Capture-culture dual-zone integrated unit array for single-cell microfluidic chips

捕获-培养双区集成的单细胞微流控芯片

¹ Department of Basic, Rocket Force University of Engineering, Xi'an 710025, China
² Science Research Institute, Chang'an University, Xi'an 710018, China

Received: 9 June 2025

Abstract

Single-cell culture has long posed a significant challenge and remained a hot topic in biomedical engineering. To address the limitations in existing microfluidic single-cell chips, where cell culture space is constrained by the capture zones——impeding long-term cultivation. This study presents a capture-culture dual-zone integrated microfluidic chip, which integrates cell capture and culture functional region within a single unit. Through optimized chip layout design, the device achieves a microsphere capture rate of 91.93% and a cell capture rate of 60.1%.This study employs mouse fibroblast L929 cells (average diameter: 10 μm) as the model system, establishing a hydrodynamic flow model of cell suspension within the chip through theoretical calculations and computational simulations. To ensure cell viability, a microchannel width of 60 μm achieves optimal capture efficiency. To validate the chip performance, under conditions of a cell (or microsphere) input density of 1×10⁵ cell/mL and a flow velocity of 300 μm/s, the cells exhibited stable post-entry conditions. With a microchannel width of 60 μm, the chip achieved both high capture efficiency and single-cell isolation rates, yielding a robust single-cell microfluidic device with stable bonding. This chip demonstrates a simple structure and straightforward fabrication process, serving as a novel tool for single-cell analysis and research.

摘要

单细胞培养一直以来都是生物工程的难点和热点。针对现有单细胞微流控芯片中存在的细胞培养空间受制于细胞捕获空间, 导致细胞无法长期培养的问题, 设计了一种捕获-培养双区集成芯片。在单个单元中集成细胞捕获与细胞培养2种功能区域, 设计出简单高效的芯片布置方式, 实现了91.93%的微球捕获率和60.1%的细胞捕获率。以小鼠成纤维细胞L929(细胞系平均直径10 μm)为研究对象, 通过理论计算和模拟仿真建立细胞悬液在芯片中的流动模型。在保证细胞活性的前提下, 微通道宽度60 μm时可以得到较好的捕获效果。为了验证芯片性能, 在细胞(微球)进样密度为1×10⁵ cell/mL, 进样流速为300 μm/s时, 细胞进入芯片后状态良好; 芯片微通道宽度为60 μm时, 可获得捕获率和单细胞率较好且键合稳定的单细胞微流控芯片。文中设计的芯片结构简单、制备方便, 有望为单细胞研究提供新工具。