Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification
Abstract The current single-cell analysis technologies such as fluorescence-activated cell sorting (FACS) and fluorescence-activated droplet sorting (FADS) could decipher the cellular heterogeneity but were constrained by low sorting performance and cell viability. Here, an ultra-sensitive single-ce...
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2025-01-01
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| Series: | Microsystems & Nanoengineering |
| Online Access: | https://doi.org/10.1038/s41378-024-00861-8 |
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| author | Long Chen Yi Xu Lele Zhou Ding Ma Rong Zhang Yifan Liu Xianqiang Mi |
| author_facet | Long Chen Yi Xu Lele Zhou Ding Ma Rong Zhang Yifan Liu Xianqiang Mi |
| author_sort | Long Chen |
| collection | DOAJ |
| description | Abstract The current single-cell analysis technologies such as fluorescence-activated cell sorting (FACS) and fluorescence-activated droplet sorting (FADS) could decipher the cellular heterogeneity but were constrained by low sorting performance and cell viability. Here, an ultra-sensitive single-cell sorting platform has been developed by integrating the FADS technology with Tetramer-HCR-EvaGreen (THE) fluorescence signal amplification. The THE system produced much higher fluorescence signal than that of the single Tetramer or Tetramer-HCR signal amplification. Upon application to target MCF-7 cells, the platform exhibited high efficacy and selectivity while maintaining more than 95% cell viability. The THE-FADS achieved sorting efficiencies of 55.5% and 50.3% with purities of 91% and 85% for MCF-7 cells in PBS solutions and simulated serum samples, respectively. The sorted MCF-7 cells showed similar proliferation together with CK19 and EGFR mRNA expression compared with the control cells. The established THE-FADS showed the promising prospects to cellular heterogeneity understanding and personalized medicine. |
| format | Article |
| id | doaj-art-055d13ea99204e01812d01e3324c6d98 |
| institution | Kabale University |
| issn | 2055-7434 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Microsystems & Nanoengineering |
| spelling | doaj-art-055d13ea99204e01812d01e3324c6d982025-01-19T12:27:07ZengNature Publishing GroupMicrosystems & Nanoengineering2055-74342025-01-0111111310.1038/s41378-024-00861-8Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplificationLong Chen0Yi Xu1Lele Zhou2Ding Ma3Rong Zhang4Yifan Liu5Xianqiang Mi6Shanghai Institute of Microsystem and Information Technology, Chinese Academy of SciencesShanghai Institute of Microsystem and Information Technology, Chinese Academy of SciencesSchool of Physical Science and Technology, ShanghaiTech UniversityShanghai Institute of Microsystem and Information Technology, Chinese Academy of SciencesSchool of Physical Science and Technology, ShanghaiTech UniversitySchool of Physical Science and Technology, ShanghaiTech UniversityShanghai Institute of Microsystem and Information Technology, Chinese Academy of SciencesAbstract The current single-cell analysis technologies such as fluorescence-activated cell sorting (FACS) and fluorescence-activated droplet sorting (FADS) could decipher the cellular heterogeneity but were constrained by low sorting performance and cell viability. Here, an ultra-sensitive single-cell sorting platform has been developed by integrating the FADS technology with Tetramer-HCR-EvaGreen (THE) fluorescence signal amplification. The THE system produced much higher fluorescence signal than that of the single Tetramer or Tetramer-HCR signal amplification. Upon application to target MCF-7 cells, the platform exhibited high efficacy and selectivity while maintaining more than 95% cell viability. The THE-FADS achieved sorting efficiencies of 55.5% and 50.3% with purities of 91% and 85% for MCF-7 cells in PBS solutions and simulated serum samples, respectively. The sorted MCF-7 cells showed similar proliferation together with CK19 and EGFR mRNA expression compared with the control cells. The established THE-FADS showed the promising prospects to cellular heterogeneity understanding and personalized medicine.https://doi.org/10.1038/s41378-024-00861-8 |
| spellingShingle | Long Chen Yi Xu Lele Zhou Ding Ma Rong Zhang Yifan Liu Xianqiang Mi Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification Microsystems & Nanoengineering |
| title | Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification |
| title_full | Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification |
| title_fullStr | Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification |
| title_full_unstemmed | Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification |
| title_short | Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification |
| title_sort | ultra sensitive fluorescence activated droplet single cell sorting based on tetramer hcr evagreen amplification |
| url | https://doi.org/10.1038/s41378-024-00861-8 |
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