Ultra-sensitive fluorescence-activated droplet single-cell sorting based on Tetramer-HCR-EvaGreen amplification.

IF 7.3 1区 工程技术 Q1 INSTRUMENTS & INSTRUMENTATION
Long Chen, Yi Xu, Lele Zhou, Ding Ma, Rong Zhang, Yifan Liu, Xianqiang Mi
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引用次数: 0

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.

基于四聚体- hcr - evgreen扩增的超灵敏荧光激活液滴单细胞分选。
目前的单细胞分析技术,如荧光激活细胞分选(FACS)和荧光激活液滴分选(FADS),虽然可以解释细胞的异质性,但受分选性能和细胞活力低等限制。本研究将FADS技术与四聚体- hcr - evgreen (the)荧光信号放大技术相结合,开发了一个超灵敏的单细胞分选平台。该系统比单一四聚体或四聚体- hcr信号放大产生更高的荧光信号。应用于MCF-7细胞后,该平台在保持95%以上的细胞存活率的同时表现出较高的效率和选择性。The - fads在PBS溶液和模拟血清样品中对MCF-7细胞的分选效率分别为55.5%和50.3%,纯度分别为91%和85%。与对照细胞相比,分选后的MCF-7细胞在CK19和EGFR mRNA表达方面具有相似的增殖能力。建立的The - fads在了解细胞异质性和个性化医疗方面具有良好的前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Microsystems & Nanoengineering
Microsystems & Nanoengineering Materials Science-Materials Science (miscellaneous)
CiteScore
12.00
自引率
3.80%
发文量
123
审稿时长
20 weeks
期刊介绍: Microsystems & Nanoengineering is a comprehensive online journal that focuses on the field of Micro and Nano Electro Mechanical Systems (MEMS and NEMS). It provides a platform for researchers to share their original research findings and review articles in this area. The journal covers a wide range of topics, from fundamental research to practical applications. Published by Springer Nature, in collaboration with the Aerospace Information Research Institute, Chinese Academy of Sciences, and with the support of the State Key Laboratory of Transducer Technology, it is an esteemed publication in the field. As an open access journal, it offers free access to its content, allowing readers from around the world to benefit from the latest developments in MEMS and NEMS.
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