Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy
Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy
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DOI：
10.1073/pnas.1315858110
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发表时间：
2013-12-24
影响因子：
11.1
通讯作者：
Enderlein, Joerg
中科院分区：
文献类型：
--
作者：
Schulz, Olaf;Pieper, Christoph;Enderlein, Joerg
我们演示了如何传统的共焦旋转盘（CSD）显微镜可以转换成一个双分辨率的图像扫描显微镜（ISM）系统，而不改变其光学或机械元件的任何部分。利用CSD显微镜的固有特性，我们照亮频闪，产生一个阵列的激发焦点，通过改变频闪激发和旋转盘的旋转之间的相位移动整个样品。然后ISM生成分辨率几乎翻倍的图像。使用常规荧光团，我们对核膜中的单核孔复合物和GFP缀合的Tau蛋白聚集体进行了三维成像。多色ISM显示在细胞质相关的结构蛋白和三维四色图像，包括MitoTracker和Hoechst染色。常规CSD设备的简单适应允许各种各样的细胞生物学问题的超分辨率调查。
We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.