Expansion microscopy is a sample preparation technique that enables the optical imaging of biological specimens at super-resolution owing to their physical magnification, which is achieved through water-absorbing polymers. The technique uses readily available chemicals and does not require sophisticated equipment, thus offering super-resolution to laboratories that are not microscopy-specialised. Here we present a protocol combining sample expansion with light sheet microscopy to generate high-contrast, high-resolution 3D reconstructions of whole virus-infected cells. The results are superior to those achievable with comparable imaging modalities and reveal details of the infection cycle that are not discernible before expansion. An image resolution of approximately 95 nm could be achieved in samples labelled in 3 colours. We resolve that the viral nucleoprotein is accumulated at the membrane of vesicular structures within the cell cytoplasm and how these vesicles are positioned relative to cellular structures. We provide detailed guidance and a video protocol for the optimal application of the method and demonstrate its potential to study virus-host cell interactions.

中文翻译：

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结合样品扩展和光片显微镜，对病毒感染的细胞进行超分辨率体积成像


膨胀显微镜是一种样品制备技术，由于生物样本的物理放大作用是通过吸水性聚合物实现的，因此能够以超分辨率对生物样本进行光学成像。该技术使用现成的化学品，不需要复杂的设备，从而为非显微镜专业的实验室提供超分辨率。在这里，我们提出了一种将样本扩展与光片显微镜相结合的方案，以生成整个病毒感染细胞的高对比度、高分辨率 3D 重建。结果优于可比较的成像方式所获得的结果，并揭示了在扩展之前无法辨别的感染周期的细节。用 3 种颜色标记的样品可实现约 95 nm 的图像分辨率。我们解决了病毒核蛋白在细胞质内的囊泡结构膜上积累以及这些囊泡如何相对于细胞结构定位的问题。我们为该方法的最佳应用提供详细的指导和视频协议，并展示其研究病毒与宿主细胞相互作用的潜力。