Development of a microfluidic cell-based biosensor integrating a millisecond chemical pulse generator

Area of Science:

• Biomedical Engineering
• Cellular Biosensing
• Microfluidics

Background:

• Cell-based biosensors face limitations due to agonist-induced receptor desensitization.
• Continuous stimulation leads to reduced cell-surface receptor responsiveness.

Purpose of the Study:

• To develop a microfluidic cell-based biosensor (μCBB) for ATP detection.
• To overcome receptor desensitization issues in cell-based sensing.
• To enable sensitive detection of pulsatile signals in liquid environments.

Main Methods:

• Development of a microfluidic chip with hydrodynamic gated injection.
• Integration of a millisecond chemical pulse generator for pulsatile sample introduction.
• Utilizing NIH-3T3 cells expressing P2Y receptors as the sensing element.
• Stimulation with millisecond ATP pulses (100 ms every 50s).

Main Results:

• The μCBB successfully detected ATP in liquid environments.
• Pulsatile ATP stimulation (100 ms pulses every 50s) minimized receptor desensitization.
• Sustained amplitude of calcium spikes indicated stable receptor function.
• The μCBB demonstrated sensitivity for ATP detection.
• Cellular responses to millisecond pulses were investigated.

Conclusions:

• The developed μCBB effectively minimizes receptor desensitization.
• Pulsatile stimulation allows for extended use times of cell-based biosensors.
• The μCBB is suitable for detecting pulsatile samples.
• This technology advances cell-based biosensing for dynamic analyte monitoring.