Using soft X-ray tomography for rapid whole-cell quantitative imaging of SARS-CoV-2-infected cells.
Using soft X-ray tomography for rapid whole-cell quantitative imaging of SARS-CoV-2-infected cells.
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DOI：
10.1016/j.crmeth.2021.100117
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发表时间：
2021-11-22
期刊：
影响因子：
--
通讯作者：
Weinhardt V
中科院分区：
文献类型：
--
作者：
Loconte V;Chen JH;Cortese M;Ekman A;Le Gros MA;Larabell C;Bartenschlager R;Weinhardt V
宿主细胞超微结构的高分辨率和快速成像可以产生对病毒疾病机制的见解，例如严重急性呼吸道综合征冠状病毒-2（SARS-CoV-2）感染。在这里，我们采用全旋转软X射线断层扫描（SXT）检查SARS-CoV-2诱导的细胞器重塑在全细胞水平具有高空间分辨率和吞吐量。大多数当前的SXT系统由于使用平面样品支撑和由于缺失数据而产生的伪影而遭受受限的视场。在这种方法中，使用圆柱形样品架，在不到10分钟的时间内完成人肺上皮细胞的全旋转断层扫描。我们通过观察SARS-CoV-2病毒粒子的聚集体和病毒诱导的细胞内改变来证明SXT成像的潜力。这种快速的全细胞成像方法使我们能够以定量的方式可视化病毒感染后细胞器的时空变化。软X射线断层扫描可实现细胞器形态的无标记成像圆柱形样品架可捕获大细胞的全旋转断层图像10分钟内快速获得断层图像SARS-CoV-2诱导改变的全细胞定量表型分析感染细胞的3D结构分析对于理解病毒性疾病的机制至关重要。荧光和电子显微镜技术，特别是聚焦离子束扫描电子显微镜（FIB-SEM）已广泛用于对细胞的3D结构进行成像。然而，尽管FIB-SEM实现了上级空间分辨率，但该方法的吞吐量仍然很低：8 nm各向同性体素尺寸的整个哺乳动物细胞需要大约一周的时间来成像，并产生大量的数据，这些数据难以分析。为了克服这些限制，需要一种具有最少样品制备和快速图像采集的全细胞成像技术来定量分析病毒感染细胞中诱导的结构变化。需要高分辨率，快速成像技术来分析3D细胞结构，以了解病毒性疾病的机制。在这里，Loconte等人。使用软X射线断层扫描技术对感染SARS-CoV-2的全细胞进行快速成像，为分析病毒-细胞相互作用和抗病毒药物的有效性开辟了途径。
High-resolution and rapid imaging of host cell ultrastructure can generate insights toward viral disease mechanism, for example for a severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. Here, we employ full-rotation soft X-ray tomography (SXT) to examine organelle remodeling induced by SARS-CoV-2 at the whole-cell level with high spatial resolution and throughput. Most of the current SXT systems suffer from a restricted field of view due to use of flat sample supports and artifacts due to missing data. In this approach using cylindrical sample holders, a full-rotation tomogram of human lung epithelial cells is performed in less than 10 min. We demonstrate the potential of SXT imaging by visualizing aggregates of SARS-CoV-2 virions and virus-induced intracellular alterations. This rapid whole-cell imaging approach allows us to visualize the spatiotemporal changes of cellular organelles upon viral infection in a quantitative manner. Soft X-ray tomography enables label-free imaging of organelle morphology Cylindrical sample holders enable capture of a full-rotation tomogram of large cells Tomogram obtained rapidly within 10 min Whole-cell quantitative phenotypic analysis of SARS-CoV-2-induced alterations The analysis of 3D architecture of infected cells is critical for understanding the mechanisms of viral diseases. Fluorescence and electron microscopy techniques, in particular, focused ion beam scanning electron microscopy (FIB-SEM) have been extensively used to image the 3D structure of cells. However, although FIB-SEM achieves superior spatial resolution, the method remains low throughput: an entire mammalian cell at 8 nm isotropic voxel size takes about a week to image and generates an enormous amount of data that is daunting to analyze. To overcome these limitations, a whole-cell imaging technique with minimal sample preparation and rapid image acquisition is required for quantitative analysis of structural changes induced in virus-infected cells. High-resolution, rapid imaging techniques are needed to analyze 3D cell architecture for understanding viral disease mechanisms. Here, Loconte et al. use soft X-ray tomography to rapidly image SARS-CoV-2-infected whole cells, opening avenues to analyze virus-cell interactions and efficacy of antiviral drugs in statistically significant numbers.
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影响因子：
5.6
作者：
Dolskiy AA;Grishchenko IV;Yudkin DV
通讯作者：
Yudkin DV
影响因子：
4
作者：
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2.2
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--
作者：
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通讯作者：
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3
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