School of Biomedical Engineering, Faculty of Medicine, Dalian University of Technology, No. 2, Linggong Rd., Dalian, 116024, China.
School of Biomedical Engineering, Faculty of Medicine, Dalian University of Technology, No. 2, Linggong Rd., Dalian, 116024, China.
Talanta. 2024 Jan 15;267:125172. doi: 10.1016/j.talanta.2023.125172. Epub 2023 Sep 7.
Microfluidic-based analyses of single-cell dynamics in response to dynamic biochemical signals are emerging as pivotal approaches for investigating the effects of extracellular microenvironmental biochemical factors on cellular structure, function, and behavior. However, current devices often fail to consistently apply identical dynamic biochemical signals to trapped cells. In this study, we introduce a novel radially distributed single-cell trapping microfluidic array, designed to quantitatively and consistently apply identical biochemical stimulating signals to each trapped cell. Numerical simulations were employed to optimize microchannel geometry, enhancing trapping efficiency while minimizing signal distortion. Experimental validation demonstrated the trapping success rate and the single-cell trapping efficiency exceeding 99% and 85%, respectively. The microarray's capability to deliver identical dynamic biochemical stimulating signals, with various waveforms, to each unit was confirmed through fluorescein transport tests. Furthermore, we examined the intracellular calcium dynamics of U-2 OS human osteosarcoma cells in response to dynamic ATP signals, observing both single-peak calcium responses and calcium oscillations, which were modelled by a second-order system with a natural frequency of 1.6 mHz. Overall, our proposed microfluidic array offers a robust and valuable framework for advancing the understanding of single-cell dynamics.
基于微流控的单细胞动力学分析，用于响应动态生化信号，正在成为研究细胞外微观环境生化因素对细胞结构、功能和行为影响的关键方法。然而，目前的设备往往无法始终如一地将相同的动态生化信号应用于被困的细胞。在这项研究中，我们引入了一种新型的径向分布的单细胞捕获微流控阵列，旨在定量且一致地将相同的生化刺激信号应用于每个捕获的细胞。数值模拟被用来优化微通道的几何形状，提高了捕获效率，同时最小化了信号失真。实验验证表明，捕获成功率和单细胞捕获效率分别超过 99%和 85%。通过荧光素运输测试证实了微阵列向每个单元提供相同的动态生化刺激信号的能力，包括各种波形。此外，我们研究了 U-2 OS 人骨肉瘤细胞对动态 ATP 信号的细胞内钙动力学，观察到单峰钙响应和钙振荡，这些响应由具有自然频率为 1.6 mHz 的二阶系统建模。总的来说，我们提出的微流控阵列为深入了解单细胞动力学提供了一个强大而有价值的框架。
