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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
Published on: May 23, 2018
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spinDrop: a droplet microfluidic platform to maximise single-cell sequencing information content.
Joachim De Jonghe1,2, Tomasz S Kaminski1,3, David B Morse4
1Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
Nature Communications
|August 8, 2023
Summary
This study introduces a novel droplet microfluidic method enhancing single-cell sequencing. It improves gene detection rates and reduces background noise for more accurate molecular atlases.
Area of Science:
- Biotechnology
- Genomics
- Microfluidics
Background:
- Droplet microfluidics enables high-throughput single-cell sequencing but suffers from increased background noise and lower RNA capture efficiency, especially with challenging samples.
- Current methods lack strategies for enriching high-quality material or specific cell types during encapsulation and efficient multi-step enzymatic processes for improved RNA capture.
Purpose of the Study:
- To address the limitations of droplet microfluidics by developing a method that enriches for viable single cells and enhances RNA capture efficiency.
- To improve the sensitivity and accuracy of single-cell profiling for generating high-quality molecular atlases.
Main Methods:
- Utilized fluorescence-activated droplet sorting to enrich droplets containing single viable cells, intact nuclei, fixed cells, or target cell types.
- Implemented reagent addition via picoinjection into droplets to perform multi-step lysis and reverse transcription.
Main Results:
- Achieved a fivefold increase in gene detection rates.
- Reduced background noise by up to 50%.
- Successfully generated high-quality molecular atlases of mouse brain development and nascent RNA transcription during organogenesis from damaged input material.
Conclusions:
- The developed methodology overcomes key bottlenecks in droplet-based single-cell sequencing, significantly enhancing sensitivity and accuracy.
- This approach is broadly applicable to various droplet-based workflows, offering a cost-effective solution for sensitive and accurate single-cell profiling.
