Real-time measurement of thrombin generation using continuous droplet microfluidics

Area of Science:

• Biomedical Engineering
• Clinical Chemistry
• Cardiovascular Research

Background:

• Thrombin plays a critical role in blood coagulation and is a significant biomarker for hemostasis and cardiovascular disease (CVD).
• Accurate and dynamic monitoring of thrombin generation is crucial for understanding hemostasis and diagnosing CVD.
• Existing methods may lack the real-time, continuous monitoring capabilities needed for dynamic assessment.

Purpose of the Study:

• To develop and validate a novel droplet microfluidic system for continuous, real-time monitoring of individual thrombin generation.
• To assess the system's capability in measuring thrombin generation curves dynamically using plasma samples.
• To investigate the potential for in vivo applications in rapid CVD diagnosis.

Main Methods:

• Utilized droplet microfluidic technology to perform thrombin generation assays based on a fluorogenic substrate.
• Measured real-time thrombin generation curves from plasma samples activated by tissue factor.
• Incorporated inhibitor injection to study thrombin self-inhibition dynamics.
• Integrated the microfluidic system with a microdialysis probe for potential in vivo application.

Main Results:

• Successfully demonstrated continuous, real-time monitoring of individual thrombin generation within microfluidic droplets.
• Obtained dynamic thrombin generation curves reflecting real-time sample conditions.
• Developed a method to assay inhibited curves, providing insights into thrombin self-inhibition.
• Integrated a microdialysis probe, paving the way for future in vivo measurements.

Conclusions:

• The developed droplet microfluidic system offers a powerful tool for dynamic and real-time thrombin generation analysis.
• This technology has significant potential for advancing hemostasis research and enabling rapid CVD diagnosis.
• The integration with microdialysis opens new avenues for in vivo monitoring in living animals.