Correlative light-electron microscopy using small gold nanoparticles as single probes.
Correlative light-electron microscopy using small gold nanoparticles as single probes.
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DOI：
10.1038/s41377-023-01115-4
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发表时间：
2023-03-29
影响因子：
19.4
通讯作者：
Verkade, Paul
中科院分区：
文献类型：
--
作者：
Pope, Iestyn;Tanner, Hugh;Masia, Francesco;Payne, Lukas;Arkill, Kenton Paul;Mantell, Judith;Langbein, Wolfgang;Borri, Paola;Verkade, Paul
相关光电子显微镜（CLEM）需要在光学和电子显微镜下都可见的稳健探针的可用性。在这里，我们展示了一个CLEM的方法，使用小的金纳米粒子作为一个单一的探针。结合到表皮生长因子蛋白的单个金纳米颗粒通过使用共振四波混频（FWM）的光学显微镜以纳米精度在人类癌细胞中无背景地定位，并以高精度相关映射到相应的透射电子显微镜图像。我们使用了10 nm和5 nm半径的纳米颗粒，并在大于10 µm尺寸的区域内显示出低于60 nm的相关精度，而无需额外的基准标记。通过减小系统误差，相关精度提高到40 nm以下，而定位精度低于10 nm。偏振分辨的四波混频与纳米颗粒的形状相关，有望在未来的应用中通过形状识别进行复用。由于金纳米粒子的光稳定性和FWM显微镜对活细胞的适用性，FWM-CLEM开辟了一个强大的替代荧光为基础的方法。相关光学电子显微镜使用人类细胞中的单个金纳米颗粒作为单探针，通过四波混合成像在光学显微镜中直接可见，具有高定位精度。
Correlative light-electron microscopy (CLEM) requires the availability of robust probes which are visible both in light and electron microscopy. Here we demonstrate a CLEM approach using small gold nanoparticles as a single probe. Individual gold nanoparticles bound to the epidermal growth factor protein were located with nanometric precision background-free in human cancer cells by light microscopy using resonant four-wave mixing (FWM), and were correlatively mapped with high accuracy to the corresponding transmission electron microscopy images. We used nanoparticles of 10 nm and 5 nm radius, and show a correlation accuracy below 60 nm over an area larger than 10 µm size, without the need for additional fiducial markers. Correlation accuracy was improved to below 40 nm by reducing systematic errors, while the localisation precision is below 10 nm. Polarisation-resolved FWM correlates with nanoparticle shapes, promising for multiplexing by shape recognition in future applications. Owing to the photostability of gold nanoparticles and the applicability of FWM microscopy to living cells, FWM-CLEM opens up a powerful alternative to fluorescence-based methods. Correlative light electron microscopy using individual gold nanoparticles in human cells as single probes directly visible in light microscopy by four-wave mixing imaging with high localisation precision.
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影响因子：
14.8
作者：
通讯作者：
--
影响因子：
4.6
作者：
Johnson E;Seiradake E;Jones EY;Davis I;Grünewald K;Kaufmann R
通讯作者：
Kaufmann R
影响因子：
4.6
作者：
Tuijtel, Maarten W.;Koster, Abraham J.;Sharp, Thomas H.
通讯作者：
Sharp, Thomas H.
影响因子：
4.6
作者：
Miles BT;Greenwood AB;Benito-Alifonso D;Tanner H;Galan MC;Verkade P;Gersen H
通讯作者：
Gersen H
DOI：
10.1007/978-1-0716-0783-1_2
发表时间：
2021-01-01
期刊：
CRYOPRESERVATION AND FREEZE-DRYING PROTOCOLS, 4 EDITION
影响因子：
--
作者：
Fahy, Gregory M.;Wowk, Brian
通讯作者：
Wowk, Brian