Rapid and stain-free quantification of viral plaque via lens-free holography and deep learning

• URL: http://arxiv.org/abs/2207.00089v2
• Date: Thu, 22 Jun 2023 20:29:10 GMT
• Title: Rapid and stain-free quantification of viral plaque via lens-free holography and deep learning
• Authors: Tairan Liu, Yuzhu Li, Hatice Ceylan Koydemir, Yijie Zhang, Ethan Yang, Merve Eryilmaz, Hongda Wang, Jingxi Li, Bijie Bai, Guangdong Ma, Aydogan Ozcan
• Abstract summary: This device captures 0.32 Giga-pixel/hour phase information of the objects per test well, covering an area of 30x30 mm2, in a label-free manner. Using a neural network, this device automatically detected the first cell lysing events due to the VSV viral replication as early as 5 hours after the incubation.
• Score: 2.0185810603595686
• License: http://arxiv.org/licenses/nonexclusive-distrib/1.0/
• Abstract: We present a rapid and stain-free quantitative viral plaque assay using lensfree holographic imaging and deep learning. This cost-effective, compact, and automated device significantly reduces the incubation time needed for traditional plaque assays while preserving their advantages over other virus quantification methods. This device captures ~0.32 Giga-pixel/hour phase information of the objects per test well, covering an area of ~30x30 mm^2, in a label-free manner, eliminating staining entirely. We demonstrated the success of this computational method using vesicular stomatitis virus (VSV), herpes simplex virus (HSV-1) and encephalomyocarditis virus (EMCV). Using a neural network, this stain-free device automatically detected the first cell lysing events due to the VSV viral replication as early as 5 hours after the incubation, and achieved >90% detection rate for the VSV plaque-forming units (PFUs) with 100% specificity in <20 hours, providing major time savings compared to the traditional plaque assays that take at least 48 hours. Similarly, this stain-free device reduced the needed incubation time by ~48 hours for HSV-1 and ~20 hours for EMCV, achieving >90% detection rate with 100% specificity. We also demonstrated that this data-driven plaque assay offers the capability of quantifying the infected area of the cell monolayer, performing automated counting and quantification of PFUs and virus-infected areas over a 10-fold larger dynamic range of virus concentration than standard viral plaque assays. This compact, low-cost, automated PFU quantification device can be broadly used in virology research, vaccine development, and clinical applications.

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