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Cryogenic Super-Resolution Fluorescence and Electron Microscopy Correlated at the Nanoscale
Annual Review of Physical Chemistry ( IF 11.7 ) Pub Date : 2021-04-20 , DOI: 10.1146/annurev-physchem-090319-051546
Peter D Dahlberg 1 , W E Moerner 1
Affiliation  

We review the emerging method of super-resolved cryogenic correlative light and electron microscopy (srCryoCLEM). Super-resolution (SR) fluorescence microscopy and cryogenic electron tomography (CET) are both powerful techniques for observing subcellular organization, but each approach has unique limitations. The combination of the two brings the single-molecule sensitivity and specificity of SR to the detailed cellular context and molecular scale resolution of CET. The resulting correlative data is more informative than the sum of its parts. The correlative images can be used to pinpoint the positions of fluorescently labeled proteins in the high-resolution context of CET with nanometer-scale precision and/or to identify proteins in electron-dense structures. The execution of srCryoCLEM is challenging and the approach is best described as a method that is still in its infancy with numerous technical challenges. In this review, we describe state-of-the-art srCryoCLEM experiments, discuss the most pressing challenges, and give a brief outlook on future applications.

中文翻译:


低温超分辨率荧光和电子显微镜在纳米尺度上相关

我们回顾了超分辨低温相关光学和电子显微镜 (srCryoCLEM) 的新兴方法。超分辨率 (SR) 荧光显微镜和低温电子断层扫描 (CET) 都是观察亚细胞组织的强大技术,但每种方法都有其独特的局限性。两者的结合将 SR 的单分子敏感性和特异性带到了 CET 的详细细胞环境和分子尺度分辨率。由此产生的相关数据比其部分的总和更具信息性。相关图像可用于以纳米级精度在 CET 的高分辨率环境中查明荧光标记蛋白质的位置和/或识别电子密集结构中的蛋白质。srCryoCLEM 的执行具有挑战性,这种方法最好被描述为一种仍处于起步阶段的方法,面临着许多技术挑战。在这篇综述中,我们描述了最先进的 srCryoCLEM 实验,讨论了最紧迫的挑战,并对未来的应用进行了简要展望。

更新日期:2021-04-21
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