Single-cell level point mutation analysis of circulating tumor cells through droplet microfluidics

• Published in: Chinese chemical letters 2022-05, Vol.33 (5), p.2701-2704
• Main Authors: Qiu, Shihui, Shen, Chuanjie, Jian, Xiaoyu, Lu, Yunxing, Tong, Zhaoduo, Wu, Zhenhua, Mao, Hongju, Zhao, Jianlong
• Format: Article
• Language: English
• Subjects: CTCs; EGFR; Microfluidics; Point mutation; Single cell
• ISSN: 1001-8417; 1878-5964
• DOI: 10.1016/j.cclet.2021.08.128

Tumor heterogeneity plays a critical role in the determination of appropriate anticancer therapy. As circulating tumor cells (CTCs) contain all tumor-related information, the genetic changes on CTCs could help us choose the appropriate treatments for different patients. Single-base mutations are very common in tumor genetic changes which may result in drug resistance. Here, we introduce a single-cell mutation detection platform based on droplet microfluidics. This platform integrates cell capsulation, cell lysis, polymerase chain reaction (PCR) and the observation process. The droplets’ generation speed is over 6000 per minute and more than 600 cells could be encapsulated in one second. To verify the performance of our platform in practical use, we performed the mutation analysis of 4 kinds of cells with our platform and noted that the genetic status of each single cell was clearly discriminated. Moreover, these results agreed with those from direct sequencing. Compared with other forms of single-cell mutation detection techniques, our platform has high throughput, short experimental time and less experimental operations. Graphical Abstract [Display omitted] .