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Nat Protocols：利用低温光学和电子显微镜联合技术观察并研究活体病毒

2017-01-08 佚名 生物谷

活体病毒
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图片来源：www.phys.org 光镜和电镜研究领域的巨大进步目前正在改善科学家们对多种病毒可视化研究的能力，比如HIV、呼吸道合胞体病毒、麻疹病毒、流感病毒以及寨卡病毒等，近日，一项刊登在国际杂志Nature Protocols上的研究报告中，来自埃默里大学医学院等研究机构的研究人员通过研究开发出了一种新型的低温相关的光镜和电镜工作流（工作站，cryo-CLEM）。 此前研究中，研究

图片来源：www.phys.org

光镜和电镜研究领域的巨大进步目前正在改善科学家们对多种病毒可视化研究的能力，比如HIV、呼吸道合胞体病毒、麻疹病毒、流感病毒以及寨卡病毒等，近日，一项刊登在国际杂志Nature Protocols上的研究报告中，来自埃默里大学医学院等研究机构的研究人员通过研究开发出了一种新型的低温相关的光镜和电镜工作流（工作站，cryo-CLEM）。

此前研究中，研究者对纯化的病毒进行研究能够获取多种已知病毒电子显微镜图像，然而病毒纯化的过程常常会使得包膜病毒的结构发生改变，因此研究者Wright及同事就对当前技术进行了改善来研究病毒的结构特性，因此如今研究者就能够清楚观察到病毒进入细胞并在细胞中组装的信息了。研究者表示，我们非常想知道某些病毒如何在细胞中复制，如今我们在光镜和电镜之间建立了一种“桥梁”，我们希望通过这种联合技术能够更加清楚地观察病毒的作用机制。

文章中，研究者首先在脆性碳覆盖的黄金网格中对病毒感染或转染的细胞进行培养，随后进行玻璃化操作，即对细胞快速冷却以免冰晶形成，一旦细胞被冷却研究人员就能够利用低温荧光显微镜和低温电子断层扫描术对细胞进行研究，低温（负150度以下）对于这两种技术而言非常必要，而且样品玻璃化后进行光学显微镜检查还能够抑制细胞继续生长以及位置发生改变。

研究者指出，利用低温电子断层扫描术获得的数据还能够帮助我们在高分辨率下获取单一完整病毒和病毒蛋白的图像，这项研究中研究者对呼吸道合胞体病毒进行了研究，他们发现，候选减毒活疫苗在结构上同呼吸道合胞体病毒结构非常相似。这种名为cryo-CLEM的新技术能够对扁平生长的细胞进行研究，因为标准的光束并不能够穿透厚度大于1微米的细胞，而哺乳动物的细胞通常有几微米宽，而诸如HIV等病毒大约为0.1微米。

本文研究中，研究人员开发的新型cryo-CLEM技术或许还能够帮助研究包括神经细胞或细菌生物被膜在内的多个系统的功能。

原始出处

Cheri M Hampton, Joshua D Strauss, Zunlong Ke, Rebecca S Dillard, Jason E Hammonds, Eric Alonas, Tanay M Desai, Mariana Marin, Rachel E Storms, Fredrick Leon, Gregory B Melikyan, Philip J Santangelo, Paul W Spearman & Elizabeth R Wright.Correlated fluorescence microscopy and cryo-electron tomography of virus-infected or transfected mammalian cells.Nat Protocols.2016

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    2017-12-08 12498561m39(暂无昵称)

    #protocol#

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    2017-01-18 tulenzi

    #显微镜#

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    2017-02-16 12498f17m66暂无昵称

    #电子显微镜#

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    2017-11-26 liye789132251

    #Nat#

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    2017-01-10 一叶知秋

    #PRO#

    0

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