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

Area of Science:

• Biomedical Engineering
• Cell Biology
• Biochemistry

Background:

• Understanding intracellular calcium (Ca2+) signaling is crucial for cell function.
• Dynamic biochemical stimulation is essential for studying cellular responses.
• Single-cell resolution is needed to capture cellular heterogeneity.

Purpose of the Study:

• To develop and validate a microfluidic array for trapping single cells.
• To deliver identical dynamic biochemical stimuli to individual cells.
• To investigate Ca2+ signaling dynamics at single-cell resolution.

Main Methods:

• Design and fabrication of a microfluidic array using stagnation point flow and physical barriers.
• Numerical simulations and trajectory analysis for device validation.
• Fluorescent experiments to assess biochemical concentration profiles and Ca2+ dynamics.

Main Results:

• The microfluidic array effectively traps single cells and provides identical dynamic stimulation.
• Stimulation parameters (flow rate, frequency) influence biochemical concentration profiles.
• Consistent intracellular Ca2+ dynamics and cellular heterogeneity were observed in K562 cells upon Adenosine Triphosphate (ATP) stimulation.

Conclusions:

• The developed microfluidic array enables precise control over single-cell stimulation.
• The device facilitates the study of cellular responses to dynamic environments at single-cell resolution.
• This technology can be applied to investigate complex cellular signaling pathways.