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Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification
Long Chen1,2,3, Yi Xu1,2, Lele Zhou4
1Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China.
Microsystems & Nanoengineering
|January 17, 2025
Summary
A new ultra-sensitive single-cell sorting platform, THE-FADS, enhances fluorescence detection for improved cellular heterogeneity analysis. This technology maintains high cell viability and sorting efficiency, advancing personalized medicine applications.
Area of Science:
- Biotechnology
- Cell Biology
- Analytical Chemistry
Background:
- Current single-cell analysis methods like FACS and FADS have limitations in sorting performance and cell viability.
- Deciphering cellular heterogeneity is crucial for biological understanding and medical advancements.
Purpose of the Study:
- To develop an ultra-sensitive single-cell sorting platform with enhanced performance and cell viability.
- To improve the detection and isolation of specific cell populations for further analysis.
Main Methods:
- Integration of FADS technology with Tetramer-HCR-EvaGreen (THE) fluorescence signal amplification.
- Application of the THE-FADS platform to sort target MCF-7 cells from various sample types.
Main Results:
- The THE system demonstrated significantly higher fluorescence signal amplification compared to existing methods.
- The THE-FADS platform achieved high sorting efficiencies (55.5% and 50.3%) and purities (91% and 85%) for MCF-7 cells.
- Sorted cells maintained over 95% viability and exhibited normal proliferation and gene expression.
Conclusions:
- The developed THE-FADS platform offers superior performance for single-cell sorting.
- This technology shows significant promise for advancing the understanding of cellular heterogeneity.
- The platform has potential applications in personalized medicine and diagnostics.
