Three-dimensional multi-color optical nanoscopy at sub-10-nm resolution based on small-molecule organic probes.
Three-dimensional multi-color optical nanoscopy at sub-10-nm resolution based on small-molecule organic probes.
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DOI：
10.1016/j.crmeth.2023.100556
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发表时间：
2023-09-25
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
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Achieving nanometer-scale resolution remains challenging in expansion microscopy due to photon loss. To address this concern, here we develop a multi-color expansion stimulated emission depletion technique based on small-molecule probes to realize high labeling density and intensity. Our method substantially lowers the barrier to visualizing diverse intracellular proteins and their interactions in three dimensions. It enables us to achieve sub-10-nm resolution in structures such as microfilaments, lysosomes, and mitochondria, providing new insights into cell biology. Small-molecule organic probes provide high labeling density and intensity Small marker sizes decrease imaging artifacts caused by probe displacement Adaptive optics configuration in STED compensates for sample-induced aberrations The approach reveals subcellular structures and interactions at sub-10-nm resolution Combining expansion microscopy with other super-resolution techniques poses challenges due to the reduction in fluorescence density and intensity in expanded samples. Although employing multiple and post-expansion labeling of fluorescent proteins or antibodies can enhance the labeling density, attaining sub-10-nm resolution in super-resolution microscopy requires careful consideration of marker size. To address this issue, we have developed a multi-color 3D nanoscopy technique that employs small-molecule organic probes. The utilization of small-molecule organic probes serves to enhance both the density and intensity of labeling while reducing measurement error. Achieving isotropic nanometer-scale resolution remains challenging when combining STED with expansion microscopy. Han et al. establish a 3D multi-color nanoscopy technique based on the high labeling density and intensity of small-molecule organic probes, revealing details of the structures and interactions of actin filaments, lysosomes, and mitochondria.
长期/超分辨率荧光成像中活体标本的标记策略
影响因子：
5.5
作者：
Han Y;Zhang Z;Liu W;Yao Y;Xu Y;Liu X;Kuang C;Hao X
通讯作者：
Hao X