Rapid and stain-free quantification of viral plaque via lens-free holography and deep learning.
Rapid and stain-free quantification of viral plaque via lens-free holography and deep learning.
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DOI：
10.1038/s41551-023-01057-7
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发表时间：
2023-08
影响因子：
28.1
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中科院分区：
文献类型：
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空斑试验是测量具有复制能力的裂解病毒体浓度的金标准方法，需要染色，通常需要48小时以上的运行时间。在这里，我们展示了无透镜全息成像和深度学习可以结合起来，以加快和自动化分析。该紧凑型成像设备以每孔每小时约0.32千兆像素的速率捕获无标记的相位信息，覆盖约30 × 30 mm2的面积和比标准检测大10倍的病毒浓度动态范围，并定量感染面积和空斑形成单位的数量。对于水泡性口炎病毒，自动空斑试验检测到的第一个细胞裂解事件所造成的病毒复制早在5小时后孵育，并在不到20小时，它检测到的空斑形成单位的比率高于90%，在100%的特异性。此外，它将单纯疱疹病毒1型的潜伏期缩短了约48小时，将脑心肌炎病毒的潜伏期缩短了约20小时。免染色检测方法可用于病毒学研究、疫苗开发和临床诊断。利用无透镜全息成像和深度学习的自动化噬菌斑检测快速准确地检测由病毒复制引起的细胞裂解事件。
A plaque assay—the gold-standard method for measuring the concentration of replication-competent lytic virions—requires staining and usually more than 48 h of runtime. Here we show that lens-free holographic imaging and deep learning can be combined to expedite and automate the assay. The compact imaging device captures phase information label-free at a rate of approximately 0.32 gigapixels per hour per well, covers an area of about 30 × 30 mm2 and a 10-fold larger dynamic range of virus concentration than standard assays, and quantifies the infected area and the number of plaque-forming units. For the vesicular stomatitis virus, the automated plaque assay detected the first cell-lysing events caused by viral replication as early as 5 h after incubation, and in less than 20 h it detected plaque-forming units at rates higher than 90% at 100% specificity. Furthermore, it reduced the incubation time of the herpes simplex virus type 1 by about 48 h and that of the encephalomyocarditis virus by about 20 h. The stain-free assay should be amenable for use in virology research, vaccine development and clinical diagnosis. An automated plaque assay leveraging lens-free holographic imaging and deep learning rapidly and accurately detects the cell-lysing events caused by viral replication.
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影响因子：
2.1
作者：
Basak, Swarnendu;Kang, Hae-Ji;Chu, Ki-Back;Oh, Judy;Quan, Fu-Shi
通讯作者：
Quan, Fu-Shi
影响因子：
--
作者：
Abdelmageed, Alaa A;Ferran, Maureen C
通讯作者：
Ferran, Maureen C
DOI：
10.1007/978-1-61779-108-6_6
发表时间：
2011-01-01
期刊：
MAMMALIAN CELL VIABILITY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ke, Ning;Wang, Xiaobo;Abassi, Yama A.
通讯作者：
Abassi, Yama A.
影响因子：
19.4
作者：
Kim, Geon;Ahn, Daewoong;Kang, Minhee;Park, Jinho;Ryu, DongHun;Jo, YoungJu;Song, Jinyeop;Ryu, Jea Sung;Choi, Gunho;Chung, Hyun Jung;Kim, Kyuseok;Chung, Doo Ryeon;Yoo, In Young;Huh, Hee Jae;Min, Hyun-seok;Lee, Nam Yong;Park, YongKeun
通讯作者：
Park, YongKeun
影响因子：
7
作者：
Isil, Cagatay;de Haan, Kevin;Ozcan, Aydogan
通讯作者：
Ozcan, Aydogan