A microfluidic platform with pneumatically switchable single-cell traps for selective intracellular signals probing

摘要

To investigate rapid suspension cell signaling, a microfluidic platform was urgently needed for flexibly manipulation of single cells and simultaneous generation of controllable chemical signals to stimulate single cells, In this paper, a microfluidic biosensor was developed to monitor intracellular calcium signal, integrated with single-cell trapping, chemical stimulation and releasing. Selective entrapment and discharge of individual cell were achieved by controlling the deformable membrane with pneumatic traps. The activation of intracellular calcium signal was qualitatively and quantitatively investigated by high-controllable chemical single-cell stimulation based on flexible hydrodynamic gating. And performing chemical stimulation and control assay in the same channel would improve the experimental robustness and effectiveness. Further investigation of the cellular responses to ATP pulses of varying concentrations and durations indicated that 20 mu M ATP pulses with duration as short as 200 ms resulted in the same level of Ca2+ response induced by sustained stimulations. Washing with buffer for 30 s was sufficient for single cell to recover from receptor desensitization caused by ATP stimulation. In addition, the responses of cells to ATP stimulation were heterogeneous. The developed microfluidic method opens up a new avenue for intracellular signaling studies and drug screening.