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 sort...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.展开更多
基金funded by National Key Research and Development Program of China(grant number 2022YFC3502002)CAS-NSTDA Joint Research Project(2024)(grant number 101GJHZ2024017MI).
文摘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.
