A Microfluidic Chip for Efficient Circulating Tumor Cells Enrichment, Screening and Single-cell RNA Sequencing.

Single-cell RNA sequencing on circulating tumor cells (CTCs) has been proved useful to study mechanisms of tumor heterogeneity, metastasis and drug resistance. Currently, single-cell RNA sequencing of CTCs usually takes three prerequisite steps, enrichment of CTCs from whole blood, characterization of captured cells by immunostaining and microscopic imaging, and single-cell isolation through micromanipulation. However, multiple pipetting and transferring steps could easily cause the loss of rare CTCs. To address this issue, we developed a novel integrated microfluidic chip for sequential enrichment, isolation and characterization of CTCs at single-cell level. And single CTC lysis was achieved on the same chip. The microfluidic chip included functions of blood clot filtration, single-cell isolation, identification, and target single-cell lysate collection. By spiking tumor cells into whole blood, it was validated that this microfluidic chip could effectively conduct single-cell CTCs RNA sequencing. Our approach laid a solid foundation for the analysis of RNA expression profiling of single-cell CTCs. This article is protected by copyright. All rights reserved.