Single‐cell RNA sequencing on circulating tumor cells (CTCs) proves 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 can easily cause the loss of rare CTCs. To address this issue, a novel integrated microfluidic chip for sequential enrichment, isolation, and characterization of CTCs at single‐cell level, is developed. And, single CTC lysis is achieved on the same chip. The microfluidic chip includes functions of blood clot filtration, single‐cell isolation, identification, and target single‐cell lysate collection. By spiking tumor cells into whole blood, it is validated that this microfluidic chip can effectively conduct single‐cell CTCs RNA sequencing. The approach lays a solid foundation for the analysis of RNA expression profiling of single‐cell CTCs.

中文翻译：

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用于高效循环肿瘤细胞富集、筛选和单细胞 RNA 测序的微流控芯片

循环肿瘤细胞 (CTC) 的单细胞 RNA 测序证明对研究肿瘤异质性、转移和耐药性机制很有用。目前，CTC 的单细胞 RNA 测序通常需要三个先决步骤：从全血中富集 CTC，通过免疫染色和显微成像表征捕获的细胞，以及通过显微操作分离单细胞。然而，多次移液和转移步骤很容易导致稀有 CTC 的损失。为了解决这个问题，开发了一种用于在单细胞水平上连续富集、分离和表征 CTC 的新型集成微流控芯片。并且，单个 CTC 裂解是在同一芯片上实现的。微流控芯片具有血凝块过滤、单细胞分离、鉴定、目标单细胞裂解液收集等功能。通过将肿瘤细胞掺入全血，验证了这种微流控芯片可以有效地进行单细胞 CTCs RNA 测序。该方法为分析单细胞 CTC 的 RNA 表达谱奠定了坚实的基础。