用无标记动态成像实现病毒斑点的连续量化，提速近一天半并提升精度。
Continuous quantification of viral plaque dynamics using ultra-large-area label-free imaging enables rapid antiviral susceptibility testing
- 结合超大面积传感器与深度学习，实现培养箱内无标记自动追踪病毒斑点动态。
- 在感染后60小时内完成药物效力评估，比传统方法快约26小时且零误报。
- 可揭示药物浓度对病毒扩散时间延迟的差异化影响，适合高通量药筛与临床诊断。
斑块减少试验（PRA）仍是抗病毒敏感性检测的金标准，通过测量斑块形成单位（PFU）的减少来评估药物效力。然而传统PRA耗时长、人工操作多、易出错，且依赖破坏性固定和化学染色，仅提供静态终点观测，掩盖了剂量依赖性病毒抑制的时间动态。本文提出一种无标记、时间分辨的PRA平台，将传统方法转变为连续的高维病毒感染动力学测量。系统整合紧凑型无透镜成像装置与定制的超大面积（100 cm²）薄膜晶体管（TFT）图像传感器，配合深度学习算法，在培养箱内自主量化PFU动态。以阿昔洛韦处理的单纯疱疹病毒1型（HSV-1）验证，该平台与化学染色基准结果一致，零误报，并将读取时间缩短约26小时。关键发现：药物浓度升高会引发不同时间延迟并抑制新PFU形成，可在感染后约60小时内得出明确药物有效性结论。这一可扩展的无标记框架，将抗病毒检测重构为快速、时间分辨、信息丰富的测量体系，适用于病毒学研究、高通量药物筛选及临床诊断。
原文摘要 · Abstract (English)
The plaque reduction assay (PRA) remains the gold standard for antiviral susceptibility testing, evaluating drug potency by measuring reductions in plaque-forming units (PFUs). However, the traditional PRA is time-consuming, labor-intensive, prone to manual counting errors, and offers limited scalability. Moreover, its reliance on destructive fixation and chemical staining reduces the assay to a static, endpoint observation, obscuring the dynamic, time-resolved kinetics of dose-dependent viral inhibition. Here, we introduce a label-free, time-resolved PRA platform that transforms the conventional assay into a continuous, high-dimensional measurement of viral infection dynamics. Our system integrates a compact lens-free imaging setup with a custom-designed ultra-large-area (100 cm^2) thin-film transistor (TFT) image sensor and deep learning-based algorithms to autonomously quantify PFU dynamics within an incubator. Validated using herpes simplex virus type-1 (HSV-1) treated with acyclovir, the platform matched chemically-stained ground truth measurements with zero false positives while accelerating readout by ~26 hours. Crucially, our system revealed that increasing drug concentrations induce temporally distinct delays and suppress new PFU formation, enabling conclusive drug efficacy evaluations within ~60 hours post-infection. This scalable, label-free framework redefines antiviral susceptibility testing as a rapid, time-resolved and information-rich measurement framework, providing a generalizable platform for virology research, high-throughput drug screening, and clinical diagnostics.
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