Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules.
Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules.
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DOI：
10.1038/s41467-020-20863-0
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发表时间：
2021-01-22
影响因子：
16.6
通讯作者：
Stefani FD
中科院分区：
文献类型：
--
作者：
Szalai AM;Siarry B;Lukin J;Williamson DJ;Unsain N;Cáceres A;Pilo-Pais M;Acuna G;Refojo D;Owen DM;Simoncelli S;Stefani FD
单分子定位显微镜能够实现横向分辨率在10至20纳米范围内的远场成像，利用单分子图像的中心位置可以比该图像本身的大小高得多的精度来确定。然而，在轴向（第三）维度上获得相同水平的分辨率仍然具有挑战性。在这里，我们提出了超临界照明显微镜光度z-本地化与增强分辨率（SIMPLER），一种光度法解码的轴向位置的单分子在全内反射荧光显微镜。SIMPLER不需要对传统的全内反射荧光显微镜进行任何硬件修改，并补充任何2D单分子定位显微镜方法，以提供具有近各向同性纳米分辨率的3D图像。性能示例包括具有亚20 nm轴向定位精度的核孔复合物的SIMPLER直接随机光学重建显微镜图像，以及通过SIMPLER-DNA点累积的微管横截面可视化，用于具有亚10 nm轴向定位精度的纳米级形貌成像。实现高轴向分辨率是单分子定位显微镜的挑战。在这里，作者提出了一种光度法解码的轴向位置的单分子在全内反射荧光显微镜没有硬件修改，并显示近各向同性的纳米分辨率。
Single-molecule localization microscopy enables far-field imaging with lateral resolution in the range of 10 to 20 nanometres, exploiting the fact that the centre position of a single-molecule’s image can be determined with much higher accuracy than the size of that image itself. However, attaining the same level of resolution in the axial (third) dimension remains challenging. Here, we present Supercritical Illumination Microscopy Photometric z-Localization with Enhanced Resolution (SIMPLER), a photometric method to decode the axial position of single molecules in a total internal reflection fluorescence microscope. SIMPLER requires no hardware modification whatsoever to a conventional total internal reflection fluorescence microscope and complements any 2D single-molecule localization microscopy method to deliver 3D images with nearly isotropic nanometric resolution. Performance examples include SIMPLER-direct stochastic optical reconstruction microscopy images of the nuclear pore complex with sub-20 nm axial localization precision and visualization of microtubule cross-sections through SIMPLER-DNA points accumulation for imaging in nanoscale topography with sub-10 nm axial localization precision. Achieving high axial resolution is challenging in single-molecule localization microscopy. Here, the authors present a photometric method to decode the axial position of single molecules in a total internal reflection fluorescence microscope without hardware modification, and show nearly isotropic nanometric resolution.
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DOI：
10.1111/j.1365-2818.1994.tb03426.x
发表时间：
1994-01-01
期刊：
JOURNAL OF MICROSCOPY-OXFORD
影响因子：
--
作者：
BURMEISTER, JS;TRUSKEY, GA;REICHERT, WM
通讯作者：
REICHERT, WM
影响因子：
35
作者：
Chizhik, Alexey I.;Rother, Jan;Enderlein, Joerg
通讯作者：
Enderlein, Joerg
影响因子：
56.9
作者：
Balzarotti, Francisco;Eilers, Yvan;Hell, Stefan W.
通讯作者：
Hell, Stefan W.
影响因子：
1.6
作者：
Barabas, Federico M.;Masullo, Luciano A.;Stefani, Fernando D.
通讯作者：
Stefani, Fernando D.
影响因子：
48
作者：
Franke, Christian;Sauer, Markus;van de Linde, Sebastian
通讯作者：
van de Linde, Sebastian