Yakimovich Artur, Andriasyan Vardan, Witte Robert, Wang I-Hsuan, Prasad Vibhu, Suomalainen Maarit, Greber Urs F
Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
PLoS One. 2015 Sep 28;10(9):e0138760. doi: 10.1371/journal.pone.0138760. eCollection 2015.
Classical plaque assay measures the propagation of infectious agents across a monolayer of cells. It is dependent on cell lysis, and limited by user-specific settings and low throughput. Here, we developed Plaque2.0, a broadly applicable, fluorescence microscopy-based high-throughput method to mine patho-biological clonal cell features. Plaque2.0 is an open source framework to extract information from chemically fixed cells by immuno-histochemistry or RNA in situ hybridization, or from live cells expressing GFP transgene. Multi-parametric measurements include infection density, intensity, area, shape or location information at single plaque or population levels. Plaque2.0 distinguishes lytic and non-lytic spread of a variety of DNA and RNA viruses, including vaccinia virus, adenovirus and rhinovirus, and can be used to visualize simultaneous plaque formation from co-infecting viruses. Plaque2.0 also analyzes clonal growth of cancer cells, which is relevant for cell migration and metastatic invasion studies. Plaque2.0 is suitable to quantitatively analyze virus infections, vector properties, or cancer cell phenotypes.
经典蚀斑测定法用于测量感染因子在单层细胞中的传播。它依赖于细胞裂解，且受用户特定设置和低通量的限制。在此，我们开发了Plaque2.0，这是一种广泛适用的、基于荧光显微镜的高通量方法，用于挖掘病理生物学克隆细胞特征。Plaque2.0是一个开源框架，可通过免疫组织化学或RNA原位杂交从化学固定细胞中提取信息，或从表达绿色荧光蛋白转基因的活细胞中提取信息。多参数测量包括单个蚀斑或群体水平上的感染密度、强度、面积、形状或位置信息。Plaque2.0可区分多种DNA和RNA病毒（包括痘苗病毒、腺病毒和鼻病毒）的裂解性和非裂解性传播，还可用于可视化共感染病毒同时形成蚀斑的情况。Plaque2.0还可分析癌细胞的克隆生长，这与细胞迁移和转移侵袭研究相关。Plaque2.0适用于定量分析病毒感染、载体特性或癌细胞表型。
