An AI-Enhanced Microfluidic Platform for Dynamic Monitoring and Quantitative Evaluation of Single-Cell Migration

Cell migration plays a pivotal role in critical biological processes, including immune response, wound healing, embryonic development, and cancer metastasis. However, current technologies exhibit significant limitations in the real-time, dynamic monitoring and quantitative analysis of single-cell migration, particularly in high-throughput tracking of cellular behavior. To address this challenge, we developed an innovative single-cell migration analysis platform by combining microfluidics and the TOPICTrack multi-object tracking algorithm. Using AG490 to inhibit NIH/3T3 cell migration as a demonstration, we highlighted the platform’s effectiveness in automating parallel monitoring and analysis of single-cell migration. Experimental results show that as AG490 concentration increased from 0 <italic>μ</italic>M to 64 <italic>μ</italic>M, the average migration distance of cells significantly decreased from 159.44 <italic>μ</italic>m to 37.97 <italic>μ</italic>m, revealing both single-cell heterogeneity and population characteristics. Additionally, the platform reduced reagent consumption to 1/55 of traditional methods, significantly lowering experimental costs, while shortening experimental time from 48 hours to 12 hours. The platform supports single-cell migration analysis under multiple concentrations and field-of-view conditions, providing a rapid solution for high-throughput drug screening. Furthermore, through intelligent algorithms, the platform minimizes human intervention, enhancing accuracy and reproducibility. This platform represents a significant methodological breakthrough, enabling real-time parallel monitoring and analysis of single-cell behavior with minimal reagent consumption. It not only expands the tools for cell dynamics research but also offers strong technical support and application potential for pharmacological mechanism analysis, precision medicine, and personalized drug screening.