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

Abstract

A microfluidic array was constructed for trapping single cell and loading identical dynamic biochemical stimulation for gain a better understanding of Ca2+ signalling in single cells by applying extracellular dynamic biochemical stimulus. 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 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 array chip, the captured single cells in each trapping channels were able to receive identical extracellular dynamic biochemical stimuli which being transmitted from the entrance at 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 in single cells to temporally varying environments by modulating the stimulation signal in terms of concentration, pattern, and duration of exposure.

Signalling dynamics, cell decisions, and homeostatic control in health and disease

“…The use of biosensors is somewhat established and gradually reaching an increasing number of non-specialist laboratories. Integrating biochemical imaging with microfluidics also facilitates the study of cell signalling with real-time control of both stimuli and responses [ 63 , 64• , 65 , 66 , 67•• , 68 ].…”

A sensitive electrochemical platform integrated with a 3D graphene aerogel for point-of-care testing for tumor markers

“…[14][15][16] Owing to the miniaturization of the detection platform, microfluidic biochips can provide relatively larger interactive areas and thus realize the trace detection of various biochemical molecules, including circulating tumor cells (CTCs), enzymes, Ca 2+ , etc. [17][18][19][20] Despite efforts to miniaturize electrodes, the lack of high-sensitivity electrodes greatly limits the practicality of tumor marker detection. In order to improve detection sensitivity, several portable, fast and ultrasensitive selfpowered biosensors were developed for the detection of micro-RNA and L-Cys.…”