VirusMapper: open-source nanoscale mapping of viral architecture through super-resolution microscopy.
VirusMapper: open-source nanoscale mapping of viral architecture through super-resolution microscopy.
复制标题
DOI：
10.1038/srep29132
复制
发表时间：
2016-07-04
影响因子：
4.6
通讯作者：
Henriques R
中科院分区：
文献类型：
--
作者：
Gray RD;Beerli C;Pereira PM;Scherer KM;Samolej J;Bleck CK;Mercer J;Henriques R
哺乳动物病毒在其感染性生命周期中的纳米级分子组装仍然知之甚少。它们的小尺寸，通常低于光学显微镜的300 nm衍射极限，限制了大多数电子显微镜的成像研究。超分辨率（SR）光学显微镜的最新发展现在允许以数十纳米的分辨率可视化病毒结构。此外，这些技术提供了额外的好处，分子特异性标记和使用活细胞显微镜研究病毒结构动力学的能力。然而，缺乏强大的分析工具，允许在感染环境中精确绘制病毒结构。在这里，我们提出了一个开源的分析框架，它结合了超分辨率成像和朴素的单粒子分析，以生成无偏的分子模型。该工具，VirusMapper，是一个高通量，用户友好，基于ImageJ的软件包，允许保守的多分子结构，如病毒亚结构或完整病毒的自动统计映射。我们证明了VirusMapper的可用性，将其应用到SIM和STED图像的牛痘病毒在隔离和与宿主细胞时。VirusMapper允许生成准确的、高含量的、分子特异性病毒体模型，并检测病毒结构中的纳米级变化。
The nanoscale molecular assembly of mammalian viruses during their infectious life cycle remains poorly understood. Their small dimensions, generally bellow the 300nm diffraction limit of light microscopes, has limited most imaging studies to electron microscopy. The recent development of super-resolution (SR) light microscopy now allows the visualisation of viral structures at resolutions of tens of nanometers. In addition, these techniques provide the added benefit of molecular specific labelling and the capacity to investigate viral structural dynamics using live-cell microscopy. However, there is a lack of robust analytical tools that allow for precise mapping of viral structure within the setting of infection. Here we present an open-source analytical framework that combines super-resolution imaging and naïve single-particle analysis to generate unbiased molecular models. This tool, VirusMapper, is a high-throughput, user-friendly, ImageJ-based software package allowing for automatic statistical mapping of conserved multi-molecular structures, such as viral substructures or intact viruses. We demonstrate the usability of VirusMapper by applying it to SIM and STED images of vaccinia virus in isolation and when engaged with host cells. VirusMapper allows for the generation of accurate, high-content, molecular specific virion models and detection of nanoscale changes in viral architecture.