Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Oxford, United Kingdom; Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany; Centre for Structural Systems Biology, Hamburg, Germany.
Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Oxford, United Kingdom; Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany; Centre for Structural Systems Biology, Hamburg, Germany; Department of Chemistry, University of Hamburg, Hamburg, Germany.
Methods Cell Biol. 2021;162:253-271. doi: 10.1016/bs.mcb.2020.10.021. Epub 2020 Dec 19.
The combination of super-resolution fluorescence microscopy and electron microscopy at ambient temperatures has become an established technique and a broad variety of modalities are now available to the cell biology community. In contrast, correlative cryogenic super-resolution fluorescence and electron microscopy (super-resolution cryo-CLEM) is just emerging. Aside from technical challenges, one of the major issues is the risk of devitrification of the specimen caused by the laser intensities required for super-resolution imaging. Cryo-SOFI (cryogenic super-resolution optical fluctuation imaging) allows the reconstruction of super-resolution images at particularly low laser intensities. It is fully compatible with the standard sample preparation for cryogenic electron microscopy (cryo-EM) and fairly easy to implement in any standard cryogenic fluorescence microscope.
在常温条件下，将超分辨率荧光显微镜与电子显微镜相结合已经成为一种成熟的技术，现在细胞生物学领域有多种模式可供选择。相比之下，低温条件下的相关超分辨率荧光和电子显微镜技术（低温相关超分辨率荧光和电子显微镜技术，super-resolution cryo-CLEM）才刚刚出现。除了技术挑战之外，主要问题之一是由于超分辨率成像所需的激光强度，导致标本发生非晶化的风险。Cryo-SOFI（低温超分辨率光波动成像）允许在特别低的激光强度下重建超分辨率图像。它与低温电子显微镜（cryo-EM）的标准样品制备完全兼容，并且在任何标准低温荧光显微镜中都非常容易实现。
