Birefringent Fourier filtering for single molecule coordinate and height super-resolution imaging with dithering and orientation.
Birefringent Fourier filtering for single molecule coordinate and height super-resolution imaging with dithering and orientation.
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DOI：
10.1038/s41467-020-19064-6
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发表时间：
2020-10-20
影响因子：
16.6
通讯作者：
Alonso MA
中科院分区：
文献类型：
--
作者：
Curcio V;Alemán-Castañeda LA;Brown TG;Brasselet S;Alonso MA
基于单分子定位的超分辨率成像允许以高精度访问生物样品中的纳米尺度信息。然而，包括分子取向在内的完整测量仍然具有挑战性。在显微镜成像中，取向与位置本质上是耦合的，并且在图像积分时间期间的分子摆动可以使取向测量偏置。提供3D分子取向和取向波动将提供新的方法来评估蛋白质结构的对齐程度，这不能通过纯定位来监测。在这里，我们证明，通过在成像路径的傅立叶平面中将偏振控制添加到相位控制，可以从单个分子中明确地确定所有参数：3D空间位置、3D取向和摆动或抖动角度。该方法应用于附着在单个肌动蛋白丝上的荧光标记，提供了数十纳米的位置和几度的方向精度。在超分辨率成像中确定单分子的取向是具有挑战性的。这里，通过在成像路径的傅立叶平面中将偏振控制添加到相位控制，可以从单个分子确定诸如3D空间位置、3D取向和摆动或抖动角的参数。
Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging. Orientation is intrinsically coupled to position in microscopy imaging, and molecular wobbling during the image integration time can bias orientation measurements. Providing 3D molecular orientation and orientational fluctuations would offer new ways to assess the degree of alignment of protein structures, which cannot be monitored by pure localization. Here we demonstrate that by adding polarization control to phase control in the Fourier plane of the imaging path, all parameters can be determined unambiguously from single molecules: 3D spatial position, 3D orientation and wobbling or dithering angle. The method, applied to fluorescent labels attached to single actin filaments, provides precisions within tens of nanometers in position and few degrees in orientation. Determining the orientation of single molecules in super resolution imaging is challenging. Here, by adding polarization control to phase control in the Fourier plane of the imaging path, parameters such as 3D spatial position, 3D orientation and wobbling or dithering angle can be determined from single molecules.
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影响因子：
48
作者：
Dempsey, Graham T.;Vaughan, Joshua C.;Chen, Kok Hao;Bates, Mark;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei
影响因子：
3.6
作者：
Backer, Adam S.;Backlund, Mikael P.;Moerner, W. E.
通讯作者：
Moerner, W. E.
DOI：
10.1364/josaa.36.001346
发表时间：
2019-08-01
影响因子：
1.9
作者：
Chandler, Talon;Shroff, Hari;La Riviere, Patrick
通讯作者：
La Riviere, Patrick
DOI：
10.1364/josab.20.000554
发表时间：
2003-03-01
影响因子：
1.9
作者：
Böhmer, M;Enderlein, J
通讯作者：
Enderlein, J
影响因子：
10.8
作者：
Deutsch, Bradley;Hillenbrand, Rainer;Novotny, Lukas
通讯作者：
Novotny, Lukas