Dynamic video recognition for cell-encapsulating microfluidic droplets.

Droplet microfluidics is a highly sensitive and high-throughput technology extensively utilized in biomedical applications, such as single-cell sequencing and cell screening. However, its performance is highly influenced by the droplet size and single-cell encapsulation rate (following random distribution), thereby creating an urgent need for quality control. Machine learning has the potential to revolutionize droplet microfluidics, but it requires tedious pixel-level annotation for network training. This paper investigates the application software of the weakly supervised cell-counting network (WSCApp) for video recognition of microdroplets. We demonstrated its real-time performance in video processing of microfluidic droplets and further identified the locations of droplets and encapsulated cells. We verified our methods on droplets encapsulating six types of cells/beads, which were collected from various microfluidic structures. Quantitative experimental results showed that our approach can not only accurately distinguish droplet encapsulations (micro-F1 score > 0.94), but also locate each cell without any supervised location information. Furthermore, fine-tuning transfer learning on the pre-trained model also significantly reduced (>80%) annotation. This software provides a user-friendly and assistive annotation platform for the quantitative assessment of cell-encapsulating microfluidic droplets.

WSCNet: Biomedical Image Recognition for Cell Encapsulated Microfluidic Droplets.

Microfluidic droplets accommodating a single cell as independent microreactors are frequently demanded for single-cell analysis of phenotype and genotype. However, challenges exist in identifying and reducing the covalence probability (following Poisson's distribution) of more than two cells encapsulated in one droplet. It is of great significance to monitor and control the quantity of encapsulated content inside each droplet. We demonstrated a microfluidic system embedded with a weakly supervised cell counting network (WSCNet) to generate microfluidic droplets, evaluate their quality, and further recognize the locations of encapsulated cells. Here, we systematically verified our approach using encapsulated droplets from three different microfluidic structures. Quantitative experimental results showed that our approach can not only distinguish droplet encapsulations (F1 score > 0.88) but also locate each cell without any supervised location information (accuracy > 89%). The probability of a "single cell in one droplet" encapsulation is systematically verified under different parameters, which shows good agreement with the distribution of the passive method (Residual Sum of Squares, RSS < 0.5). This study offers a comprehensive platform for the quantitative assessment of encapsulated microfluidic droplets.