A microfluidic array device for single cell capture and intracellular Ca2+ response analysis induced by dynamic biochemical stimulus

A microfluidic array was constructed for trapping single cell and loading identical dynamic biochemical stimulation for gain a better understanding of Ca2+ signaling at single cell resolution in the present study. This microfluidic array consists of multiple radially aligned flow channels with equal intersection angles, which was designed by a combination of stagnation point flow and physical barrier. Numerical simulation results and trajectory analysis have shown the effectiveness of this single cell trapping device. Fluorescent experiment results demonstrated the effects of flow rate and frequency of dynamic stimulus on the profiles of biochemical concentration which exposed on captured cells. In this microarray, the captured single cells in each trapping channels were able to receive identical extracellular dynamic biochemical stimuli which being transmitted from the entrance in the middle of the microfluidic array. Besides, after loading dynamic Adenosine Triphosphate (ATP) stimulation on captured cells by this device, consistent average intracellular Ca2+ dynamics phase and cellular heterogeneity were observed in captured single K562 cells. Furthermore, this device is able to be used for investigating cellular respond on single cell resolution to temporally varying environments by modulating the stimulation signal in terms of concentration, pattern, and duration of exposure.

Automated summary

In this study, a microfluidic array was developed to trap single cells and deliver consistent dynamic biochemical stimulation, which enhanced the understanding of Ca2+ signaling at single cell resolution. The effectiveness of the device was demonstrated through numerical simulation, trajectory analysis, and fluorescent experiments. The device enabled investigation of cellular response to dynamic stimuli at a single cell level.