Stable and High-Throughput Single-Cell Sorting of Food Bacteria Using Spatiotemporal Video-Enhanced Raman Tweezers

Rapid detection of foodborne pathogenic and spoilage microorganisms is critical for ensuring food safety and quality in liquid matrices. While Raman tweezers spectroscopy (RTS) enables label-free single-cell analysis, its application in high-throughput inline inspection faces a fundamental bottleneck: high flow rates required for efficiency induce severe motion blur and low signal-to-noise ratios (SNR), which blind automated control systems and destabilize optical trapping. To overcome this, we present a Spatiotemporal Video-Enhanced Raman Tweezers (SVERT) system integrating a deceleration-optimized microfluidic chip with a deep learning-based visual feedback loop. We propose a Local–Global Unified Denoising Network (LGU-Net) tailored to recover high-fidelity bacterial structures from low-SNR video streams, achieving a deterministic processing latency of ~0.49 ms. Experimental results demonstrate that SVERT improves the optical trapping success rate from 21.27% ± 2% to 91.47% ± 1.8% compared to raw video input, enabling a four-fold increase in spectral acquisition efficiency. Leveraging the acquired high-quality dataset, we achieved a classification accuracy of 96.74% across four bacterial species of relevance to food safety and quality. Crucially, we validated the system’s practical robustness by successfully isolating and tracking trace E. coli in an unpurified commercial beverage. This capability to effectively mitigate natural background interference demonstrates the system’s promising potential to be expanded for broader applications in liquid food safety screening.