
Conclusions:
- The developed DMF-based dPCR system offers a robust and scalable solution for single-cell genetic analysis.
- This integrated platform provides real-time visualization and precise control, advancing applications in clinical diagnostics and personalized medicine.
- The system holds significant potential for research and clinical settings requiring high-throughput, sensitive nucleic acid quantification.
