Yang Canfeng, He Chunhua, Zhuo Huasheng, Wang Jianxin, Yong Tuying, Gan Lu, Yang Xiangliang, Nie Lei, Xi Shuang, Liu Zhiyong, Liao Guanglan, Shi Tielin
State Key Laboratory of Digital Manufacturing Equipment and Technology, School of Mechanical Science & Engineering, Huazhong University of Science and Technology, Wuhan 430074, China.
National Engineering Research Center for Nanomedicine, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
Lab Chip. 2025 Feb 11;25(4):698-713. doi: 10.1039/d4lc00900b.
Microfluidic flow cytometry (MFCM) is considered to be an effective substitute for traditional flow cytometry, because of its advantages in terms of higher integration, smaller device size, lower cost, and higher cell sorting activity. However, MFCM still faces challenges in balancing parameters such as sorting throughput, viability, sorting efficiency, and cost. Here, we demonstrate a cost-effective and high-performance microfluidic cytometry cell sorting system, along with a customized microfluidic chip that integrates hydrodynamic focusing, droplet encapsulation, and sorting for precise cell manipulation. An innovative photon incremental counting-based fluorescence detection method is proposed, which requires only one-fiftieth of the data compared to traditional methods. This significantly simplifies the structure of the system and substantially reduces costs. The system exhibits detection recoveries exceeding 95% across sample solution flow rates ranging from 10 to 80 μL min. Moreover, it accurately achieves individual droplet deflections at a droplet generation frequency of 1600 Hz. Ultimately, our cell sorting system offers an impressive sorting efficiency of 90.7% and a high cell viability of 94.3% when operating at a droplet generation frequency of 1316 Hz, highlighting its accuracy and gentleness throughout the entire process. Our work will enhance advances in the life sciences, thereby creating a boom in great applications in single-cell cloning, single-cell analysis, drug screening,
微流控流式细胞术（MFCM）因其在更高集成度、更小设备尺寸、更低成本和更高细胞分选活性方面的优势，被认为是传统流式细胞术的有效替代品。然而，MFCM在平衡分选通量、活力、分选效率和成本等参数方面仍面临挑战。在此，我们展示了一种经济高效且高性能的微流控细胞术细胞分选系统，以及一种定制的微流控芯片，该芯片集成了流体动力聚焦、液滴包裹和分选功能，用于精确的细胞操作。我们提出了一种基于光子增量计数的创新荧光检测方法，与传统方法相比，该方法仅需传统方法五十分之一的数据量。这显著简化了系统结构并大幅降低了成本。该系统在10至80μL min的样品溶液流速范围内，检测回收率超过95%。此外，它能在1600 Hz的液滴生成频率下精确实现单个液滴的偏转。最终，我们的细胞分选系统在1316 Hz的液滴生成频率下运行时，分选效率高达90.7%，细胞活力高达94.3%，凸显了其在整个过程中的准确性和温和性。我们的工作将推动生命科学的进步，从而在单细胞克隆、单细胞分析、药物筛选等众多应用领域引发热潮。
