Angular reconstitution-based 3D reconstructions of nanomolecular structures from superresolution light-microscopy images
Angular reconstitution-based 3D reconstructions of nanomolecular structures from superresolution light-microscopy images
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DOI：
10.1073/pnas.1704908114
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发表时间：
2017-08-29
影响因子：
11.1
通讯作者：
Nollmann, Marcelo
中科院分区：
文献类型：
--
作者：
Salas, Desiree;Le Gall, Antoine;Nollmann, Marcelo
超分辨光学显微镜允许在超过经典衍射极限的分辨率下对标记的超分子组装体进行成像。超分辨方法的一个严重限制是样本的异质性和标记过程的随机性。为了提高超分辨结果的再现性和分辨率，我们采用了多元统计分析方法和最初为单粒子低温电子显微镜开发的3D重建方法。这些方法允许从二维超分辨率投影图像的纳米分子结构的无参考3D重建。由于这些2D投影图像都显示了光学显微镜的高分辨率方向上的结构，因此所得到的3D重建在所有方向上都具有最佳的各向同性分辨率。
Superresolution light microscopy allows the imaging of labeled supramolecular assemblies at a resolution surpassing the classical diffraction limit. A serious limitation of the superresolution approach is sample heterogeneity and the stochastic character of the labeling procedure. To increase the reproducibility and the resolution of the superresolution results, we apply multivariate statistical analysis methods and 3D reconstruction approaches originally developed for cryogenic electron microscopy of single particles. These methods allow for the reference-free 3D reconstruction of nanomolecular structures from two-dimensional superresolution projection images. Since these 2D projection images all show the structure in high-resolution directions of the optical microscope, the resulting 3D reconstructions have the best possible isotropic resolution in all directions.